PDF(Pubmed)
Abstract：
Gingival fibroblasts (GFs) can differentiate into osteoblast-like cells and induce osteoclast precursors to differentiate into osteoclasts. As it is unclear whether these two processes influence each other, we investigated how osteogenic differentiation of GFs affects their osteoclast-inducing capacity. To establish step-wise mineralization, GFs were cultured in four groups for 3 weeks, without or with osteogenic medium for the final 1, 2, or all 3 weeks. The mineralization was assessed by ALP activity, calcium concentration, scanning electron microscopy (SEM), Alizarin Red staining, and quantitative PCR (qPCR). To induce osteoclast differentiation, these cultures were then co-cultured for a further 3 weeks with peripheral blood mononuclear cells (PBMCs) containing osteoclast precursors. Osteoclast formation was assessed at different timepoints with qPCR, enzyme-linked immunosorbent assay (ELISA), TRAcP activity, and staining. ALP activity and calcium concentration increased significantly over time. As confirmed with the Alizarin Red staining, SEM images showed that the mineralization process occurred over time. Osteoclast numbers decreased in the GF cultures that had undergone osteogenesis. TNF-α secretion, a costimulatory molecule for osteoclast differentiation, was highest in the control group. GFs can differentiate into osteoblast-like cells and their degree of differentiation reduces their osteoclast-inducing capacity, indicating that, with appropriate stimulation, GFs could be used in regenerative periodontal treatments.
摘要：
牙龈成纤维细胞（GFs）可以分化为成骨细胞样细胞，并诱导破骨细胞前体分化为破骨细胞。由于不清楚这两个过程是否相互影响，我们研究了GFs成骨分化如何影响其破骨细胞诱导能力。为了建立逐步的矿化，四组GFs培养3周,在最后1、2或全部3周内不使用或使用成骨培养基。通过ALP活性评估矿化，钙浓度，扫描电子显微镜（SEM），茜素红染色,和定量PCR(qPCR)。诱导破骨细胞分化，然后将这些培养物与含有破骨细胞前体的外周血单核细胞(PBMC)共培养3周.用qPCR在不同时间点评估破骨细胞形成，酶联免疫吸附测定(ELISA),TRAcP活性,和染色。ALP活性和钙浓度随时间显著增加。如茜素红染色所证实，SEM图像显示矿化过程随时间发生。在经历成骨的GF培养物中，破骨细胞数量减少。TNF-α分泌，破骨细胞分化的共刺激分子，对照组最高。GFs可以分化为成骨细胞样细胞，其分化程度降低其破骨细胞诱导能力，表明,在适当的刺激下，GFs可用于再生牙周治疗。
