Poliani de Oliveira

Low-Level Laser Therapy Induces Differential Expression of Osteogenic Genes During Bone Repair in Rats

OBJECTIVES The aim of this study was to measure the temporal pattern of the expression of osteogenic genes after low-level laser therapy during the process of bone healing. We used quantitative real-time polymerase chain reaction (qPCR) along with histology to assess gene expression following laser irradiation on created bone defects in tibias of rats. MATERIAL AND METHODS The animals were randomly distributed into two groups: control or laser-irradiated group. Noncritical size bone defects were surgically created at the upper third of the tibia. Laser irradiation started 24 h post-surgery and was performed for 3, 6, and 12 sessions, with an interval of 48 h. A 830 nm laser, 50 J/cm(2), 30 mW, was used. On days 7, 13, and 25 post-injury, rats were sacrificed individually by carbon dioxide asphyxia. The tibias were removed for analysis. RESULTS The histological results revealed intense new bone formation surrounded by highly vascularized connective tissue presenting slight osteogenic activity, with primary bone deposition in the group exposed to laser in the intermediary (13 days) and late stages of repair (25 days). The quantitative real-time PCR showed that laser irradiation produced an upregulation of BMP-4 at day 13 post-surgery and an upregulation of BMP4, ALP, and Runx 2 at day 25 after surgery. CONCLUSION Our results indicate that laser therapy improves bone repair in rats as depicted by differential histopathological and osteogenic genes expression, mainly at the late stages of recovery.

Comparative effects of low-intensity pulsed ultrasound and low-level laser therapy on injured skeletal muscle.

OBJECTIVE The main purpose of this study was to compare the effects of low-intensity pulsed ultrasound (US) and low-level laser therapy (LLLT) on injured skeletal muscle after cryolesion by means of histopathological analysis and immunohistochemistry for cyclo-oxygenase-2 (COX-2). BACKGROUND AND METHODS Thirty-five male Wistar rats were randomly distributed into four groups: intact control group with uninjured and untreated animals; injured control group with muscle injury and no treatment; LLLT-treated group with muscle injury treated with 830-nm laser; and US-treated group with muscle injury treated with US. Treatments started 24 h postsurgery and were performed during six sessions. RESULTS LLLT-treated animals presented minor degenerative changes of muscle tissue. Exposure to US reduced tissue injuries induced by cryolesion, but less effectively than LLLT. A large number of COX-2 positive cells were found in untreated injured rats, whereas COX-2 immunoexpression was lower in both LLLT- and US-treated groups. CONCLUSION This study revealed that both LLLT and US therapies have positive effects on muscle metabolism after an injury in rats, but LLLT seems to produce a better response.

Effect of low-level laser therapy (660 nm) on the healing of second-degree skin burns in rats

The aim of this study was to investigate the effects of 660 nm laser on the healing of burn wounds made on the backs of rats. Thirty-two Wistar male rats were used. The animals were randomly distributed into 2 groups of 16 animals each: control group (burned rats without treatment) and laser-treated group (burned rats treated with laser therapy). Each group was divided into two different subgroups, euthanized in different periods (subgroup A: 7 days post-surgery and subgroup B: 14 days post-surgery). Histopathological analysis revealed a significant decrease in the necrotic area in the laser-treated group compared to the controls at days 7 and 14 post-injury. COX-2 positive cells were found in a strong pattern in the group submitted to laser therapy after 7 days. Regarding VEGF immunomarker, a significant VEGF immunoexpression was detected in the laser-exposed group after 14 days when compared to the negative control group. Taken together, our results demonstrate that laser therapy is able to promote skin repair of burned rats as a result of decreasing necrotic area and an up-regulation of COX-2 and VEGF immunoexpression.

Comparação dos efeitos do laser de baixa potência e do ultrassom de baixa intensidade no processo de reparo ósseo em tíbia de rato

BACKGROUND: Electrophysical agents such as Ultrasound (US) and low-level laser therapy (LLLT) have been increasingly used in physical therapy practice. Studies suggest that these devices are able to stimulate osteoblast proliferation and osteogenesis at the fracture site, resulting in a greater deposition of bone mass and speeding up the consolidation process. OBJECTIVE: The aim of this study was to analyze the effects of US and LLLT on the bone healing process, through biomechanical and histological analysis of the bone callus. METHODS: A total of 30 rats were randomly allocated into three groups: control group fracture without treatment (GC); fracture group treated with pulsed US, burst 1.5 MHz, 200us, 1KHz, 30 mW/cm2 (GUS) and fracture group treated with laser 830nm, 100mW, 120J/cm² (GL). Bone defects were performed with a circular drill of 2mm in diameter in the animals tibias. The treatments were carried out after surgery consisting of 7 applications every 48 hours. After 14 days the animals were sacrificed and the tibias were removed to perform the analysis, being the right tibia designated for biomechanical analysis, while the left tibia for histological analysis. RESULTS: The biomechanical analysis showed no statistically significant difference between biomechanical properties of the CG, CL and GUS. In morphometric analysis, both GUS and GL showed a significantly higher woven bone tissue area compared to the control group. However, when the two treatment modalities were compared, there were no statistical differences between them. CONCLUSION: Both devices used in this study were able to accelerate the bone healing process in rats.

