Enhancement of periodontal tissue regeneration by transplantation of osteoprotegerin-engineered periodontal ligament stem cells.
Enhancement of periodontal tissue regeneration by transplantation of osteoprotegerin-engineered periodontal ligament stem cells.
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DOI：
10.1186/s13287-015-0023-3
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发表时间：
2015-03-12
影响因子：
7.5
通讯作者：
Liu HC
中科院分区：
文献类型：
--
作者：
Su F;Liu SS;Ma JL;Wang DS;E LL;Liu HC
本研究的目的是评估人骨保护素（hOPG）转染的牙周膜干细胞（PDLSC）的组织工程复合物接种在β-磷酸三钙（β-TCP）上再生新西兰兔牙槽骨缺损的能力。从兔牙周膜组织中分离PDLSC，并在体外扩增以丰富PDLSC数量，并在特定诱导条件下评估其增殖活性和分化能力。利用Gateway技术构建含有hOPG和增强型绿色荧光蛋白（EGFP）的慢病毒载体，并转染兔PDLSCs。通过实时定量逆转录聚合酶链反应和蛋白质印迹法测定hOPG的表达。移植前将含有或不含工程化 hOPG 的 PDLSC 接种在 β-TCP 支架上。通过扫描电子显微镜进行细胞和材料的形态表征。 20只患有牙槽骨缺损的兔子被随机分为四组，分别移植β-TCP、PDLSCs/β-TCP和hOPG转染的PDLSCs/β-TCP，或不予治疗作为对照。术后12周处死动物进行组织学观察和组织形态学分析。 PDLSC 表达 STRO-1 和 vementin，并有利于条件培养基中的成骨和脂肪生成。慢病毒载体转染PDLSC后，hOPG的表达显着上调。 PDLSCs在β-TCP上附着和铺展良好，hOPG转染组和未转染组之间PDLSCs在β-TCP上的生长没有显着差异。组织学观察和组织形态学分析表明，与对照、β-TCP和PDLSCs/β-TCP复合物相比，hOPG转染的PDLSCs/β-TCP复合物在支架内表现出更早的矿化和更多的骨形成。在圆偏振光显微镜下通过 EGFP 基因表达确定，hOPG 转染的 PDLSC 的植入有助于新骨形成。本研究证明了β-TCP支架在体外和体内原代PDLSC培养和hOPG基因表达的可行性，并且hOPG转染的PDLSC可以作为牙周骨再生的潜在细胞来源，这可能揭示全身hOPG基因治疗与PDLSC组织工程相结合作为牙周组织工程牙槽骨再生的良好候选者的潜力。
The objective of the present study was to evaluate the capacity of a tissue-engineered complex of human osteoprotegerin (hOPG)-transfected periodontal ligament stem cells (PDLSCs) seeding on beta-tricalcium phosphate (β-TCP) to regenerate alveolar bone defects in New Zealand rabbits. PDLSCs were isolated from rabbit periodontal ligament tissues and expanded in vitro to enrich PDLSC numbers, and their proliferative activities and differentiation capability were evaluated under specific induction conditions. Lentiviral vector containing hOPG and enhanced green fluorescent protein (EGFP) was constructed by using Gateway technology and transfected into rabbit PDLSCs. The expression of hOPG was determined with quantitative real-time reverse transcription-polymerase chain reaction and Western blot. The PDLSCs with or without engineered hOPG were seeded on β-TCP scaffolds prior to transplantation. Morphological characterization of cells and materials was done by scanning electron microscope. Twenty rabbits with alveolar bone defects were randomly allocated into four groups and transplanted with β-TCP, PDLSCs/β-TCP, and hOPG-transfected PDLSCs/β-TCP or were left untreated as a control. Animals were sacrificed 12 weeks after operation for histological observation and histomorphometric analysis. PDLSCs expressed STRO-1 and vementin and favored osteogenesis and adipogenesis in conditioned media. Expressions of hOPG were significantly upregulated after transfection of the lentiviral vector into PDLSCs. PDLSCs attached and spread well on β-TCP, and there was no significant difference in growth of PDLSCs on β-TCP between the hOPG transfection group and the non-transfection group. The histological observation and histomorphometric analysis showed that the hOPG-transfected PDLSCs/β-TCP complex exhibited an earlier mineralization and more bone formation inside the scaffold than control, β-TCP, and PDLSCs/β-TCP complexes. Implantation of hOPG-transfected PDLSCs contributed to new bone formation as determined by EGFP gene expression under circularly polarized light microscopy. The present study demonstrated the feasibility of β-TCP scaffolds for primary PDLSC culture and expression of hOPG gene in vitro and in vivo, and hOPG-transfected PDLSCs could serve as a potential cell source for periodontal bone regeneration, which may shed light on the potential of systemic hOPG gene therapy in combination with PDLSC tissue engineering as a good candidate in periodontal tissue engineering for alveolar bone regeneration.
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影响因子：
14
作者：
Kondo, N;Ogose, A;Endo, N
通讯作者：
Endo, N
影响因子：
7.6
作者：
Mogi, M;Otogoto, J;Togari, A
通讯作者：
Togari, A
影响因子：
7.6
作者：
Kanzaki, H;Chiba, M;Mitani, H
通讯作者：
Mitani, H
影响因子：
4
作者：
McMahon, JM;Conroy, S;O'Brien, T
通讯作者：
O'Brien, T
影响因子：
1.9
作者：
Honda, Masaki J;Imaizumi, Mari;Morsczeck, Christian
通讯作者：
Morsczeck, Christian