Research Of Rabbit Tendinopathy Of Adipose Derived Stem Cells Platelet-rich Plasma

Objective: How to treat the tendinopathy is a clinical difficulty. The applicationof Adipose derived stem cells (ADSCs) in treating tendinopathy is a new method whichhas already leaded to some positive results. ADSCs as one kind of MSCs that have thepotentials of multi-differentiation and could be divided into various cell types, but asboth of the mechanism of differentiation and the role ADSCs playing in the therapeuticprocess are uncertain and because of which the scientific and Clinical application ofADSCs has been restricted. We were trying to build an indirect coculture system toinvestigate differentiation of ADSCs induced by indirect coculturing with tenocytes.Comparison was also made between the simple coculture group and the coculture groupwith platelet rich plasma added to study the effects of platelet-rich plasma to ADSCs onsecretion of extracellular matrix when being inducted by indirect coculture withtenocytes. On this basis, we could explore the differentiation mechanism of ADSCs andmake a reference for Clinical application of it in treatment of tendinopathy.Methods: Culture the isolatedly tenocytes of New Zealand white rabbits withtissue culture method. ADSCs were collected in vitro and be expanded. Rabbit bloodwas prepared and had plasma activated to get the plasma extraction. An indirectcoculture system was established by using coculture kit of0.4m to ensure circulationof culture fluid and avoid direct contact of cells. With well-grown ADSCs(P3）andtenocytes (P3) collected, ADSCs were placed in the outer chamber and tenocytes in theinner chamber of this system. An additional PRP was added and indirect coculturegroup was set up. The corresponding protein levels of TNMD and type I collagen bytechnique of immunohistochemistry at various time points (3d,7d,14d) to study thesecretion of extra cellular matrix and differentiation of ADSCs induced by the cocultureenvironment.Use the prostaglandin E2to establish the tendinopathy model, have the ADSCs compounded PRP reinjected and analysis the effect of its restoration.Results: ADSCs and Tendon Cells were isolated by using explant cuture and wereproved to be tenocytes. Neither significant TNMD nor type I collagen expression wasobserved3days after coculture. Some of the Type I collagen and TNMD expression byADSCs in coculture environment were P+positive staining7days later. On the14th dayafter coculture, type I collagen and TNMD expression by ADSCs in cocultureenvironment and ADSCs of experimental groups were widely stained byimmunohistochemical staining. Mean while, the results of immunohistochemicalstaining were measured for statistical analysis. The statistical data showed thatexperimental groups had a notable higher type I collagen and TNMD expression thanthe negative control but lower than the positive control. Have the ADSCs compoundedPRP reinjected and analysis the effect of its restoration. Data of experiment shows thatrestoration effects of the experimental group are better than the control group.Conclusions: Through explant culture method the tenocytes and ADSCs of NewZealand white rabbits could be successfully isolated. The type I Collagen and TNMDexpression of ADSCs could be induced by the environment of beingcocultured with tenocytes。There is some significant effect of PRP on the secretionof extra cellular matrix of ADSCs in the environment of coculture. Evidence proves thatADSCs could differentiate into tenocytes after being induced by the environment ofcoculture.Result of the animal reinjection shows that ADSCs compounded platelet-richplasma helps repair the tendinopathy.