Ozone therapy improves early stages of osseointegration in ovariectomized rats.

OBJECTIVE: the aim of this study was to analyze the influence of ozone therapy (OZN) on peri-implant bone repair in critical bones by installing osseointegrated implants in the tibia of ovariectomized rats. METHODOLOGY: ovariectomy was performed on 30 Wistar rats, aged six months (Rattus novergicus), and, after 90 days, osseointegrated implants were installed in each tibial metaphysis. The study groups were divided into the animals that received intraperitoneal ozone at a concentration of 700 mcg/kg - OZ Group (n=15) - and a control group that received an intraperitoneal saline solution and, for this reason, was named the SAL group (n=15). The applications for both groups occurred during the immediate post-operative period on the 2nd, 4th, 6th, 8th, 10th, and 12th day post-surgery. At various stages (14, 42, and 60 days), the animals were euthanized, and tests were performed on their tibiae. These tests include histomorphometric and immunohistochemical analyses, computerized microtomography, sampling in light-cured resin for calcified sections, and confocal microscopy. The obtained data were then analyzed using One-way ANOVA and the Shapiro-Wilk, Kruskal-Wallis, and student t-tests (P<0.05). RESULTS: our findings indicate that the OZ group (3.26±0.20 mm) showed better cellular organization and bone neoformation at 14 days (SAL group, 0.90±1.42 mm) (P=0.001). Immunohistochemistry revealed that osteocalcin labeling was moderate in the OZ group and mild in the SAL group at 14 and 42 days post-surgery. The data from the analysis of calcified tissues (microtomography, histometric, and bone dynamism analysis) at 60 days showed no statistically significant differences between the groups (P=0.32). CONCLUSION: it was concluded that ozone therapy anticipated the initial phases of the peri-implant bone repair process.

Polydioxanone Enhances Bone Regeneration After Resection and Reconstruction of Rat Femur with rhBMP2.

The aim of this study was to assess the bone regeneration potential of a polydioxanone (PDO) scaffold together with recombinant human bone morphogenetic protein-2 (rhBMP-2) for the reconstruction of large bone defect. In total, 24 male rats (6 months old) were subjected to bilateral femoral stabilization using titanium plates to create a 2 mm gap, and reconstruction using rhBMP-2 (Infuse®; 3.25 µg). The bone defects were covered with PDO (PDO group), or with titanium mesh (Ti group). Animals were euthanized on days 14 and 60. Simultaneously, 16 rats received PDO and Ti in their dorsum for the purpose of biocompatibility analysis at 3, 5, 7, and 10 days postoperatively. X-ray densitometry showed a higher density in the PDO group on day 14. On day 60, coverage of the bone defect with PDO showed a larger quantity of newly formed bone than that found for the Ti group, a lower inflammatory infiltrate value, and a more significant number of blood vessels on day 14. By immunohistochemical assessment, runt-related transcription factor 2 (RUNX2) and osteocalcin (OCN) showed higher labeling on day 14 in the PDO group. On day 60, bone morphogenetic protein-2 (BMP-2) showed higher labeling in the PDO group, whereas Ti showed higher labeling for osteoprotegerin, nuclear factor kappa B ligand-activating receptor, RUNX2, and OCN. Furthermore, biocompatibility analysis showed a higher inflammatory response in the Ti group. The PDO scaffold enhanced bone regeneration when associated with rhBMP-2 in rat femur reconstruction. Impact statement Regeneration of segmental bone defects is a difficult task, and several techniques and materials have been used. Recent advances in the production of synthetic polymers, such as polydioxanone (PDO), produced by three-dimensional printing, have shown distinct characteristics that could improve tissue regeneration even in an important bone defect. The present preclinical study showed that PDO membranes used as scaffolds to carry recombinant human bone morphogenetic protein-2 (rhBMP-2) improved bone tissue regeneration by more than 8-fold when compared with titanium mesh, suggesting that PDO membranes could be a feasible and useful material for use in guided bone regeneration. (In English, viable is only used for living creatures capable of sustaining life.

