Strategic Design of Delivery Systems for Nutraceuticals

Chapter 4

Strategic Design of Delivery Systems for Nutraceuticals S. Lee Khalifa University of Science, Technology, and Research, Abu Dhabi, United Arab Emirates

1. INTRODUCTION Until recently, food scientist and engineers limited the analysis of food to its flavor such as sensory taste and texture or its nutritional value such as composition of carbohydrates, fats, proteins, water, vitamins, and minerals. However, increasing evidence demonstrates that some chemical components of food may play an integral role in health (Hardy, 2000). The chemical components including probiotics, antioxidants, vitamins, essential minerals, and phytochemicals are derived from plant, food, and microbial sources and they provide medicinal benefits for long-term health (Roudebush et al., 2008; Varker et al., 2012). The term nutraceuticals is a combination of the words nutrition and pharmaceuticals and was coined in 1989 by Stephen L. DeFelice, MD, the Foundation for Innovation in Medicine (New York, USA) (Curry et al., 1999). Nutraceuticals or functional foods are defined as foods or part of foods that provide medical or health benefits including the prevention and treatment of chronic diseases as well as basic nutrition (Curry et al., 1999; Zeisel, 1999). Nutraceuticals range from isolated nutrients, purified food, dietary supplements, herbal products, and specific diets to genetically engineered foods such as vitamins, minerals, cereals, herbals, milk, soups, and beverages (Zhao, 2007). The link between food and health was established long ago. “Let food be thy medicine and medicine be thy food” said Hippocrates, the father of modern medicine, around 2500 years ago. In Asia and Africa, the natural herbs and spices were used as folk medicine or traditional medicine for centuries and traditional medicines include traditional Chinese medicine, traditional Korean medicine, Ayurveda, Siddha medicine, Unani, Islamic medicine, ancient Iranian medicine, Muti, Ifá, and traditional African medicine.

2. NUTRACEUTICALS Nutraceuticals or functional foods are currently in the limelight as an alternative medicine, as the dramatically longer life span of people during the 20th century has resulted in a much greater number of individuals suffering from agingassociated chronic diseases such as cardiovascular diseases (CVDs), diabetes mellitus type II, cancers, and neurodegenerative diseases. Aging and disease processes induce inflammation that decreases physical function and increases the risk for aging-associated diseases (Chung et al., 2009a; Grivennikov et al., 2010). Inflammation is the first response of the immune system to infection or irritation, such as infectious agents, physical injury, hypoxia, or disease processes (Lu et al., 2006a). Inflammation involves four symptoms: redness, heat, pain, and swelling (Yan et al., 2014). All inflammatory diseases also share common inflammation characteristics on the cellular or molecular level, including oxidative stress, remodeling of the extracellular matrix, angiogenesis, and fibrosis (Kolluru et al., 2012). Nutraceuticals with anti-inflammatory properties hold promise for slowing down the process of aging and disease progression. One example is phytochemical nutraceuticals that are used as a powerful tool in maintaining health and to act against acute and chronic diseases, thereby promoting optimal health, longevity, and quality of life (Ho et al., 2012).

Nanotechnology Applications in Food. http://dx.doi.org/10.1016/B978-0-12-811942-6.00004-2 Copyright © 2017 Elsevier Inc. All rights reserved.

65

66

Nanotechnology Applications in Food

Nutraceuticals with medicinal value include natural foods with antioxidants or vitamins, essential minerals, functional foods, phytochemicals, prebiotics, and probiotics.

2.1 Antioxidants Reactive oxygen species (ROS) are produced as by-products of normal cellular metabolism, mainly in the mitochondria (Finkel and Holbrook, 2000). ROS are highly reactive chemical forms of oxygen such as superoxide, hydrogen peroxide, and hydroxyl radical, and they can oxidize DNA, proteins, and lipids. ROS are very useful in destroying biological pathogens by damaging their proteins, lipids, and DNA (Hultqvist et al., 2009; Wu and Cederbaum, 2003). However, ROS are not pathogen specific and overproduction of ROS is toxic to even host cells if not enough antioxidants are present. Antioxidants are substances that protect cells against ROS (Mates, 2000). ROS can be scavenged in vivo by antioxidant enzymes such as superoxide dismutase (SOD), catalase, and peroxidase (Loprasert et al., 1996; Greenwald, 1990). For example, metabolic enzymes such as nicotinamide adenine dinucleotide phosphate (NADPH) oxidase and xanthine oxidase are involved in the conversion of oxygen into superoxide (Jaimes et al., 2004). Antioxidant enzymes can convert ROS into less reactive and nontoxic compounds such as water and oxygen. SOD transforms superoxide to hydrogen peroxide (H2O2) and then catalase/glutathione peroxidase (GPx) converts H2O2 to water (Finaud et al., 2006). Fig. 4.1 demonstrates the list of ROS, chronic diseases, and antioxidants. Oxidative stress is the steady-state level of oxidative damage in a cell, tissue, or organ that is caused by ROS and the stress increases with aging or external sources such as tobacco smoke and radiation exposure (Wei, 1998). Oxidative stress is linked to many diseases including cancer, arthritis, atherosclerosis, Alzheimer disease (AD), and diabetes (Pham-Huy et al., 2008). Oxidative stress can be alleviated by increasing the uptake of both antioxidant enzymes and nonenzymatic antioxidants such as vitamin A, vitamin C, vitamin E, polyphenols, carotenoids, and flavonoids such as catechin, quercetin, and anthocyanidin (Bouayed and Bohn, 2010). Young people naturally produce the antioxidant enzymes such as SOD, catalase, and GPx to help protect against free radicals produced during normal physiological processes (Flora, 2007; Pham-Huy et al., 2008). Levels of SOD, catalase, and other antioxidant enzymes decline with age, contributing to the scourge of age-related diseases. For example, the levels of antioxidant enzymes in young people (22 healthy males, age ¼ 28  12 years) are catalase ¼ 394.9  22.8 IU/g Hb and SOD ¼ 5981  375 IU/g Hb (Ferreira et al., 1999). However, the levels of antioxidant enzymes in old people (1780 healthy males, age ¼ 55e69 years) decrease to catalase ¼ 52.9  17.1 IU/g Hb and SOD ¼ 995  145 IU/g Hb (Mulholland et al., 1999). Most vegetables and fruits are a good source of antioxidants (Miller et al., 2000). GPx is found in almost all fruits and vegetables such as asparagus, broccoli, spinach, tomato, watermelon, and avocado. Some of the foods that contain SOD include barley grass, wheatgrass, broccoli, brussels sprouts, cabbage, and cantaloupe (Lalles et al., 2011). Beef liver, potatoes, and avocados contain large amounts of catalase enzyme (Murthy et al., 1981; Reid et al., 1981). Vitamin E (a-tocopherol) can neutralize the unpaired electron of free radicals and it becomes a-tocoquinone (Bolt and Stewart, 2012; Gulcin, 2012). Vitamin C can scavenge free radicals by donating a single electron and it forms ascorbyl radical that is more stable than other radical species because of resonance delocalization (Bolt and Stewart, 2012; Gulcin, 2012). Citrus fruits, green peppers, broccoli, green leafy vegetables, strawberries, raw cabbage, and potatoes are important sources of vitamin C (Wannamethee et al., 2006; Watzl and Bub, 2003). Vitamin E is rich in wheat germ, nuts, seeds, whole grains, green leafy vegetables, vegetable oil, and fish-liver oil (Atalay et al., 2000; Farwer et al., 1994). Food sources for flavonoids include cranberries, kale, beets, berries, red and black grapes, oranges, lemons, grapefruits, and green tea (Lu et al., 2006b; Hughes, 1978).

FIGURE 4.1 List of reactive oxygen species (ROS), chronic diseases, and antioxidants. ROS are chemically reactive oxygen compounds, and overproduction of ROS, oxidative stress, is responsible for chronic diseases. Both enzymatic and nonenzymatic antioxidants can scavenge ROS before they can damage cells or induce chronic diseases.

ROS Superoxide anion (O2∙–) Hydroxyl radical (OH∙) Hydrogen peroxide (H2O2) Singlet oxygen (1O2) Peroxynitrite (ONOO–) Hypochlorous acid (HOCl) Lipid peroxide (ROO–)

Enzymatic Antioxidants SOD Catalase Glutathione peroxidase

Nonenzymatic Antioxidants Vitamin A, Vitamin C, Vitamin E, Coenzyme Q, Cysteine, Flavonoids, β-Carotene

Chronic Diseases Cardiovascular Diseases Diabetes Alzheimer Disease

Strategic Design of Delivery Systems for Nutraceuticals Chapter | 4

67

2.2 Vitamins Vitamins are organic compounds that cannot be synthesized by humans and therefore must be ingested to prevent metabolic disorders (Fletcher and Fairfield, 2002). Vitamins are classified as either fat soluble (vitamins A, D, E, and K) or water soluble (vitamins B and C) (Beisel, 1982). Classic vitamin deficiency syndromes are scurvy, beriberi, and pellagra; however, many chronic diseases are also strongly related to vitamin deficiency (Fairfield and Fletcher, 2002). Vitamin A is a group of unsaturated organic compounds including retinol, retinal, and retinoic acid, and several provitamin A carotenoids. Owing to its antioxidant effects, it helps prevent diseases such as CVD and cancers (Palace et al., 1999). Vitamin A is rich in carrots, sweet potatoes, dark leafy greens, winter squashes, lettuce, dried apricots, cantaloupe, bell peppers, fish, and liver (van Jaarsveld et al., 2005). The B vitamins are a class of water-soluble compounds that play important roles in cell metabolism. Vitamin B1, thiamine, plays a central role in the generation of energy from carbohydrates (Guilland, 2013). Vitamin B2, riboflavin, is also involved in energy production. Vitamin B3, niacin, plays an important role in energy transfer reactions in the metabolism of glucose, fat, and alcohol. Vitamin B5, pantothenic acid, is involved in the oxidation of fatty acids and carbohydrates. Vitamin B6, pyridoxine, pyridoxal, and pyridoxamine, participates in more than 100 enzymatic reactions and is needed for protein metabolism. Vitamin B7, biotin, plays a key role in the metabolism of lipids, proteins, and carbohydrates. Vitamin B9, folic acid, is involved in the transfer of single-carbon units in the metabolism of nucleic acids and amino acids. Vitamin B12, cobalamin, acts as a coenzyme in fat and carbohydrate metabolism, protein synthesis, and hematopoiesis (Fairfield and Fletcher, 2002). The deficiency of vitamin B may result in many diseases such as beriberi, ariboflavinosis, acne, impaired growth and neurological disorders in infants, and macrocytic anemia (Mahalle et al., 2013; Kumar et al., 2009; Svenson, 2007). The richest food sources of B vitamins are dark-green leafy vegetables, whole-grain cereals, fortified grain products, and animal products such as beef liver, trout, salmon, and tuna. Vitamin C, ascorbic acid or L-ascorbate, is water soluble and acts as a cofactor in hydroxylation reactions, which are required for collagen synthesis. Vitamin C also promotes hormone synthesis, wound healing, and iron absorption (Padh, 1991). It is also a strong antioxidant. Vitamin C is important to prevent diseases such as scurvy, cancer, cataract, and CVDs (Heo et al., 2013; Padayatty et al., 2003). Food sources of vitamin C include citrus fruits, strawberries, melons, tomatoes, broccoli, and peppers. Vitamin D refers to a group of fat-soluble vitamins [vitamin D2 (ergocalciferol), vitamin D3 (cholecalciferol), and alfacalcidol] that are responsible for enhancing intestinal absorption of calcium, iron, magnesium, phosphate, and zinc (Christakos et al., 2011; Fleet and Schoch, 2010). Vitamin D is not a true vitamin, as humans are able to synthesize it with adequate sunlight exposure. Vitamin D may also be ingested in the diet in the form of vitamin D3. People who have low levels of vitamin D are more likely to have diabetes (Pittas et al., 2007). Vitamin D deficiency is also associated with rickets in children, whereas in adults, it leads to secondary hyperparathyroidism, bone loss, osteopenia, osteoporosis, and increased fracture risk (Luboshitzky and Hardoff, 1997). Food sources for vitamin D include fortified milk, saltwater fish, and fish-liver oil. Vitamin E is a group of fat-soluble compounds such as tocopherols and tocotrienols. a-Tocopherol is the most abundant form in foods and is generally used in supplements. Vitamin E is an antioxidant that can scavenge free radicals and is also involved in immune function (van Acker et al., 1993). Vitamin E deficiency is rare and is seen primarily in special situations resulting in fat malabsorption, including cystic fibrosis, chronic cholestatic liver disease, abetalipoproteinemia, and short bowel syndrome (Sitrin et al., 1987). Major dietary sources of vitamin E include salad oils, margarine, legumes, and nuts. Vitamin K is a fat-soluble vitamin that plays an important role in blood clotting and building strong bones (Hart, 1987). Major forms of vitamin K are vitamin K1 and vitamin K2. Vitamin K1, phylloquinone, phytomenadione, or phytonadione, is synthesized by plants and is involved in the production of blood-clotting proteins. Animals may also convert it to vitamin K2. Vitamin K2 (menaquinones or MKs) can prevent bone loss and fractures. Vitamin K deficiency may result in clotting disorders, osteoporosis, vascular calcification, and CVD (Szulc and Meunier, 2001). The major food sources of vitamin K1 are leafy greens and some other vegetables. Vitamin K2 is largely obtained from meats, cheese, and eggs, and is also synthesized by bacteria.

