Establishment and characterization of novel epithelial-like cell lines derived from human periodontal ligament tissue in vitro

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.Electronic supplementary materialThe online version of this article (doi:10.1007/s13577-017-0173-y) contains supplementary material, which is available to authorized users.

Mechanical stress alters protein O‐GlcNAc in human periodontal ligament cells

“…PDL may consist of fibroblasts, endo‐ and epithelial cells, osteoblasts and stem cells. However, fibroblast‐like cells are predominant in the cell cultures derived from PDL . The regulation of O‐GlcNAc in osteoblasts was investigated in a few studies, showing that osteoblast differentiation is influenced by O‐GlcNAc .…”

Los restos de Malassez, vida y destino en los tejidos odontogénicos. Una revisión de la literatura

“…Nos referimos específicamente a que los REM presentan organelos de unión específicos como tonofilamentos, hemidesmosomas, desmosomas y uniones estrechas cuya función es facilitar el paso de agua e iones. Los tonofilamentos pueden estar agrupados dentro del citoplasma y se denominan tonofibrillas, las cuales no tienen contacto con el cemento ( 1,3,15 ). La distancia entre el cemento y los REM fue medida en los tres tercios de la raíz siendo en el tercio apical de 21 micrones; en el tercio medio, de 33 micrones, y en el tercio cervical, de 41 micrones, lo que indica que se encuentran más alejados cuando se aproximan a la corona ( 25).…”

“…El tejido donde se encuentran los REM abarca las áreas que están alrededor de la raíz, sobre todo las regiones supracrestales, es decir, los tejidos que contienen colágeno y que están por encima de la cresta alveolar ósea, por lo que su participación en la regeneración periodontal es cada vez más evidente. Por tanto, se deberán realizar más estudios y análisis para investigar las ubicaciones específicas y sus funciones especializadas ( 15,16 ).…”