LOW-INTENSITY PULSED ULTRASOUND PRODUCED AN INCREASE OF OSTEOGENIC GENES EXPRESSION DURING THE PROCESS OF BONE HEALING IN RATS

The aim of this study was to measure the temporal expression of osteogenic genes during the process of bone healing in low-intensity pulsed ultrasound (LIPUS) treated bone defects by means of histopathologic and real-time polymerase chain reaction (PCR) analysis. Animals were randomly distributed into two groups (n = 30): control group (bone defect without treatment) and LIPUS treated (bone defect treated with LIPUS). On days 7, 13 and 25 postinjury, 10 rats per group were sacrificed. Rats were treated with a 30 mW/cm(2) LIPUS. The results pointed out intense new bone formation surrounded by highly vascularized connective tissue presenting a slight osteogenic activity, with primary bone deposition was observed in the group exposed to LIPUS in the intermediary (13 days) and late stages of repair (25 days) in the treated animals. In addition, quantitative real-time polymerase chain reaction (RT-qPCR) showed an upregulation of bone morphogenetic protein 4 (BMP4), osteocalcin and Runx2 genes 7 days after the surgery. In the intermediary period, there was no increase in the expression. The expression of alkaline phosphatase, BMP4 and Runx2 was significantly increased at the last period. Our results indicate that LIPUS therapy improves bone repair in rats and upregulated osteogenic genes, mainly at the late stages of recovery.

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.

Effects of phototherapy on cartilage structure and inflammatory markers in an experimental model of osteoarthritis.

Abstract. The aim of this study was to evaluate the effects of laser phototherapy on the degenerative modifications on the articular cartilage after the anterior cruciate ligament transection (ACLT) in the knee of rats. Eighty male rats (Wistar) were distributed into four groups: intact control group (IG), injured control group (CG), injured laser treated group at 10  J/cm2 (L10), and injured laser treated group at 50  J/cm2 (L50). Animals were distributed into two subgroups, sacrificed in 5 and 8 weeks postsurgery. The ACLT was used to induce knee osteoarthritis in rats. After 2 weeks postsurgery, laser phototherapy initiated and it was performed for 15 and 30 sessions. The histological findings revealed that laser irradiation, especially at 10  J/cm2, modulated the progression of the degenerative process, showing a better cartilage structure and lower number of condrocytes compared to the other groups. Laser phototherapy was not able to decrease the degenerative process measured by Mankin score and prevent the increase of cartilage thickness related to the degenerative process. Moreover, it did not have any effect in the biomodulation of the expression of markers IL1β, tumor necrosis factor-α, and metalloprotein-13. Furthermore, laser irradiated animals, at 50  J/cm2 showed a lower amount of collagen type 1.

Comparative study of the effects of low-level laser and low-intensity ultrasound associated with Biosilicate® on the process of bone repair in the rat tibia

Objective: Verify the effects of the association between Biosilicate® and ultrasound and, Biosilicate® and laser in bone consolidation process of rats, through the biomechanical and histological analysis. Methods: Forthy male rats were used. The animals were randomized into four groups (n=10): control group fracture no treated (CGF); group treated with Biosilicate® (BG); group treated with Biosilicate® and laser (BLG); group treated with Biosilicate® and ultrasound (BUG). Results: The biomechanical analysis showed no significant difference among any groups after 14 days post-surgery. In the morphometric analysis, the control group showed moderate presence of new formed bone tissue inside the defects areas and the Biosilicate® group showed similar results. Despite those facts, the biomaterial osteogenic potential was demonstrated by the great amount of cells and bone tissue around the particles. Curiously, the Biosilicate® plus laser or ultrasound groups showed lower amounts of bone tissue deposition when compared with control fracture and Biosilicate® groups. Conclusion: The data from this study can conclude that Biosilicate® was able to accelerate and optimized the bone consolidation, through the modulation of the inflammatory process and the stimulation of new bone formation. However, when resources were associated, there are no positive results.

Comparação dos efeitos do laser de baixa potência e do ultrassom de baixa intensidade associado ao Biosilicato® no processo de reparo ósseo em tíbias de ratos

OBJECTIVE: Verify the effects of the association between Biosilicate® and ultrasound and, Biosilicate® and laser in bone consolidation process of rats, through the biomechanical and histological analysis. METHODS: Forthy male rats were used. The animals were randomized into four groups (n=10): control group fracture no treated (CGF); group treated with Biosilicate® (BG); group treated with Biosilicate® and laser (BLG); group treated with Biosilicate® and ultrasound (BUG). RESULTS: The biomechanical analysis showed no significant difference among any groups after 14 days post-surgery. In the morphometric analysis, the control group showed moderate presence of new formed bone tissue inside the defects areas and the Biosilicate® group showed similar results. Despite those facts, the biomaterial osteogenic potential was demonstrated by the great amount of cells and bone tissue around the particles. Curiously, the Biosilicate® plus laser or ultrasound groups showed lower amounts of bone tissue deposition when compared with control fracture and Biosilicate® groups. CONCLUSION: The data from this study can conclude that Biosilicate® was able to accelerate and optimized the bone consolidation, through the modulation of the inflammatory process and the stimulation of new bone formation. However, when resources were associated, there are no positive results.