Purpose: The purpose of this study was to investigate the effects of the immunosuppressants FK506 and cyclosporin A (CsA) on the osteogenic differentiation of rat mesenchymal stem cells (MSCs). Methods: The effect of FK506 and CsA on rat MSCs was assessed in vitro. The MTT assay was used to determine the deleterious effect of immunosuppressants on stem cell proliferation at 1, 3, and 7 days. Alkaline phosphatase (ALP) activity was analyzed on days 3, 7, and 14. Alizarin red S staining was done on day 21 to check mineralization nodule formation. Real-time polymerase chain reaction (RT-PCR) was also performed to detect the expressions of bone tissue-specific genes on days 1 and 7. Results: Cell proliferation was promoted more in the FK506 groups than the control or CsA groups on days 3 and 7. The FK506 groups showed increased ALP activity compared to the other groups during the experimental period. The ALP activity of the CsA groups did not differ from the control group in any of the assessments. Mineralization nodule formation was most prominent in the FK506 groups at 21 days. RT-PCR results of the FK506 groups showed that several bone-related genes-osteopontin, osteonectin, and type I collagen (Col-I)-were expressed more than the control in the beginning, but the intensity of expression decreased over time. Runx2 and Dlx5 gene expression were up-regulated on day 7. The effects of 50 nM CsA on osteonectin and Col-I were similar to those of the FK506 groups, but in the 500 nM CsA group, most of the genes were less expressed compared to the control. Conclusions: These results suggest that FK506 enhances the osteoblastic differentiation of rat MSCs. Therefore, FK506 might have a beneficial effect on bone regeneration when immunosuppressants are needed in xenogenic or allogenic stem cell transplantation to treat bone defects.
This study aimed to evaluate the protective effect of Orostachys malacophyllus (OM) and fermented O. malacophyllus (FOM) in Sprague-Dawley rats who had been intoxicated with 1% (w/w) orotic acid (OA) for 10 days. The activities of several hepatic enzymes, aspartate aminotransferase (AST), alanine aminotransferase (ALT), alkaline phosphatase (ALP), lactate dehydrogenase (LDH), and cholinesterase, were increased when OA was given, but these parameters were significantly decreased by FOM treatment. In addition, OA treatment resulted in an increased lipid peroxidative index (thiobarbituric acid-reactive substances, TBARS). A worsened antioxidant status (reduced glutathione) in the liver and serum was also observed. FOM treatment improved the antioxidant status of OA-induced fatty-liver rats, which was evaluated by decreased levels of the lipid peroxidative index and improved antioxidant status in the liver and serum. The contents of liver non-heme iron were increased with OA treatment and significantly decreased with FOM treatment, which suggested that the lipid peroxidation contents were inversely correlated with liver non-heme iron content. Based on these results, FOM is considered a material with significant potential for development into a functional health food that can improve fatty-liver conditions.
Polycaprolactone (PCL) and biphasic calcium phosphate (BCP) have been considered as useful materials for orthopedic devices and osseous implants because of their biocompatibility and bone-forming activity. However, PCL-based scaffolds have hydrophobic surfaces reducing initial cell adhesion or proliferation. To overcome the limitation, we fabricated surface-modified PCL/BCP nanofibers using gamma-irradiation for bone tissue engineering. PCL/BCP nanofibers were prepared by electrospinning and then we supplemented hydrophilicity by introducing acrylic acid (AAc) through gamma-irradiation. We confirmed the surface of nanofibers by SEM, and then the initial viability of MG63 was significantly increased on the AAc grafted nanofibers, and alkaline phosphatase activity($1.239{\pm}0.226nmole/{\mu}g/min$) improved on the modified nanofibers than that on the non-modified nanofibers($0.590{\pm}0.286nmole/{\mu}g/min$). Therefore, AAc-grafted nanofibers may be a good tool for bone tissue engineering applications.