Ozone therapy improves early stages of osseointegration in ovariectomized rats

Abstract Objective the aim of this study was to analyze the influence of ozone therapy (OZN) on peri-implant bone repair in critical bones by installing osseointegrated implants in the tibia of ovariectomized rats. Methodology ovariectomy was performed on 30 Wistar rats, aged six months (Rattus novergicus), and, after 90 days, osseointegrated implants were installed in each tibial metaphysis. The study groups were divided into the animals that received intraperitoneal ozone at a concentration of 700 mcg/kg — OZ Group (n=15) — and a control group that received an intraperitoneal saline solution and, for this reason, was named the SAL group (n=15). The applications for both groups occurred during the immediate post-operative period on the 2nd, 4th, 6th, 8th, 10th, and 12th day post-surgery. At various stages (14, 42, and 60 days), the animals were euthanized, and tests were performed on their tibiae. These tests include histomorphometric and immunohistochemical analyses, computerized microtomography, sampling in light-cured resin for calcified sections, and confocal microscopy. The obtained data were then analyzed using One-way ANOVA and the Shapiro-Wilk, Kruskal-Wallis, and student t-tests (P<0.05). Results our findings indicate that the OZ group (3.26±0.20 mm) showed better cellular organization and bone neoformation at 14 days (SAL group, 0.90±1.42 mm) (P=0.001). Immunohistochemistry revealed that osteocalcin labeling was moderate in the OZ group and mild in the SAL group at 14 and 42 days post-surgery. The data from the analysis of calcified tissues (microtomography, histometric, and bone dynamism analysis) at 60 days showed no statistically significant differences between the groups (P=0.32). Conclusion it was concluded that ozone therapy anticipated the initial phases of the peri-implant bone repair process.

Feasible low bone density condition for assessing bioactivity in ex-in vivo and in vivo studies.

OBJECTIVE: To choose a critical animal model for assessments of bone repair with implant installation by comparing senile rats (SENIL) to young ovariectomized rats (OXV). METHODOLOGY: For the ex-in vivo study, the femurs were precursors for bone marrow mesenchymal stem cells. Cellular responses were performed, including cell viability, gene expression of osteoblastic markers, bone sialoprotein immunolocalization, alkaline phosphatase activity, and mineralized matrix formation. For the in vivo study, the animals received implants in the region of the bilateral tibial metaphysis for histometric, microtomography, reverse torque, and confocal microscopy. RESULTS: Cell viability showed that the SENIL group had lower growth than OVX. Gene expression showed more critical responses for the SENIL group (p<0.05). The alkaline phosphatase activity obtained a lower expression in the SENIL group, as for the mineralization nodules (p<0.05). The in vivo histological parameters and biomechanical analysis showed lower data for the SENIL group. The confocal microscopy indicated the presence of a fragile bone in the SENIL group. The microtomography was similar between the groups. The histometry of the SENIL group showed the lowest values (p<0.05). CONCLUSION: In experimental studies with assessments of bone repair using implant installation, the senile model promotes the most critical bone condition, allowing a better investigation of the properties of biomaterials and topographic changes.

Single intraoperative infrared laser optimized bone repair in rat femoral osteotomies with experimentally induced osteoporosis.

This study aimed to evaluate the effect of infrared laser (IRL) on bone repair in ovariectomized rats subjected to femoral osteotomies. Of 32 rats, half underwent bilateral ovariectomy (OVX) and the other half underwent sham ovariectomy (SHAM). A period of 3 months was defined to observe the presence of osteoporosis. The rats were subjected to osteotomies in the femurs and then fixed with a miniplate and 1.5-mm system screws. Thereafter, half of the rats from both SHAM and OVX groups were not irradiated, and the other half were irradiated by IRL using the following parameters: wavelength, 808 nm; power, 100 mW; 60 s for each point; 6 J/point; and a total of 5 points of bone gap. All animals were euthanized 60 days after surgery. The femur gap was scanned using micro-computed tomography (micro-CT). The samples were then examined under a confocal laser microscope to determine the amounts of calcein and alizarin red. The slides were stained with alizarin red and Stevenel's blue for histometric analysis. In the micro-CT analysis, the OVX groups had the lowest bone volume (P < 0.05). When the laser was applied to the OVX groups, bone turnover increased (P < 0.05). New bone formation (NBF) was comparable between SHAM and OVX/IR (P > 0.05) groups; however, it was less in the OVX groups (P < 0.05). In conclusion, the results encourage the use of IRL intraoperatively as it optimizes bone repair, mainly in animals with low bone mineral density.

