Hueliton Wilian Kido

Histopathological, cytotoxicity and genotoxicity evaluation of Biosilicate® glass–ceramic scaffolds

This study evaluated the biocompatibility of Biosilicate® scaffolds by means of histopathological, cytotoxicity, and genotoxicity analysis. The histopathologic analysis of the biomaterial was performed using 65 male rats, distributed into the groups: control and Biosilicate®, evaluated at 7, 15, 30, 45, and 60 days after implantation. The cytotoxicity analysis was performed by the methyl thiazolyl tetrazolium (MTT) assay, with various concentrations of extracts from the biomaterial in culture of osteoblasts and fibroblasts after 24, 72, and 120 h. The genotoxicity analysis (comet assay) was performed in osteoblasts and fibroblasts after contact with the biomaterial during 24, 72, and 96 h. In the histopathology analysis, we observed a foreign body reaction, characterized by the presence of granulation tissue after 7 days of implantation of the biomaterial, and fibrosis connective tissue and multinucleated giant cells for longer periods. In the cytotoxicity analysis, extracts from the biomaterial did not inhibit the proliferation of osteoblasts and fibroblasts, and relatively low concentrations (12.5% and 25%) stimulated the proliferation of both cell types after 72 and 120 h. The analysis of genotoxicity showed that Biosilicate® did not induce DNA damage in both lineages tested in all periods. The results showed that the Biosilicate® scaffolds present in vivo and in vitro biocompatibility.

Biocompatibility of a porous alumina ceramic scaffold coated with hydroxyapatite and bioglass

This study aimed to evaluate the osteointegration and genotoxic potential of a bioactive scaffold, composed of alumina and coated with hydroxyapatite and bioglass, after their implantation in tibias of rats. For this purpose, Wistar rats underwent surgery to induce a tibial bone defect, which was filled with the bioactive scaffolds. Histology analysis (descriptive and morphometry) of the bone tissue and the single-cell gel assay (comet) in multiple organs (blood, liver, and kidney) were used to reach this aim after a period of 30, 60, 90, and 180 days of material implantation. The main findings showed that the incorporation of hydroxyapatite and bioglass in the alumina scaffolds produced a suitable environment for bone ingrowth in the tibial defects and did not demonstrate any genotoxicity in the organs evaluated in all experimental periods. These results clearly indicate that the bioactive scaffolds used in this study present osteogenic potential and still exhibit local and systemic biocompatibility. These findings are promising once they convey important information about the behavior of this novel biomaterial in biological system and highlight its possible clinical application.

Characterization and biocompatibility of a fibrous glassy scaffold

Bioactive glasses (BGs) are known for their ability to bond to living bone and cartilage. In general, they are readily available in powder and monolithic forms, which are not ideal for the optimal filling of bone defects with irregular shapes. In this context, the development of BG‐based scaffolds containing flexible fibres is a relevant approach to improve the performance of BGs. This study is aimed at characterizing a new, highly porous, fibrous glassy scaffold and evaluating its in vitro and in vivo biocompatibility. The developed scaffolds were characterized in terms of porosity, mineralization and morphological features. Additionally, fibroblast and osteoblast cells were seeded in contact with extracts of the scaffolds to assess cell proliferation and genotoxicity after 24, 72 and 144 h. Finally, scaffolds were placed subcutaneously in rats for 15, 30 and 60 days. The scaffolds presented interconnected porous structures, and the precursor bioglass could mineralize a hydroxyapatite (HCA) layer in simulated body fluid (SBF) after only 12 h. The biomaterial elicited increased fibroblast and osteoblast cell proliferation, and no DNA damage was observed. The in vivo experiment showed degradation of the biomaterial over time, with soft tissue ingrowth into the degraded area and the presence of multinucleated giant cells around the implant. At day 60, the scaffolds were almost completely degraded and an organized granulation tissue filled the area. The results highlight the potential of this fibrous, glassy material for bone regeneration, due to its bioactive properties, non‐cytotoxicity and biocompatibility. Future investigations should focus on translating these findings to orthotopic applications. Copyright

Effects of low level laser therapy on inflammatory and angiogenic gene expression during the process of bone healing: A microarray analysis

The process of bone healing as well as the expression of inflammatory and angiogenic genes after low level laser therapy (LLLT) were investigated in an experimental model of bone defects. Sixty Wistar rats were distributed into control group and laser group (830nm, 30mW, 2,8J, 94seg). Histopathological analysis showed that LLLT was able to modulate the inflammatory process in the area of the bone defect and also to produce an earlier deposition of granulation tissue and newly formed bone tissue. Microarray analysis demonstrated that LLLT produced an up-regulation of the genes related to the inflammatory process (MMD, PTGIR, PTGS2, Ptger2, IL1, 1IL6, IL8, IL18) and the angiogenic genes (FGF14, FGF2, ANGPT2, ANGPT4 and PDGFD) at 36h and 3days, followed by the decrease of the gene expression on day 7. Immunohistochemical analysis revealed that the subjects that were treated presented a higher expression of COX-2 at 36h after surgery and an increased VEGF expression on days 3 and 7 after surgery. Our findings indicate that LLLT was efficient on accelerating the development of newly formed bone probably by modulating the inflammatory and angiogenic gene expression as well as COX2 and VEGF immunoexpression during the initial phase of bone healing.

