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Effect of moderate exercise on bone metabolism in Wistar rats
Abstracts d per os + 1.5 μg/kg/w sc. Eight rats were sham operated. Histomorphometric and histopathological studies were performed on tibias obtained after 30 days of treatment. OVX decreased trabecular bone (%BV/TV), and Z prevented and raised it (Sham: 35.3 ± 4.7, control: 21.5 ± 1.5*, MFP: 17.1 ± 1.0*, Z: 64.3 ± 3.5*, MFP ± Z: 52.2 ± 3.2*, * p < 0.05 vs sham, mean ± SE, ANOVA). Histological examination showed, in the sham group, an adequate bone-hemopoietic (HT) tissue relationship and mature bone trabeculae (Tb). The control group showed higher HT, immature Tb, fewer osteoblasts (Ob) and higher osteoclasts (Oc) compared with the sham group. The MFP group showed abundant HT and thin Tb with abundant lining cells (LC) and Ob. In Z, anastomosed and immature Tb with invasive tendency on the marrow space were observed. A decrease in Ob and HT was also observed. In contrast, the MFP + Z group showed mature Tb with well defined marrow spaces. The trabecular edge was covered by LC and Ob. We conclude that Z treatment, alone or combined with MFP, prevented the bone loss caused by OVX. Its combination with MFP normalized the histological features of bone. The MFP, by stimulating osteoblast activity, could modulate bone remodeling by producing a better quality bone. This article is part of a Special Issue entitled AAOMM 2010 Abstracts. doi:10.1016/j.bone.2011.03.725
Novel methodology for the determination of alkaline phosphatase isoenzymes in plasma circulating C. Guglielmi, L.R. Brun, A. Rigalli Bone Biology Laboratory, Rosario, Argentina E-mail address: [email protected] Alkaline phosphatase (AP) exists in plasma as isoenzymes: bone (bAP), liver (lAP) and intestinal (iAP). The activity of bAP is particularly useful as a measure of bone formation. The objectives of this work was to characterize the activity of the AP isoenzymes of rat serum on the basis of the sensitivity of temperature (T), pH and concentration of phenylalanine (Phe) and develop a methodology to measure individual activities in a mixture of them. A mathematical model is proposed, which is based on a set of linear algebraic equations, where the activities of bAP, lAP and iAP in plasma are the unknown values. The activity of FA and the percentages of activity of each isoenzyme at a given value of T, pH, and Phe are the independent terms and the coefficients, respectively. The activity of each isoenzyme was measured at pH = 8, T = 37 °C, Phe = 0 mM and varying the incubation medium: a) pH = 6– 10, b) T = 10–45 °C; c) Phe = 0–16 mM. The percentages of activity of each isoenzyme for the range of each variable were calculated considering 100% the value of activity measured at pH = 8, T = 37 °C and Phe = 0 mM. In a mixture of the three isoenzymes at a given value, the total activity of the AP is the sum of the activities for each isoenzyme multiplied by its percentage of activity. The solution of the system is the values of activities of bAP, lAP and iAP in plasma. The activity of each isoenzyme was measured in an artificial mixture of the three purified isoenzymes (90% FAi, 5% FAo, 5% FAh). The values obtained were: FAi: 3.15 pNFF pmol/L.seg (68.12%); FAo: 0.75 (16.18%) and FAh: 0.73 (15.70%). Although they are preliminary results, the proposed methodology has acceptable values. This article is part of a Special Issue entitled AAOMM 2010 Abstracts.
