Abstract
In this study, novel human-derived epithelial-like cells (hEPLCs) lines were established from periodontal ligament (PDL) tissues, which were composed of a variety of cell types and exhibited complex cellular activities. To elucidate the putative features distinguishing these from epithelial rest of Malassez (ERM), we characterized hEPLCs based on cell lineage markers and tight junction protein expression. The aim of this study was, therefore, to establish and characterize hEPLCs lines from PDL tissues. The hEPLCs were isolated from PDL of third molar teeth. Cellular morphology and cell organelles were observed thoroughly. The characteristics of epithelial–endothelial-mesenchymal-like cells were compared in several markers by gene expression and immunofluorescence, to ERM and human umbilical-vein endothelial cells (HUVECs). The resistance between cellular junctions was assessed by transepithelial electron resistance, and inflammatory cytokines were detected by ELISA after infecting hEPLCs with periodontopathic bacteria. The hEPLCs developed into small epithelial-like cells in pavement appearance similar to ERM. However, gene expression patterns and immunofluorescence results were different from ERM and HUVECs, especially in tight junction markers (Claudin, ZO-1, and Occludins), and endothelial markers (vWF, CD34). The transepithelial electron resistance indicated higher resistance in hEPLCs, as compared to ERM. Periodontopathic bacteria were phagocytosed with upregulation of inflammatory cytokine secretion within 24 h. In conclusion, hEPLCs that were derived using the single cell isolation method formed tight multilayers colonies, as well as strongly expressed tight junction markers in gene expression and immunofluorescence. Novel hEPLCs lines exhibited differently from ERM, which might provide some specific functions such as metabolic exchange and defense mechanism against bacterial invasion in periodontal tissue.
Highlights
IntroductionElectronic supplementary material The online version of this article (doi:10.1007/s13577-017-0173-y) contains supplementary material, which is available to authorized users
Electronic supplementary material The online version of this article contains supplementary material, which is available to authorized users.Periodontal ligament (PDL) tissue is a specialized and complex connective tissue of ectomesenchymal origin
In this study, novel human-derived epithelial-like cells lines were established from periodontal ligament (PDL) tissues, which were composed of a variety of cell types and exhibited complex cellular activities
Summary
Electronic supplementary material The online version of this article (doi:10.1007/s13577-017-0173-y) contains supplementary material, which is available to authorized users. Periodontal ligament (PDL) tissue is a specialized and complex connective tissue of ectomesenchymal origin. PDL attaches tooth root cementum to alveolar bone for reciprocal support of the periodontium and tooth. PDL plays a pivotal roles in homeostasis, maintenance, and regeneration of periodontium. The heterogeneous cell population in PDL tissue is comprised of various cell types including fibroblasts, endothelial cells, epithelial cells, PDL progenitor cells, and inflammatory cells. The differentiation capacity and differentiation stage of the cell populations of PDL tissue depend on the expedient environment [1,2,3,4]. The PDL cell populations and their functions have not yet been completely characterized
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