Collagen Membranes Functionalized with 150 Cycles of Atomic Layer Deposited Titania Improve Osteopromotive Property in Critical-Size Defects Created on Rat Calvaria.

The membranes used in bone reconstructions have been the object of investigation in the field of tissue engineering, seeking to improve their mechanical strength and add other properties, mainly the osteopromotive. This study aimed to evaluate the functionalization of collagen membranes, with atomic layer deposition of TiO2 on the bone repair of critical defects in rat calvaria and subcutaneous biocompatibility. A total of 39 male rats were randomized into four groups: blood clot (BC), collagen membrane (COL), COL 150-150 cycles of titania, and COL 600-600 cycles of titania. The defects were created in each calvaria (5 mm in diameter) and covered according to each group; the animals were euthanized at 7, 14, and 28 days. The collected samples were assessed by histometric (newly bone formed, soft tissue area, membrane area, and residual linear defect) and histologic (inflammatory cells and blood cells count) analysis. All data were subjected to statistical analysis (p < 0.05). The COL150 group showed statistically significant differences compared to the other groups, mainly in the analysis of residual linear defects (1.5 ± 0.5 × 106 pixels/µm2 for COL 150, and around 1 ± 0.5 × 106 pixels/µm2 for the other groups) and newly formed bone (1500 ± 1200 pixels/µm for COL 150, and around 4000 pixels/µm for the others) (p < 0.05), demonstrating a better biological behavior in the chronology of defects repair. It is concluded that the collagen membrane functionalized by TiO2 over 150 cycles showed better bioactive potential in treating critical size defects in the rats' calvaria.

Feasible low bone density condition for assessing bioactivity in ex-in vivo and in vivo studies

Abstract Objective To choose a critical animal model for assessments of bone repair with implant installation by comparing senile rats (SENIL) to young ovariectomized rats (OXV). Methodology For the ex-in vivo study, the femurs were precursors for bone marrow mesenchymal stem cells. Cellular responses were performed, including cell viability, gene expression of osteoblastic markers, bone sialoprotein immunolocalization, alkaline phosphatase activity, and mineralized matrix formation. For the in vivo study, the animals received implants in the region of the bilateral tibial metaphysis for histometric, microtomography, reverse torque, and confocal microscopy. Results Cell viability showed that the SENIL group had lower growth than OVX. Gene expression showed more critical responses for the SENIL group (p<0.05). The alkaline phosphatase activity obtained a lower expression in the SENIL group, as for the mineralization nodules (p<0.05). The in vivo histological parameters and biomechanical analysis showed lower data for the SENIL group. The confocal microscopy indicated the presence of a fragile bone in the SENIL group. The microtomography was similar between the groups. The histometry of the SENIL group showed the lowest values (p<0.05). Conclusion In experimental studies with assessments of bone repair using implant installation, the senile model promotes the most critical bone condition, allowing a better investigation of the properties of biomaterials and topographic changes.

Miniplates coated by plasma electrolytic oxidation improve bone healing of simulated femoral fractures on low bone mineral density rats.

The treatment of polytrauma patients represents a great challenge in the maxillofacial and orthopedic surgery fields. Therefore, this study tested the hypothesis that the use of a bioactive coating (by plasma electrolytic oxidation, PEO) on titanium microplates could improve the fracture healing of low bone mineral density (BMD) rats. Thirty female rats underwent bilateral ovariectomy surgery (OVX), and 35 rats underwent fake surgery (SHAM). Three months later, animals were subjected to femoral fracture simulation and were fixed with either non-coated (CONV) or coated (PEO) titanium miniplates. Eight weeks postoperatively, microplate/bone complexes were analyzed through computed microtomography, histometric, confocal microscopy, molecular, and biomechanical analysis. Bioactive elements (Ca and P) were incorporated on the PEO microplate and the surface was modified in a volcano-like structure. In the microCT analysis the OVX/PEO group had greater values for Tb.Th (bone trabecular thickness), Tb.Sp (separation of bone trabeculae) and Tb.N (number of trabeculae) parameters compared to the OVX/CONV group. According to histometric analysis, the OVX/PEO group showed significantly higher new bone formation than the OVX/CONV group (P < 0.05). For the fluorochrome area, the OVX groups (PEO and CONV) showed greater values for calcein precipitation (old bone) than alizarin red (new bone). Molecular results showed greater values for proteins related to the final phase of bone formation (P < 0.05) in the OVX/PEO group. The OVX/PEO group showed higher bone/miniplate system resilience compared to the others (P < 0.05). It was concluded that PEO coating optimizes bone healing on simulated femoral fractures in low bone mineral density rats. This sheds new light in the treatment of osteoporotic patients with bone fractures.

