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Nanomaterials and the skin
Abstracts / Nanomedicine: Nanotechnology, Biology, and Medicine 2 (2006) 269–312 97
Sunday, September 10th (3:40) Concurrent Symposium XVIII: Oncology and Experimental Nanomedicine
Dissecting the disruptive nature of nanomedicine Best RG, Khushf G, Department of Obstetrics and Gynecology, University of South Carolina, Columbia, South Carolina, USA, Department of Philosophy and Center for Bioethics, University of South Carolina, Columbia, South Carolina, USA Nanotechnology is often described as a disruptive technology. Presumably this means that some people, companies or even industries will be undermined by the technological developments made possible by nanotechnology. With the development of specific objectives for nanomedicine through the National Nanotechnology Initiative, NIH Nanomedicine Roadmap and others, the application of nanotechnologies in health care is increasingly seen as integral to our visions of future health care. Within the context of the health care system, we consider two ways that nanotechnologies generally, and nanomedicine in particular, might be disruptive. One which considers primarily business and economic aspects of disruption (low-end disruptions), and the other which considers the broader societal issues of disruption at the level of the health care system (high-end disruptions). While the former is primarily dependent on technological developments, the latter is fundamentally dependent upon societal developments. In contrast to low end disruptions, which are extremely difficult to anticipate, high end disruptions can be anticipated. In fact, they depend on explicit recognition of the joint technology/grid systems. Since these are accessible to upstream ethical and regulatory reflection in a way that low end disruptions are not, these should be the focus of attention when we consider the disruptive character of nanomedicine. We seek to provide the foundation for a more comprehensive analysis of the promises and challenges associated with the development of nanomedicine in order to more fully anticipate and address the ethical and societal challenges that lie ahead. Robert G. Best, Ph.D, FACMG, University of South Carolina, School of Medicine. BS (biochemistry) Lehigh University 1981; MS (toxicology) 1983 and Ph.D (genetics and toxicology) 1987 North Carolina State University, Raleigh. Diplomate of the American Board of Medical Genetics (PhD Medical Genetics & Clinical Cytogenetics) 1991. Founding Fellow — American College of Medical Genetics & American Academy of Nanomedicine. Current: Professor, Dept of Ob/Gyn, and Director, Div. Genetics. Research interests: ethical, legal, and social aspects of nanotechnology and nanomedicine; complex genetic traits; birth defect prevention. Certified Master Teacher (2006). Editorial Board — Nanomedicine: Nanotechnology, Biology, and Medicine (Founding Member). Guest editor — Journal of Law, Medicine & Ethics 2006. doi:10.1016/j.nano.2006.10.108
98
Sunday, September 10th (2:00) Concurrent Symposium XIX: Toxicology Nanomedicine
Nanomaterials and the skin Monteiro-Riviere NA, Center for Chemical Toxicology Research and Pharmacokinetics, North Carolina State University, Raleigh, North Carolina, USA A significant route for environmental and occupational exposure to nanomaterials is through the skin. The focus of our research is to define the biological interactions between different types of nanomaterials and the skin, both as a route of exposure as well as a target organ. Studies of the interactions and skin penetration of multi-wall carbon nanotubes (MWCNT), fullerene based amino acids and quantum dots (QD) with
303
human epidermal keratinocytes (HEK) were conducted. Biological effects of these nanomaterials were evaluated by transmission electron microscopy (TEM), confocal microscopy, protein expression, viability and cytokine assays. MWCNT were primarily localized in intracytoplasmic vacuoles, and induced the release of proinflammatory cytokines from keratinocytes in a time dependent manner. Patterns of altered protein expression suggest involvement with vesicular trafficking, cell signaling and cytoskeletal elements, a finding consistent with the TEM data. QD with different surface characteristics had altered irritation and penetration profiles. These data show that nanomaterials, not derivatized nor optimized for biological applications, are capable of intracellular localization within HEK and can initiate an irritation response. We have also demonstrated that QD of different sizes and surface chemistries are capable of penetrating into skin. A thorough understanding of these basic exposure factors is needed before development of targeted therapeutics and for risk assessment in occupational or environmental scenarios. (Supported by USEPA-STAR Program #RD-83171501–0 and The National Academies Keck Futures Initiative) Nancy Ann Monteiro-Riviere, Ph.D. is a Professor of Investigative Dermatology and Toxicology at the Center for