doi:10.1016/j.bone.2011.03.726
Effect of moderate exercise on bone metabolism in Wistar rats A. Pietrelli, G. Bryk, N. Paglia, R. Orzuza, S. Zeni, N. Basso School of Medicine, Department of Basic Science Research, UCES, Argentina Institute of Cell Biology and Neuroscience “Prof. E. De Robertis”, UBA, CONICET, Argentina Medical Osteopathies Section, Clinical Hospital, UBA, CONICET, Argentina Institute of Cardiovascular Research “Prof. A. Taquini”, UBA, CONICET, Argentina Institute of Cardiovascular Physiology, UBA-CONICET, Argentina Physical activity is an important factor to reach an optimal peak mass during youth and to reduce the rate of bone loss later in life. Skeletal tissue responds to dynamic loading that initiate osteogénesis. Physical exercise has favorable effects on glucose and lipid profile. The aim of this study was to assess the effect of chronic aerobic training on total and regional BMD in male Wistar rats from pre-pubertal period to adulthood. Aeróbic training (EA) rats were trained on the treadmill from the juvenile period to adulthood (2 to 18 months) following an EA routine and compared to the sedentary control (SC) group. At the end of the experience body composition, BMD and BMC were assessed by DXA (Small Animal Software, Lunar DPX) and serum calcium (Ca), phosphorous (P), bone alkaline phosphatase (BAP), total cholesterol (T. Chol), glucose (Glu) and triglycerides (TGL) were determined by the habitual methods. The EA group had a significantly higher BMC: 19%; area: 16% and lean mass: 16% and a significantly lower fat percentage: 148% and serum T. Chol: 5% vs. CS group (p < 0.01) without significant differences on the other studied parameters. Chronic aerobic exercise increased bone mass and lean tissue, and induced a marked decrease in fatty tissue
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without changes in body weight suggesting a negative relationship between bone content and fatty mass. Grant: Asociación de Fomento a la Investigación Científica (AFIC). This article is part of a Special Issue entitled AAOMM 2010 Abstracts.
doi:10.1016/j.bone.2011.03.727
Calcium deficiency as a risk factor for overweight and obesity development. Experimental study C. Marotte, A. Weisstaub, M.M.S. Gonzales Chaves, G.G. Pellegrini, M. Olguin, D.M. Lucero, L. Schreier, M.L. Portela, S.N. Zeni Osteopathies Section, Hosp Clínicas, Cát Nut, Fac Farm Bioq, UBA, UNRosario, Lab Líp y lipop, Dep Bioq Clín, Fac Farm Bioq, UBA, CONICET, Argentina NHANES III and Nutr. and Health Nat. Survey (Arg) showed an inverse relationship between Ca intake and overweight/obesity incidence. Objective: To evaluate the influence of Ca intake in body composition (BC), lipidic/glucidic parameters in genetically obese male rats (IIMbb). Rats were fed isocaloric diets varying only in Ca content: 0.9 % (H), 0.5% (N), and 0.3% (L) from the beginning of pregnancy. At weaning, male pups continued receiving their mother's diets until day 50. At the end, BC, ashes, Ca and P contents; serum BGP, CTX, insulin, glucose, cholesterol (Chol) and triglycerides (TGL) were evaluated. Results (X ± SE): H, N and L, respectively: Body Weight (BW) (g) 195 ± 9a, 227 ± 13b, 262 ± 7c; Ashes (g/ 100 g BW) 2.4 ± 0.1a, 2.2 ± 0.1a, 1.5 ± 0.1b; Ca (mg/100 g BW) 720 ± 34a, 640 ± 22b, 394 ± 40c; P (mg/100 g BW) 532 ± 22a, 520 ± 31a, 882 ± 15b; %Fat 14.2 ± 0.9a, 13.7 ± 0.9a, 15.9 ± 0.6b; BGP (ng/dl) 271 ± 39a, 381 ± 40b, 478 ± 32c; CTX (ng/dl) 31.1 ± 0.3a, 57.0 ± 3.7b, 69.7 ± 2.3c; Chol (mg/dl) 67.4 ± 8.0a, 78.2 ± 5.4b, 64.5 ± 3.1a; TGL (mg/dl) 272.6 ± 39.9a, 205.0 ± 35.2b, 216.4 ± 26.8b; Glucose (mg/dl) 111.8 ± 25.5a, 151.9 ± 21.9b, 207.0 ± 11.0c; Insulin (ng/ml) 1.92 ± 0.75a, 4.07 ± 0.93b, 6.87 ± 2.24c. Different letters mean a p < 0.05. BW, Ca, BGP, CTX, glucose and insulin were significantly related to Ca intake. BW, %fat, P, BGP, sCTX, glucose and insulin were significantly higher and %ashes and Ca content significantly lower in LG. Conclusions: During growth, the low Ca intake could affect not only body Ca content, but also BC and bone remodeling in IIMbb. Surprisingly, the low Ca intake induced an increase in body fat pads and insulin resistance. UBACYT (2008–10) B091. This article is part of a Special Issue entitled AAOMM 2010 Abstracts.