Changes in serum glutamic oxaloacetic transaminase (s-GOT) activities, serum alkaline phosphatase activities and serum total protein amounts were investigated on seven Korean native cows having normal estrus cycle of 18~24 days after normal parturition, dividing estrus cycle into estrus (0~1 days), metestrus (2~6 days), diestrus (7~16 days), proestrus (17~20 days). The results were as follows. 1. Serum GOT activities at estrus ranged from 73.5 to 121.5 Ku with a mean of $89.14{\pm}17.16$ Ku, at metestrus 57 to 89 Ku with a mean of $67.01{\pm}10.81$ Ku, at diestrus 54 to 89.5 Ku with a mean of $67.05{\pm}10.05$ Ku and at proestrus 53 to 112 Ku with a mean of $73.00{\pm}20.05$ Ku. The activities were significantly increased at the estrus comparing with other stages (P<0.01). 2. Serum ALP activities at estrus ranged from 8.0 to 10.4 K-Au with a mean of $8.74{\pm}0.83$ K-Au, at metestrus 6.5 to 9.2 K-Au with a mean of$7.74{\pm}0.76$ K-Au, at diestrus 5.6 to 9.0 K-Au with a mean of $7.67{\pm}1.13$ K-Au, at proestrus 6.2 to 9.3 K-Au with a mean of $7.12{\pm}1.18$ K-Au. The significance was not recognized among the stages of estrus cycle. 3. Serum total protein amounts at estrus ranged from 6.45 to 8.0g/10dl with a mean of $7.25{\pm}0.57/100dl$, at metestrus, 6.37 to 7.9g/100dl with a mean of $7.65{\pm}0.50g/100dl$, fat diestrus, 6.56 to 8.67g/100dl, with a mean of $7.53{\pm}0.55g/100dl$ and at proestrus 5.94 to 7.71g/100dl with a mean of $6.54{\pm}0.65g/dl$. There was not significance among the stages of estrus cycle.
Objectives : This study was performed to compare the preventive effect of water extracts from Saponariae Vaccariae Semen(SVS) and Melandryi Firmi Herba(MFH) on hyperlipidemia and liver damage induced by alcohol. Methods : Except for the normal group, I fed rat on 25% alcohol for 55 days. And each extract was administrated for the same period. I measured the serum component in rat's blood, body weight and weight of liver. Results : At first, I observed effects of SVS and MFH on hyperlipidemia induced by alcohol. SVS and MFH group didn't show the significant decrease of total cholesterol in comparison with those of the control group. SVS and MFH group showed the significant increase of HDL-cholesterol in comparison with those of the control group. SVS group showed the significant decrease of triglyceride in comparison with those of the control group, but MFH group didn't show the significant decrease of triglyceride in comparison with those of the control group. SVS group showed the significant increase of body weight in comparison with those of the control group at 4weeks and 8weeks. But, MFH group didn't show the significant changes of body weight in comparison with those of the control group at 4weeks and 8weeks. By the way, SVS group showed the significant decrease of triglyceride in comparison with those of MFH group. SVS group showed the significant increase of body weight in comparison with those of MFH group at 8weeks. At second, I observed effects of SVS and MFH on liver damage induced by alcohol. SVS and MFH group showed the significant decrease of AST(aspartate aminotransferase), ALT(alanine aminotransferase), ALP(alkaline phosphatase) and LDH(lactate dehydrogenase) in comparison with those of the control group. SVS group showed the significant increase of liver weight in comparison with those of the control group, but MFH group didn't show the significant increase of liver weight in comparison with those of the control group. By the way, SVS group showed the significant increase of liver weight in comparison with those of MFH group. Conclusions : Reviewing these experimental results, it suggests that water extracts from SVS and MFH have pharmaceutical preventive efficacy on hyperlipidemia and liver damage induced by alcohol. Therefore further additional study should be conducted to elucidate the pharmaceutical efficacy of these in depth.
