Proliferation of rabbit bone marrow stromal stem cells on varying degrees of decalcified bone matrix materials / 中国组织工程研究

ABSTRACT

BACKGROUND: Demineralized bone matrix (DBM) has natural mesh pores, good plasticity and biocompatibility. However, the decalcification time in preparation of DBM remains controversial.OBJECTIVE: To compare effects of varying degrees of decalcification with DBM on the proliferation of bone marrow stromal stem cells (BMSCs) so as to provide the best DBM scaffold for cartilage tissue engineeringMETHODS: Rabbit iliac bones were prepared into strips, defatted, followed by 6,12 and 24 hours of decalcification and 2 days of soaking in alcohol to prepare DBM. DBMs were placed in 24-well plates. The third passage of BMSCs at a density of 5×10~9/L were incubated on 24-well plate with DBMs. The DBM porosity and pore size were observed by scanning electron microscopy;BMSCs proliferation on the DBM was determined by MTT, and cell attachment on DBM was observed by scanning electron microscopy.RESULTS AND CONCLUSION: DBM displayed natural high-density porous grid structure, in the presence of bone trabecula,trabecular space and bone luminal system. The porosity and pore size of DBM decalcified for 6,12 and 24 hours were similar (P > 0.05). Compared with decalcified for 12 and 24 hours, BMSCs attached to DBM decalcified for 6 hours more closely and the DBM showed better compatibility. SEM observation showed the BMSCs on DBMs decalcified for 6 hours proliferated stably after 8 days and applicable for transplantation. Moreover, the number of cells were significantly more than DBMs decalcified for 12 and24 hours (P< 0.01).