2.3 Essential Minerals Essential minerals are the chemical elements required by living organisms, other than the four elements carbon, hydrogen, nitrogen, and oxygen. There are seven major essential minerals: calcium, potassium, sodium, and magnesium as four major cations and sulfur, phosphorus, and chlorine as three major anions. Trace essential minerals include iron, cobalt, copper, zinc, manganese, selenium, iodine, bromine, and molybdenum.

68

Nanotechnology Applications in Food

Calcium is the most abundant mineral found in the body. About 99% of the calcium in the body is in bones and teeth. The remaining 1% is found in blood and soft tissues. Calcium plays an important role in blood clotting, stimulating muscle contraction, and regulating the permeability of cell membranes. Vitamin D and parathyroid hormone (PTH) help regulate the amount of calcium that is absorbed and that is eliminated by the kidneys (Chung et al., 2009b). By these various biological roles and other mechanisms, calcium is involved in reducing the risk of osteoporosis, hypertension, colon cancer, breast cancer, kidney stones, and lead intoxication (Park et al., 2015). Major food sources of calcium are dairy products, dark-green vegetables, and legumes. Potassium is the major cation of intracellular fluid that maintains electrolyte balance and pH level. Potassium controls the rhythm of heart beats and muscle function (Difrancesco, 2006), and it is also required for glycogen and protein synthesis and the metabolic breakdown of glucose. The kidneys help in maintaining potassium at a normal level. Hyperkalemia, high concentration of Kþ in blood, may occur in people with advanced stages of chronic kidney disease. Extreme hyperkalemia is a medical emergency due to the risk of potentially fatal abnormal heart rhythms. In contrast, severe hypokalemia, low concentration of Kþ in blood, may cause muscle weakness, myalgia, tremor, and muscle cramps (Mandal, 1997). Food sources of potassium are meats, dairy products, fruits, and vegetables. Sodium serves as a major monovalent ion of extracellular fluids. The principal role of sodium is to regulate osmotic pressure, keep the nerves functioning, and maintain acidebase balance. The kidneys control sodium balance by increasing or decreasing sodium levels in the urine (Otsuka et al., 1979). Sodium deficiency may cause decreased blood pressure, nausea, vomiting, dizziness, poor memory and impaired concentration, somnolence, and muscle weakness. A major source of both sodium and chloride is salt. Magnesium is important in protein synthesis, cellular energy production and storage, cell growth and reproduction, DNA and RNA synthesis, and stabilization of mitochondrial membranes (Song and Hunt, 1988). Magnesium also plays a significant role in glucose and insulin metabolism. Signs of magnesium deficiency are numbness, tingling, muscle contractions and cramps, seizures, personality changes, abnormal heart rhythms, and coronary spasms as magnesium levels decrease. Severe deficiency, hypomagnesemia, may lead to poorly controlled diabetes mellitus and chronic malabsorptive problems such as Crohn disease, gluten-sensitive enteropathy, and regional enteritis (Hessov, 1990). Dietary sources of magnesium include nuts (almonds, cashews, and peanuts), vegetables (spinach, broccoli, squash, and green leafy vegetables), whole grains (rice and wheat bran), and milk. Sulfur performs a number of functions in enzyme reactions and protein synthesis (Jacob et al., 2003). It is necessary for formation of collagen, the protein found in connective tissue in our bodies. As sulfate, sulfur is an essential component of heparin, chondroitin, fibrinogen, and taurine. Sulfur deficiency could lead to reduced protein synthesis, cell damage, and joint pain or disease. Sulfur is readily available in protein foodsdmeats, fish, poultry, eggs, milk, and legumes are all good sources. Phosphorus (P) is an essential mineral for proper cell functioning and regulation of calcium levels. It is also an essential component of human bones and teeth. In nature, phosphorus is found as PO4 that is an essential component of nucleic acids such as DNA and RNA, phospholipids, and adenosine triphosphate (ATP). High serum P levels, hyperphosphatemia, may be one of the novel cardiovascular and death risk factor and deficiency of P, hypophosphatemia, may cause muscle dysfunction and weakness, white cell dysfunction, and instability of cell membranes (Mathew et al., 2008). Phosphorus is found in almost every food; therefore, its deficiency is rare. High-phosphorus foods are seeds, cheese, fish, nuts, and beef. Chlorine is the main monovalent anion of extracellular fluids. Chlorine plays a key role in the maintenance of acidebase balance and the transport of oxygen (Chutorian, 1985). Deficiency of chloride is rare; however, the symptoms of chloride deficiency are loss of appetite, muscle weakness, and dehydration. Foods high in chloride are salt, seaweed, olives, and rye. Trace minerals are elements that are needed in smaller amounts, 1e100 mg/day by adults. They support nutrient metabolism and cellular function throughout the body. These minerals are the essential component of enzymes and are the important enzyme cofactors for numerous processes (Martin De Portela, 1982). In the human body, iron can form complexes with oxygen in hemoglobin and myoglobin that carries oxygen throughout the body. It is also a key component of redox enzymes such as peroxidases, catalase, and xanthine oxidase. Iron deficiency anemia symptoms may include extreme fatigue, pale skin, weakness, and shortness of breath (Hadzhieva et al., 2014; Armengou and Davalos, 2002). High-iron foods include clams, liver, sunflower seeds, nuts, beef, lamb, beans, and whole grains. Zinc is an essential component of more than 80 metalloenzymes and it serves as a cofactor in many enzyme systems (Tubek et al., 2008; Park et al., 2004). Chronic diseases associated with zinc are bronchial asthma, rheumatoid arthritis, and AD. Oysters contain more zinc per serving than any other food; however, red meat and poultry provide the majority of zinc.

Strategic Design of Delivery Systems for Nutraceuticals Chapter | 4

69

Manganese functions in the body as an enzyme activator for those enzymes that mediate phosphate group transfer (Pastor and Pastor, 2000). Deficiency symptoms of manganese are ataxia, fainting, hearing loss, and weak tendons and ligaments. Rich sources of manganese include whole grains, nuts, leafy vegetables, and teas. Copper is an essential component of numerous oxidationereduction enzyme systems. Symptoms of copper deficiency include fatigue, anemia, and a decreased number of white blood cells (Ghosh et al., 2014; Nath, 1997). Excellent food sources of copper are leafy greens, including turnip greens, spinach, and mustard greens. Cobalt is an integral component of cyanocobalamin (vitamin B12). Symptoms of cobalt deficiency include fatigue and weakness in the limbs. Good food sources of cobalt include fish, nuts, and green leafy vegetables such as broccoli and spinach. Iodine is an integral component of the thyroid hormones, thyroxine and triiodothyronine. Iodine deficiency leads to fatigue, cold hands and feet, brain fog, and increased need for sleep (Triggiani et al., 2009). Seaweeds such as kelp are the most well-known natural source of iodine. Egg and dairy products can also be good sources. Selenium is an essential component of the enzyme GPx (Perona et al., 1990). Trivalent chromium is an integral component of the glucose tolerance factor. Selenium deficiency affects the immune system and signs of selenium deficiency include fatigue, muscle weakness, muscle wasting, and heart problems. Good food sources of selenium are Brazil nuts, sunflower seeds, fish, shell fish, meat, and poultry. Function, food sources, and related diseases of essential minerals are summarized in Table 4.1.

2.4 Functional Foods One of the definitions of functional foods is “any food or food ingredient that provides a health benefit in addition to the traditional nutrients it contains.” Functional food is a natural or processed food containing biologically active compounds, and it is important in the prevention, management, and treatment of chronic diseases (Abuajah et al., 2015). Soybeans: Lipids and proteins are the two major components in soybean. Soybean oil is low in saturated fat, rich in essential fatty acids, and an excellent source of vitamin E. Increasing evidence demonstrate that soybean can prevent CVD, cancer, and osteoporosis and alleviate menopausal symptoms (Chon, 2013). Oats: Oats have the physiological benefits of reducing hyperglycemia, hyperinsulinemia, and hypercholesterolemia (Rasane et al., 2015). The functional components of oats are b-glucan and phytochemicals named saponin. The soluble fiber b-glucan lowers the levels of total cholesterol and low-density lipoprotein (LDL). Therefore, it reduces the risk of coronary heart disease. Flaxseed: Flaxseed is a leading source of the omega-3 fatty acid, a-linolenic acid, and phenolic compounds known as lignans. Flaxseed also reduces total cholesterol and LDL cholesterol levels, which results in cardioprotective and hypocholesterolemic effect. It also has beneficial renal function. There have been several reports demonstrating that ingestion of flaxseed elicited several hormonal changes associated with reduced breast cancer risk (Kajla et al., 2015). Tomatoes and tomato products: Many tomato products are good sources of potassium, folate, and the vitamins A, C, and E. Tomatoes also contain valuable phytochemicals, including carotenoids and polyphenols, such as the red-pigmented lycopene, b-carotene, a provitamin A compound (Viuda-Martos et al., 2014). Tomato phytochemicals contribute to the reduced risk of CVD and prostate cancer. Lycopene reduces the risk of cancers such as prostate, breast, colon, and lung cancers and prevents heart diseases. Lycopene is a powerful antioxidant and the most efficient quencher of singlet oxygen in biological systems. Leafy greens: Leafy greens are spinach, kale, collard greens, broccoli, broccoli rabe, broccoli sprouts, and arugula. They contain phytochemicals such as carotenoids, sulforaphanes, apigenin, and lutein/zeaxanthin (Znidarcic et al., 2011). Carotenoids act as an antioxidant and they block carcinogens from entering cells. Sulforaphanes and apigenin, a cancer chemopreventive agent, provide heart protection, antioxidant, and anti-inflammatory activities. Lutein reduces blindness in the elderly and zeaxanthin enhances immune function. Garlic: Garlic has numerous physiologically active organosulfur components such as allicin, allylic sulfides. Garlic has been shown to have a modest blood pressureelowering effect in clinical studies. Garlic consumption reduces certain cancer incidences in the stomach and colon (Fukushima et al., 1997). Fish: Functional components of fish are omega-3 fatty acids such as docosahexaenoic acid (DHA) and eicosapentaenoic acid (EPA) that are polyunsaturated fatty acids derived primarily from fish oil (Kaur et al., 2014). Omega-3 fatty acids may play an important role in CVDs. Grape juice and wine: Grape juice and wine, particularly red wine, is a rich source of polyphenols (Gollucke, 2010). Polyphenols include phenolic acids, flavonoids, tannins, lignans, and stilbenes. Polyphenols appear to play a role in

70

Nanotechnology Applications in Food

protecting against CVD. Phytochemicals in wine inhibit LDL oxidation as well as platelet aggregation. Increasing attention has been focused on resveratrol, which is found in high amounts in grape skins and red wines and in lower amounts in grape juice, mulberries, and peanuts. Resveratrol is a type of antibiotic produced by plants to help defend against diseases and it has been used to treat a variety of conditions including inflammation and CVD as an antioxidant.