Chemical Toxicology Research and Pharmacokinetics, North Carolina State University (NCSU), and a Professor in the Joint NCSU/UNCChapel Hill Biomedical Engineering Faculty and Professor of Dermatology, School of Medicine at UNC Chapel Hill. She received her M.S. and Ph.D. from Purdue University. She completed post-doctoral training at Chemical Industry Institute of Toxicology in Research Triangle Park, NC. Nancy was past— President of the Dermal Toxicology and In Vitro Toxicology Specialty Sections. She is also a Fellow in The Academy of Toxicological Sciences and in the American College of Toxicology. She serves on several toxicology editorial boards, and national panels, including many in nanotoxicology. Current research focuses is on the cutaneous absorption and toxicity of engineered nanomaterials on human health. doi:10.1016/j.nano.2006.10.109
99
Sunday, September 10th (2:25) Concurrent Symposium XIX: Toxicology Nanomedicine
In vivo toxicity of nanoparticles for gene therapy in the eye Lutty GA, Kim S, Bhutto I, McLeod DS, Grebe R, Prow TW, Wilmer Eye Institute, The Johns Hopkins Hospital, Baltimore, Maryland, USA These studies evaluated the toxicity of several kinds of nanoparticles for delivery of genes in the rabbit eye. Chitosan (CHI), PCEP, and magnetic (MNP) nanoparticles were chosen because they successfully transfected cells in vitro in preliminary studies. Rabbits were injected with nanoparticles either intravitreally (IV) or sub-retinally (SR) and maintained for 7 days after injection. At sacrifice, the eyes were enucleated, dissected and evaluated for vitreous clarity, retinal traction or degeneration, fibrosis, retinal pigment epithelium dysfunction, or inflammation. Chitosan, PCEP, and MNP were made as described in Leong et al. 1998, Wen et al. 2004, and Prow et al. Mol Vis. 2006 respectively. All eyes were cryopreserved for histopathological analysis and gene transfection (GFP expression). Each of the nanoparticles tested were non-toxic in vitro but showed varying levels of toxicity in vivo. All of the nanoparticles were able to transfect cells in vitro and in vivo. Chitosan, on the other hand, caused inflammation in 12/13 eyes and a hazy vitreous in 38% of the eyes. Both the PCEP and MNP were not inflammatory and did not induce retinal dysfunction in IV eyes. In PCEP and MNP SR eyes, MNP were the least toxic. PCEP-injected SR injection induced RPE dysfunction in 11% and retinal degeneration in 15%. PCEP was relatively nontoxic but yielded poor transfection in vivo. Therefore, the best nanoparticle evaluated for our in vivo studies is the least toxic
Sunday, September 10th (3:40) Concurrent Symposium XVIII: Oncology and Experimental Nanomedicine
Dissecting the disruptive nature of nanomedicine Best RG, Khushf G, Department of Obstetrics and Gynecology, University of South Carolina, Columbia, South Carolina, USA, Department of Philosophy and Center for Bioethics, University of South Carolina, Columbia, South Carolina, USA Nanotechnology is often described as a disruptive technology. Presumably this means that some people, companies or even industries will be undermined by the technological developments made possible by nanotechnology. With the development of specific objectives for nanomedicine through the National Nanotechnology Initiative, NIH Nanomedicine Roadmap and others, the application of nanotechnologies in health care is increasingly seen as integral to our visions of future health care. Within the context of the health care system, we consider two ways that nanotechnologies generally, and nanomedicine in particular, might be disruptive. One which considers primarily business and economic aspects of disruption (low-end disruptions), and the other which considers the broader societal issues of disruption at the level of the health care system (high-end disruptions). While the former is primarily dependent on technological developments, the latter is fundamentally dependent upon societal developments. In contrast to low end disruptions, which are extremely difficult to anticipate, high end disruptions can be anticipated. In fact, they depend on explicit recognition of the joint technology/grid systems. Since these are accessible to upstream ethical and regulatory reflection in a way that low end disruptions are not, these should be the focus of attention when we consider the disruptive character of nanomedicine. We seek to provide the foundation for a more comprehensive analysis of the promises and challenges associated with the development of nanomedicine in order to more fully anticipate and address the ethical and societal challenges that lie ahead. Robert G. Best, Ph.D, FACMG, University of South Carolina, School of Medicine. BS (biochemistry) Lehigh University 1981; MS (toxicology) 1983 and Ph.D (genetics and toxicology) 1987 North Carolina State University, Raleigh. Diplomate of the American Board of Medical Genetics (PhD Medical Genetics & Clinical Cytogenetics) 1991. Founding Fellow — American College of Medical Genetics & American Academy of Nanomedicine. Current: Professor, Dept of Ob/Gyn, and Director, Div. Genetics. Research interests: ethical, legal, and social aspects of nanotechnology and nanomedicine; complex genetic traits; birth defect prevention. Certified Master Teacher (2006). Editorial Board — Nanomedicine: Nanotechnology, Biology, and Medicine (Founding Member). Guest editor — Journal of Law, Medicine & Ethics 2006. doi:10.1016/j.nano.2006.10.108
