Myat Nyan

Evaluation of the osteoconductivity of α-tricalcium phosphate, β-tricalcium phosphate, and hydroxyapatite combined with or without simvastatin in rat calvarial defect†

The purpose of this study is to evaluate the osteoconductivity of three different bone substitute materials: α-tricalcium phosphate (α-TCP), (β-TCP), and hydroxyapatite (HA), combined with or without simvastatin, which is a cholesterol synthesis inhibitor stimulating BMP-2 expression in osteoblasts. We used 72 Wistar rats and prepared two calvarial bone defects of 5 mm diameter in each rat. Defects were filled with the particles of 500-750 μm diameter combined with or without simvastatin at 0.1 mg dose for each defect. In the control group, defects were left empty. Animals were divided into seven groups: α-TCP, β-TCP, HA, α-TCP with simvastatin, β-TCP with simvastatin, HA with simvastatin, and control. The animals were sacrificed at 6 and 8 weeks. The calvariae were dissected out and analyzed with micro CT. The specimens were evaluated histologically and histomorphometrically. In α-TCP group, the amount of newly formed bone was significantly more than both HA and control groups but not significantly yet more than β-TCP group. Degradation of α-TCP was prominent and β-TCP showed slower rate while HA showed the least degradation. Combining the materials with Simvastatin led to increasing in the amount of newly formed bone. These results confirmed that α-TCP, β-TCP, and HA are osteoconductive materials acting as space maintainer for bone formation and that combining these materials with simvastatin stimulates bone regeneration and it also affects degradability of α-TCP and β-TCP. Conclusively, α-TCP has the advantage of higher rate of degradation allowing the more bone formation and combining α-TCP with simvastatin enhances this property.

Effects of the combination with α-tricalcium phosphate and simvastatin on bone regeneration

BACKGROUND Although local application of statins stimulates bone formation, high dose of simvastatin induces inflammation. OBJECTIVE A study was conducted to test the hypothesis that maximum bone regeneration with less inflammation would be achieved by combining an optimal dose of simvastatin with alpha-tricalcium phosphate (alpha-TCP), which is an osteoconductive biomaterial capable of releasing the drug gradually. MATERIAL AND METHODS Bilateral 5-mm-diameter calvarial defects were created in adult Wistar rats and filled with preparations of different doses of simvastatin (0, 0.01, 0.1, 0.25 and 0.5 mg) combined with alpha-TCP particles or left empty. The animals were sacrificed at 2, 4 and 8 weeks and analyzed radiologically and histologically. Half of the animals of 4 and 8 weeks were labeled with fluorescence dyes and histomorphometrically analyzed. RESULTS Simvastatin doses of 0.25 and 0.5 mg caused inflammation of the soft tissue at the graft site whereas control and other doses did not. The micro-CT analysis revealed that the alpha-TCP with 0.1 mg simvastatin (TCP-0.1) group yielded significantly higher bone volumes than untreated control group at all three time points (249%, 227% and 266% at 2, 4 and 8 weeks, respectively). The percentage of defect closure, bone mineral content and bone mineral density were also higher in the TCP-0.1 group than in the other groups. CONCLUSION When combined with alpha-TCP particles, 0.1 mg simvastatin is the optimal dose for stimulation of the maximum bone regeneration in rat calvarial defects without inducing inflammation and it could be applied as an effective bone graft material.

Molecular and tissue responses in the healing of rat calvarial defects after local application of simvastatin combined with alpha tricalcium phosphate

We have previously reported that healing of rat calvarial defects was enhanced by application of alpha tricalcium phosphate (alphaTCP) combined with simvastatin, a cholesterol synthesis inhibitor. The purpose of the present study was to investigate the cellular and molecular mechanisms in this phenomenon. Rat calvarial defects were grafted with alphaTCP with or without simvastatin or left untreated. Animals were sacrificed on 3, 7, 10, 14, and 21 days postoperatively and histological changes in the defect region were assessed. Gene expression patterns were examined by RT-PCR. Proliferation and migration of osteoprogenitor cells from the dura mater were increased in simvastatin group from day 3 to day 10 (p < 0.01). New bone formation was significantly increased in simvastatin group on day 14 and day 21 (p < 0.01). BMP-2 expression was significantly higher in simvastatin group on day 3 and day 14 (p < 0.05) and maintained until day 21. Increased upregulation of TGF-beta1 was also observed in the simvastatin group on day 7 (p < 0.05) which was maintained until day 14. These findings suggest that the proliferation and recruitment of osteoprogenitor cells were critical steps in early stage of bone healing and that these steps were enhanced by TGF-beta1 and BMP-2, which were stimulated by simvastatin.

