摘要
牙周治疗的最终目标是牙周组织的完全再生和形成牙周新附着，因牙周病而丧失的牙周组织，其修复与重建的难度很大，传统的牙周治疗和引导性组织再生技术虽取得一定的临床效果，但总的看来，尚不能达到牙周组织完全再生的目的。组织工程技术为牙周病的治疗带来了新的希望。组织工程学研究的主要内容包括种子细胞的来源、生物载体支架的选择以及组织工程化组织的构建等三个方面。种子细胞的研究是牙周组织工程的重要内容。自体牙周膜细胞(PDLCs)可作为牙周组织工程较为理想的种子细胞，但必须拔除一定量的自体牙才能获得，临床应用受到很大限制，不易开展。骨髓基质细胞(MSCs)具有与PDLCs相似的多向分化潜能，较PDLCs来源丰富、取材简单、对机体的损伤小的优势。本研究体外分离培养了犬MSCs，并与支架材料复合修复犬人工牙周组织缺损，通过组织学观察、免疫组化及示踪方法评价MSCs对牙周组织再生的作用。初步探讨了MSCs作为牙周组织工程种子细胞的可行性，并为牙周病的治疗提供新的思路和途径。
方法：穿刺抽吸犬髂骨骨髓，进行体外培养，采用细胞形态学观察、生长曲线测定、碱性磷酸酶(ALP)染色、钙染色等方法观察MSCs的生物学特性；以自体MSCs复合乌贼骨转化羟基磷灰石材料(CBHA)修复犬人工牙周组织缺损，通过组织学观察、免疫组化方法评价MSCs对牙周组织再生的作用；
第四军医大学硕士学位论文
采用示踪实验观察MSCs在新生牙周组织的分布。
结果:犬MSCs可以在体外适宜条件下培养，经诱导有成
骨特性;MSCs复合CBHA可促进牙周组织再生;MSCs分布
在新生牙槽骨、新生结缔组织及新生牙骨质中。
结论:MSCs来源丰富、取材方便、损伤小、可参与牙周
组织修复，有可能成为牙周组织工程的种子细胞。
The ultimate aim of periodontal treatment is periodontal regeneration or forming new attachment apparatus which is one of the hotspot of periodontology. Tissue engineering has been introduced into the field of periodontology, which contribute a new vista to periodontal regeneration. One of the important parts of tissue engineering is finding proper seeded cells. Autogenous periodontal ligament cells (PDLCs) is expected to be the ideal seeded cells. However, there are also some disadvantages for PDLCs to be seeded cells, for example , extraction is necessary to obtain PDLCs, which is difficult to carry out in clinic. Marrow stromal cell (MSCs) has potential of multiple differentiation like PDLCs. Being seeded cells of periodontal tissue engineering MSCs has some advantages including abundant source, easy to obtain, less injury to body compared with PDLCs. In this study, to evaluate the effects of MSCs in periodontal regeneration, MSCs of dogs were cultured in vitro and were implanted to artificial furcation d
efects combined with cuttlebone-transformed-hydroxyapatite (CBHA). The feasibility of MSCs as seeded cells of periodontal tissue engineering was discussed.
This study may provide a new method of periodontal regeneration.
Method: MSCs of dogs obtained from dogs' iliac crest were cultured. During the culture, the growth and biological characteristic of MSCs were observed by cell morphological survey, cell growth curve, alkaline phosphatase(ALP) stain and Von Kossa stain. MSCs were placed into artificial furcation defects combined with CBHA. Immunohistochemical stain and histological observation were carried to evaluate the effects of MSCs in periodontal regeneration. The distributing of MSCs in regenerate tissue was observed by cell labeling.
Result: MSCs of dogs obtained from dogs' iliac crest could be cultured in vitro under certain condition and express osteoblastic characteristic after being cultured in induced culture media. Periodontal regeneration was promoted by MSCs combined with CBHA. MSCs were distributing in regenerate tissue including alveolar bone, cementum, periodontal ligament.
Conclusion: MSCs can be seeded cells of periodontal tissue engineering.
引文
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