MiR-10a-5p Regulates Osteogenic Differentiation Of Human Bone Marrow Mesenchymal Stem Cells

Objective h BMSCs are a kind of multilineages potential stem cell, which can differentiate to diverse mesenchymal tissues, including bone, fat, and neuron cells [1]. h BMSCs are easily separated, cultured and generated, which have potential ability of differentiate to bone tissue after passaged. However, they also have other characterisetics, such as low immunogenicity, imunoregulation and being transfated or expressed the exogenous genes easily. For those reasons above, h BMSCs may be applied to bone tissue regeneretion engineering[2], repaiering root resorption and alveolar bone loss. micro RNAs(mi RNAs) are short-noncoding RNAs and act as important regulators in diverse biological processes. Growing evidences showed that mi RNAs played a significant role in proliferation and differentiation of stem cells, including osteogenesis process. Our study verified the biological functions of mi R-10a-5p in h BMSCs by in vitro and in vivo, aiming at providing the theoretical bases to improve regenerative capacity of h BMSCs.Methods 1. The origin and differentiative capacity of h BMSCs were confirmed by the immunocytochemical and osteogenic/adipogenic differentiation potential staining. 2. We cultured h BMSC in human mesenchymal stem cell basal medium and changed mi R-10a-5p expression of h BMSCs by transient transfection. With those procedures done, q-PCR and Western Blot were applied to analysis the difference of osteoblastic differentiative ablity after changing the endogenous mi R-10a-5p expression. 3. HA/TCP scaffolds were co-culutured with h BMSCs after transfection, and transplanted into the dorsal surfaces of BABL/c nude mice. We sacrificed the mice to make tissue sections to verify the mineral characteristics in vivo by Goldner's trichrome staining after 8 weeks.Results 1. The h BMSCs' differentiation abilities were indicated by osteogenic/adipogenic staining. And the immunocytochemical staining confired that the h BMSCs were positive for the mesenchymal stem cell marker CD44, and negative for the hematopoietic stem cell maker CD34. 2. Increasing the expression level of mi R-10a-5p, q-PCR and Western blot results showed that h BMSCs' osteogensis differentiation were inhibited. However, decreasing the expression of mi R-10a-5p promoted this process. 3. The Goldner`s thrisome staining of the speices shared the similar results with the in vitro, the mimic group was in early stage of mineralization process. However, the inhibitor group formulated large mount of mineralized bone like tissue than other groups.Conclusions 1. h BMSCs has multilingeages differentiation potential and origin from mesenchymal tissue, not hematopoietic. 2. mi R-10a-5p down regulates h BMSCs in osteogenic process in vitro and in vivo.