TABLE 4.1 List of Major or Trace Essential Minerals: Their Functions, Food Sources, and Related Diseases Major Essential Minerals

Major Functions

Dietary Sources

Related Diseases

Calcium (Ca)

Bone and tooth formation

Dark-green vegetables, legumes

Osteoporosis, hypertension, colon cancer, breast cancer

Potassium (K)

Maintaining electrolyte balance and pH level

Meat, dairy products, fruits, vegetables

Muscle cramps, abnormal heart rhythms, tremor

Sodium (Na)

Regulation of osmotic pressure, nerve function, acidebase balance

Salt

Muscle weakness, decreased blood pressure

Magnesium (Mg)

Protein synthesis, cellular energy production and storage, cell growth and reproduction

Nuts, vegetables, whole grains

Muscle contractions and cramps, seizures, personality changes, abnormal heart rhythms

Sulfur (S)

Enzyme reactions and protein synthesis

Meat, fish, poultry, eggs, milk, legumes

Reduced protein synthesis, cell damage, joint pain

Phosphorus (P)

Essential components of bones, teeth, and cartilage

Seeds, cheese, fish, nuts, beef

Muscle dysfunction and weakness, white cell dysfunction

Chloride (Cl)

Transport of oxygen and carbon dioxide in the blood, maintenance of digestive juice pH

Salt, seaweed, olive, rye

Muscle weakness, loss of appetite, dehydration

Trace Essential Minerals

Major Functions

Dietary Sources

Related Diseases

Iron (Fe)

Component of hemoglobin and myoglobin

Clams, liver, sunflower seeds, nuts, beef, lamb, beans, whole grains

Fatigue, pale skin, weakness, shortness of breath

Zinc (Zn)

Component of more than 80 metalloenzymes

Oysters, red meat, poultry

Alzheimer disease, rheumatoid arthritis bronchial asthma

Manganese (Mn)

Enzyme activator/cofactor

Whole grains, nuts, leafy vegetables, tea

Ataxia, fainting, hearing loss, weak tendons and ligaments

Copper (Cu)

Component of numerous oxidationereduction enzyme systems

Leafy greens, including turnip greens, spinach, mustard greens

Fatigue, anemia, decreased number of white blood cells

Cobalt (Co)

Integral component of cyanocobalamin (vitamin B12)

Fish, nuts, green leafy vegetables such as broccoli and spinach

Fatigue, weakness in limbs such as arms and legs

Iodine (I)

Integral component of thyroid hormone

Seaweeds, eggs, dairy products

Fatigue, cold hands and feet, brain fog, increased need for sleep

Selenium (Se)

Component of the enzyme glutathione peroxidase

Brazil nuts, sunflower seeds, fish, shell fish, meat, poultry

Fatigue, muscle weakness, muscle wasting, heart problems

Strategic Design of Delivery Systems for Nutraceuticals Chapter | 4

71

Cranberry: Cranberry is most commonly used for prevention and treatment of urinary tract infections. Cranberry juice inhibits the adherence of Escherichia coli to uroepitheial cells (Ruel et al., 2008). Cranberry contains significant amounts of salicylic acid, which is an important ingredient in aspirin. Salicylic acid can reduce swelling, can prevent blood clots, and has antitumor effects. Tea: Green tea provides antioxidant, anti-inflammatory, antimicrobial, and antimutagenic properties (Cabrera et al., 2006). The active ingredients in green tea are polyphenols that consist of 30% of the total dry weight of fresh tea leaves. Among all tea polyphenols, catechins are the most significant. The four catechins in green tea are epicatechin, epigallocatechin-3-gallate, epigallocatechin, and epicatechin-3-gallate. Traditionally it was used in the management of chronic systemic diseases including cancer. Tea consumption may also reduce the risk of CVD.

2.5 Phytochemicals Phytochemicals are secondary metabolic compounds that are naturally produced by plants (“phyto” means “plant” in Greek) (Kennedy and Wightman, 2011). Plants produce these chemicals to protect themselves from ultraviolet radiation or aggression by pathogens; however, recent research demonstrates that many phytochemicals can also protect humans against diseases. Health benefits may also be provided by more than 25,000 phytonutrients found in plant-based foods such as fruits, vegetables, beans, and grains. Phytochemicals act as substrates for biochemical reactions, cofactors or inhibitors of enzyme reactions, and ligands that agonize or antagonize cell receptors. Possible health benefits of phytochemicals include preventing cancers, coronary heart disease, diabetes, high blood pressure, and microbial/viral infection. Phytochemicals can be grouped by their chemical nature into polyphenols, flavonoids, tannins, phenolic acids, alkaloids, terpenoids, essential oils, and saponins; the dietary phytochemicals are classified in Fig. 4.2. Polyphenols are antioxidant phytochemicals containing more than one phenolic hydroxyl group. Polyphenols are the most abundant antioxidants in diet (Halliwell, 2008). Their natural antioxidant role helps avoid cell oxidation and hence prevents cell aging. Polyphenols can also modulate the activity of a wide range of enzymes and cell receptors. This is crucial for the prevention and treatment of cancers and inflammatory, cardiovascular, and neurodegenerative diseases. The most important food sources are commodities widely consumed in large quantities such as fruits and vegetables, green tea, black tea, red wine, coffee, chocolate, olives, and extra virgin olive oil. Some polyphenols are specific to particular foods (e.g., flavanones in citrus fruits, isoflavones in soya, phloridzin in apples), whereas others such as quercetin are found in all plant products such as fruits, vegetables, cereals, leguminous plants, tea, and wine. There are several families among which the most famous are flavonoids and tannins. Flavonoids (from the Latin word flavus meaning yellow, which is their natural color) are water-soluble polyphenolic molecules that have a common chemical structure. They have the general structure of a 15-carbon skeleton, which consists of two phenyl rings (A and B) and a heterocyclic ring (C). The chemical structures of flavonoids are shown in Fig. 4.3. Many studies have found that flavonoids have antioxidant activity as well as antiallergic, anticancer, anti-inflammatory, and antiviral activities (Siddiqui et al., 2015; Somani et al., 2015). Dietary sources of flavonoids include tea, red wine, fruit, vegetables, and legumes. The flavonoids are divided into six subgroups; flavonols, flavanones, flavones, isoflavones, flavanols, and anthocyanidins (proanthocyanidins and catechins).

Phytochemicals

Polyphenols

Alkaloids

Terpenoids

Essential oils Saponins

Flavonoids Tannins Phenolic acids Stilbenes Lignans

Flavonols Flavones Isoflavones Flavanones Anthocyanidins Flavanols

FIGURE 4.2 Classification of dietary phytochemicals. Phytochemicals have a range of different biochemical and physiological effects.

72

Nanotechnology Applications in Food

(A)

(B)

(C)

(E)

(F)

OH

(D) OH

OH

+

FIGURE 4.3 Chemical structures of flavonoids. Flavonoids can be divided into various classes based on their chemical structures: (A) flavonols, (B) flavones, (C) flavanones (D) flavanols, (E) isoflavones, and (F) anthocyanidins.

Flavonols are a class of flavonoids that have the 3-hydroxyflavone backbone (3-hydroxy-2-phenylchromen-4-one). Flavonols include quercetin, kaempferol, myricetin, and isorhamnetin that are widely distributed in yellow onions, scallions, kale, broccoli, apples, berries, and tea. Flavonols provide anti-inflammatory benefits to cells (Leo and Woodman, 2015). Flavones are a class of flavonoids based on the backbone of 2-phenylchromen-4-one (2-phenyl-1-benzopyran-4-one). Natural flavones include apigenin, luteolin, and tangeretin in parsley, thyme, celery, and hot peppers. Flavones are antioxidants with potential chemo-protective benefits (Woodman and Chan, 2004). Soy flavonoids or isoflavones (also referred to as phytoestrogens) have similar structure to human estrogen. Examples of isoflavones are daidzein, genistein, and glycitein in soybeans, soy foods, and legumes. Isoflavones can protect against osteoporosis and alleviate the symptoms of menopause (Castelo-Branco and Soveral, 2013). Flavanone is a class of flavonoid ketones, many of which occur in nature as glycosides. Flavanones include hesperetin, naringenin, and eriodictyol in citrus fruits and oranges or juices of these fruits. Flavanones can lower the risk of ischemic stroke (Chanet et al., 2012). Anthocyanins are a very large group of redeblue plant pigments (Iwashina, 2015). Many anthocyanins are red at acidic conditions and turn blue at less acid conditions. They are all based on a single basic core structure, the flavylium ion. Examples of anthocyanins are cyanidin, delphinidin, malvidin, pelargonidin, peonidin, and petunidin in red, blue, and purple berries; red and purple grapes; and red wine. Anthocyanidins dilate blood vessels, induce cancer cell death, and improve insulin sensitivity (Vendrame and Klimis-Zacas, 2015). Flavanols (also referred to as flavan-3-ols) are derivatives of flavans that use the 2-phenyl-3,4-dihydro-2H-chromen-3ol skeleton. Monomers of flavanols are catechin, epicatechin, epigallocatechin, epicatechin, and gallate in tea, chocolates, grapes, berries, and apples. Dimers and polymers of flavanols are theaflavins, thearubigins, and proanthocyanidins in tea, chocolates, berries, red grapes, and red wines. Naturally occurring flavanols in cocoa improved the memory and cognitive health of adults by enhancing brain blood flow (Scholey and Owen, 2013). Tannins (commonly referred to as tannic acid) are water-soluble polyphenols that bind proteins and promote the tanning of leather. They are odorless, yellowish or brownish, bitter or astringent tasting organic substances found in seeds, leaves, and fruit skins. Tannin is a polymer of gallic acid molecules and glucose, and it hydrolyzes into glucose and gallic or ellagic acid units. Tannins have demonstrated antioxidant and anticancer properties (Marzouk et al., 2007). It also functions as protecting agents in the urinary tract and the cardiovascular and immune system. Phenolic acids and derivatives are organic compounds containing a carboxylic acid function and a phenolic ring such as benzoic acid derivatives (gallic acid, vanillic acid) and cinnamic acid derivatives (chlorogenic acid or 3-dicafeoylquinique). Gallic acid is found in tea and grape seeds. Coffee contains caffeic acid and chlorogenic acid. Blueberries, kiwis, plums, cherries, and apples contain large amounts of caffeic acid. Red wine and citrus fruits contain cinnamic acid. Phenolic compounds are essential for the growth and reproduction of plants, and studies demonstrate that phenolic acids protect against diseases causing oxidative damage, such as coronary heart disease, stroke, and cancers (Aboul-Enein et al., 2013). Alkaloids (alkali-like) are a group of naturally occurring organic nitrogen-containing bases. Well-known alkaloids include morphine, strychnine, quinine, ephedrine, and nicotine. Caffeine, cocaine, codeine and nicotine are water soluble (with a solubility of 1 g/L), whereas others, including morphine and yohimbine, are highly water soluble (0.1e1 g/L). The alkaloid content in plants is usually a few percent and is inhomogeneous over the plant tissues. Depending on the type

Strategic Design of Delivery Systems for Nutraceuticals Chapter | 4

73

of plants, the maximum concentration is observed in the leaves (black henbane), fruits or seeds (strychnine tree), root (Rauwolfia serpentina), or bark (cinchona). Pure alkaloid extracts are usually bitter, colorless solids, and alkaloids must be extracted from plants before they can be used. After the plants have been dried and crushed, chemical reagents such as alcohol and dilute acids are used to extract the alkaloids. The primary use of alkaloids is in medicine, because they can act quickly on specific areas of the nervous system (Vohora et al., 1984). Alkaloids are the active components of many anesthetics, sedatives, stimulants, relaxants, and tranquilizers. Natural plant alkaloids are also used by the pharmaceutical industry for the development of antimalarial agents (quinine and chloroquinine), anticancer agents (taxol, vinblastine, and vincristine), and agents promoting blood circulation in the brain (vincamine) (Lu et al., 2012). They are taken by mouth and administered by injection. Except under a physician’s supervision, use of alkaloids is dangerous, because most are habit-forming (for example, almost all narcotics are alkaloids) and large doses can be poisonous. Plants containing alkaloids do not feature strongly in herbal medicine, but they have always been important in the allopathic system where dosage is strictly controlled and in homoeopathy where the dose rate is so low as to be harmless. Terpene is any of a class of isomeric hydrocarbons, C10H16, contained chiefly in the essential oils of coniferous plants. Terpenoids are compounds related to terpenes, which may include some oxygen functionality or some rearrangement (Gonzalez-Burgos and Gomez-Serranillos, 2012). Terpenoids are aromatic compounds synthesized by thousands of plants and they contribute the scents and various flavors. Examples of well-known terpenoids are ginkgolide and bilobalide in Ginkgo biloba, cannabinoids in cannabis, and curcuminoids in turmeric and mustard seed. The physiological functions of terpenes are diverse, such as hormones and pigments. Other pharmaceutical functions are under investigation. Essential oils are a concentrated hydrophobic liquid containing volatile aromatic compounds from plant materials (flowers, buds, seeds, leaves, twigs, bark, herbs, wood, fruits, and roots) that are extracted by steam or solvent distillation (Guenther, 1948). In the recent decades, interest in essential oils has revived with the popularity of aromatherapy, an alternative medicine that claims that essential oils and other aromatic compounds have curative effects. Essential oils are slightly enriched in terpenoids (Guenther, 1948), possess antiseptic and antimicrobial activity (Abed, 2007; Bansod, 2008), and enhance the body’s ability to fight off a range of infections. They have anti-inflammatory and antispasmodic effects (chamomile and yarrow), have anticancer effects (Zu et al., 2010), act as expectorants (thyme and hyssop), are used in tonics to enhance the appetite and the digestion and absorption of food (rosemary, fennel, and marjoram), and stimulate the heart and circulatory system (ginger, rosemary, and thyme). Saponins are plant-derived surfactants and they are used as natural emulsifiers and foaming agents in food and beverage, pharmaceutical, ore processing, and other industries. Saponins, a class of plant steroids, are structurally amphipathic glycosides having one or more hydrophilic glycoside moieties combined with a lipophilic triterpene derivative. Ginsenosides are found in ginseng or red ginseng and gypenosides in Gynostemma pentaphyllum. Saponins can also be found in soapberry, soapnut, maples, horse chestnuts, and soapbark. Saponins have many health benefits such as cholesterol reduction, antitumor or antimutagenic activity, antioxidant activity, protective role on bone loss, and stimulation of the immune system (Shi et al., 2004; Shibata, 2001).

