Preparation Of Acellular Bovince Cancellous Bone And Biocompatibility With Co-cultured Rabbit Bone Marrow Stromal Cells Study In Vitro

The ideal scafold materials of tissue engineering should have the good biocompatibility, proper biodegradation, porous structure and proper mechanical function, which will meet the need of the cells adhesiveness,growth, proliferation and differentiation. Now biomaterial research focus on how to make acellular tissue matrix. Acellular bone extracellular matrix(ABECM) as scafold materials which had been proven excellent in bone tissue engineering.The scaffold provides the base of cell adhesion, growth and diferentiation, also the scaffold is the structure that fixes growth factors in cell components and maintains three dimension spatial configuration to create new tissue. Although someone has narrated cell framework of acellular membrane,no ideal preparation of ABCB or histomoiphometry of bone lacunas have been reported in details. This experiment prepares Acellular Bovince Cancellous Bone (ABCB) by removing cell components and carries morphologic study to provide foundation for new natural supporting material of bone tissue engineering.Bone marrow stromal stem cell(BMSCs) can easily expand in vitro,and can give rise to a broad spectrum of fully differentiated connective tissues under appropriate perimental conditions. BMSCs have a numher of potential therapeutic applications. Their applications in the tissue engineering of bone are not enough. We used BMSCs as seed cells and cultured the cells combined with ABCB in vitro, and observed theintegration of the donor cells.This provided the morphologic foundation for new natural supporting scafold of tissue engineering of bone.1.ABCB preparationbovince upper humerus cancellous bone was made of 15mm×10mm×3mm sizes.It was approached acellularly by using 0.5%sodium chloride,10% sodium chloride and TritonX-100 according to analytic factorial experiment.Speciments were stained with HE, Massion and observed with naked eye,microscopy and scanning electron microscopy,to find the best mathod of ABCB.2.The culture and induction of BMSCs in vitroThe primary culture of New Zealand Rabbit's BMSCs, which were collected from the femur and tibia marrow of rabbit about a month old by PBS, centrifugalized in the rate of 2000r/min, mixed with DMEM +20% NBCS, cultured in the condition of 37℃,5%CO2,,and refreshed the DMEM every three day. Harvested them after they formed one layer, the cells were washed,trypsinized for 3 min, and suspended in...