98
Sunday, September 10th (2:00) Concurrent Symposium XIX: Toxicology Nanomedicine
Nanomaterials and the skin Monteiro-Riviere NA, Center for Chemical Toxicology Research and Pharmacokinetics, North Carolina State University, Raleigh, North Carolina, USA A significant route for environmental and occupational exposure to nanomaterials is through the skin. The focus of our research is to define the biological interactions between different types of nanomaterials and the skin, both as a route of exposure as well as a target organ. Studies of the interactions and skin penetration of multi-wall carbon nanotubes (MWCNT), fullerene based amino acids and quantum dots (QD) with
303
human epidermal keratinocytes (HEK) were conducted. Biological effects of these nanomaterials were evaluated by transmission electron microscopy (TEM), confocal microscopy, protein expression, viability and cytokine assays. MWCNT were primarily localized in intracytoplasmic vacuoles, and induced the release of proinflammatory cytokines from keratinocytes in a time dependent manner. Patterns of altered protein expression suggest involvement with vesicular trafficking, cell signaling and cytoskeletal elements, a finding consistent with the TEM data. QD with different surface characteristics had altered irritation and penetration profiles. These data show that nanomaterials, not derivatized nor optimized for biological applications, are capable of intracellular localization within HEK and can initiate an irritation response. We have also demonstrated that QD of different sizes and surface chemistries are capable of penetrating into skin. A thorough understanding of these basic exposure factors is needed before development of targeted therapeutics and for risk assessment in occupational or environmental scenarios. (Supported by USEPA-STAR Program #RD-83171501–0 and The National Academies Keck Futures Initiative) Nancy Ann Monteiro-Riviere, Ph.D. is a Professor of Investigative Dermatology and Toxicology at the Center for Chemical Toxicology Research and Pharmacokinetics, North Carolina State University (NCSU), and a Professor in the Joint NCSU/UNCChapel Hill Biomedical Engineering Faculty and Professor of Dermatology, School of Medicine at UNC Chapel Hill. She received her M.S. and Ph.D. from Purdue University. She completed post-doctoral training at Chemical Industry Institute of Toxicology in Research Triangle Park, NC. Nancy was past— President of the Dermal Toxicology and In Vitro Toxicology Specialty Sections. She is also a Fellow in The Academy of Toxicological Sciences and in the American College of Toxicology. She serves on several toxicology editorial boards, and national panels, including many in nanotoxicology. Current research focuses is on the cutaneous absorption and toxicity of engineered nanomaterials on human health. doi:10.1016/j.nano.2006.10.109
99
Sunday, September 10th (2:25) Concurrent Symposium XIX: Toxicology Nanomedicine
In vivo toxicity of nanoparticles for gene therapy in the eye Lutty GA, Kim S, Bhutto I, McLeod DS, Grebe R, Prow TW, Wilmer Eye Institute, The Johns Hopkins Hospital, Baltimore, Maryland, USA These studies evaluated the toxicity of several kinds of nanoparticles for delivery of genes in the rabbit eye. Chitosan (CHI), PCEP, and magnetic (MNP) nanoparticles were chosen because they successfully transfected cells in vitro in preliminary studies. Rabbits were injected with nanoparticles either intravitreally (IV) or sub-retinally (SR) and maintained for 7 days after injection. At sacrifice, the eyes were enucleated, dissected and evaluated for vitreous clarity, retinal traction or degeneration, fibrosis, retinal pigment epithelium dysfunction, or inflammation. Chitosan, PCEP, and MNP were made as described in Leong et al. 1998, Wen et al. 2004, and Prow et al. Mol Vis. 2006 respectively. All eyes were cryopreserved for histopathological analysis and gene transfection (GFP expression). Each of the nanoparticles tested were non-toxic in vitro but showed varying levels of toxicity in vivo. All of the nanoparticles were able to transfect cells in vitro and in vivo. Chitosan, on the other hand, caused inflammation in 12/13 eyes and a hazy vitreous in 38% of the eyes. Both the PCEP and MNP were not inflammatory and did not induce retinal dysfunction in IV eyes. In PCEP and MNP SR eyes, MNP were the least toxic. PCEP-injected SR injection induced RPE dysfunction in 11% and retinal degeneration in 15%. PCEP was relatively nontoxic but yielded poor transfection in vivo. Therefore, the best nanoparticle evaluated for our in vivo studies is the least toxic