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.
Injectable Platelet Rich Fibrin (i-PRF) is a popular periodontal regenerative therapy in dentistry. i-PRF centrifugation resulted in the fractionation of the yellow layer and buffy coat with a red layer. The red i-PRF contains a greater number of Growth Factor carrier cells than yellow layer, besides that liquid-PRF has a suitable consistency so it easy to combine with other biomaterials. Another biomaterial that can be combined with i-PRF is collagen-chitosan with hydrogel preparations. This study aims to determine the effect of a combination of collagen-chitosan hydrogel formulation with injection of platelet-rich fibrin fractionation on osteoblast cell migration and proliferation. This type of research was an experimental laboratory divided into 3 groups, which were given a combination of the collagen-chitosan hydrogel and i-PRF red layer, a combination of collagen-chitosan hydrogel and i-PRF yellow layer, and only a combination of collagen-chitosan hydrogel. Chitosan collagen hydrogel was prepared with a collagen/chitosan ratio of 25/75, then mixed with i-PRF fractionation with a ratio of 1:1. The cell proliferation test used CCK-8, while the cell migration test used scratch wound healing. Data were tested with One way ANOVA for cell migration and Two way ANOVA for cell proliferation, followed by a Post Hoc test. The results showed that migration of osteoblast cells after 24 hours in the combination group of chitosan collagen hydrogel and i-PRF red layer was higher than the other groups with significance (p<0.05). The results of osteoblast cell proliferation on days 1, 3, and 5 in the combination group of chitosan collagen hydrogel and i-PRF red layer were also higher than the other groups with significance (p<0.05). This study concluded that the combination of chitosan collagen hydrogel with injectable platelet-rich fibrin red layer had a more effect on increasing the migration and proliferation of osteoblast cells than the other groups.
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