Combination of etodolac and dexamethasone improves preemptive analgesia in third molar surgery: a randomized study.

OBJECTIVE: This randomized, controlled, triple-blind, crossover clinical trial aimed to investigate the use of dexamethasone (DEX) and etodolac (ETO) as preemptive analgesia before mandibular third molar extraction. METHODS: Patients were divided into three groups (n = 20 teeth each) based on the drug administered: DEX 8 mg (DEX); DEX 8 mg plus ETO 300 mg (DEX + ETO), and ETO 300 mg (ETO). Paracetamol (750 mg) tablets were administered as rescue analgesics. Pain was evaluated using the visual analog scale (VAS) at 6, 12, 24, 48, and 72 h and 7 days postoperatively. Edema and trismus were assessed 48 and 72 h postoperatively. All data were subjected to statistical analysis, where a P value < .05 indicated statistical significance. RESULTS: VAS scores and the number of rescue analgesics taken were lower in the DEX + ETO group than in the other groups (P < .001 and P = .014, respectively). At 48 h, trismus was similar among all groups; however, the ETO group showed the highest trismus 7 days postoperatively (P < .05). Edema was similar among all groups at all time points (P > .05). CONCLUSION: The combined use of the anti-inflammatory drugs, DEX and ETO, resulted in better pain control and the need for fewer rescue analgesics than the use of either drug alone, which indicated their effectiveness in mandibular third molar extractions preoperatively. CLINICAL RELEVANCE: This drug combination can lead to less pain, edema, and trismus and reduce the use of rescue analgesics in the postoperative period.

Author Correction: Plasma Electrolytic Oxidation as a Feasible Surface Treatment for Biomedical Applications: an in vivo study.

An amendment to this paper has been published and can be accessed via a link at the top of the paper.

Plasma Electrolytic Oxidation as a Feasible Surface Treatment for Biomedical Applications: an in vivo study.

OBJECTIVES: In this in vivo animal study, we evaluated the effect of plasma electrolytic oxidation (PEO) coating on the topographic and biological parameters of implants installed in rats with induced osteoporosis and low-quality bones. MATERIALS AND METHODS: In total 44 Wistar rats (Rattus novergicus), 6 months old, were submitted to ovariectomy (OXV group) and dummy surgery (SHAM group). After 90 days, the ELISA test was performed and the ovariectomy effectiveness was confirmed. In each tibial metaphysis, an implant with PEO coating containing Ca2+ and P5+ molecules were installed, and the other tibia received an implant with SLA acid etching and blasting (AC) (control surface). After 42 days, 16 rats from each group were euthanized, their tibias were removed for histological and immunohistochemical analysis (OPG, RANKL, OC and TRAP), as well as reverse torque biomechanics. Data were submitted to One-way ANOVA or Kruskal-Wallis tests, followed by a Tukey post-test; P < 0.05. Histological analyses showed higher bone neoformation values among the members of the PEO group, SHAM and OVX groups. Immunohistochemical analysis demonstrated equilibrium in all groups when comparing surfaces for TRAP, OC and RANKL (P > 0.05), whereas OPG showed higher PEO labeling in the OVX group (P < 0.05). Biomechanical analysis showed higher reverse torque values (N.cm) for PEO, irrespective of whether they were OVX or SHAM groups (P < 0.05). CONCLUSION: The results indicated that the PEO texturing method favored bone formation and showed higher bone maturation levels during later periods in osteoporotic rats.