Allogeneic Transplantation of Periodontal Ligament-Derived Multipotent Mesenchymal Stromal Cell Sheets in Canine Critical-Size Supra-Alveolar Periodontal Defect Model.
Allogeneic Transplantation of Periodontal Ligament-Derived Multipotent Mesenchymal Stromal Cell Sheets in Canine Critical-Size Supra-Alveolar Periodontal Defect Model.
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DOI：
10.1089/biores.2015.0043
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发表时间：
2016
影响因子：
--
通讯作者：
Izumi Y
中科院分区：
文献类型：
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作者：
Tsumanuma Y;Iwata T;Kinoshita A;Washio K;Yoshida T;Yamada A;Takagi R;Yamato M;Okano T;Izumi Y
牙周炎是一种慢性炎症性疾病，会导致牙齿支持组织的破坏，然后是牙齿脱落。虽然已有几种方法应用于牙周再生，但尚未完全实现牙周再生。使用细胞和支架相结合的组织工程被认为是一种可行的替代策略。我们已经证明了牙周膜来源的多能间充质基质细胞(PDL-MSC)片的自体移植在犬模型中再生了牙周组织。然而，自体细胞移植在临床上的适应症是有限的。因此，本研究使用犬水平牙周缺损模型评价了同种异体PDL-MSC片移植的安全性和有效性。用增强型绿色荧光蛋白(EGFP)标记犬PDL-MSCs，在温度敏感型培养皿上培养。将三层细胞片自体或异体移植到裸露的牙根表面。将β-磷酸三钙和胶原凝胶的混合物放置在骨缺损处。移植后8周，对犬实施安乐死，并进行显微计算机断层扫描和组织学分析。从石蜡切片中提取RNA和DNA，以验证移植部位是否存在EGFP。用酶联免疫吸附试验对外周血中的炎症标志物进行定量。自体移植组和同种异体移植组均观察到牙周再生。同种异体移植组新生牙骨质再生尤为明显，这是牙周再生的关键。外周血炎性标志物水平在自体和同种异体组之间差异不大。同种异体组石蜡切片中可检测到EGFP扩增产物。这些结果表明，同种异体PDL-MSC片可促进牙周组织再生，且无副作用。因此，同种异体PDL-MSC片移植有可能成为牙周再生的替代策略。
Periodontitis is a chronic inflammatory disease that induces the destruction of tooth-supporting tissues, followed by tooth loss. Although several approaches have been applied to periodontal regeneration, complete periodontal regeneration has not been accomplished. Tissue engineering using a combination of cells and scaffolds is considered to be a viable alternative strategy. We have shown that autologous transplantation of periodontal ligament-derived multipotent mesenchymal stromal cell (PDL-MSC) sheets regenerates periodontal tissue in canine models. However, the indications for autologous cell transplantation in clinical situations are limited. Therefore, this study evaluated the safety and efficacy of allogeneic transplantation of PDL-MSC sheets using a canine horizontal periodontal defect model. Canine PDL-MSCs were labeled with enhanced green fluorescent protein (EGFP) and were cultured on temperature-responsive dishes. Three-layered cell sheets were transplanted around denuded root surfaces either autologously or allogeneically. A mixture of β-tricalcium phosphate and collagen gel was placed on the bone defects. Eight weeks after transplantation, dogs were euthanized and subjected to microcomputed tomography and histological analyses. RNA and DNA were extracted from the paraffin sections to verify the presence of EGFP at the transplantation site. Inflammatory markers from peripheral blood sera were quantified using an enzyme-linked immunosorbent assay. Periodontal regeneration was observed in both the autologous and the allogeneic transplantation groups. The allogeneic transplantation group showed particularly significant regeneration of newly formed cementum, which is critical for the periodontal regeneration. Serum levels of inflammatory markers from peripheral blood sera showed little difference between the autologous and allogeneic groups. EGFP amplicons were detectable in the paraffin sections of the allogeneic group. These results suggest that allogeneic PDL-MSC sheets promoted periodontal tissue regeneration without side effects. Therefore, allogeneic transplantation of PDL-MSC sheets has a potential to become an alternative strategy for periodontal regeneration.