2.6 Probiotics and Prebiotics Probiotics, derived from the Greek and meaning “life,” are defined as live microorganisms such as bacteria and yeasts that help your digestive system stay healthy. Probiotic bacteria in fermented foods can strengthen the body’s immune system (Perdigon et al., 1995). Several probiotic strains enhance phagocytic activity and natural killer (NK) cell activity. Several functions of the intestinal microbiota may be beneficially influenced by probiotics. The gastrointestinal (GI) microflora (microbiota) is an extremely complex ecosystem that coexists in equilibrium with the host. In the human body, microbiota constitutes approximately 1014 bacterial cells that is 10 times greater than the number of human cells (Sekirov et al., 2010). Because of the low intragastric pH (1.3), only a small number of bacterial species [0e103 colony-forming unit (CFU)/mL] can survive in the stomach. Predominant species in the stomach are streptococci, staphylococci, lactobacilli, enterobacteriaceae, and yeasts. More than 500 different bacterial species live in the intestinal microbiota. The number of bacteria increases from 0e105 CFU/g in the duodenum to 108 CFU/g in the ileum and 1010e1012 CFU/g in the colon (Zilberstein et al., 2007). A healthy microbiota has been considered to comprise significant numbers of bifidobacteria and lactobacilli (Gritz and Bhandari, 2015). The genera Bifidobacterium and Lactobacillus do not have any known pathogenic species, and they are primarily carbohydrate-fermenting bacteria. A breast-fed baby has a microbiota containing proportionately higher numbers of bifidobacteria, which is believed to be part of the baby’s defense against pathogenic microorganisms and which may be important primers for their immune system (Guaraldi and Salvatori, 2012). The feces of breast-fed babies contain 99% bifidobacteria in their microbiota, whereas in formula-fed babies, there is a much more variable gut microflora.

74

Nanotechnology Applications in Food

Breast milk has oligosaccharides and additional materials, termed bifidus factors, which encourage the viability and activity of bifidobacteria. Lactic acideproducing bacteria, such as bifidobacteria and lactobacilli, are believed to play a significant role in the maintenance of colonization resistance by a variety of mechanisms. The products of carbohydrate fermentation, principally short-chain fatty acids (i.e., acetate, propionate, and butyrate), are also beneficial to host health. Among the short-chain fatty acids, butyrate is a major metabolite from bacterial fermentation of dietary fiber (Pryde et al., 2002). Butyrate can be a food source for intestinal epithelial cells (colonocytes). It induces the proliferation and differentiation of colonic cells and also has many health benefits. Butyrate reduces inflammation and oxidative stress and prevents colorectal cancer by inhibiting the IFN-g/STAT1 (interferon-g/signal transducer and activator of transcription 1) signaling (Klampfer et al., 2004). Yogurt is one of the most familiar sources of probiotics, and fermented cheese or fermented vegetables also may act as a carrier for probiotics. This microbiota is nurtured by oligosaccharides in breast milk, which can be considered to be the original prebiotics. Prebiotics are nondigestible oligosaccharides that act as food for probiotics. Generally, oligosaccharides are combinations of sugars with a different degree of polymerization (Crittenden and Playne, 1996). Examples of prebiotics are fructooligosaccharides, galactooligosaccharides, inulin, lactulose, and maltooligosaccharides. Prebiotics are found in several vegetables and fruits such as asparagus, Jerusalem artichokes, bananas, oatmeal, red wine, honey, maple syrup, and legumes. Prebiotic oligosaccharides can also be manufactured by three different methods: (1) isolation from plant resources, (2) microbiological production or enzymatic synthesis, and (3) enzymatic degradation of polysaccharide. The oligosaccharides in human breast milk are considered the prototypic prebiotics because they facilitate the preferential growth of bifidobacteria and lactobacilli in the colon in exclusively breast-fed babies. Synbiotics, the mixture of probiotics and prebiotics, are often used to take advantage of their synergic effects in application to food products. Prebiotics such as RaftilosesP95 when combined with Bifidobacterium spp., Lactobacillus casei, Lactobacillus acidophilus, and Lactobacillus rhamnosus improve their viabilities at 4 C for 4 weeks of storage. Owing to their immunomodulating properties, probiotics, prebiotics, and synbiotics may constitute valuable tools to treat and prevent immune disorders such as allergy.

2.7 Dietary Supplements Dietary supplements are concentrated sources of nutrients or other substances with a nutritional or physiological effect whose purpose is to supplement the normal diet. Amino acids are biologically important organic compounds composed of amine (eNH2) and carboxylic acid (eCOOH) functional groups, along with a side chain specific to each amino acid. Amino acids can be divided into three categories: essential amino acids, nonessential amino acids, and conditional amino acids. Essential amino acids cannot be made by the body and must be supplied by food. Nonessential amino acids are made by the body from essential amino acids or by the normal breakdown of proteins. Conditional amino acids are usually not essential, except in times of illness, stress, or for someone challenged with a lifelong medical condition. The nine amino acids humans cannot synthesize are phenylalanine, valine, threonine, tryptophan, methionine, leucine, isoleucine, lysine, and histidine. Foods rich in essential amino acids are eggs, meats, soybeans, and quinoa. The deficiency of essential amino acids affects all the organs and many systems including the immune system, the brain and brain function, gut mucosal function and permeability, and kidney function (Ha and Zemel, 2003). Essential fatty acids such as a-linolenic acid (an omega-3 fatty acid) and linoleic acid (an omega-6 fatty acid) are fatty acids that humans and other animals must ingest because the body requires them for good health but cannot synthesize them (Riediger et al., 2009). Major food sources of essential fatty acids are nuts, soybeans, walnuts, cold water fatty fish (salmon, cod, tuna, and blue fish), and flaxseed oil. Essential fatty acids regulate blood pressure, immune responses and liver function, as well as help with blood clotting and breaking down cholesterol (Simopoulos, 1999). They also help you look good, as a diet low in these fatty acids has been shown to create skin problems, including eczema, dandruff, split nails, and brittle hair. Deficiency in these fatty acids leads to a host of symptoms and disorders including abnormalities in the liver and the kidneys, reduced growth rate, decreased immune function, depression, and dryness of the skin. Taurine, or 2-aminoethanesulfonic acid, is a major constituent of bile and the biological roles of taurine are conjugation of bile acids, antioxidation, osmoregulation, membrane stabilization, and modulation of calcium signaling (Chesney, 1985). Mammalian taurine synthesis occurs in the pancreas via the cysteine sulfinic acid pathway. Taurine is regularly used as an ingredient in energy drinks and the major food sources of taurine include fish and meat. Taurine is essential and beneficial for cardiovascular function and the development and function of the skeletal muscles, retina, and central nervous system (CNS).

Strategic Design of Delivery Systems for Nutraceuticals Chapter | 4

75

Chondroitin sulfate is a sulfated glycosaminoglycan composed of a chain of alternating sugars (N-acetylgalactosamine and glucuronic acid) (Sugahara et al., 2003). Chondroitin sulfate is an important structural component of cartilage and provides much of its resistance to compression. The food sources of chondroitin sulfate are bovine (cow), porcine (pork), chicken and shark. Health benefits of chondroitin sulfate include protection against osteoarthritis, heart disease, weak bones (osteoporosis), and high cholesterol levels. Although high intake of dietary supplements may have risks and side effects, other dietary supplements such as carnitine, creatine, and glucosamine are also very beneficial to your health.

3. DESIGN OF DELIVERY VEHICLES FOR NUTRACEUTICALS Delivery systems using nanotechnology such as hydrogel, nanoparticles, or microparticles hold promise in the field of food and pharmaceutical sciences, as they offer tools and nanostructured materials that enhance the efficacy of nutraceuticals or pharmaceutical drugs. For example, as much as 60% of probiotic bacteria cannot survive in the gastric environment and the stability of anthocyanidins, a flavonoid, is dependent on the pH of the GI tract. Therefore, a delivery system for nutraceuticals is greatly needed to protect probiotic bacteria or nutraceuticals from the harsh gastric environment.

3.1 Polymers To protect nutraceuticals from the harsh conditions of the GI tract, polymers can be designed and used as delivery vehicles, allowing nutraceuticals to be incorporated into the polymeric particles. Polymers for the delivery of nutraceuticals can be natural or synthetic polymers consisting of nano- or microparticles (Vorhies and Nemunaitis, 2009).

3.1.1 Biodegradable Polymers Owing to their biocompatibility and biodegradability, biodegradable polymers, both natural and synthetic, have been widely considered for medical applications, especially drug or nutraceutical delivery and tissue engineering (Nicolas et al., 2013). Biodegradable polymers degrade in the body by chemical or biological processes, and most synthetic biodegradable polymers are designed to degrade by hydrolysis. Nutraceuticals encapsulated in biodegradable polymers are released by degradation. It is important that the degradation products should also be biologically acceptable.

3.1.2 Natural Biodegradable Polymers Although natural polymers have several drawbacks such as inadequate biomechanical properties and structural complexity, natural polymers are still attractive primarily because they are commercially available, capable of chemical modifications, potentially degradable and compatible because of to their biological origin, and the host can metabolize and clear the delivery system successfully. Natural polymers include proteins (e.g., collagen, gelatin, albumin, globulin, gliadin, zein, elastin, and casein) and polysaccharides (e.g., carrageenan, alginate, dextran, chitin, chitosan, and hyaluronic acid). 3.1.2.1 Proteins Proteins are biological macromolecules consisting of one or more long chains of amino acids, and they are essential for the human body. In addition to the basic nutrition, many proteins are used as delivery vehicles for nutraceuticals by coating or encapsulating nutraceuticals. As delivery vehicles, various kinds of animal proteins have been investigated, including gelatin (Franz et al., 1998; Payne et al., 2002), collagen (Alex, 1990; Swatschek et al., 2002; Rossler et al., 1995), casein (Latha et al., 2000, 1995), albumin (Sokoloski and Royer, 1984; Tomlinson and Burger, 1985), and whey protein (Beaulieu et al., 2002; Picot, 2004; Rosenberg and Young, 1993), in addition to plant proteins such as soy glycinin (Lazko et al., 2004), zein (Liu et al., 2005), and wheat gliadin (Ezpeleta et al., 1996). Collagen and gelatin have been widely used in drug or nutraceutical delivery and tissue engineering because of their biodegradability and nonimmunogenicity. Collagen can be formulated in different forms such as particles, gels, sponge, and films and can deliver nutraceuticals, antibiotics, and anti-inflammatory drugs (Friess, 1998). Albumin, another protein polymer, also has great potential for drug or nutraceutical delivery because it is nontoxic, nonimmunogenic, biocompatible, and biodegradable. Albumin nanoparticles were already used in the delivery of anticancer drugs such as noscapine and doxorubicin for treating breast cancer or of anti-inflammatory drugs such as SB202190 for inhibiting p38 mitogenactivated protein kinase and the secretion of multiple inflammatory cytokines (Sebak et al., 2010; Bae et al., 2012). Gliadin and zein, a group of prolamin proteins used for storage by plants, have been developed as oral delivery vehicles. Prolamin proteins include gliadin from wheat, hordein from barley, and zein from corn. Antioxidant proteins such

76

Nanotechnology Applications in Food

as catalase and SOD have a great therapeutic potential. However, in vivo application is limited, as antioxidant proteins are easily degraded in vivo by the harsh conditions of the GI tract, which has a low pH and protein-degrading enzymes such as pepsin and trypsin. Antioxidant proteins were encapsulated in either gliadin or zein, and prolamin proteins successfully protect antioxidant proteins from the harsh GI tract environment (Lee et al., 2013). 3.1.2.2 Polysaccharides Polysaccharides are polymeric carbohydrate molecules composed of long chains of monosaccharide units, and their function in living organisms is usually either structure or storage related. Polysaccharides can be obtained from plants (e.g., pectin, inulin, fiber, and starch) and animals (e.g., chitosan, glycogen, and chondroitin sulfate). Polysaccharides are broken down into smaller components by the colonic microflora. Polysaccharide-based delivery systems protect nutraceuticals from the harsh conditions of the GI tract. They are hydrolyzed when they arrive in the colon and the delivery system releases nutraceuticals into colon. The main application of polysaccharide-based delivery systems is to deliver probiotics such as bifidobacteria and lactobacilli.

