Adriana Luisa Gonçalves de Almeida

Poly(vinylidene-trifluoroethylene)/barium titanate composite for in vivo support of bone formation

In this study, we evaluated the effect of poly(vinylidene fluoride-trifluoroethylene)/barium titanate (P(VDF-TrFE)/BT) membrane on in vivo bone formation. Rat calvarial bone defects were implanted with P(VDF-TrFE)/BT and polytetrafluoroethylene (PTFE) membranes, and at 4 and 8 weeks, histomorphometric and gene expression analyses were performed. A higher amount of bone formation was noticed on P(VDF-TrFE)/BT compared with PTFE. The gene expression of RUNX2, bone sialoprotein, osteocalcin, receptor activator of nuclear factor-kappa B ligand, and osteoprotegerin indicates that P(VDF-TrFE)/BT favored the osteoblast differentiation compared with PTFE. These results evidenced the benefits of using P(VDF-TrFE)/BT to promote new bone formation, which may represent a promising alternative to be employed in guided bone regeneration.

Effect of collagen sponge and fibrin glue on bone repair

ABSTRACT The ability of hemostatic agents to promote bone repair has been investigated using in vitro and in vivo models but, up to now, the results are inconclusive. Objective In this context, the aim of this study was to compare the potential of bone repair of collagen sponge with fibrin glue in a rat calvarial defect model. Material and Methods Defects of 5 mm in diameter were created in rat calvariae and treated with either collagen sponge or fibrin glue; untreated defects were used as control. At 4 and 8 weeks, histological analysis and micro-CT-based histomorphometry were carried out and data were compared by two-way ANOVA followed by Student-Newman-Keuls test when appropriated (p≤0.05). Results Three-dimensional reconstructions showed increased bone formation in defects treated with either collagen sponge or fibrin glue compared with untreated defects, which was confirmed by the histological analysis. Morphometric parameters indicated the progression of bone formation from 4 to 8 weeks. Additionally, fibrin glue displayed slightly higher bone formation rate when compared with collagen sponge. Conclusion Our results have shown the benefits of using collagen sponge and fibrin glue to promote new bone formation in rat calvarial bone defects, the latter being discreetly more advantageous.

Early peri-implant endosseous healing of two implant surfaces placed in surgically created circumferential defects. A histomorphometric and fluorescence study in dogs.

OBJECTIVE Several implant surfaces are being developed, some in the nanoscale level. In this study, two different surfaces had their early healing properties compared in context of circumferential defects of various widths. MATERIAL AND METHODS Six dogs had the mandibular premolars extracted. After 8 weeks, four implants were placed equicrestally in each side. One acted as control, while the others were inserted into sites with circumferential defects of 1.0, 1.5 and 2.0 mm wide and 5 mm deep. A nano-modified surface was used on one side and a micro-rough on the other. Bone markers were administered on the third day after implant placement and then after 1, 2, 4 weeks to investigate the bone formation dynamic through fluorescence analysis. Ground sections were prepared from 8-week healing biopsies and histomorphometry was performed. RESULTS The fluorescence evaluation of the early healing showed numerically better results for the nano-modified group; however this trend was not followed by the histomorphometric evaluation. A non-significant numerical superiority of the micro-rough group was observed in terms of vertical bone apposition, defect bone fill, bone-to-implant contact and bone density. In the intra-group analysis, the wider defects showed the worse results while the control sites showed the best results for the different parameters, but without statistical relevance. CONCLUSION Both surfaces may lead to complete fill of circumferential defects, but the gap width has to be considered as a challenge. The nano-scale modification was beneficial in the early stages of bone healing, but the micro-rough surface showed numerical better outcomes at the 8-week final period.

Short-Term Evaluation of Grafts Fixed With Either N-Butyl-2-Cyanocrylate or Screws