doi:10.1016/j.bone.2011.03.728
Daily amount of vitamin D (D) would be given an important factor to equal the effectiveness of cholecalciferol and ergocalciferol M.M.S. Gonzales Chaves, C. Marotte, G.G. Pellegrini, S.M. Friedman, S.N. Zeni Osteopathy Medical Section, University Hospital, UBA, CONICET, Argentina Fac. Dentistry, UBA, Argentina Some authors consider that daily D2 and D3 in equivalent doses have the same effectiveness in rising and maintaining 25OHD levels. However, others suggest that D3 is more effective than D2. Objective: To determine in a model of D insufficiency and established osteopenia if the effectiveness of D2 and D3 varies according to the daily amount received. Female Wistar rats were ovariectomized (OVX) (N = 64) or sham operated (SHAM) (N = 64). During 15 days post surgery all received a commercial diet (Granave SA, BA). After that, they were fed a semisynthetic diet without D (0 UI%) during 45 days to become D insufficiency. At day-60 they were divided into 2 groups that received: A: 100 IU% and B: 200 IU% of D2 or D3, respectively. The 25ODH (125I RIA Kit, DiaSorin, Stillwater, MN, USA) levels were determined at baseline, 60, 85 and 105 days of experience. Results (x ± ES): Baseline 25OHD: 19.2 ± 0.3 ng/ml. T = 60; T = 85; T = 105, respectively. Group A: OVXD3; 6.0 ± 0.7a; 12.9 ± 2.5c; 14.5 ± 2c; OVXD2; 6.4 ± 0.5a; 8.9 ± 1.0b; 8.7 ± 0.9b; SHAMD3; 7.0 ± 0.4a; 11.6 ± 1.0c; 14.0 ± 1.9c; SHAMD2; 6.0 ± 0.5a; 7.7 ± 0.9b; 8.1 ± 0.7b. Group B: OVXD2; 5.0 ± 0.9a; 16.6 ± 1.5b; 15.5 ± 0.3b; OVXD3; 5.0 ± 0.6a; 15.1 ± 0.6b; 15.9 ± 1.0b; SHAMD2; 4.8 ± 0.5a; 14.7 ± 1.4b; 15.5 ± 0.9b; SHAMD3; 6.0 ± 1.2a; 16.9 ± 2.1b; 18.0 ± 1.3b. Different letters indicate p < 0.05. At T = 60 there were no differences between groups. In B 25OHD was similar in rats feeding D2 or D3 from day 85 until the end of the experience, whereas in A the 25OHD was significantly higher in rats feeding D3. Conclusions: Under our experimental conditions, D3 and D2 could have different effectiveness depending on the amount supplied daily. PIP 6483. This article is part of a Special Issue entitled AAOMM 2010 Abstracts.
doi:10.1016/j.bone.2011.03.729
Novel methodology for the determination of alkaline phosphatase isoenzymes in plasma circulating C. Guglielmi, L.R. Brun, A. Rigalli Bone Biology Laboratory, Rosario, Argentina E-mail address: [email protected] Alkaline phosphatase (AP) exists in plasma as isoenzymes: bone (bAP), liver (lAP) and intestinal (iAP). The activity of bAP is particularly useful as a measure of bone formation. The objectives of this work was to characterize the activity of the AP isoenzymes of rat serum on the basis of the sensitivity of temperature (T), pH and concentration of phenylalanine (Phe) and develop a methodology to measure individual activities in a mixture of them. A mathematical model is proposed, which is based on a set of linear algebraic equations, where the activities of bAP, lAP and iAP in plasma are the unknown values. The activity of FA and the percentages of activity of each isoenzyme at a given value of T, pH, and Phe are the independent terms and the coefficients, respectively. The activity of each isoenzyme was measured at pH = 8, T = 37 °C, Phe = 0 mM and varying the incubation medium: a) pH = 6– 10, b) T = 10–45 °C; c) Phe = 0–16 mM. The percentages of activity of each isoenzyme for the range of each variable were calculated considering 100% the value of activity measured at pH = 8, T = 37 °C and Phe = 0 mM. In a mixture of the three isoenzymes at a given value, the total activity of the AP is the sum of the activities for each isoenzyme multiplied by its percentage of activity. The solution of the system is the values of activities of bAP, lAP and iAP in plasma. The activity of each isoenzyme was measured in an artificial mixture of the three purified isoenzymes (90% FAi, 5% FAo, 5% FAh). The values obtained were: FAi: 3.15 pNFF pmol/L.seg (68.12%); FAo: 0.75 (16.18%) and FAh: 0.73 (15.70%). Although they are preliminary results, the proposed methodology has acceptable values. This article is part of a Special Issue entitled AAOMM 2010 Abstracts.