3.1.3 Synthetic Biodegradable Polymers As nutrient carriers, the use of synthetic biodegradable polymers could increase the efficiency of delivery by sustaining the release of the encapsulated nutraceuticals or bioactive compounds over several days to weeks. In comparison, natural polymers have a relatively short duration of drug release and are in general limited by the use of organic solvents and relatively harsher formulation conditions (Coelho et al., 2010). Synthetic polymers have the advantage over natural polymers in being able to control or sustain the release of the bioactive compounds over several days to weeks (Coelho et al., 2010). However, synthetic polymers could also cause toxicity and chronic inflammation (Coelho et al., 2010). Therefore, when synthetic polymers are used for delivery vehicles, their toxicity and immunogenicity should also be considered. 3.1.3.1 Polymers Based on Ester, Anhydride, and Amide Bonds Synthetic biodegradable polymers include polylactic acid (PLA), polyglycolic acid (PGA), poly(lactic-co-glycolic) acid (PLGA), polyanhydrides, polyorthoesters, and polyamide. Ester-based polymers such as PLA, PGA, and PLGA have been widely used (Makadia and Siegel, 2011). The chemical structures of synthetic biodegradable polymers are shown in Table 4.2. These polymers have many significant applications in the field of biomedical science as drug delivery carriers, resorbable sutures, and artificial-tissue materials (Makadia and Siegel, 2011). PLGA is a copolymer composed of lactic acid and glycolic acid in which a different composition of two monomers determines the polymer properties. PGA composed of glycolic acid only is a hard, tough, and crystalline polymer. PGA has excellent fiber-forming properties but is insoluble in most common polymer solvents that limits its application for drug carriers, as it cannot be made into films, rods, or capsules (Makadia and Siegel, 2011). PLA composed of lactic acid only is a thermoplastic biodegradable polymer and is degraded by hydrolysis. PLGA is used for delivering anti-inflammatory drugs such as dexamethasone and releases drugs over a long period to suppress chronic inflammatory diseases (Makadia and Siegel, 2011). Biocompatible polyanhydrides can be formulated into disks, coatings, microspheres, and tubes for controlled delivery of drugs or nutraceuticals. Nonsteroidal anti-inflammatory drugs (NSAIDs) such as salicylic acid or antibiotics such as ampicillin were encapsulated in polyanhydrides and released as the polymers degrade (Griffin et al., 2011). Polyorthoesters are stable at room temperature under anhydrous conditions. Polyorthoesters release NSAIDs by surface erosion. One advantage of these polymers is that drug release rates from polyorthoester nano-/microspheres can be varied from a few days to months (Engesaeter et al., 1992). Although polyamide is biodegradable, the application of polyamide has been limited by its immunogenicity and poor mechanical properties.

3.1.4 Stimuli-Responsive Polymers: Smart Polymers 3.1.4.1 Temperature-Sensitive Polymers Temperature-sensitive hydrogels are smart gels that experience solegel transition at a certain temperature (Park, 1999; Shim et al., 2007; Wang et al., 2008). Polymeric hydrogels form a three-dimensional loosely cross-linked polymeric network, which absorbs significant amount of water via hydration with higher number of hydrophilic groups. Sol is a stable suspension of colloidal solid particles (0.1e1 mm) or polymers in a liquid. One of the temperature-sensitive hydrogels currently in use is poly(N-isopropylacrylamide) (PNIPAAm). PNIPAAm is soluble in water below the lower critical solution temperature (LCST) when the phase of PNIPAAm changes from a swollen hydrated state to a shrunken dehydrated

Strategic Design of Delivery Systems for Nutraceuticals Chapter | 4

77

TABLE 4.2 Synthetic Biodegradable Polymers for Drug Delivery Application Name

Polyester

PLA

>24

PGA

6e12

PLGA

1e6

Poly[1,6-bis(p-carboxyphenoxy)hexane]

12

Polyanhydrides

Chemical Structure

Degradation Rate (in Months)

Type

R=

Polyorthoesters

3

(POE(70)LA(30)): Copolymer of 70 R1 and 30 R2

R1 = (CH2)10 R2 = (CH2)10(COOCHCH3)2

PGA, polyglycolic acid; PLA, polylactic acid; PLGA, poly(lactic-co-glycolic) acid.

state that is due to the temperature being above LCST (Lutz et al., 2006). At approximately 32 C (LCST for PNIPAAm) the hydrogel changes around 90% of its volume (Plunkett et al., 2006). PNIPAAm is relevant for controlled release of drugs or nutraceuticals based on temperature, which increases in the presence of inflammation. Drugs or nutraceuticals encapsulated in PNIPAAm will be stable above LCST; however, drugs or nutraceuticals will be released at a temperature above LCST (Rejinold et al., 2014). Thermoresponsive polymers with LCST include poly(N-vinylcaprolactam) (PVCL), poly[2-(dimethylamino)ethyl methacrylate] (PDMAEMA), and poly(N,N-diethylacrylamide) (PDEAAm). Thermoresponsive polymers with the upper critical solution temperature (UCST) have an opposite phase transition to LCST hydrogels. UCST hydrogels are soluble and swell in water above the UCST. The hydrogel solution will be cloudy below the UCST. UCST hydrogels include poly(acrylic acid) (PAA) and polyacrylamide (PAAm) or poly(acrylamide-co-butyl methacrylate). Smart hydrogels are responsive to not only temperature but also other conditions such as pH, enzyme, or electrical potential based on the nature of hydrogels (Alexander, 2006; Traitel et al., 2008). 3.1.4.2 pH-Sensitive Polymers Most pH-sensitive polymers have either acidic (carboxylic or sulfonic acid) or basic (ammonium salts) functional groups to response to changes in environmental pH (Alexander, 2006; Traitel et al., 2008). Polyanions used in drug delivery include

78

Nanotechnology Applications in Food

PAA and poly(sulfonic acid) polymers. Acidic functional groups are ionized in neutral pH and alkaline or high pH leading to the polymers dissolving or swelling more because of ionization. On the other hand, polycations such as poly(N,N9diethylaminoethyl methacrylate) (PDEAEM) dissolve or swell more at low pH (Qiu and Park, 2001). Polycationic hydrogels have been used for drug delivery in the stomach as they swell more under acidic conditions in the stomach. This type of hydrogels would be ideal for delivering antibiotics, such as amoxicillin and metronidazole, in the stomach for the treatment of Helicobacter pylori infection. Polyketals, a new family of acid-sensitive polymers, are synthetic polymers with ketal linkages in their backbone. For drug or nutraceutical delivery, polyketals are designed to hydrolyze, after phagocytosis by macrophages, in the acidic environment of the phagosome and enhance the intracellular delivery of phagocytosed therapeutic drugs. Poly(cyclohexane-1,4-diyl acetone dimethylene ketal) (PCADK) microparticles dramatically improved the ability of SOD, an enzyme scavenging superoxide, to remove ROS produced by macrophages (Lee et al., 2007). PK3, another polyketal, efficiently delivered tumor necrosis factor (TNF)-a small interfering RNA (siRNA) in vivo to Kupffer cells and inhibited gene expression in the liver (Lee et al., 2009). Examples of stimuli-responsive polymers are listed in Table 4.3.

TABLE 4.3 Stimuli-Responsive Polymers: Smart Polymers Type

Name

Temperature-sensitive polymers

Poly(Nisopropylacrylamide)

LCST CP at 32 C

Poly(Nvinylcaprolactam)

LCST CP at 34e37 C

Poly(acrylamideacrylic acid)

UCST CP at 30e34 C

Poly(sulfonic acid)

Polyanion: ionized in neutral and alkaline pH

Polyketal (PCADK)

Degradation at acidic pH

pH-Sensitive polymers

Chemical Structure

Property

CP, cloud points; LCST, lower critical solution temperature; PCADK, poly(cyclohexane-1,4-diyl acetone dimethylene ketal); UCST, upper critical solution temperature.

Strategic Design of Delivery Systems for Nutraceuticals Chapter | 4

(A)

Hydrophilic Group

(B)

79

FIGURE 4.4 (A) Micelles and (B) liposomes as nutraceutical delivery vehicles. Micelles composed of amphiphilic molecules are widely used for delivery of hydrophobic drugs or nutraceuticals. The size of most micelles is less than 50 nm and hence they can avoid renal exclusion as well as enhance endothelial cell permeability in solid tumors by passive diffusion. Because of the hydrophobic layer and hydrophilic core, liposomes can load both hydrophobic and hydrophilic drugs or nutraceuticals. Most liposomes are less than 400 nm in size; however, the size varies greatly depending on the number of liposome layers.

Hydrophilic Group

Hydrophobic Group

Hydrophobic Group

Hydrophobic Drug

Hydrophilic Drug

3.2 Micelles Polymer micelles have been widely used to deliver hydrophobic drugs or nutraceuticals. As shown in Fig. 4.4, micelles are composed of a hydrophilic head region in contact with surrounding solvent and the hydrophobic regions in the center. Polymeric micelles have great potential for improving the efficacy of therapeutic drugs associated with CNS diseases, including drugs for AD, which are restricted by the bloodebrain barrier (BBB). Owing to the restrictive nature of the BBB, only 5% out of 7000 drugs developed for treating CNS diseases, such as depression, schizophrenia, and insomnia, showed activity (Pardridge, 2005). Almost all hydrophilic and large-molecular drugs are not active, as they cannot penetrate the BBB. Although only small molecules with high lipid solubility and a low molecular mass (Mr <400e500 Da) can cross the BBB (Pardridge, 2005), extremely hydrophobic drugs are totally insoluble in aqueous body fluids and poorly absorbed through the surface of cells. Micellar drug delivery systems for hydrophobic drugs offer obvious advantages over the free hydrophobic drugs. A highly water-bound barrier resulting from the hydrophilic head region prevents plasma proteins, known as opsonins, from recognizing hydrophobic drugs. Otherwise, opsonins can bind hydrophobic drugs and remove them from the circulation within seconds to minutes through the reticuloendothelial system (RES) (Rios-Doria et al., 2012). Nano-sized micelles (10e200 nm in diameter) retard the rate of body clearance by renal filtration and the RES (Kim et al., 2010). Surface modification of micellar drug delivery systems by specific ligands enhances the targeting efficiency and reduces the toxicity to normal cells (Gullotti and Yeo, 2009). For example, the conjugation of polyethylene glycol (PEG) to a micelle is a key component of increasing the time of a drug in the blood circulation (Gullotti and Yeo, 2009). Folate ligands are also widely used for targeting cancer cells that upregulate the folate receptor by two orders of magnitude, compared to normal tissue. Conjugation of folate ligands to micelles enhances the efficacy of drug delivery to cancer cells and has the potential to improve treatment outcomes (Gullotti and Yeo, 2009). Although micelles are good drug delivery vehicles for hydrophobic drugs, there are still two major obstacles for micelles to be an effective drug delivery system, namely, low drug loading efficiency and low serum stability (Kim et al., 2010). Low serum stability of polymer micelles is due to micelles being diluted by the blood after systemic injection. Micelles become unstable below a critical micelle concentration, which is the lowest concentration of polymer required for micelles to form.

3.3 Liposome A liposome is a lipid bilayer composed of phosphatidylcholine-enriched phospholipids. Liposome may also contain lipid or lipid chains such as cholesterol, sphingolipids, long-chain fatty acids, and phosphatidylethanolamine (van Meer et al., 2008).

80

Nanotechnology Applications in Food

Liposome is a small vesicle of spherical shape with an internal aqueous environment and hydrophobic membrane layer in the middle of the bilayer (van Meer et al., 2008). Table 4.4 is a list of micelles or liposomes that can load a great variety of molecules including small molecules, hydrophobic drugs, therapeutic proteins, antisense nucleotides, siRNA, and oligodeoxynucleotides (van Meer et al., 2008; Wang et al., 2010). As liposomes are cleared by the mononuclear phagocyte system (MPS) in vivo, liposomes can naturally target cells of the MPS, especially macrophages and deliver drugs to MPS with high efficiency (Immordino et al., 2006). Liposomes can be pegylated to enhance their long blood circulating times and their reduced clearance by the RES (Immordino et al., 2006). As a result, pegylation increases the ability of liposomes to deliver drugs. Liposomes can also be conjugated with a targeting compound such as an antibody or a ligand and can target specific type of cells or tissue (Immordino et al., 2006). Liposomes are used for delivering therapeutic drugs for CNS diseases such as brain tumors, ischemia, and brain infection (Blasi et al., 2009; Chakraborty et al., 2009). Therapeutic drugs contained within liposomes can be administrated intravenously, intraventricularly, or intracerebrally. Liposomes are one of the most widely investigated delivery systems because of their low immunogenicity and biocompatibility; however, there are shortcomings such as low stability, rapid removal by RES after intravenous injection, short shelf life, and high-cost to produce (Luisa Corvo et al., 2002).