PURPOSE N-butyl-2-cyanoacrylate (NB-Cn) is an alternative method for onlay graft fixation and might be efficient for preserving the graft volume. Our aim was to analyze the gene expression and mineralized tissue variations of calvarial bone grafting fixed in the mandible with either NB-Cn or a titanium screw (TiS). MATERIALS AND METHODS New Zealand rabbits had bilateral calvarial grafts fixed at both sides of the mandible with either NB-Cn or a TiS. The rabbits were sacrificed at 4 and 8 days, and micro-computed tomography analysis was performed. For molecular analysis, the gene expression of interleukin-6, interleukin-10, and tumor necrosis factor-α was assessed. Quantification using real-time polymerase chain reaction was performed. Statistical analysis was performed using the paired Student t test (P < .05). RESULTS Bone graft fixation with NB-Cn promoted superior volume and density preservation. The percentage of mineralized tissue at the center portion and border of the graft was very similar (NB-Cn, 50.6% ± 8.3% and 50.3% ± 10.6%, respectively) and superior than in the TiS group (32.5% ± 3.5% and 33.8% ± 6%, respectively). Genes from the NB-Cn group were upregulated compared with those in the TiS group at the initial phases of bone healing (4 days), with the profile reversed at the 8-day point. At day 8, the osteoclastogenesis-related genes were upregulated in the TiS group. CONCLUSIONS Onlay bone grafts fixed with screws induced more inflammation during the initial remodeling process than did NB-Cn. The differences in the incorporation into the host bed suggest that the use of adhesives for graft fixation will promote superior volume and density preservation.

Buccal bone plate remodeling after immediate implants with or without synthetic bone grafting and flapless surgery: a histomorphometric and fluorescence study in dogs

OBJECTIVE The aim of this study was to evaluate the buccal bone plate remodeling after immediate implantation using the flapless approach with or without bone graft into the gap between the implant and the buccal bone. MATERIAL AND METHODS Eight dogs had the mandibular bicuspids extracted without flaps, and four implants were installed on each side, totaling eight implants per animal. Randomly, in one side, the implants were positioned at the bone crest level (equicrestal), and on the opposite side, the implants were positioned 2 mm subcrestal. All the implants were positioned 2.0 mm from the buccal bone plate (gap) and associated or not with grafting material. Therefore, the following treatments were performed: implants subcrestal test (SCTG) with bone graft and control (SCCG) without bone graft, and equicrestal test (ECTG) with bone graft and control (ECCG) without bone graft. One week following the surgeries, metallic prostheses were installed. Bone markers were administered 1, 2, 4, and 12 weeks after implant placement for fluorescence analysis. Ground sections were prepared from 12-week healing biopsies, and histomorphometry was performed. RESULTS The histomorphometric evaluation presents significant better results for the ECTG in the vertical crestal bone resorption, but the other parameters showed better results for the SCCG. The fluorescence evaluation in adjacent areas showed numerically different results between groups with a small decrease at 12 weeks, except for the SCCG, which was higher at this time. The distant area showed a continuous increase in the marked bone. CONCLUSION The equicrestally placed implants presented little or no loss of the buccal bone wall. The subcrestally positioned implants presented loss of buccal bone, regardless of the use of bone graft. However, the buccal bone was always coronal to the implant shoulder. Both the equicrestal and subcrestal groups were benefited in the early stages of bone healing as evidenced by the fluorescence analysis.

Participation of MicroRNA-34a and RANKL on bone repair induced by poly(vinylidene-trifluoroethylene)/barium titanate membrane

Abstract The poly(vinylidene-trifluoroethylene)/barium titanate (PVDF) membrane enhances in vitro osteoblast differentiation and in vivo bone repair. Here, we hypothesized that this higher bone repair could be also due to bone resorption inhibition mediated by a microRNA (miR)/RANKL circuit. To test our hypothesis, the large-scale miR expression of bone tissue grown on PVDF and polytetrafluoroethylene (PTFE) membranes was evaluated to identify potential RANKL-targeted miRs modulated by PVDF. The animal model used was rat calvarial defects implanted with either PVDF or PTFE. At 4 and 8 weeks, the bone tissue grown on membranes was submitted to a large-scale analysis of miRs by microarray. The expression of miR-34a and some of its targets, including RANKL, were evaluated by real-time polimerase chain reaction and osteoclast activity was detected by tartrate-resistant acid phosphatase (TRAP) staining. Among more than 250 miRs, twelve, including miR-34a, were simultaneously higher expressed (≥2 fold) at 4 and 8 weeks on PVDF. The higher expression of miR-34a was concomitant with a reduced expression of all its evaluated targets, including RANKL. Additionally, more TRAP-positive cells were observed in bone tissue grown on PTFE compared with PVDF in both time points. In conclusion, our results suggest that the higher bone formation induced by PVDF could be, at least in part, triggered by a miR-34a increase and RANKL decrease, which may inhibit osteoclast differentiation and activity, and bone resorption.

Association of mesenchymal stem cells and osteoblasts for bone repair.

