Abstract:

Objective To construct functional tissue-engineered bone with cell sheet technology and method of traditional bone tissue engineering. Methods Canine bone marrow mesenchymal stem cells（BMSCs）were isolated with the method of density gradient centrifugation and cultured. BMSCs were induced to differentiate into osteoblasts and cultured in temperature-responsive culture dishes at 37 ℃, 5%CO2 and saturated humidity. BMSCs cell sheet was prepared when temperature was changed to 20 ℃. Demineralized bone matrix（DBM） and platelet-rich plasma（PRP） were prepared, and complex of DBM/PRP/BMSCs cell sheet/BMSCs was construsted and implanted under the left latissimus dorsi muscle. Complex of DBM/PRP/BMSCs was implanted under the right latissimus dorsi muscle. Results When temperature dropped at 20 ℃, BMSCs detached automatically from the temperature-responsive culture dishes and formed an intact cell sheet. The osteogenesis of the DBM/PRP/BMSCs cell sheet/BMSCs group was better than that of the DBM/PRP/ BMSCs group. Conclusion Cell sheet technology combined with traditional bone tissue provides a new way for construction of ideal functional tissue-engineered bone.

Key words: bone marrow mesenchymal stem cells, cell sheet technology, tissue-engineered bone