Effect of Collagen Chitosan Hydrogel with Injectable Platelet-Rich Fibrin on Alkaline Phosphatase Activity and Calcium Deposition An In Vitro Study on Osteoblast-like Cell Line MG63

Abstract

Abstract Background Periodontal tissue regeneration is one of the main goals of periodontal therapies that is consisted of 4 main components e.g., growth factor such as injectable Platelet-Rich Fibrin (i-PRF) and scaffold, using collagen chitosan hydrogel which hoped to replace bone graft because its donor risk and immunogenic responses.Methods Bone formation markers such as alkaline phosphatase activity, an enzyme produced by osteoblast, and calcium deposit, late marker of osteoblast mineralization were measured using ALP staining kit (SensoLyte) and Alizarin Red S staining (Merck) in 1, 7, 14, and 21 days using MG63 osteoblast and divided into 3 groups: Hydrogel i-PRF, Bone Graft i-PRF, and Control group. The result was then analyzed using Two-way ANOVA test and Post-Hoc Tukey HSD test.Results Hydrogel i-PRF application induced significant ALP activity in day 7 and 14 compared to Bone Graft and Control group. Calcium deposits in day 7 of Hydrogel i-PRF and Bone Graft i-PRF group were found to be significantly different to Control group. Hydrogel application still holds the highest deposit in 14 days to 21 days. These results showed the ossification capability of collagen chitosan hydrogel with i-PRF compared to bone graft application. Through PDGF and BMP content in i-PRF which are bounded to collagen fibril, acceleration of osteoblast’s proliferation and migration rate, followed by the rise of differentiation and mineralization rate of osteoblast by BMP.Conclusion collagen chitosan hydrogel with i-PRF application enhanced ALP activity and calcium deposit compared to bone graft with i-PRF.