| Objective:to articular cartilage repair after injury is always difficult clinical problem, this and cartilage cells regeneration, poor ability of local repair factor and low concentration repair stents are related. In recent years the development of cartilage tissue engineering, for this kind of damage provides a feasible way. This experiment in vitro extraction rabbit bone marrow mesenchymal stem cells, ChuanDai and induction into cartilage cells, take with rabbit ears cartilage cells made off cartilage (decellularization cartilage, DC), and with the nano beta3phosphoric acid calcium (beta tricalcium phosphate, beta TCP) composite, join repair factor implanted damaged cartilage model. To observe the repair ability, for the clinical repair such damage to provide the basis.Methods:in vitro extraction Zealand white rabbit bone marrow mesenchymal stem cells (bone marrow stromal stem cells BMSCs), the in vitro separation, training, ChuanDai, determine the growth curve to flow cytometry for its identification technology, the third generation to add induced factor induction into cartilage cells, do II collagen type immunohistochemical stains evaluations. With rabbit ears take in cartilage trypsin-dedicated casein gained legally take off cell cartilage stents, line histologic dyed and scanning electron microscopy (sem) to take off the cells and pore stents. The in vitro will beta TCP-DC combined with a complex, with negative pressure attract will induce cartilage cells in the vaccination stent. After incubation implanted Zealand white rabbit has preparation knee joint damage model, and the blank, controlled and experiment are divided into three groups. Blank group defect not join restorative materials, the control group implanted beta TCP-DC restoration, the implant beta TCP-DC and TGF γ-regularly injected, IGF-I. Four months after take repair model line macroscopic observation and histological dyeing.Results:with density gradient centrifugation can be successful separation and amplification BMSCs, after training the cells are long spindle, spiral shape growth, ChuanDai cells grow quickly, curve is good, streaming results show that the third generation of highly express CD29, CD44, not CD45expression, which show that the training of the BMSCs for a single cells, high purity. Bone marrow stromal induced stem cells after21d II collagen type immune cells and cytoplasm is tan-yellow chemical color, can form cartilage cells. Rabbit ears cartilage matrix cells off the milky, histologic dyeing and scanning electron microscopy (sem) and take off the cells by stent pore even, complete structure, still save a lot of acidic sticky glycosaminoglycan and procollagen composition. The aperture length (33.70+/-4.33μm, pore ratio (65.23+1-135)%. In vitro will beta TCP-DC form a composite stents and vaccination induction, little cartilage cells of dyeing and scanning electron microscopy (sem) and histological both good adhesion, and accompanied by a lot of matrix secretion. Complex implanted defect20weeks after, the blank group not repair defects, and the bottom for white fibrous tissue. Lack of control groupConclusion:the application of density gradient centrifugation can be successful separation and BMSCs amplification, BMSCs induction for cartilage cells with foreign cells after take off the ear cartilage have good biocompatibility, beta TCP-DC composite stents more pore structure, degradation speed, organization and cells compatibility good adhesion, TGF-beta and I igf-ii, can promote the role of cartilage cell proliferation, in new cartilage formation at the same time, beta TCP-DC layer stents gradually degradation absorption, is tissue engineering articular cartilage repair the ideal seed cells carrier. |
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