3.4 Example of Nutraceutical Delivery: Probiotics Probiotics help digestive systems stay healthy by improving microbial balance. However, as much as 60% of probiotic bacteria cannot survive in the gastric environment. Because of the low intragastric pH (1.3), only small number of bacterial species (0e103 CFU/mL) can survive in the stomach. Predominant species in the stomach are streptococci, staphylococci, lactobacilli, enterobacteriaceae, and yeasts. More than 500 different bacterial species live in the intestinal microbiota. More probiotic bacteria can survive in the small intestine and the number of bacteria increases from 0e105 CFU/g in the duodenum to 108 CFU/g in the ileum and 1010e1012 CFU/g in the colon (Zilberstein et al., 2007). Therefore, a delivery system for nutraceuticals is greatly needed to protect probiotic bacteria or nutraceuticals from the harsh gastric environment. Microencapsulation of probiotics can protect biological cells against harsh environments. Probiotic bacteria are entrapped in the gel matrix using different gel forming mechanisms. Probiotic cells can be encapsulated by extrusion, emulsion, and spray drying methods. Extrusion is the oldest common technique for probiotic formulation. For alginate capsule using an extrusion method, cell suspension is prepared by adding probiotic cells to hydrocolloid solution. The cell suspension is passed through the syringe needle to form droplets that are directly dripped into the hardening solution containing cations such as calcium. The alginate polymers in the cell suspension are crosslinked by cations in the hardening solution, which forms alginate capsule. The alginate capsule is separated and dried using an appropriate method. Emulsion is a useful tool to encapsulate lactic acid bacteria. In this method, a dispersed phase that is composed of a small volume of cell and polymer slurry is emulsified into a continuous phase that is a large volume of vegetable oil such as soy oil, sunflower, corn, and light paraffin oil. The gel formation of emulsion is done by different cross-linking methods such as ionic, enzymatic, and interfacial polymerization. Gum arabic and starches tend to form a spherical microparticle during the drying process. In the spray drying method, probiotic cells and the dissolved polymer matrix of gum arabic or starches is prepared. The spray drying method can form microparticles successfully; however, probiotic cells could be damaged during the drying process because of heat generation and physical injury to microparticles. In order to reduce the loss of probiotic cells, the inlet and outlet temperature for spray drying should be optimized and proper cryoprotectant should be used during freeze drying. The delivery of probiotic bacteria is important, as they are thought to influence the immune system. To deliver probiotic bacteria efficiently, appropriate encapsulation techniques and encapsulating materials should be selected. The probiotic bacteria should be released to changes of environmental pH, mechanical stress, temperature, enzymatic activity, time, and osmotic force. During the manufacturing processes, many factors such as heat generation should also be considered to enhance the viability of probiotic bacteria.

4. CONCLUSIONS Chronic inflammatory diseases such as atherosclerosis, cancer, AD, rheumatoid arthritis, asthma, and diabetes can be caused by various factors including oxidative stress, aging, and elevated blood sugar levels. Although not all mechanisms or treatment for inflammatory diseases are clear, it is beneficial to reduce the inflammation that is often associated with

Strategic Design of Delivery Systems for Nutraceuticals Chapter | 4

81

TABLE 4.4 Micelles and Liposomes: Chemical Structure and Drug Delivery Application Type

Name

Micelle

Pluronic

Doxorubicin, cisplatin, carboplatin

Poly(aspartic acid)-PEG

Doxorubicin, lysozyme, adriamycin

Poly(glutamic acid)-PEG

Cisplatin

Liposome

Chemical Structure

Drug Loaded

Lipofectamine (DOPE:DOSPA ¼ 1:3)

DNA, siRNA

DOPE R1=R2=C18:1 R3=

DOSPA R1=R2=C18:1

R3=

DOPE, 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine; DOSPA, 2,3-di-oleyloxy-N-[2(spermine-carboxamido)ethyl]-N,N-dimethyl-1-propanaminium.

disease processes. Anti-inflammatory drugs as well as nutraceuticals can inhibit the inflammation process. Nutraceuticals are foods that provide medical or health benefits including the prevention and treatment of chronic diseases as well as basic nutrition. Nutraceuticals with medicinal value include natural foods with antioxidants or vitamins, essential minerals, functional foods, phytochemicals, prebiotics, and probiotics. To deliver the nutraceuticals efficiently, they should be encapsulated in a safe, biocompatible, target-specific delivery system. As a delivery vehicle, polymers offer unique properties that cannot be attained by any other material. Especially biodegradable and stimuli-responsive polymers have been a useful tool to control drug release rates. Amphiphilic vehicles such as micelles and liposomes have low serum stability; however, micelles and liposomes have also been widely used for delivering small molecules, proteins, antisense nucleotides, and siRNA. Nanotechnology, including the development of biomaterials and design of delivery systems, holds promise for treating inflammatory diseases associated with aging and diverse disease efficiently.

82

Nanotechnology Applications in Food

REFERENCES Abed, K.F., 2007. Antimicrobial activity of essential oils of some medicinal plants from Saudi Arabia. Saudi J. Biol. Sci. 14, 53e60. Aboul-Enein, H.Y., Berczynsk, P., Kruk, I., 2013. Phenolic compounds: the role of redox regulation in neurodegenerative disease and cancer. Mini Rev. Med. Chem. 13, 385e398. Abuajah, C.I., Ogbonna, A.C., Osuji, C.M., 2015. Functional components and medicinal properties of food: a review. J. Food Sci. Technol. 52, 2522e2529. Alex, R.B.R., 1990. Encapsulation of water-soluble drugs by a modified solvent evaporation method. I. Effect of process and formulation variables on drug entrapment. J. Microencaps. 7, 347e355. Alexander, C., 2006. Temperature- and pH-responsive smart polymers for gene delivery. Expert Opin. Drug Deliv. 3, 573e581. Armengou, A., Davalos, A., 2002. A review of the state of research into the role of iron in stroke. J. Nutr. Health Aging 6, 207e208. Atalay, M., Laaksonen, D.E., Khanna, S., Kaliste-Korhonen, E., Hanninen, O., Sen, C.K., 2000. Vitamin E regulates changes in tissue antioxidants induced by fish oil and acute exercise. Med. Sci. Sports Exerc. 32, 601e607. Bae, S., Ma, K., Kim, T.H., Lee, E.S., Oh, K.T., Park, E.S., Lee, K.C., Youn, Y.S., 2012. Doxorubicin-loaded human serum albumin nanoparticles surface-modified with TNF-related apoptosis-inducing ligand and transferrin for targeting multiple tumor types. Biomaterials 33, 1536e1546. Bansod, S.R.M., 2008. Antifungal activity of essential oils from Indian medicinal plants against human pathogenic Aspergillus fumigatus and A. niger. World J. Med. Sci. 3, 81e88. Beaulieu, L., Savoie, L., Paquin, P., Subirade, M., 2002. Elaboration and characterization of whey protein beads by an emulsification/cold gelation process: application for the protection of retinol. Biomacromolecules 3, 239e248. Beisel, W.R., 1982. Single nutrients and immunity. Am. J. Clin. Nutr. 35, 417e468. Blasi, P., Schoubben, A., Giovagnoli, S., Rossi, C., Ricci, M., 2009. Lipid nanoparticles for drug delivery to the brain: in vivo veritas. J. Biomed. Nanotechnol. 5, 344e350. Bolt, H.M., Stewart, J.D., 2012. Antioxidant activity of food constituents: relevance for the risk of chronic human diseases. Arch. Toxicol. 86, 343e344. Bouayed, J., Bohn, T., 2010. Exogenous antioxidantsddouble-edged swords in cellular redox state: health beneficial effects at physiologic doses versus deleterious effects at high doses. Oxid. Med. Cell. Longev. 3, 228e237. Cabrera, C., Artacho, R., Gimenez, R., 2006. Beneficial effects of green teaea review. J. Am. Coll. Nutr. 25, 79e99. Castelo-Branco, C., Soveral, I., 2013. Phytoestrogens and bone health at different reproductive stages. Gynecol. Endocrinol. 29, 735e743. Chakraborty, C., Sarkar, B., Hsu, C.H., Wen, Z.H., Lin, C.S., Shieh, P.C., 2009. Future prospects of nanoparticles on brain targeted drug delivery. J. Neurooncol. 93, 285e286. Chanet, A., Milenkovic, D., Manach, C., Mazur, A., Morand, C., 2012. Citrus flavanones: what is their role in cardiovascular protection? J. Agric. Food Chem. 60, 8809e8822. Chesney, R.W., 1985. Taurine: its biological role and clinical implications. Adv. Pediatr. 32, 1e42. Chon, S.U., 2013. Total polyphenols and bioactivity of seeds and sprouts in several legumes. Curr. Pharm. Des. 19, 6112e6124. Christakos, S., Dhawan, P., Porta, A., Mady, L.J., Seth, T., 2011. Vitamin D and intestinal calcium absorption. Mol. Cell. Endocrinol. 347, 25e29. Chung, H.Y., Cesari, M., Anton, S., Marzetti, E., Giovannini, S., Seo, A.Y., Carter, C., Yu, B.P., Leeuwenburgh, C., 2009a. Molecular inflammation: underpinnings of aging and age-related diseases. Ageing Res. Rev. 8, 18e30. Chung, M., Balk, E.M., Brendel, M., Ip, S., Lau, J., Lee, J., Lichtenstein, A., Patel, K., Raman, G., Tatsioni, A., Terasawa, T., Trikalinos, T.A., 2009b. Vitamin D and calcium: a systematic review of health outcomes. Evid. Rep. Technol. Assess. (Full Rep.) 1e420. Chutorian, A.M., 1985. Dietary chloride deficiency. Pediatr. Neurol. 1, 350. Coelho, J.F., Ferreira, P.C., Alves, P., Cordeiro, R., Fonseca, A.C., Gois, J.R., Gil, M.H., 2010. Drug delivery systems: advanced technologies potentially applicable in personalized treatments. EPMA J. 1, 164e209. Crittenden, R.G., Playne, M.J., 1996. Production, properties and applications of food-grade oligosaccharides. Trends Food Sci. Technol. 71, 353e361. Curry, M., Hazard-Daniel, A., Daniel, H.J., 1999. Nutraceuticals. Science 286, 1854e1855. Difrancesco, D., 2006. Funny channels in the control of cardiac rhythm and mode of action of selective blockers. Pharmacol. Res. 53, 399e406. Engesaeter, L.B., Sudmann, B., Sudmann, E., 1992. Fracture healing in rats inhibited by locally administered indomethacin. Acta Orthop. Scand. 63, 330e333. Ezpeleta, I., Irache, J.M., Stainmesse, S., Chabenat, C., Gueguen, J., Popineau, Y., Orecchioni, A., 1996. Gliadin nanoparticles for the controlled release of all-trans-retinoic acid. Int. J. Pharm. 131, 191e200. Fairfield, K.M., Fletcher, R.H., 2002. Vitamins for chronic disease prevention in adults: scientific review. JAMA 287, 3116e3126. Farwer, S.R., Der Boer, B.C., Haddeman, E., Kivits, G.A., Wiersma, A., Danse, B.H., 1994. The vitamin E nutritional status of rats fed on diets high in fish oil, linseed oil or sunflower seed oil. Br. J. Nutr. 72, 127e145. Ferreira, A.L., Machado, P.E., Matsubara, L.S., 1999. Lipid peroxidation, antioxidant enzymes and glutathione levels in human erythrocytes exposed to colloidal iron hydroxide in vitro. Braz. J. Med. Biol. Res. 32, 689e694. Finaud, J., Lac, G., Filaire, E., 2006. Oxidative stress: relationship with exercise and training. Sports Med. 36, 327e358. Finkel, T., Holbrook, N.J., 2000. Oxidants, oxidative stress and the biology of ageing. Nature 408, 239e247. Fleet, J.C., Schoch, R.D., 2010. Molecular mechanisms for regulation of intestinal calcium absorption by vitamin D and other factors. Crit. Rev. Clin. Lab. Sci. 47, 181e195. Fletcher, R.H., Fairfield, K.M., 2002. Vitamins for chronic disease prevention in adults: clinical applications. JAMA 287, 3127e3129. Flora, S.J., 2007. Role of free radicals and antioxidants in health and disease. Cell. Mol. Biol. (Noisy-le-grand) 53, 1e2.

