Cytotoxicity Testing of a Novel Crosslinked PRF (C-Prf) Developed for Dental Pulp Tissue Engineering Purpose

Abstract

Background- Whole pulp amputation followed by pulp space disinfection and lling with an articial material causes loss of signicant amount of dentin leaving a non-vital and weakened tooth. Regenerative endodontics with its emerging eld of modern tissue engineering has demonstrated promising results using stem cells associated with scaffolds and responsive molecules. [1] Introduction- PRF was recognized as the “second generation” of this family of biomaterials. [6] PRF being tested in pulp tissue engineering by different research groups showed mixed results. (7,8) Research studies have shown that the interactions between the cells and their niche are closely related to physicochemical properties of the scaffolding materials [9, 10]. As PRF is a fragile gel its physical character needs to be improved by cross linking and thereby more longer period of liberation of its growth factors and delayed disintegration in physiological system. Aims and Objectives- Aim of our study was to prepare a very economical and autologous biomaterial for pulp tissue engineering by crosslinking of PRF with tannic acid. Our objective was to detect cytotoxic effect of tannic acid in PRF. Methods and Materials- We followed Choukroun et al. protocol to prepare PRF samples from whole venous blood collected from donors. PRF samples were then cross-linked in freshly prepared TA solution in dapendish for 10 minutes at room temperature. Concentrations of TA 1 wt% was used for preparing samples. After crosslinking, the gels were washed with normal saline for 5 min. to ensure that all excess TA was removed. The viability of cells cultured on the scaffolds was assessed through MTT assay (EZcountTM MTT cell Assay Kit, HiMedia, Mumbai, India). Observations- Both MTT Assay and Phalloidine staining showed favourable results of no clear cytotoxic effects of C-PRF. Conclusion- Based on the results of the cell viability analysis it can be concluded that none of the tannic acid crosslinked PRF created any clear cytotoxicity in the MC3T3 cells. So, C-PRF can safely be used as scaffold for dental pulp or similar tissue engineering purposes.