Osteogenic Differentiation Gingival Stromal Progenitor Cells with Platelet Rich Fibrin for Bone Remodelling (In-vitro Study)

Objectives: to analyze osteogenic differentiation ability of Gingival Stromal Progenitor Cells (GSPCs) with Platelet Rich Fibrin for bone remodeling.
Methods: true experimental with post-test only control group design. Sample groups were selected by random sampling method. GPSCs were isolated from four healthy, 250 gram, 1 month old, male rat’s (Rattus Novergicus) lower gingival tissue through gingivectomy procedure. Gingiva were minced into small fragments then cultured in 2 weeks. The culture was passaged every 4-5 days after cultured and plated. GSPCs isolated in passage 3 were characterized by CD34, CD45, CD44, CD73, CD90, CD105 using Fluorescein Isothiocyanate (FITC) Immunocytochemistry (ICC) and Flowcytometry (FC) examination. GSPCs in passage 3-5 cultured in twelve M24 culture plates (n=270; each sample marker n=6) for Day 7, Day 14, and Day 21 with 3 different cultures medium group (Control negative group:αModified Eagle Medium (αMEM); Control positive group:High Glucose-Dulbecco’s Modified Eagle Medium (DMEM-HG)+osteogenic medium; Treatment group: DMEM-HG+osteogenic medium+PRF). Osteogenic differentiation potential ability of GSPCs invitro cultured in 3 different cultures medium by evaluating several markers such as Runt-Related Transcription Factor 2 (RUNX2), Bone Alkaline Phosphatase (BALP), and Osteocalcin (OSC) examined with ICC using monoclonal antibody. Oneway Analysis of Variance (ANOVA) was performed (p<0.05) based on Saphiro-Wilk normality test and Levene's of variance Homogeneity test (p>0.05), then continued by post-hoc Least Significant Difference (LSD) test (p<0.05).
Results: GSPCs were showed +CD44, +CD73, +CD90, +CD105 and CD34, -CD45 expression as Stromal Progenitor Cells marker. Positive expressions of RUNX2, BALP, and OSC in all culture medium. Treatment group showed highest RUNX2 (14.2±1.32) and BALP (16.1±1.12) expression in Day 7 while OSC expression (13.5±1.29) in Day 21 with significant different between groups (p<0.05).
Conclusions: GSPCs cultured in PRF had potential osteogenic differentiation ability that able to accelerate bone remodeling in vitro by increased expression of RUNX2, BALP, and OSC.