Osteogenic induction of human periodontal ligament fibroblasts under two- and three-dimensional culture conditions
Osteogenic induction of human periodontal ligament fibroblasts under two- and three-dimensional culture conditions
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DOI：
10.1089/ten.2006.12.257
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发表时间：
2006-02-01
影响因子：
--
通讯作者：
Elcin, YM
中科院分区：
文献类型：
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作者：
Inanc, B;Elcin, AE;Elcin, YM
人牙周膜成纤维细胞(HPDLF)在引导组织再生过程中对牙周室的再生起着关键作用。这一特性归因于居住在该地区的祖细胞亚群。本研究的目的是探讨在二维和三维(2D和3D)培养条件下，hPDLFs在成骨诱导条件下是否可以进行成骨分化。从6名健康供者身上分离hPDFL，按标准方案进行培养和扩增。然后，建立了三种成骨培养条件(地塞米松、抗坏血酸和β-甘油磷酸)：1)二维单层培养，2)矿化聚(DL-乳酸-乙醇酸)(PLGA)支架的三维静态培养，3)在NASA批准的模拟微重力条件下的矿化PLGA支架上进行三维培养。经过21天的成骨诱导后，大多数单层培养细胞已经分化为成骨细胞系，从形态变化、矿化分析和一些表型特征来看。然而，免疫组织化学显示，支架培养物表达的成骨标记蛋白水平高于单层培养物。第二，在生物反应器中构建的hPDLF-PLGA载体在培养21d后骨桥蛋白和骨钙素的表达比静态3D培养中的表达增加。结果表明，人牙周膜含有一组细胞亚群，能够进行成骨分化，并可能有助于邻近区域骨缺损的再生。在微重力生物反应器中种植的人PDLF矿化PLGA支架可用于支持体外成骨分化。因此，该系统可能为牙周组织工程提供新的潜在益处。
Human periodontal ligament fibroblasts (hPDLF) play a key role in the regeneration of periodontal compartment during guided tissue regeneration procedures. This property is attributed to the progenitor cell subsets residing in the area. The aim of this study was to investigate whether hPDLFs could undergo an osteogenic differentiation under two- and three-dimensional (2D and 3D) culture conditions upon osteogenic induction. hPDLFs were isolated from six healthy donors, cultured, and expanded according to standard protocols. Then, three osteogenic culture conditions ( dexamethasone, ascorbic acid, and beta-glycerophosphate) were established: 1) 2D culture as single-cell monolayer, 2) 3D-static culture on mineralized poly( DL-lactic-co-glycolic acid) ( PLGA) scaffold, and 3) 3D culture on mineralized PLGA scaffold inside the NASA-approved bioreactor stimulating microgravity conditions. After 21 days of osteogenic induction, the majority of monolayer cultures had undergone differentiation toward osteogenic lineage, as indicated by morphological changes, mineralization assay, and some phenotypical properties. However, immunohistochemistry revealed that the scaffold cultures expressed higher levels of osteogenic marker proteins compared with that of the monolayers. Secondly, hPDLF-PLGA constructs in bioreactor showed an increased expression of osteopontin and osteocalcin compared with that of static 3D culture after 21 days. Results indicate that human periodontal ligament contains a subpopulation of cells capable of undergoing osteogenic differentiation and presumably contributing to regeneration of bone defects in the adjacent area. Human PDLF-seeded mineralized PLGA scaffold in microgravity bioreactor may be used to support osteogenic differentiation in vitro. Thus, this system may offer new potential benefits as a tool for periodontal tissue engineering.