Expression Of Bone Formation Markers Research Articles

Response of Bone and Periodontal Ligament Cells to Biodegradable Polymer Scaffolds In Vitro

In this in vitro study, the initial response of human periodontal ligament (PDL) cells and alveolar osteoblast-like cells (HOB) to three biodegradable polymers with varying pore size and different mechanical properties were evaluated. Scaffolds were synthesized from poly(L-lactide), [poly(LLA)], poly(L-lactide-co-1,5-dioxepan-2-one), [poly(LLA-co-DXO)], poly(L-lactide-co-ε-caprolactone), and [poly(LLA-co-CL)] with pore sizes greater or less than 90 µm by salt leaching. Cells were obtained from patients undergoing routine oral surgery. After 2—4 passages, the cells were grown on scaffolds and in culture plates (control) for 3 h (PDL cells), 3 days (PDL cells and HOB), 10 and 14 days (HOB), respectively. The cellular morphology and spreading were determined by scanning electron microscopy (SEM) and the attachment and proliferation were evaluated by MTT assays. The SEM images revealed heterogeneous cellular morphology and good spreading. Cellular attachment and proliferation were significantly higher on poly(LLA-co-DXO) and poly(LLA-co-CL) than on poly(LLA) scaffolds (p = 0.003) and highest for poly(LLA-co-DXO). Expression of bone formation markers, collagen-I (COL-I), transforming growth factor-β 1 (TGF-β1), and osteocalcin (OCN), was determined by ELISA. The expression of COL-1 was similar for HOB and PDL cells, but significantly higher for pore size >90 µm while the HOB expression of TGFβ 1 and OCN was greater on poly(LLA-co-CL) and poly(LLA-co-DXO) than on poly(LLA) scaffolds.

All-trans retinoic acid reversibly suppresses cancellous bone formation in intact female rats

While recombinant human bone morphogenetic protein (rhBMP)-2-based bone therapy presents potential osteoinductivity, it also leads concern due to transient osteoclast activation during early healing periods, ultimately limiting its clinical use. Therefore, we investigated in vivo and in vitro rhBMP-2 signaling which mediates early bone resorbing effect, depending on the dose, and attempted to inhibit this resorption phenomenon using NFAT inhibitor as a target molecule. High-dose of rhBMP-2 (20 μg/defect) enhanced osteoclast activation and the expression of bone resorption markers, compared to low dose (5 μg/defect) at one week after surgery in collagen sponge-delivered rat calvarial defect models. Interestingly, this trend was also observed in the expression of bone formation markers. In particular, rhBMP-2 upregulated RANKL expression, while it downregulated osteoprotegerin (OPG) expression, resulting in a dose-dependent increase in the ratio of RANKL to OPG. NFAT inhibitor (150 μm) treatment in vivo suppressed the high-dose effect of rhBMP-2 on both resorption and formation. In vitro results of rhBMP-2 signaling and NFAT inhibitor effects in rat mesenchymal stem cells showed similar trends as in vivo results. Microcomputer tomography-based evaluation at 4 weeks showed that combined treatment of NFAT inhibitor with 20 μg rhBMP-2 in vivo increased bone volume (BV) more than 20 μg rhBMP-2 alone, which showed little difference in BV compared to 5 μg of rhBMP-2. These results demonstrated that rhBMP-2 implantation concurrently signalized into enhanced osteoclastogenesis and osteoblastogenesis in vivo, dose-dependently. Ratio of RANKL/OPG might be an index for early bone resorbing activity of implanted rhBMP-2. A local cocktail treatment of NFAT inhibitor and high-dose rhBMP-2 might be an alternative to overcome early bone resorbing effects, thereby accelerating bone formation.