Combined Treatment with Low-Level Laser and rhBMP-2 Promotes Differentiation and Mineralization of Osteoblastic Cells under Hypoxic Stress.

BACKGROUND: The aim of this study was to evaluate the combined effect of low-level laser treatment (LLLT) and recombinant human bone morphological protein-2 (rhBMP-2) applied to hypoxic-cultured MC3T3-E1 osteoblastic cells and to determine possible signaling pathways underlying differentiation and mineralization of osteoblasts under hypoxia. METHODS: MC3T3-E1 cells were cultured under 1% oxygen tension for 72 h. Cell cultures were divided into four groups: normoxia control, low-level laser (LLL) alone, rhBMP-2 combined with LLLT, and rhBMP-2 under hypoxia. Laser irradiation was applied at 0, 24, and 48 h. Cells were treated with rhBMP-2 at 50 ng/mL. Alkaline phosphatase activity was measured at 3, 7, and 14 days to evaluate osteoblastic differentiation. Cell mineralization was determined with Alizarin red S staining at 7 and 14 days. Western blot assays were performed to evaluate whether p38/protein kinase D (PKD) signaling was involved. RESULTS: The results indicate that LLLT and rhBMP-2 synergistically increased alkaline phosphatase (ALP) activity and mineralization. Western blot analyses showed that expression of type I collagen, runt-related transcription factor 2 (RUNX2), and Osterix (Osx), increased and expression of hypoxia-inducible factor 1-alpha (HIF-1α), decreased more in the LLLT and rhBMP-2 combined group than in the rhBMP-2 or LLL alone groups. Moreover, LLLT and rhBMP-2 stimulated p38 phosphorylation and rhBMP-2 and LLLT increased Prkd1 phosphorylation. CONCLUSION: Combined treatment with rhBMP-2 and LLL induced differentiation and mineralization of hypoxic-cultured MC3T3-E1 osteoblasts by activating p38/PKD signaling in vitro.

Osseointegration of magnesium-incorporated sand-blasted acid-etched implant in the dog mandible: Resonance frequency measurements and histomorphometric analysis.

The aim of this pilot study was to investigate the bone responses of novel magnesium (Mg)-incorporated sand-blasted and acid-etched (SLA) titanium (Ti) implant in an experimental animal model. Novel Mg-incorporated SLA Ti implant was obtained via vacuum arc source ion implantation method and Mg-ions were implanted into the SLA implant surface. Control group consisted of two commercial implants; resorbable blasting media (RBM) and SLA. Twelve implants from each group were placed into the mandibles of 6 mongrel dogs. Experimental animals were divided into 2 groups of 3 animals, with 4 weeks and 8 weeks healing time points. Resonance frequency analysis was performed at the time of fixture installation, 1, 2, 4, and 8 weeks after installation. Bone to implant contact (BIC) measurements were assessed at the 4 and 8 weeks healing time points. The overall implant survival rate was 97.2%. The Mg-incorporated SLA Ti implants showed more rapid osseointegration than control group implants at follow-up periods of 4 weeks. Histomorphometric analysis showed a tendency for BIC% values of Mg-incorporated SLA Ti implant to be higher than that of other the implant groups. The results of this study suggest that Mg-incorporated SLA Ti implant may be effective in enhancing the bone responses by rapid osseointegration in early healing periods.