Calcium phosphate fibers coated with collagen: In vivo evaluation of the effects on bone repair.

The aim of this study was to assess the characteristics of the CaP/Col composites, in powder and fiber form, via scanning electron microscopy (SEM), pH and calcium release evaluation after immersion in SBF and to evaluate the performance of these materials on the bone repair process in a tibial bone defect model. For this, four different formulations (CaP powder - CaPp, CaP powder with collagen - CaPp/Col, CaP fibers - CaPf and CaP fibers with collagen - CaPf/Col) were developed. SEM images indicated that both material forms were successfully coated with collagen and that CaPp and CaPf presented HCA precursor crystals on their surface. Although presenting different forms, FTIR analysis indicated that CaPp and CaPf maintained the characteristic peaks for this class of material. Additionally, the calcium assay study demonstrated a higher Ca uptake for CaPp compared to CaPf for up to 5 days. Furthermore, pH measurements revealed that the collagen coating prevented the acidification of the medium, leading to higher pH values for CaPp/Col and CaPf/Col. The histological analysis showed that CaPf/Col demonstrated a higher amount of newly formed bone in the region of the defect and a reduced presence of material. In summary, the results indicated that the fibrous CaP enriched with the organic part (collagen) glassy scaffold presented good degradability and bone-forming properties and also supported Runx2 and RANKL expression. These results show that the present CaP/Col fibrous composite may be used as a bone graft for inducing bone repair.

Laser therapy modulates systemic inflammatory processes and muscle atrophy in an experimental model of sepsis in rats

Abstract Objective: The aim of this study was to determine the effectiveness of low-level laser therapy (LLLT) on the modulation of the systemic inflammatory processes and skeletal muscle morphology in an experimental sepsis model (cecal ligation and puncture, CLP). Study design: Seventy-two male Wistar rats were randomly divided into three groups: control group (CG); sepsis group (SG) where rats were submitted to CLP but without LLLT treatment, and the sepsis laser-treated group (SLG). Laser irradiation (GaAlAs laser, continuous wave, 808 nm, 30 mW, 48 s, 30 J/cm2, 0.028 cm2, 1.07 mW/cm2) was performed immediately after surgery and every 24 h at 4 points (on the middle of tibialis anterior and diaphragm, bilaterally), through the punctual contact technique. All sepsis animals were sacrificed at 6, 24, 48 and 72 h post-surgery. The immunohistochemistry analysis was used to verify the expression of proteins related to the regulation of muscle wasting (MuRF-1 and atrogin). In order to investigate the action of LLLT on inflammatory mediators in the rat sepsis model, two inflammatory cytokines IL-6 and IL-10, were evaluated. Results: The results showed that the laser-treated animals presented a lower IL-6 activity and decreased atrogin and MuRF-1 immunoexpression. However, no difference was observed in muscle cross-sectional area between the experimental groups. Conclusion: These results suggest that LLLT was able to decrease the systemic inflammation and muscle atrophy markers, preventing muscle protein degradation.

Mitochondrial dynamics (fission and fusion) and collagen production in a rat model of diabetic wound healing treated by photobiomodulation: comparison of 904 nm laser and 850 nm light-emitting diode (LED)

OBJECTIVE Mitochondrial dysfunction has been associated with the development of diabetes mellitus which is characterized by disorders of collagen production and impaired wound healing. This study analyzed the effects of photobiomodulation (PBM) mediated by laser and light-emitting diode (LED) on the production and organization of collagen fibers in an excisional wound in an animal model of diabetes, and the correlation with inflammation and mitochondrial dynamics. METHODS Twenty Wistar rats were randomized into 4 groups of 5 animals. Groups: (SHAM) a control non-diabetic wounded group with no treatment; (DC) a diabetic wounded group with no treatment; (DLASER) a diabetic wounded group irradiated by 904 nm pulsed laser (40 mW, 9500 Hz, 1 min, 2.4 J); (DLED) a diabetic wounded group irradiated by continuous wave LED 850 nm (48 mW, 22 s, 1.0 J). Diabetes was induced by injection with streptozotocin (70 mg/kg). PBM was carried out daily for 5 days followed by sacrifice and tissue removal. RESULTS Collagen fibers in diabetic wounded skin were increased by DLASER but not by DLED. Both groups showed increased blood vessels by atomic force microscopy. Vascular endothelial growth factor (VEGF) was higher and cyclooxygenase (COX2) was lower in the DLED group. Mitochondrial fusion was higher and mitochondrial fusion was lower in DLED compared to DLASER. CONCLUSION Differences observed between DLASER and DLED may be due to the pulsed laser and CW LED, and to the higher dose of laser. Regulation of mitochondrial homeostasis may be an important mechanism for PBM effects in diabetes.