doi:10.1016/j.bone.2011.03.726
Effect of moderate exercise on bone metabolism in Wistar rats A. Pietrelli, G. Bryk, N. Paglia, R. Orzuza, S. Zeni, N. Basso School of Medicine, Department of Basic Science Research, UCES, Argentina Institute of Cell Biology and Neuroscience “Prof. E. De Robertis”, UBA, CONICET, Argentina Medical Osteopathies Section, Clinical Hospital, UBA, CONICET, Argentina Institute of Cardiovascular Research “Prof. A. Taquini”, UBA, CONICET, Argentina Institute of Cardiovascular Physiology, UBA-CONICET, Argentina Physical activity is an important factor to reach an optimal peak mass during youth and to reduce the rate of bone loss later in life. Skeletal tissue responds to dynamic loading that initiate osteogénesis. Physical exercise has favorable effects on glucose and lipid profile. The aim of this study was to assess the effect of chronic aerobic training on total and regional BMD in male Wistar rats from pre-pubertal period to adulthood. Aeróbic training (EA) rats were trained on the treadmill from the juvenile period to adulthood (2 to 18 months) following an EA routine and compared to the sedentary control (SC) group. At the end of the experience body composition, BMD and BMC were assessed by DXA (Small Animal Software, Lunar DPX) and serum calcium (Ca), phosphorous (P), bone alkaline phosphatase (BAP), total cholesterol (T. Chol), glucose (Glu) and triglycerides (TGL) were determined by the habitual methods. The EA group had a significantly higher BMC: 19%; area: 16% and lean mass: 16% and a significantly lower fat percentage: 148% and serum T. Chol: 5% vs. CS group (p < 0.01) without significant differences on the other studied parameters. Chronic aerobic exercise increased bone mass and lean tissue, and induced a marked decrease in fatty tissue
S285
without changes in body weight suggesting a negative relationship between bone content and fatty mass. Grant: Asociación de Fomento a la Investigación Científica (AFIC). This article is part of a Special Issue entitled AAOMM 2010 Abstracts.
doi:10.1016/j.bone.2011.03.727
Calcium deficiency as a risk factor for overweight and obesity development. Experimental study C. Marotte, A. Weisstaub, M.M.S. Gonzales Chaves, G.G. Pellegrini, M. Olguin, D.M. Lucero, L. Schreier, M.L. Portela, S.N. Zeni Osteopathies Section, Hosp Clínicas, Cát Nut, Fac Farm Bioq, UBA, UNRosario, Lab Líp y lipop, Dep Bioq Clín, Fac Farm Bioq, UBA, CONICET, Argentina NHANES III and Nutr. and Health Nat. Survey (Arg) showed an inverse relationship between Ca intake and overweight/obesity incidence. Objective: To evaluate the influence of Ca intake in body composition (BC), lipidic/glucidic parameters in genetically obese male rats (IIMbb). Rats were fed isocaloric diets varying only in Ca content: 0.9 % (H), 0.5% (N), and 0.3% (L) from the beginning of pregnancy. At weaning, male pups continued receiving their mother's diets until day 50. At the end, BC, ashes, Ca and P contents; serum BGP, CTX, insulin, glucose, cholesterol (Chol) and triglycerides (TGL) were evaluated. Results (X ± SE): H, N and L, respectively: Body Weight (BW) (g) 195 ± 9a, 227 ± 13b, 262 ± 7c; Ashes (g/ 100 g BW) 2.4 ± 0.1a, 2.2 ± 