Purpose: We have previously reported that tetra-cell adhesion molecule (T-CAM) markedly enhanced the differentiation of osteoblast-like cells grown on anorganic bone mineral (ABM). T-CAM comprises recombinant peptides containing the Arg- Gly-Asp (RGD) sequence in the tenth type III domain, Pro-His-Ser-Arg-Asn (PHSRN) sequence in the ninth type III domain of fibronectin (FN), and the Glu-Pro-Asp-Ilu-Met (EPDIM) and Tyr-His (YH) sequence in the fourth fas-1 domain of ${\beta}$ig-h3. Therefore, the purpose of this study was to evaluate the cellular activity of osteoblast-like cells and the new bone formation on ABM coated with T-CAM, while comparing the results with those of synthetic cell binding peptide (PepGen P-15). Methods: To analyze the cell viability, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay was performed, andto analyze gene expression, northernblot was performed. Mineral nodule formations were evaluated using alizarin red stain. The new bone formations of each group were evaluated using histologic observation and histomorphometrc analysis. Results: Expression of alkaline phosphatase mRNA was similar in all groups on days 10 and 20. The highest expression of osteopontin mRNA was observed in the group cultured with ABM/P-15, followed by those with ABM/T-CAM and ABM on days 20 and 30. Little difference was seen in the level of expression of collagen type I mRNA on the ABM, ABM/T-CAM, and ABM/P-15 cultured on day 20. There were similar growth and proliferation patterns for the ABM/T-CAM and ABM/P-15. The halo of red stain consistent with $Ca^{2+}$ deposition was wider and denser around ABM/T-CAM and ABM/P-15 particles than around the ABM particles. The ABM/T-CAM group seemed to have bone forming bioactivity similar to that of ABM/P-15. A complete bony bridge was seen in two thirds of the defects in the ABM/T-CAM and ABM/P-15 groups. Conclusions: ABM/T-CAM, which seemed to have bone forming bioactivity similar to ABM/P-15, was considered to serve as effective tissue-engineered bone graft material.
Roberts, E.S.;Heugten, E. van;Spears, J.W.;Routh, P.A.;Lloyd, K.L.;Almond, G.W.
Asian-Australasian Journal of Animal Sciences
/
v.17
no.10
/
pp.1438-1445
/
2004
The objective of this study was to determine the effects of dietary Zn level on performance, serum Zn concentrations, alkaline phosphatase activity (ALP), and immune response of pigs inoculated with Porcine Reproductive and Respiratory Syndrome virus (PRRSv) and Mycoplasma hyopneumoniae. A $2{\times}4$ factorial arrangement of treatments was used in a randomized design. Factors included; 1) PRRSv and M. hyopneumoniae inoculation (n=36 pigs) or sham inoculation (n=36 pigs) with media when pigs entered the grower facility (d 0) at 9 weeks of age and 2) 10, 50, 150 ppm supplemental Zn sulfate (${ZnSO}_4$) from weaning until the completion of the study, or 2,000 ppm supplemental ${ZnSO}_4$for two weeks in the nursery and then supplementation with 150 ppm ${ZnSO}_4$for the remainder of the trial. The basal diet contained 34 ppm Zn. Pigs were weighed on d 0, 10, 17, 24 and 31 and blood samples were collected on d 0, 7, 14, 21 and 28. Pigs inoculated with PRRSv were serologically positive at d 28 and control pigs remained negative to PRRSv. In contrast, the M hyopneumoniae inoculation was inconsistent with 33.3% and 52.8% of pigs serologically positive at d 28 in the control and infected groups, respectively. A febrile response was observed for approximately one week after inoculation with PRRSv. Feed intake (p<0.01) and gain (p<0.1) were less in PRRSv infected pigs than control pigs for the 31 d study. However, performance did not differ among pigs in the four levels of ${ZnSO}_4$. Assessments of immune responses failed to provide unequivocal influence of either PRRSv inoculation or ${ZnSO}_4$level. These data suggest that PRRSv and M. hyopneumoniae act to produce some performance deficits and the influence of Zn supplementation of nursery age pigs does not have clear effect in grower pigs affected with disease.
Porcine embryonic stem (ES) cells have a great potential as tools for transgenic animal production and studies of regulation of differentiation genes. Although several studies showed successful derivation of porcine ES-like cells, these cells were not maintained long-term in culture. Therefore, this study was conducted to establish porcine pluripotent ES-like cells using in vivo fertilized embryos and to maintain these cells in long term culture. Porcine ES-like cells from in vivo embryos obtained by immunosurgery or whole explant culture were successfully cultured for over 56 passages. Morphology of porcine ES-like cells was flat-shaped with a monolayer type colony. These cells stained for alkaline phosphatase throughout the culture. Furthermore, porcine ES-like cells reacted with antibodies against Oct-4, SSEA-1, SSEA-4, Tra-1-60, and Tra-1-81, which are typical markers of undifferentiated stem cells. To characterize the ability of porcine ES-like cells to differentiate into three germ layers, embryoid body formation was induced. After plating of these cells, porcine ES-like cells were spontaneously differentiated into various cell types of all three germ layers. In addition, porcine ES-like cells were successfully derived from IVF blastocysts in media containing human recombinant basic fibroblast growth factor.