Strategic Design of Delivery Systems for Nutraceuticals Chapter | 4

83

Franz, J., Pokorova, D., Hampl, J., Dittrich, M., 1998. Adjuvant efficacy of gelatin particles and microparticles. Int. J. Pharm. 168, 153e161. Friess, W., 1998. Collagenebiomaterial for drug delivery. Eur. J. Pharm. Biopharm. 45, 113e136. Fukushima, S., Takada, N., Hori, T., Wanibuchi, H., 1997. Cancer prevention by organosulfur compounds from garlic and onion. J. Cell. Biochem. Suppl. 27, 100e105. Ghosh, A., Trivedi, P.P., Timbalia, S.A., Griffin, A.T., Rahn, J.J., Chan, S.S., Gohil, V.M., 2014. Copper supplementation restores cytochrome c oxidase assembly defect in a mitochondrial disease model of COA6 deficiency. Hum. Mol. Genet. 23, 3596e3606. Gollucke, A.P., 2010. Recent applications of grape polyphenols in foods, beverages and supplements. Recent Pat. Food Nutr. Agric. 2, 105e109. Gonzalez-Burgos, E., Gomez-Serranillos, M.P., 2012. Terpene compounds in nature: a review of their potential antioxidant activity. Curr. Med. Chem. 19, 5319e5341. Greenwald, R.A., 1990. Superoxide dismutase and catalase as therapeutic agents for human diseases. A critical review. Free Radic. Biol. Med. 8, 201e209. Griffin, J., Delgado-Rivera, R., Meiners, S., Uhrich, K.E., 2011. Salicylic acid-derived poly(anhydride-ester) electrospun fibers designed for regenerating the peripheral nervous system. J. Biomed. Mater. Res. A 97, 230e242. Gritz, E.C., Bhandari, V., 2015. The human neonatal gut microbiome: a brief review. Front. Pediatr. 3, 17. Grivennikov, S.I., Greten, F.R., Karin, M., 2010. Immunity, inflammation, and cancer. Cell 140, 883e899. Guaraldi, F., Salvatori, G., 2012. Effect of breast and formula feeding on gut microbiota shaping in newborns. Front. Cell. Infect. Microbiol. 2, 94. Guenther, E., 1948. The Essential Oils. D. Van Nostrand Company, Inc., New York. Guilland, J.C., 2013. Vitamin B1 (thiamine). Rev. Prat. 63, 1074e1075, 1077e8. Gulcin, I., 2012. Antioxidant activity of food constituents: an overview. Arch. Toxicol. 86, 345e391. Gullotti, E., Yeo, Y., 2009. Extracellularly activated nanocarriers: a new paradigm of tumor targeted drug delivery. Mol. Pharm. 6, 1041e1051. Ha, E., Zemel, M.B., 2003. Functional properties of whey, whey components, and essential amino acids: mechanisms underlying health benefits for active people (review). J. Nutr. Biochem. 14, 251e258. Hadzhieva, M., Kirches, E., Mawrin, C., 2014. Review: iron metabolism and the role of iron in neurodegenerative disorders. Neuropathol. Appl. Neurobiol. 40, 240e257. Halliwell, B., 2008. Are polyphenols antioxidants or pro-oxidants? What do we learn from cell culture and in vivo studies? Arch. Biochem. Biophys. 476, 107e112. Hardy, G., 2000. Nutraceuticals and functional foods: introduction and meaning. Nutrition 16, 688e689. Hart, J.E., 1987. Vitamin K-dependent blood clotting changes in female rats treated with oestrogens. Thromb. Haemost. 57, 273e277. Heo, J.H., Hyon, L., Lee, K.M., 2013. The possible role of antioxidant vitamin C in Alzheimer’s disease treatment and prevention. Am. J. Alzheimers Dis. Other Demen. 28, 120e125. Hessov, I., 1990. Magnesium deficiency in Crohn’s disease. Clin. Nutr. 9, 297e298. Ho, J.W., Cheung, M.W., Yu, V.W., 2012. Active phytochemicals from Chinese herbs as therapeutic agents for the heart. Cardiovasc. Hematol. Agents Med. Chem. 10, 251e255. Hughes, R.E., 1978. Fruit flavonoids: some nutritional implications. J. Hum. Nutr. 32, 47e52. Hultqvist, M., Olsson, L.M., Gelderman, K.A., Holmdahl, R., 2009. The protective role of ROS in autoimmune disease. Trends Immunol. 30, 201e208. Immordino, M.L., Dosio, F., Cattel, L., 2006. Stealth liposomes: review of the basic science, rationale, and clinical applications, existing and potential. Int. J. Nanomed. 1, 297e315. Iwashina, T., 2015. Contribution to flower colors of flavonoids including anthocyanins: a review. Nat. Prod. Commun. 10, 529e544. Jacob, C., Giles, G.I., Giles, N.M., Sies, H., 2003. Sulfur and selenium: the role of oxidation state in protein structure and function. Angew. Chem. Int. Ed. Engl. 42, 4742e4758. Jaimes, E.A., Demaster, E.G., Tian, R.X., Raij, L., 2004. Stable compounds of cigarette smoke induce endothelial superoxide anion production via NADPH oxidase activation. Arterioscler. Thromb. Vasc. Biol. 24, 1031e1036. Kajla, P., Sharma, A., Sood, D.R., 2015. Flaxseed-a potential functional food source. J. Food Sci. Technol. 52, 1857e1871. Kaur, N., Chugh, V., Gupta, A.K., 2014. Essential fatty acids as functional components of foodsea review. J. Food Sci. Technol. 51, 2289e2303. Kennedy, D.O., Wightman, E.L., 2011. Herbal extracts and phytochemicals: plant secondary metabolites and the enhancement of human brain function. Adv. Nutr. 2, 32e50. Kim, S., Shi, Y., Kim, J.Y., Park, K., Cheng, J.X., 2010. Overcoming the barriers in micellar drug delivery: loading efficiency, in vivo stability, and micelle-cell interaction. Expert Opin. Drug Deliv. 7, 49e62. Klampfer, L., Huang, J., Swaby, L.A., Augenlicht, L., 2004. Requirement of histone deacetylase activity for signaling by STAT1. J. Biol. Chem. 279, 30358e30368. Kolluru, G.K., Bir, S.C., Kevil, C.G., 2012. Endothelial dysfunction and diabetes: effects on angiogenesis, vascular remodeling, and wound healing. Int. J. Vasc. Med. 2012, 918267. Kumar, J., Garg, G., Sundaramoorthy, E., Prasad, P.V., Karthikeyan, G., Ramakrishnan, L., Ghosh, S., Sengupta, S., 2009. Vitamin B12 deficiency is associated with coronary artery disease in an Indian population. Clin. Chem. Lab. Med. 47, 334e338. Lalles, J.P., Lacan, D., David, J.C., 2011. A melon pulp concentrate rich in superoxide dismutase reduces stress proteins along the gastrointestinal tract of pigs. Nutrition 27, 358e363. Latha, M.S., Rathinam, K., Mohanan, P., Jayakrishnan, A., 1995. Bioavailability of theophylline from glutaraldehyde cross-linked casein microspheres in rabbits following oral administration. J. Control. Release 34, 1e7.

84

Nanotechnology Applications in Food

Latha, M.S., Lal, A.V., Kumary, T.V., Sreekumar, R., Jayakrishnan, A., 2000. Progesterone release from glutaraldehyde cross-linked casein microspheres: in vitro studies and in vivo response in rabbits. Contraception 61, 329e334. Lazko, J., Popineau, Y., Legrand, J., 2004. Soy glycinin microcapsules by simple coacervation method. Colloids Surf. B Biointerfaces 37, 1e8. Lee, S., Alwahab, N.S., Moazzam, Z.M., 2013. Zein-based oral drug delivery system targeting activated macrophages. Int. J. Pharm. 454, 388e393. Lee, S., Yang, S.C., Heffernan, M.J., Taylor, W.R., Murthy, N., 2007. Polyketal microparticles: a new delivery vehicle for superoxide dismutase. Bioconjug. Chem. 18, 4e7. Lee, S., Yang, S.C., Kao, C.Y., Pierce, R.H., Murthy, N., 2009. Solid polymeric microparticles enhance the delivery of siRNA to macrophages in vivo. Nucleic Acids Res. 37, e145. Leo, C.H., Woodman, O.L., 2015. Flavonols in the prevention of diabetes-induced vascular dysfunction. J. Cardiovasc. Pharmacol. 65, 532e544. Liu, X., Sun, Q., Wang, H., Zhang, L., Wang, J.Y., 2005. Microspheres of corn protein, zein, for an ivermectin drug delivery system. Biomaterials 26, 109e115. Loprasert, S., Vattanaviboon, P., Praituan, W., Chamnongpol, S., Mongkolsuk, S., 1996. Regulation of the oxidative stress protective enzymes, catalase and superoxide dismutase in Xanthomonasea review. Gene 179, 33e37. Lu, H., Ouyang, W., Huang, C., 2006a. Inflammation, a key event in cancer development. Mol. Cancer Res. 4, 221e233. Lu, J.J., Bao, J.L., Chen, X.P., Huang, M., Wang, Y.T., 2012. Alkaloids isolated from natural herbs as the anticancer agents. Evid. Based Complement. Altern. Med. 2012, 485042. Lu, Y., Zhang, C., Bucheli, P., Wei, D., 2006b. Citrus flavonoids in fruit and traditional Chinese medicinal food ingredients in China. Plant Foods Hum. Nutr. 61, 57e65. Luboshitzky, R., Hardoff, R., 1997. Recovery from metabolic bone disease in a girl with vitamin D deficiency rickets associated with primary hyperparathyroidism. J. Pediatr. Endocrinol. Metab. 10, 237e241. Luisa Corvo, M., Jorge, J.C., Van’t Hof, R., Cruz, M.E., Crommelin, D.J., Storm, G., 2002. Superoxide dismutase entrapped in long-circulating liposomes: formulation design and therapeutic activity in rat adjuvant arthritis. Biochim. Biophys. Acta 1564, 227e236. Lutz, J.F., Akdemir, O., Hoth, A., 2006. Point by point comparison of two thermosensitive polymers exhibiting a similar LCST: is the age of poly(NIPAM) over? J. Am. Chem. Soc. 128, 13046e13047. Mahalle, N., Kulkarni, M.V., Garg, M.K., Naik, S.S., 2013. Vitamin B12 deficiency and hyperhomocysteinemia as correlates of cardiovascular risk factors in Indian subjects with coronary artery disease. J. Cardiol. 61, 289e294. Makadia, H.K., Siegel, S.J., 2011. Poly lactic-co-glycolic acid (PLGA) as biodegradable controlled drug delivery carrier. Polymers (Basel) 3, 1377e1397. Mandal, A.K., 1997. Hypokalemia and hyperkalemia. Med. Clin. North Am. 81, 611e639. Martin De Portela, M.L., 1982. Review of present knowledge on the evaluation of the nutritional status of mineral elements. II. Trace elements. Arch. Latinoam. Nutr. 32, 439e449. Marzouk, M.S., Moharram, F.A., Mohamed, M.A., Gamal-Eldeen, A.M., Aboutabl, E.A., 2007. Anticancer and antioxidant tannins from Pimenta dioica leaves. Z. Naturforsch. C 62, 526e536. Mates, J.M., 2000. Effects of antioxidant enzymes in the molecular control of reactive oxygen species toxicology. Toxicology 153, 83e104. Mathew, S., Tustison, K.S., Sugatani, T., Chaudhary, L.R., Rifas, L., Hruska, K.A., 2008. The mechanism of phosphorus as a cardiovascular risk factor in CKD. J. Am. Soc. Nephrol. 19, 1092e1105. Miller, H.E., Rigelhof, F., Marquart, L., Prakash, A., Kanter, M., 2000. Antioxidant content of whole grain breakfast cereals, fruits and vegetables. J. Am. Coll. Nutr. 19, 312Se319S. Mulholland, C.W., Elwood, P.C., Davis, A., Thurnham, D.I., Kennedy, O., Coulter, J., Fehily, A., Strain, J.J., 1999. Antioxidant enzymes, inflammatory indices and lifestyle factors in older men: a cohort analysis. QJM 92, 579e585. Murthy, M.R., Reid 3rd, T.J., Sicignano, A., Tanaka, N., Rossmann, M.G., 1981. Structure of beef liver catalase. J. Mol. Biol. 152, 465e499. Nath, R., 1997. Copper deficiency and heart disease: molecular basis, recent advances and current concepts. Int. J. Biochem. Cell Biol. 29, 1245e1254. Nicolas, J., Mura, S., Brambilla, D., Mackiewicz, N., Couvreur, P., 2013. Design, functionalization strategies and biomedical applications of targeted biodegradable/biocompatible polymer-based nanocarriers for drug delivery. Chem. Soc. Rev. 42, 1147e1235. Otsuka, Y., Carretero, O.A., Albertini, R., Binia, A., 1979. Angiotensin and sodium balance: their role in chronic two-kidney Goldblatt hypertension. Hypertension 1, 389e396. Padayatty, S.J., Katz, A., Wang, Y., Eck, P., Kwon, O., Lee, J.H., Chen, S., Corpe, C., Dutta, A., Dutta, S.K., Levine, M., 2003. Vitamin C as an antioxidant: evaluation of its role in disease prevention. J. Am. Coll. Nutr. 22, 18e35. Padh, H., 1991. Vitamin C: newer insights into its biochemical functions. Nutr. Rev. 49, 65e70. Palace, V.P., Khaper, N., Qin, Q., Singal, P.K., 1999. Antioxidant potentials of vitamin A and carotenoids and their relevance to heart disease. Free Radic. Biol. Med. 26, 746e761. Pardridge, W.M., 2005. The bloodebrain barrier: bottleneck in brain drug development. NeuroRx 2, 3e14. Park, J.S., Choi, S.B., Rhee, Y., Chung, J.W., Choi, E.Y., Kim, D.W., 2015. Parathyroid hormone, calcium, and sodium bridging between osteoporosis and hypertension in postmenopausal korean women. Calcif. Tissue Int. 96, 417e429. Park, S.Y., Birkhold, S.G., Kubena, L.F., Nisbet, D.J., Ricke, S.C., 2004. Review on the role of dietary zinc in poultry nutrition, immunity, and reproduction. Biol. Trace Elem. Res. 101, 147e163. Park, T.G., 1999. Temperature modulated protein release from pH/temperature-sensitive hydrogels. Biomaterials 20, 517e521. Pastor, T.J., Pastor, F.T., 2000. The role of manganese(IV) compounds as oxidantsea review. Talanta 52, 959e970.

