ABSTRACT
This experiment is sought to study the contribution of different gene in osteogenous differentiation of bone marrow stromal cells (MSCs) and optimize the seed cells of bone tissue engineering. Firstly, we obtained the full length gene of BMP-2, VEGF165 and bFGF by RT-PCR, and cloned into the expression vector pcDNA3. 0. After to be transfected, the MSCs cell lines which could express each of the target protein were selected out by G418. RT-PCR and immunohistochemistry were used to confirm the exogenous gene expression in MSCs. MTT showed that almost all of the MSCs which have been transfected with exogenous gene had more strong proliferative potential than the untransfected group did. There was a notable increase of ALP activity in transfected cell compared with the control group. The concentration of OCN in cell culture medium had a increase in some degree except of VEGF group. The outstanding osteogenous differentiation could be observed in BMP-2 and bFGF transfected MSCs. The results showed that the MSCs modified by BMP-2 and bFGF would be more effective in bone tissue engineering field.
Subject(s)
Animals , Rats , Bone Marrow Cells , Cell Biology , Bone Morphogenetic Proteins , Genetics , Cell Differentiation , Cells, Cultured , Fibroblast Growth Factor 2 , Genetics , Mesenchymal Stem Cells , Cell Biology , Osteogenesis , Tissue Engineering , Methods , Transfection , Vascular Endothelial Growth Factor A , GeneticsABSTRACT
To investigate the growth and osteogenesis characteristics of cultured canine mesenchymal stem cells (cMSCs) under osteogenic induction. We found the cMSCs were isolated from adult canine using density gradient separation method. The cMSCs attachment formed soon after seeding and grew into colonies with the appearance of fibroblastic cells. The osteogenic induction compound of Dexamethasone (Dex), beta-sodium glycerphosphate (beta-GP), ascorbic acid (AA) was added to passaged cMSCs and the proliferation and osteogenic differentiation of them was studied. The morphology of cells was observed by light micrograph and transmission electron microscope. The proliferation and growth characteristics of cMSCs were observed during primary and passage cultures through MTT. The differentiation were assayed by alkaline phophatase (ALP) and osteocalcin (OCN). We found the cMSCs have an active proliferative ability in primary and passage culture, and cMSCs under osteogenic induction have the typical characteristic of a secretory cell; the osteogenic induction compound may induce cMSCs to differentiate to osteoblasts. There are higher expression of ALP and OCN in passage 3 cMSCs under osteogenic induction than that of the osteoblasts osteogenic induction condition. Our research suggest the cMSCs in our culture system are mainly undifferentiated osteoprogenitors and can differentiate to osteoblast under osteogenic induction.
Subject(s)
Animals , Dogs , Bone Marrow Cells , Cell Biology , Cell Differentiation , Physiology , Cells, Cultured , Mesenchymal Stem Cells , Cell Biology , Osteoblasts , Cell Biology , Osteogenesis , Tissue Engineering , MethodsABSTRACT
OBJECTIVE:To explore the clinical efficacy of 131I in the treatment of hyperthyroidism.METHODS:131I therapy for 784 cases of hyperthyroidism was analyzed retrospectively.The dose of 131I(?Ci) was calculated with formula:?Ci=the weight of thyroid(g)?80-120(?Ci?g-1)/the highest absorbed iodine rate(%).Clinical efficacy was evaluated according to the results.RESULTS:394 cases were relieved completely after once therapy,523 cases after twice therapies and 547 cases after three times of therapies.577 patients were visited at random and 94.80% of them were cured.Early-stage toxic reaction had been found in 46 cases with incidence of 7.97%.Hyperthyroidism crisis had not been developed.Temporary hypothyroidism had been found in 68 cases with incidence of 11.79%,while permanent hypothyroidism had been found in 146 cases with incidence of 25.30%.CONCLUSION:131I has an obvious effect on hyperthyroidism with one disadvantage of high incidence of hyperthyroidism.However,131I is an effective therapy which is safe,economical,simple and convenient.
ABSTRACT
There are three key factors in tissue engineering: seeding cells, scaffold and their interaction. Although mesenchymal stem cells (MSCs) are potential seeding cells, the problem of what phase MSCs should be used is not yet solved. On the other hand, degradable porous scaffolds which have good mechanics and good biocompatibility are preferred. To choose the optimum seeding cells and the suitable ratio of beta-TCP/PLLA porous scaffold, we observed the phenotype of the male SD rat's osteoblastic MSCs and detected the amount of alkaline phosphatase, osteocalcin and type I collagen secreted by the osteoblastic rMSCs in different phase. About 10, 14 and 20 days after induction, the induced cells came into proliferative phase, matrix synthesis phase and mineralization phase, respectively. Then we chose the suitable cells and seeded them on beta-TCP/PLLA composite scaffolds with different ratios (beta-TCP/PLLA = 1:1; beta-TCP/PLLA = 1:2; and beta-TCP/PLLA = 2:1). Fluorescence microscope, scanning electron microscope and MTT assay were used to observe and to detect the biocompatibility of the scaffolds. The results indicated that all of these materials have biocompatibility to some extent. Cells can grow well on all of the scaffolds. However, scaffold beta-TCP/PLLA = 2:1 seems to be a more suitable tissue engineering scaffold on account of its minimal influence on cell growth and differentiation.
Subject(s)
Animals , Male , Rats , Biocompatible Materials , Calcium Phosphates , Pharmacology , Cell Differentiation , Cells, Cultured , Coculture Techniques , Lactic Acid , Pharmacology , Mesenchymal Stem Cells , Cell Biology , Osteoblasts , Cell Biology , Polyesters , Polymers , Pharmacology , Porosity , Rats, Sprague-Dawley , Tissue EngineeringABSTRACT
To establish and observe rabbit hyperlipemia and atherosclerosis model, we combined the method of high-lipid diet and immuoreactive injure. We divided 45 New Zealand rabbit into control group and high-lipid group. According to the time (4, 8, 12 weeks) of established model, the control group and high-lipid group were divided into 3 groups respectively. The blood-lipid, the hemodynamics parameter and vascular intima were determined and observed. The results showed: (1) After feeding 4, 8, 12 weeks, TC and LDL-C of the high-lipid group in the serum increased obviously. After feeding 8 week, TG of the high-lipid group began to increase obviously. HDL-C of the high-lipid group was higher than control group, but with a descendent trend. (2) Blood viscosity of the high-lipid group increased obviously under the 0.512 S(-1) and 5.96 S(-1) at 12 week. Blood flow increased obviously at 8 and 12 week. SBP increased evidently at 8 and 12 week. Alteration of the plasma viscosity and vascular diameter were not obvious. (3) By observing with optical microscope the intima of the control group is smooth. The intima of the high-lipid group has a light incrassation at 4 week. The intima of the high-lipid group has a obvious incrassation at 8 and 12 week. By observing with through transmission electron microscopy (TEM), the lipid vacuole is under the endothelium cell. (4) By adopting the immunohistochemistry, the foam cell that derived from smooth muscle cell were determined. We concluded that the blood lipid can have a prefigurative role in atherosclerosis; blood flow and blood pressure and blood viscosity increase at low shear rate in the course of the atherosclerosis; vascular intima is incrassate and the composition of the AS plaque is variational continually.