0.1a, 1.5 ± 0.1b; Ca (mg/100 g BW) 720 ± 34a, 640 ± 22b, 394 ± 40c; P (mg/100 g BW) 532 ± 22a, 520 ± 31a, 882 ± 15b; %Fat 14.2 ± 0.9a, 13.7 ± 0.9a, 15.9 ± 0.6b; BGP (ng/dl) 271 ± 39a, 381 ± 40b, 478 ± 32c; CTX (ng/dl) 31.1 ± 0.3a, 57.0 ± 3.7b, 69.7 ± 2.3c; Chol (mg/dl) 67.4 ± 8.0a, 78.2 ± 5.4b, 64.5 ± 3.1a; TGL (mg/dl) 272.6 ± 39.9a, 205.0 ± 35.2b, 216.4 ± 26.8b; Glucose (mg/dl) 111.8 ± 25.5a, 151.9 ± 21.9b, 207.0 ± 11.0c; Insulin (ng/ml) 1.92 ± 0.75a, 4.07 ± 0.93b, 6.87 ± 2.24c. Different letters mean a p < 0.05. BW, Ca, BGP, CTX, glucose and insulin were significantly related to Ca intake. BW, %fat, P, BGP, sCTX, glucose and insulin were significantly higher and %ashes and Ca content significantly lower in LG. Conclusions: During growth, the low Ca intake could affect not only body Ca content, but also BC and bone remodeling in IIMbb. Surprisingly, the low Ca intake induced an increase in body fat pads and insulin resistance. UBACYT (2008–10) B091. This article is part of a Special Issue entitled AAOMM 2010 Abstracts.
doi:10.1016/j.bone.2011.03.728
Daily amount of vitamin D (D) would be given an important factor to equal the effectiveness of cholecalciferol and ergocalciferol M.M.S. Gonzales Chaves, C. Marotte, G.G. Pellegrini, S.M. Friedman, S.N. Zeni Osteopathy Medical Section, University Hospital, UBA, CONICET, Argentina Fac. Dentistry, UBA, Argentina Some authors consider that daily D2 and D3 in equivalent doses have the same effectiveness in rising and maintaining 25OHD levels. However, others suggest that D3 is more effective than D2. Objective: To determine in a model of D insufficiency and established osteopenia if the effectiveness of D2 and D3 varies according to the daily amount received. Female Wistar rats were ovariectomized (OVX) (N = 64) or sham operated (SHAM) (N = 64). During 15 days post surgery all received a commercial diet (Granave SA, BA). After that, they were fed a semisynthetic diet without D (0 UI%) during 45 days to become D insufficiency. At day-60 they were divided into 2 groups that received: A: 100 IU% and B: 200 IU% of D2 or D3, respectively. The 25ODH (125I RIA Kit, DiaSorin, Stillwater, MN, USA) levels were determined at baseline, 60, 85 and 105 days of experience. Results (x ± ES): Baseline 25OHD: 19.2 ± 0.3 ng/ml. T = 60; T = 85; T = 105, respectively. Group A: OVXD3; 6.0 ± 0.7a; 12.9 ± 2.5c; 14.5 ± 2c; OVXD2; 6.4 ± 0.5a; 8.9 ± 1.0b; 8.7 ± 0.9b; SHAMD3; 7.0 ± 0.4a; 11.6 ± 1.0c; 14.0 ± 1.9c; SHAMD2; 6.0 ± 0.5a; 7.7 ± 0.9b; 8.1 ± 0.7b. Group B: OVXD2; 5.0 ± 0.9a; 16.6 ± 1.5b; 15.5 ± 0.3b; OVXD3; 5.0 ± 0.6a; 15.1 ± 0.6b; 15.9 ± 1.0b; SHAMD2; 4.8 ± 0.5a; 14.7 ± 1.4b; 15.5 ± 0.9b; SHAMD3; 6.0 ± 1.2a; 16.9 ± 2.1b; 18.0 ± 1.3b. Different letters indicate p < 0.05. At T = 60 there were no differences between groups. In B 25OHD was similar in rats feeding D2 or D3 from day 85 until the end of the experience, whereas in A the 25OHD was significantly higher in rats feeding D3. Conclusions: Under our experimental conditions, D3 and D2 could have different effectiveness depending on the amount supplied daily. PIP 6483. This article is part of a Special Issue entitled AAOMM 2010 Abstracts.
doi:10.1016/j.bone.2011.03.729