应用牙周膜干细胞—牙囊干细胞复合细胞膜片同种异体移植修复比格犬牙周组织缺损的研究

The study of the application of the periodontal membrane stem cells-dental follicle stem cells composite membrane allograft in the repairment of Beagle canine periodontal tissue defect

目的拟构建比格犬牙周缺损模型,观察牙周膜干细胞—牙囊干细胞复合细胞膜片修复牙周组织缺损的效果。方法分离培养比格犬牙囊干细胞、牙周膜干细胞,制备牙囊干细胞细胞膜片、牙周膜干细胞细胞膜片以及以上2种细胞的复合细胞膜片,将膜片混合支架材料同种异体植入成年雄性健康比格犬牙周缺损模型,对照组只植入支架材料。分别于移植后1周、2周、4周、6周、8周、12周检测探诊深度、牙龈退缩、附着丧失情况。12周后观察分析牙周再生的组织形态学变化。结果牙周临床检查及组织学形态学分析显示,3个膜片移植组牙周再生效果较对照组效果明显,其中复合细胞膜片移植组明显高于其余两个单一膜片移植组(P<0.05)。结论牙周膜干细胞—牙囊干细胞复合膜片同种异体移植更有利于修复比格犬牙周缺损。

Objective To construct the Beagle canine periodontal defect model and to observe the effect of periodontal tissue defect repairment of the periodontal membrane stem cells-dental follicle stem cells composite membrane. Methods Dog follicle stem cells（ d DFSCs） and dog periodontal ligament stem cells（ d PDLSCs） were isolated enzymatically and expanded in vitro. The d DFSCs, d PDLSCs and the mixed cells were used to make cell sheets separately. Then, three types of cell sheets were transplanted to dental root surfaces having three-wall periodontal defects in an allogeneic manner, and bone defects were filled with hydroxyaptite-tricalcium phosphatel（ HA-TCP）. The group only filled with HA-TCP was set as the control group. The periodontal clinical indices were evaluated at the time of 1 week and 2, 4, 6, 8 and 12 weeks after surgery. 12 weeks after surgery, the animals were sacrificed and decalcified specimens were prepared. Healing of the periodontal defects was evaluated histologically and histometrically. Results Periodontal clinical examination, histological and histometrical analysis indicate that the periodontal healing of cell sheets groups were superior to the control group, and the mixed cells sheets group was superior to the other two groups（ P 0. 05）. Conclusion Allogeneic composite cell sheets of d DFSCs and d PDLSCs shows better effects in the repairment of Beagle periodontal tissue defect.