Attachment and proliferation of human gingival fibroblasts seeded on barrier membranes using Wharton's jelly‐derived stem cells conditioned medium: An in vitro study

Abstract

AbstractThe effect of Wharton's jelly mesenchymal stem cells conditioned medium (WJMSCs‐CM) and zinc oxide nanoparticles (ZnO‐NPs) on cultured human gingival fibroblasts on various barrier membranes was investigated in this study. In this study, human gingival fibroblasts were prepared and cultured on three membranes: collagen membrane, acellular dermal matrix (ADM) with ZnO‐NPs, and ADM without ZnO‐NPs. WJMSCs‐CM was given to the testing groups, while control groups received the same membranes without WJMSCs‐CM. Following 48 and 72 h, 3‐(4,5‐Dimethylthiazol‐2‐yl)‐2,5‐diphenyltetrazolium bromide (MTT) tests were performed to assess cell survival. Cell proliferation on the membranes was also evaluated using 4′,6‐diamidino‐2‐phenylindole (DAPI) staining after 48 and 72 h. Field emission scanning electron microscopy was used to determine membrane surface structure and cell adhesion. Nanoparticles were also subjected to an energy‐dispersive x‐ray analysis to identify their chemical structure. Two‐way analysis of variance was used to conduct the statistical analysis. The p‐value ≤.05 was considered significant. When ADM‐ZnO‐NPs were combined with CM, fibroblast viability, and adhesion significantly differed from ADM‐ZnO‐NPs alone. DAPI results confirmed cell proliferation in all six groups on both experiment days. The abundance and concentrated distribution of cells during cell proliferation were found in CM‐containing membranes, specifically the ADM‐ZnO‐NPs membrane, demonstrating the improved biocompatibility of the ADM‐ZnO‐NPs membrane for cell proliferation. The other groups did not significantly differ from one another. WJMSCs‐CM positively affected the viability and proliferation of gingival fibroblasts, but only marginally. Under certain conditions, ZnO‐NPs below a specific concentration increased the biocompatibility of the membranes.