AIM We tested the hypothesis that the association of bone marrow mesenchymal stem cells (MSCs) and osteoblasts (OBs) optimize bone repair. MATERIALS & METHODS MSCs were cultured in growth or osteogenic medium and seeded into gelatin sponge prior to implantation. Defects were created into rat calvariae and implanted with gelatin sponge without cells, with MSCs, with OBs and with association of MSCs and OBs. Histological analysis and micro-CT-based histomorphometry were carried out after 4 weeks. RESULTS Increased bone formation was observed in defects treated with cells and bone volume was greater in defects treated with either OBs or MSCs/OBs. CONCLUSION Association of MSCs and OBs did not increase the process of bone repair compared with cell-based therapy using either MSCs or OBs alone.

Influence of periodontal tissue thickness on buccal plate remodelling on immediate implants with xenograft

AIM To evaluate the influence of gingival thickness and bone grafting on buccal bone plate remodelling after immediate implant placement in sockets with thin buccal bone, using a flapless approach. MATERIALS AND METHODS The gingiva of eight dogs was thinned at one side of the mandible, mandibular premolars were extracted without flaps, and four implants were installed on each side at 1.5 mm from the buccal bone. The sites were randomly assigned into: TG (test group) = thin gingiva; TG + GM (TG with grafting material); CG (control group) = normal gingiva; and CG + GM (CG with grafting material). After 12 weeks the dogs were sacrificed and the samples were processed for histological analysis. RESULTS All animals exhibited a thin buccal bone initially. In all the experimental groups the buccal gap was filled with newly formed bone and the buccal bone level was slightly apical to the implant shoulder. There were no statistically significant differences among the groups for the histomorphometric parameters. CONCLUSIONS The thickness of the buccal bone was a fundamental factor in buccal bone plate resorption, even with flapless implantation. The gingival thickness or the addition of a biomaterial in the gap did not influence the results.

Bone Formation in a Local Defect around Dental Implants Coated with Extracellular Matrix Components

PURPOSE The coating of implant surfaces with components of the extracellular matrix offers an approach to influence peri-implant bone healing. In this study, bone healing around coated implants is analyzed in a peri-implant defect model. MATERIALS AND METHODS Eight months after extraction of the premolar teeth, six dogs received 48 implants (eight per animal) in the mandible. Implant surfaces were sandblasted and acid-etched, and some were additionally coated with collagen type II and chondroitin sulfate (collagen/CS). On each side of the mandible, implants either had no peri-implant defect (control side) or a vertical defect of 5 mm in depth and 0.5, 1.0, or 2.0 mm in width. Implants healed submerged for 8 weeks. Fluorochrome staining, histology, and histomorphometry were used to analyze implant osseointegration. RESULTS Fluorochrome labels showed an increased mineralization around collagen/CS-coated surfaces at 4 weeks (p = .031). Histomorphometry generally showed lower vertical and horizontal bone apposition with increasing gap size for both surface types. In gapless sites and 0.5-mm gaps, collagen/CS coated implants showed increased bone volume in areas directly adjacent to the implant, in comparison with uncoated implants (p < .05). CONCLUSION The width of the peri-implant gap influences peri-implant bone formation. Complete filling of all gaps by newly formed bone could not be observed around either surface. In proximity to the surface, implant surface coating by collagen/CS positively influenced bone formation.

The effect of a flapless alveolar ridge preservation procedure with or without a xenograft on buccal bone crest remodeling compared by histomorphometric and microcomputed tomographic analysis

OBJECTIVES This study evaluated buccal bone crest remodeling, socket composition after healing, and dimensional ridge preservation after flapless tooth extraction procedures with or without a xenograft comparing histomorphometric and microcomputed tomographic (micro-CT) data. MATERIAL AND METHODS The mandibular premolars of eight dogs were extracted without flaps. One socket on each side received a grafting material (test group), and the other remained only with a blood clot (control group). Twelve weeks after treatment, buccal bone crest, alveolar ridge dimensions, and composition were analyzed by histomorphometry and micro-CT. RESULTS Two- and three-dimensional evaluations showed better results for the grafted group when compared to the non-grafted group. CONCLUSION The flapless alveolar ridge preservation procedure with deproteinized bovine bone material enhanced buccal bone crest, alveolar ridge dimensions and bone formation when compared to sockets with the blood clot only, as observed by histomorphometric and micro-CT analysis.