Exploitation of a novel polysaccharide nanogel cross-linking membrane for guided bone regeneration (GBR).

Cholesterol‐bearing pullulan (CHP) nanogel is a synthetic degradable biomaterial for drug delivery with high biocompatibility. Guided bone regeneration (GBR) is a bone augmentation technique in which a membrane is used to create and keep a secluded regenerative space. The purpose of the present study was to evaluate the effects of the novel CHP nanogel membrane in GBR. Thirty‐six adult Wistar rats were used and bilaterally symmetrical full‐thickness parietal bone defects of 5 mm diameter were created with a bone trephine burr. Each defect was covered with the collagen membrane or the CHP nanogel membrane or untreated without any membrane. The animals were sacrificed at 2, 4 and 8 weeks and analysed radiologically and histologically. Furthermore, after incubating human serum with CHP nanogel or collagen, the amount of PDGF in the serum was measured using ELISA. New bone formation in terms of bone volume was higher in the nanogel group than in the control or collagen groups at 2 and 4 weeks. At 8 weeks, both membrane groups showed higher bone volumes than the control group. Notably, the newly‐formed bone in the bone defect in the nanogel group was uniform and histologically indistinguishable from the original bone, whereas in the collagen group the new bone showed an irregular structure that was completely different from the original bone. After incubating with CHP nanogel, the amount of PDGF in the serum decreased significantly. CHP nanogel GBR membrane favourably stimulated bone regeneration, in which a unique characteristic of CHP nanogel, the storage of endogenous growth factors, was likely implicated. Copyright

Implantation of green tea catechin α-tricalcium phosphate combination enhances bone repair in rat skull defects

UNLABELLED The purpose of the present study is to investigate effects of the combination of epigallocatechin-3-gallate (EGCG) and α-tricalcium phosphate (α-TCP) on bone regenerative capacity in a bilateral rat calvarial bone defect model. MATERIALS AND METHODS Bilateral 5-mm-diameter calvarial defects were created in adult male Wistar rats and filled with preparations of EGCG (0, 0.1, 0.2, 0.4 mg) combined with α-TCP particles. This was done by dissolving EGCG in 100% ethanol (50 μL/14 mg) and dropping under sterile condition. The control group was left unfilled (n = 8). The animals were sacrificed at 2 and 4 weeks. Radiological images were taken, and histological analysis was done. Six animals from control (0 mg EGCG + α-TCP) group and (0.2 mg EGCG+ α-TCP) group were labeled with fluorescent dyes and histomorphometrically analyzed (n = 6) at 2 and 4 weeks. RESULTS Histomorphometric analysis revealed that the combination of EGCG and α-TCP at doses of 0.1 and 0.2 mg yielded significantly more new bone formation than untreated control group at 2 and 4 weeks (p > 0.05). Mineral apposition rate at 0.2-TCP group was enhanced compared with the one of the positive control α-TCP group at 4 weeks (p > 0.05). CONCLUSION The combination of α-TCP particles and 0.2 mg EGCG stimulates maximum bone regeneration in rat calvarial defects, and this combination would be potentially effective as bone graft material.

Accelerated and Enhanced Bone Formation on Novel Simvastatin-Loaded Porous Titanium Oxide Surfaces

BACKGROUND With increasing application of dental implants in poor-quality bones, the need for implant surfaces ensuring accelerated osseointegration and enhanced peri-implant bone regeneration is increased. PURPOSE A study was performed to evaluate the osseointegration and bone formation on novel simvastatin-loaded porous titanium oxide surface. MATERIALS AND METHODS Titanium screws were treated by micro-arc oxidation to form porous oxide surface and 25 or 50 μg of simvastatin was loaded. The nontreated control, micro-arc oxidized, and simvastatin-loaded titanium screws were surgically implanted into the proximal tibia of 16-week-old male Wistar rats (n = 36). Peri-implant bone volume, bone-implant contact, and mineral apposition rates were measured at 2 and 4 weeks. Data were analyzed by one-way analysis of variance followed by Tukeys post hoc test. RESULTS New bone was formed directly on the implant surface in the bone marrow cavity in simvastatin-loaded groups since 2 weeks. Bone-implant contact values were significantly higher in simvastatin-loaded groups than control and micro-arc oxidized groups at both time points (p < .05). Peri-implant bone volume and mineral apposition rate of simvastatin-loaded groups were significantly higher than control and micro-arc oxidized groups at 2 weeks (p < .05). CONCLUSIONS These data suggested that simvastatin-loaded porous titanium oxide surface provides faster osseointegration and peri-implant bone formation and it would be potentially applicable in poor-quality bones.