A Preliminary Study About The Effect Of Platelet-rich Plasma On Bone Marorw Stromal Cells In Femoral Head Necrosis Micro-environment In Vitro

Objective：To observe the effects of femoral head necrosis (FHN) micro-environment on the proliferative and osteogenetic activity of bone marrow stromalcells (BMSCs)．In addition, the effect of platelet-rich plasma(PRP) on BMSCs inFHN micro-environment.Methods：The dog models of FHN were estblished by using freezing methodwith liquid nitrogen. The crushed tissue of FHN was immersed in DMEM. TheDMEM with5%soak solution was were divided into5groups：blank group(addednormal DMEM),group A(added DMEM with5%soak solution),group B(addedDMEM with5%soak solution and5%PRP),group C(added DMEM with5%soaksolution and10%PRP) and group D(added DMEM with5%soak solution and15%PRP). The effect of soak solution or soak solution+PRP on proliferation ofBMSCs was examined by the MTT assay at3,6and8days. Furthermore,Extracellular alkaline phosphatase (ALP) assay(at7days),ALP staining(at10days),and the calcium staining(at19days) were used to evaluate the effect of soak solutionor soak solution+PRP on osteogenic differatiation of BMSCs.Results：Proliferation: at3,6and8days,MTT method showed that theproliferative activity of the group A was obviously stronger than that of the blankgroup(P<0.05), the group B,C,D was obviously stronger than the group A(P<0.05).There was no significant difference in group B,group C and group D at3days. Thegroup D was obviously stronger than the group B,C(P<0.05) at6days, but there wasno significant difference between group B and group C. At8days, there wassignificant difference in group B,group C,group D(P<0.05), the group D show thestrongest proliferative activity, Secondly group B, and thirdly group C. Osteogenicdifferatiation: Extracellular ALP assay,ALP staining, and the calcium stainingshow：there was no significant difference between group A and blank group. Theosteogenic differatiative of the group B was obviously stronger than groups A (P<0.05), but there was no significant difference in groups C,groups D and groups A.Conclusion：the femoral head necrosis micro-environment exhibits promotive effect on the prolfieration of BMSCs,but can`t exhibits promotive effect on theosteogenic differatiation．In femoral head necrosis micro-environment, PRP exhibitspromotive effect on the prolfieration, and proliferation strength of BMSCs wascorrelated to the density of PRP. BMSC proliferation would be accelerated moremarkedly by the higher density of PRP. PRP could promote BMSC osteogenicdifferatiation markedly and differatiation strength of BMSCs was correlated to thedensity of PRP. BMSC osteogenic differatiation would be accelerated by the lowdensity of PRP.