Comparison of the Effects of Xenograft Bone Granules Containing and Lacking Hyaluronic Acid on the Viability, Proliferation, and Differentiation of MG-63 Osteoblast-Like Cells: An In Vitro Study

Abstract Background: The therapeutic goal in periodontology is to preserve teeth and restore hard and soft tissues lost to periodontal disease. High–molecular–weight hyaluronic acid (HA) can stimulate bone formation during the wound-healing phase. One emerging approach in maxillofacial regenerative therapy is combining HA with xenogeneic bone grafts. This in vitro study compared the biological effects of an HA-enriched bovine bone xenograft (Cerabone® Plus) with those of a non-HA bovine bone xenograft (Cerabone® Granulate) on MG‑63 osteoblast-like cell viability, proliferation, migration, and osteogenic differentiation. Methods: Two bovine bone xenograft types—Cerabone® Plus (0.5–1 mm granule, HA-enriched) and Cerabone® Granulate (0.5–1 mm, without HA)—were tested. Extracts from each sample were prepared after 24 h and 72 h of incubation in culture medium. Cell viability and proliferation were assessed via a methyl-thiazolyl tetrazolium (MTT) assay after exposure to undiluted, half, and quarter dilutions. Migration was examined via the scratch wound-healing assay at 24 h and 48 h. Osteogenic differentiation was evaluated viaquantitative real-time PCR for alkaline phosphatase (ALP), osteopontin (OP), and osteocalcin (OC). Statistical analysis was performed via one-way ANOVA with Tukey's post hoc test via GraphPad Prism (p<0.05). Results: At 24 h of exposure, no significant differences in viability were detected among the examined extract dilutions from either xenograft and the control (p>0.05). After 72 h of exposure to the extracts for 24 h, Cerabone® Plus maintained significantly greater viability than Cerabone® Granulate did (p<0.05). After 72 h of exposure to the extracts, viability was significantly greater for Cerabone® Granulate at the half- and quarter-fold dilutions (p<0.05). At 48 h,scratch closure, migration and proliferation were lower in the Cerabone® Granulate group than in the Cerabone® Plus and control groups. Gene expression analysis revealed no significant difference in ALP between the groups, but OP and OC were significantly greater in the Cerabone® Plus group (p<0.05), suggesting accelerated osteoblastic maturation. Conclusions: Within the constraints of this in vitro design, HA-enriched bovine xenografts (Cerabone® Plus) promoted faster migration and greater late-stage osteogenic differentiation of MG‑63 cells thannon-HA xenografts did.