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The purpose spine fracture patients is often complicated by the supraspinous ligament, interspinous ligament injury. Ligament injury does not heal or scar healing, often leading to significant loss of joint function and pain. Despite ligament injury can heal, but the healing ligament ultrastructure and biochemical composition of great change, even in a number of years after the healing of the ligament and its biomechanical properties than normal ligament. At home and abroad in recent years, researchers have tried using biological methods to promote the healing of ligaments. Its main cytokine therapy, gene therapy and tissue engineering methods. Bone marrow mesenchymal stem cells (Mesenchymal stem cells, MSCs). Potential as a stem cell, with a multi-directional differentiation. In this experiment, the MSCs as seed cells, tissue engineering methods to promote the healing of ligaments. Observe the methods to promote rat supraspinous, interspinous ligament healing effect. Method of MSCs in vitro culture the rat femur MSCs by density gradient centrifugation get. Inoculated to a density of 2.0 × 10 5 / mL of DMEM medium. Primary cell proliferation to approximately 80% of the covered bottom ratio of 1:3, to be passaged. Second, cells - Preparation of the composite body of the collagen sponge in the third generation cell culture medium was added to 3mg / L BrdU cultured for 24 hours to be marked. And then digested with trypsin, and cell suspension diluted to a concentration of 10 × 10 6 / mL of DMEM culture medium, dripping the UV disinfection has been used on the collagen sponge, 5% CO 2 incubation boxes at 37 ° C or incubated for 24 hours so that the cells were adsorbed to the collagen sponge made of cells - collagen sponge composite. Animal model cut rat L4-L5 supraspinous, interspinous ligament. Experimental group cells - collagen sponge composite grafts in the lesion, the control group, only transplanted collagen sponge. , HE staining 4 weeks after transplantation animals were killed, cut specimens were taken for histological examination. Each the specimens selected a slice HE staining. Immunohistochemical staining again select the slices of each specimen, do immunohistochemical staining. Nucleus brownish yellow by positive staining cells. Results of MSCs in vitro culture can be seen under an inverted microscope 3 days after inoculation, MSCs that some cells adherent. Cultured for 12-14 days of cell growth into a single layer. After passage, MSCs proliferation 9-10 days significantly accelerated both fused into a single layer. Second, the results of the animal models of 20 animals survived well. Third, the experimental group after 4 weeks of the results of histological observation visible six cases of transplantation ligament-like tissue is formed at the visible shape, arrangement direction close to the normal tissue, fibroblast cells, similar to the normal ligament tissue. The control group formed by the newly formed tissue containing more fat tissue and cell disorder, collagen fibers were loose network of filamentous. Between the two groups healing superior, as the percentage difference was significant (P <0.05). Immunohistochemical results in the experimental group slices visible nucleus tan-positive cells. The control group showed positive staining cells. Discussion the MSCs as ideal seed cells. Use of collagen type I is more reasonable as a carrier. Part of the tissue to repair damaged ligaments from the transplanted MSCs. MSCs without the addition of exogenous growth factors, can repair ligament defect in the body effectively. Experimental group ligament recovered well, relatively poor recovery of the control group. Conclusion tissue engineering technology to MSCs repair supraspinous, interspinous ligament injury is feasible. The method can promote the healing of the ligament, and a healing effect in the natural healing effect of the ligament.
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