The Study Of The In Vitro Comparative Experiment About Osteogenic Differentiation Of The BMSCs And AMSCs In Rabbits And The Effect Of OGP On AMSCs Of Rabbits In Osteogenic Differentiation

Chapter 1 The comparison of proliferation and bone induction about bone marrow mesenchymal stem cells and adipose mesenchymal stem cells of rabbits【Objective】Isolating the bone marrow mesenchymal stem cells and adipose mesenchymal stem cells,comparing the characteristics of two cells,providing the evidence for the adipose mesenchymal stem cells as the substitude of bone marrow mesenchymal stem cells in bone tissue engineering.【Method】The inguinal fat pads and iliac bone marrow were obtained with sterile technique.By the adherence,we isolate the bone marrow mesenchymal stem cells and adipose mesenchymal stem cells,to compare the characteristics of the two cells,including the morphos of the cells,the rate of adherence,cell kinetics,cell surface marker,ALP solution and Von kossa staining,quantity detection of ALP and the caicium in induced cells.To analyse the data.【Results】(1)The morphous of two cells are chord shape,like fibroblast cells.(2)) By quantity detection of adherence rate and analysis of variance with repeated measurement data,no significant difference exist between two groups of the adherence rate(F=2.905,P＞0.05).The adherences of the two cells are similar at different time. (3)The reproductive activity are fairly between the adipose mesenchymal stem cells and bone marrow mesenchymal stem cells,the postdigestive 2^nd and 5^th generation cells have 6～9 days log phase growth after the latent period about 24～48 hours,then transit to the platform period.(4)By immunocytochemistry,adipose mesenchymal stem cells and bone marrow mesenchymal stem cells express CD44 positive.(5)The ALP and Von kossa staining of the two cells are both positive.(6) By quantity detection of alkaline phosphatase and analysis of variance with repeated measurement data,no significant difference exist between two groups of the quantity of ALP(F=3.067,P＞0.05).The quantity of ALP of the two cells are similar at different time.(7) By quantity detection of accumulation of calcium and analysis of variance with repeated measurement data,no significant difference exist between two groups of the quantity of accumulation of calcium(F=3.067,P＞0.05).The accumulation of calcium of the two cells are similar at different time【Conclusion】(1)Similar adipose mesenchymal stem cells,like bone marrow mesenchymal stem cells,can be isolated from the lipose tissue in rabbit.(2)No significant difference exist about the capacity of bone induction between adipose mesenchymal stem cells and bone marrow mesenchymal stem cells.(3) Adipose mesenchymal stem cells can work as the seeds of bone tissue engineering instead of bone marrow mesenchymal stem cells.Chapter 2 The study of the effect of osteogenic growth peptide on adipose mesenchymal stem cells of rabbits in proliferation and osteogenic differentiation【Objective】To value the effect of osteogenic growth peptide on the proliferation and osteogenic differentiation of adipose mesenchymal stem cells in rabbits.【Method】By the adherence,to isolate the adipose mesenchymal stem cells,the cells were purified through passaging and frequent replacing of medium.In order to obtain the most efficient concentration of OGP to promote the proliferation of the AMSCs,passage 2 cells were detected by MTT analysis,In order to obtain the most efficient concentration of OGP of promoting the osteogenic differentiation of the AMSCs,Passage 3 cells were planted in 6-well plate at the concentration of 10⁵/well.We also compared the ability of promoting the osteogenic differentiation of the AMSCs among the OGP at the most efficient concentration,10^-8mol/L dexamethasone and the integrating application of OGP at the most efficient concentration and 10^-8mol/L dexamethason【Results】OGP can promote the activity of proliferation of adipose mesenchymal stem cells in the concentration range of 10^-11-10^-7mol/L.The most best concentration of OGP is 10^-11mol/L.OGP at any concentration can promote the ability of the osteogenic differentiation of the AMSCs(P＜0.05),and the most efficient concentration is 10^-9mol/L(P＜0.05).10^-9mol/LOGP and the 10^-8mol/L dexamethasone have the same ability of promoting the osteogenic differentiation of the AMSCs(P＞0.05).The integrating application of OGP at the most efficient concentration and 10^-8mol/L dexamethason have the positive coopertive effect(P＜0.05).【Conclusion】OGP can promote the proliferation and osteogenic differentiation of adipose mesenchymal stem cells of rabbits in the model of these cells cultured in vitro and it hinted the promising prospect in bone tissue engineering.