Strategic Design of Delivery Systems for Nutraceuticals Chapter | 4

85

Payne, R.G., Yaszemski, M.J., Yasko, A.W., Mikos, A.G., 2002. Development of an injectable, in situ crosslinkable, degradable polymeric carrier for osteogenic cell populations. Part 1. Encapsulation of marrow stromal osteoblasts in surface crosslinked gelatin microparticles. Biomaterials 23, 4359e4371. Perdigon, G., Alvarez, S., Rachid, M., Aguero, G., Gobbato, N., 1995. Immune system stimulation by probiotics. J. Dairy Sci. 78, 1597e1606. Perona, G., Schiavon, R., Guidi, G.C., Veneri, D., Minuz, P., 1990. Selenium dependent glutathione peroxidase: a physiological regulatory system for platelet function. Thromb. Haemost. 64, 312e318. Pham-Huy, L.A., He, H., Pham-Huy, C., 2008. Free radicals, antioxidants in disease and health. Int. J. Biomed. Sci. 4, 89e96. Picot, A.L.C., 2004. Encapsulation of bifidobacteria in whey protein-based microcapsules and survival in simulated gastrointestinal conditions and in yoghurt. Int. Dairy J. 14, 505e515. Pittas, A.G., Lau, J., Hu, F.B., Dawson-Hughes, B., 2007. The role of vitamin D and calcium in type 2 diabetes. A systematic review and meta-analysis. J. Clin. Endocrinol. Metab. 92, 2017e2029. Plunkett, K.N., Zhu, X., Moore, J.S., Leckband, D.E., 2006. PNIPAM chain collapse depends on the molecular weight and grafting density. Langmuir 22, 4259e4266. Pryde, S.E., Duncan, S.H., Hold, G.L., Stewart, C.S., Flint, H.J., 2002. The microbiology of butyrate formation in the human colon. FEMS Microbiol. Lett. 217, 133e139. Qiu, Y., Park, K., 2001. Environment-sensitive hydrogels for drug delivery. Adv. Drug Deliv. Rev. 53, 321e339. Rasane, P., Jha, A., Sabikhi, L., Kumar, A., Unnikrishnan, V.S., 2015. Nutritional advantages of oats and opportunities for its processing as value added foods - a review. J. Food Sci. Technol. 52, 662e675. Reid 3rd, T.J., Murthy, M.R., Sicignano, A., Tanaka, N., Musick, W.D., Rossmann, M.G., 1981. Structure and heme environment of beef liver catalase at 2.5 A resolution. Proc. Natl. Acad. Sci. U.S.A. 78, 4767e4771. Rejinold, N.S., Baby, T., Chennazhi, K.P., Jayakumar, R., 2014. Dual drug encapsulated thermo-sensitive fibrinogen-graft-poly(N-isopropyl acrylamide) nanogels for breast cancer therapy. Colloids Surf. B Biointerfaces 114, 209e217. Riediger, N.D., Othman, R.A., Suh, M., Moghadasian, M.H., 2009. A systemic review of the roles of n-3 fatty acids in health and disease. J. Am. Diet. Assoc. 109, 668e679. Rios-Doria, J., Carie, A., Costich, T., Burke, B., Skaff, H., Panicucci, R., Sill, K., 2012. A versatile polymer micelle drug delivery system for encapsulation and in vivo stabilization of hydrophobic anticancer drugs. J. Drug Deliv. 2012, 951741. Rosenberg, M., Young, S.L., 1993. Whey proteins as microencapsulating agents. Microencapsulation of anhydrous milkfatdstructure evaluation. Food Struct. 12, 31e41. Rossler, B., Kreuter, J., Scherer, D., 1995. Collagen microparticles: preparation and properties. J. Microencapsul. 12, 49e57. Roudebush, P., Schoenherr, W.D., Delaney, S.J., 2008. An evidence-based review of the use of nutraceuticals and dietary supplementation for the management of obese and overweight pets. J. Am. Vet. Med. Assoc. 232, 1646e1655. Ruel, G., Pomerleau, S., Couture, P., Lemieux, S., Lamarche, B., Couillard, C., 2008. Low-calorie cranberry juice supplementation reduces plasma oxidized LDL and cell adhesion molecule concentrations in men. Br. J. Nutr. 99, 352e359. Scholey, A., Owen, L., 2013. Effects of chocolate on cognitive function and mood: a systematic review. Nutr. Rev. 71, 665e681. Sebak, S., Mirzaei, M., Malhotra, M., Kulamarva, A., Prakash, S., 2010. Human serum albumin nanoparticles as an efficient noscapine drug delivery system for potential use in breast cancer: preparation and in vitro analysis. Int. J. Nanomed. 5, 525e532. Sekirov, I., Russell, S.L., Antunes, L.C., Finlay, B.B., 2010. Gut microbiota in health and disease. Physiol. Rev. 90, 859e904. Shi, J., Arunasalam, K., Yeung, D., Kakuda, Y., Mittal, G., Jiang, Y., 2004. Saponins from edible legumes: chemistry, processing, and health benefits. J. Med. Food 7, 67e78. Shibata, S., 2001. Chemistry and cancer preventing activities of ginseng saponins and some related triterpenoid compounds. J. Korean Med. Sci. (Suppl. 16), S28eS37. Shim, W.S., Kim, J.H., Kim, K., Kim, Y.S., Park, R.W., Kim, I.S., Kwon, I.C., Lee, D.S., 2007. pH- and temperature-sensitive, injectable, biodegradable block copolymer hydrogels as carriers for paclitaxel. Int. J. Pharm. 331, 11e18. Siddiqui, J.A., Singh, A., Chagtoo, M., Singh, N., Godbole, M.M., Chakravarti, B., 2015. Phytochemicals for breast cancer therapy: current status and future implications. Curr. Cancer Drug Targets 15, 116e135. Simopoulos, A.P., 1999. Essential fatty acids in health and chronic disease. Am. J. Clin. Nutr. 70, 560Se569S. Sitrin, M.D., Lieberman, F., Jensen, W.E., Noronha, A., Milburn, C., Addington, W., 1987. Vitamin E deficiency and neurologic disease in adults with cystic fibrosis. Ann. Intern Med. 107, 51e54. Sokoloski, T.D., Royer, G.P., 1984. Microspheres and Drug Therapy. Elsevier. Somani, S.J., Modi, K.P., Majumdar, A.S., Sadarani, B.N., 2015. Phytochemicals and their potential usefulness in inflammatory bowel disease. Phytother. Res. 29, 339e350. Song, M.K., Hunt, J.A., 1988. Specific role of manganese and magnesium on RNA synthesis in rabbit bone marrow erythroid cell nuclei. Biol. Trace Elem. Res. 16, 203e219. Sugahara, K., Mikami, T., Uyama, T., Mizuguchi, S., Nomura, K., Kitagawa, H., 2003. Recent advances in the structural biology of chondroitin sulfate and dermatan sulfate. Curr. Opin. Struct. Biol. 13, 612e620. Svenson, J., 2007. Neurologic disease and vitamin B12 deficiency. Am. J. Emerg. Med. 25 (987), e3ee4. Swatschek, D., Schatton, W., Muller, W., Kreuter, J., 2002. Microparticles derived from marine sponge collagen (SCMPs): preparation, characterization and suitability for dermal delivery of all-trans retinol. Eur. J. Pharm. Biopharm. 54, 125e133.

86

Nanotechnology Applications in Food

Szulc, P., Meunier, P.J., 2001. Is vitamin K deficiency a risk factor for osteoporosis in Crohn’s disease? Lancet 357, 1995e1996. Tomlinson, E., Burger, J.J., 1985. Incorporation of water-soluble drugs in albumin microspheres. Methods Enzymol. 112, 27e43. Traitel, T., Goldbart, R., Kost, J., 2008. Smart polymers for responsive drug-delivery systems. J. Biomater. Sci. Polym. Ed. 19, 755e767. Triggiani, V., Tafaro, E., Giagulli, V.A., Sabba, C., Resta, F., Licchelli, B., Guastamacchia, E., 2009. Role of iodine, selenium and other micronutrients in thyroid function and disorders. Endocr. Metab. Immune Disord. Drug Targets 9, 277e294. Tubek, S., Grzanka, P., Tubek, I., 2008. Role of zinc in hemostasis: a review. Biol. Trace Elem. Res. 121, 1e8. van Acker, S.A., Koymans, L.M., Bast, A., 1993. Molecular pharmacology of vitamin E: structural aspects of antioxidant activity. Free Radic. Biol. Med. 15, 311e328. van Jaarsveld, P.J., Faber, M., Tanumihardjo, S.A., Nestel, P., Lombard, C.J., Benade, A.J., 2005. Beta-carotene-rich orange-fleshed sweet potato improves the vitamin A status of primary school children assessed with the modified-relative-dose-response test. Am. J. Clin. Nutr. 81, 1080e1087. van Meer, G., Voelker, D.R., Feigenson, G.W., 2008. Membrane lipids: where they are and how they behave. Nat. Rev. Mol. Cell Biol. 9, 112e124. Varker, K.A., Ansel, A., Aukerman, G., Carson 3rd, W.E., 2012. Review of complementary and alternative medicine and selected nutraceuticals: background for a pilot study on nutrigenomic intervention in patients with advanced cancer. Altern. Ther. Health Med. 18, 26e34. Vendrame, S., Klimis-Zacas, D., 2015. Anti-inflammatory effect of anthocyanins via modulation of nuclear factor-kappaB and mitogen-activated protein kinase signaling cascades. Nutr. Rev. 73, 348e358. Viuda-Martos, M., Sanchez-Zapata, E., Sayas-Barbera, E., Sendra, E., Perez-Alvarez, J.A., Fernandez-Lopez, J., 2014. Tomato and tomato byproducts. Human health benefits of lycopene and its application to meat products: a review. Crit. Rev. Food Sci. Nutr. 54, 1032e1049. Vohora, S.B., Kumar, I., Khan, M.S., 1984. Effect of alkaloids of Solanum melongena on the central nervous system. J. Ethnopharmacol. 11, 331e336. Vorhies, J.S., Nemunaitis, J.J., 2009. Synthetic vs. natural/biodegradable polymers for delivery of shRNA-based cancer therapies. Methods Mol. Biol. 480, 11e29. Wang, B., Xu, X.D., Wang, Z.C., Cheng, S.X., Zhang, X.Z., Zhuo, R.X., 2008. Synthesis and properties of pH and temperature sensitive P(NIPAAm-coDMAEMA) hydrogels. Colloids Surf. B Biointerfaces 64, 34e41. Wang, J., Lu, Z., Wientjes, M.G., Au, J.L., 2010. Delivery of siRNA therapeutics: barriers and carriers. AAPS J. 12, 492e503. Wannamethee, S.G., Lowe, G.D., Rumley, A., Bruckdorfer, K.R., Whincup, P.H., 2006. Associations of vitamin C status, fruit and vegetable intakes, and markers of inflammation and hemostasis. Am. J. Clin. Nutr. 83, 567e574. Quiz 726e7. Watzl, B., Bub, A., 2003. Fruit and vegetable concentrate or vitamin supplement? J. Nutr. 133, 3725. Author reply 3726. Wei, Y.H., 1998. Oxidative stress and mitochondrial DNA mutations in human aging. Proc. Soc. Exp. Biol. Med. 217, 53e63. Woodman, O.L., Chan, E., 2004. Vascular and anti-oxidant actions of flavonols and flavones. Clin. Exp. Pharmacol. Physiol. 31, 786e790. Wu, D., Cederbaum, A.I., 2003. Alcohol, oxidative stress, and free radical damage. Alcohol Res. Health 27, 277e284. Yan, J., Fu, Q., Cheng, L., Zhai, M., Wu, W., Huang, L., Du, G., 2014. Inflammatory response in Parkinson’s disease (review). Mol. Med. Rep. 10, 2223e2233. Zeisel, S.H., 1999. Regulation of “nutraceuticals”. Science 285, 1853e1855. Zhao, J., 2007. Nutraceuticals, nutritional therapy, phytonutrients, and phytotherapy for improvement of human health: a perspective on plant biotechnology application. Recent Pat. Biotechnol. 1, 75e97. Zilberstein, B., Quintanilha, A.G., Santos, M.A., Pajecki, D., Moura, E.G., Alves, P.R., Maluf Filho, F., De Souza, J.A., Gama-Rodrigues, J., 2007. Digestive tract microbiota in healthy volunteers. Clinics (Sao Paulo) 62, 47e54. Znidarcic, D., Ban, D., Sircelj, H., 2011. Carotenoid and chlorophyll composition of commonly consumed leafy vegetables in Mediterranean countries. Food Chem. 129, 1164e1168. Zu, Y., Yu, H., Liang, L., Fu, Y., Efferth, T., Liu, X., Wu, N., 2010. Activities of ten essential oils towards Propionibacterium acnes and PC-3, A-549 and MCF-7 cancer cells. Molecules 15, 3200e3210.