Effect on Cellular Vitality In Vitro of Novel APRF-Chlorhexidine Treated Membranes

J Funct Biomater. 2022 Nov 7;13(4):226. doi: 10.3390/jfb13040226.

Abstract

Introduction: Chlorhexidine (CHX) has been used for some time in clinical practice as a local antiseptic agent with excellent efficacy. The combination of CHX with APRF (Advanced-platelet rich fibrin) membrane has the potential to stimulate tissue regeneration and to provide a bactericidal effect. We hypothesize that this may reduce the rate of infections development and protect cell viability.

Aim: The aim of this study was two-fold-to create a stable APRF membrane treated with different concentrations of CHX (0.01% and 0.02%) and to monitor its effect on the viability of PDL cells in vitro. This benefits the introduction of a new protocol for APRF membrane production -CHX-PRF and enriches the available evidence on the effect of this antiseptic agent on PDL (Periodontal ligament) cells.

Materials and methods: APRF membranes were prepared by the addition of two concentrations (0.01% and 0.02%) of CHX. Membranes without the antiseptic were also prepared and used as control samples. PDL cells were cultivated on the membranes for 72 h. Cell number and vitality were examined by fluorescent cell viability assays.

Results: Our results demonstrated that a concentration of 0.01% CHX allowed the production of a stable APRF membrane. This concentration slightly reduced the viability of PDL cells to 96.7%, but significantly decreased the average number of cells attached to the membrane-149 ± 16.5 cells/field compared to controls -336 ± 26.9 cells/field. APRF-CHX 0.02% membranes were unstable, indicating a dose-dependent cytotoxic effect of CHX.

Conclusions: The introduced novel protocol leads to the production of a new type of APRF membrane-CHX-PRF. The incorporation of an antiseptic into the APRF membrane can improve its bactericidal activity and might serve as an important step for the prevention of postoperative infections.

Keywords: APRF membrane; Chlorhexidine; PDL cells; vitality.

Grants and funding

This research was funded by Medical University of Plovdiv, grant number 04/04.2019 and by project National University Complex for Biomedical and Applied Research, with participation in NUCBAR-BBMRI.BG, grant number D01-285/17.12.2019 and DO1-395/18.12.2020, within the National Road Map for Research Infrastructure.