An experiment was conducted to study the effect of different dietary protein levels on the performance, nutrient balances, blood biochemical parameters and thyroid hormones of crossbred calves. Thirty crossbred (Bos taurus${\times}$Bos indicus) calves aged 3-5 months were divided into 3 equal groups of 10 each and fed graded levels of crude protein, namely 100 (NP), 75 (LP) and 125 (HP) percent of the Kearl recommendations for 105 d. The calves had access to ad libitum oat hay as the basal roughage. A metabolism trial of 6 d duration was conducted at 90 d of the study. Blood collection and its analysis for various hematological and biochemical parameters as well as thyroid hormones was done both during the pre- and post-experimental periods. The fortnightly body weight changes and the net gain did not differ significantly due to dietary variation. The average daily gain was $367{\pm}21.6$, $347{\pm}22.9$ and $337{\pm}26.4g$ in calves fed NP, LP and HP diets, respectively. Averaged across the feeding trial, oat hay intake was higher (p<0.05) in NP animals than HP or LP fed groups. The dry matter (DM) intake showed no significant difference between the 3 groups but the DM digestibility was higher (p<0.05) in the HP fed animals. The digestibility of crude protein, organic matter, crude fiber and nitrogen-free extract was significantly higher (p<0.05) on HP diets compared to LP or NP diets. The calves on all 3 diets were in positive nitrogen (N) balance, however the N retention was higher (p<0.05) in HP than in LP fed calves. The intake and retention of calcium and phosphorus were similar between the treatments. The blood biochemical profile revealed no significant influence of the dietary treatments on hemoglobin, packed cell volume as well as serum levels of glucose, total protein, albumin, globulin, Ca, P, and alkaline phosphatase. Serum levels of the circulating thyroid hormones ($T_3$ and $T_4$) tended to be lower (p>0.05) on feeding of the LP diet besides showing an increasing trend with the advancement of age. Considering the similar performance and metabolic profile, it could be concluded that crossbred calves can be satisfactorily reared on 25% lower protein level as recommended by Kearl for developing countries, which would not only economize the cost of production but also help to reduce environmental pollution attributable to livestock production.
Objective: Astragalus membranaceus root is a well-known traditional Chinese herbal medicine with many biological active constituents. This study was conducted to examine the effects of Astragalus membranaceus root powder (AMP) on growth performance, serum antioxidant and immune response in finishing lambs. Methods: A total of thirty-six Guangling fat-tailed ram lambs (body weight = 19±2 kg, mean±standard deviation) were randomly assigned to one of six treatments for a 40 d feeding period, with the first 10 d for adaptation. Treatments consisted of the lambs' basal diets with addition of 0, 5, 10, 15, 20, and 30 g/kg of diet of AMP. Results: Response to supplementation level of AMP was quadratic (p≤0.032) for final weight and ADG with the greatest at 10 g/kg of diet, but dry matter intake was not affected (p≥0.227) by treatments. The increase of AMP supplementation resulted in a quadratic response in contents of triglyceride and creatinine (p<0.05), with the lowest values for 10 and 20 g/kg of diet, respectively. A linear and quadratic decrease was observed in activity of alkaline phosphatase in serum of lambs. As the AMP supplementation increased, the activities of total superoxide dismutase and total antioxidant capacity increased linearly (p≤0.018) and hydroxyl radical (OH-) decreased linearly (p = 0.002). For catalase (CAT) and malondialdehyde (MDA), quadratic (p≤0.001) effects were observed among treatments, with the greatest CAT and lowest MDA values at 10 g/kg AMP. Additionally, supplementing AMP up to a level of 10 or 15 g/kg of diet quadratically increased immunoglobulin and interleukin contents in the serum. Conclusion: The results indicated that AMP can be used as natural feed additive in the ration of lambs to improve ADG, antioxidant status, and immune functions, and the optimal dose was 10 g/kg of diet under the condition of this experiment.
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