Electromagnetic stimulation increases mitochondrial function in osteogenic cells and promotes bone fracture repair.

Bone fracture is a growing public health burden and there is a clinical need for non-invasive therapies to aid in the fracture healing process. Previous studies have demonstrated the utility of electromagnetic (EM) fields in promoting bone repair; however, its underlying mechanism of action is unclear. Interestingly, there is a growing body of literature describing positive effects of an EM field on mitochondria. In our own work, we have previously demonstrated that differentiation of osteoprogenitors into osteoblasts involves activation of mitochondrial oxidative phosphorylation (OxPhos). Therefore, it was reasonable to propose that EM field therapy exerts bone anabolic effects via stimulation of mitochondrial OxPhos. In this study, we show that application of a low intensity constant EM field source on osteogenic cells in vitro resulted in increased mitochondrial membrane potential and respiratory complex I activity and induced osteogenic differentiation. In the presence of mitochondrial inhibitor antimycin A, the osteoinductive effect was reversed, confirming that this effect was mediated via increased OxPhos activity. Using a mouse tibial bone fracture model in vivo, we show that application of a low intensity constant EM field source enhanced fracture repair via improved biomechanical properties and increased callus bone mineralization. Overall, this study provides supporting evidence that EM field therapy promotes bone fracture repair through mitochondrial OxPhos activation.

Is an anodizing coating associated to the photobiomodulation able to optimize bone healing in ovariectomized animal model?

This in vivo study investigated whether the bioactivity of anodizing coating, produced by plasma electrolytic oxidation (PEO), on mini-plate in femur fracture could be improved with the association of photobiomodulation (PBM) therapy. From the 20 ovariectomized Wistar female rats, 8 were used for model characterization, and the remaining 12 were divided into four groups according to the use of PBM therapy by diode laser (808 nm; power: 100 mW; energy: 6.0 J; energy density: 212 J/cm2; power density: 3.5 W/cm2) and the type of mini-plate surface (commercially pure titanium mini-plate -cpTi- and PEO-treated mini-plate) as follow: cpTi; PEO; cpTi/PBM; and PEO/PBM. After 60 days of surgery, fracture healing underwent microstructural, bone turnover, histometric, and histologic adjacent muscle analysis. Animals of groups with PEO and PBM showed greater fracture healing than cpTi control group under histometric and microstructural analysis (P < 0.05); however, bone turnover was just improved in PBM's groups (P < 0.05). there was no difference between cpTi and PEO without PBM (P > 0.05). Adjacent muscle analysis showed no metallic particles or muscle alterations in all groups. PEO and PBM are effective strategies for bone repair in fractures, however their association does not provide additional advantages.

Effects of photobiomodulation in the treatment of fractures: a systematic review and meta-analysis of randomized clinical trials.

Several therapeutic strategies have been proposed to optimize the conventional treatment of fractures. Photobiomodulation (PBM) appears to help reduce pain and control inflammation, and it also accelerates bone repair. This systematic review aimed to evaluate the effectiveness and safety of PBM with low-level laser therapy (LLLT) in the bone fracture healing process. We included randomized controlled trials (RCTs) comparing the effects of PBM with those of any other intervention in adults with lower or upper limb bone fractures. The primary outcomes investigated were pain reduction, radiographic healing, and adverse events. The searches were conducted in October 2018. Two RCTs were included that compared PBM to the placebo. A meta-analysis showed significant difference in favor of PBM for pain reduction (MD 1.19, 95% CI [0.61 to 1.77], 106 participants, two RCTs), but this difference was not clinically significant. One RCT (50 participants) showed a clinical and statistical improvement in physical function (MD - 14.60, 95% CI [- 21.39 to - 7.81]) and no difference in radiographic healing, regarding absence of fracture line (RR 1.00, 95% CI [0.93 to 1.08]) and visible bone callus (RR 0.33, 95% CI [0.01 to 7.81]). The certainty of evidence was classified as low to very low. Based on the evidence of low to very low certainty, PBM seems to be associated with the improvement of pain and function. Therefore, new RCTs are required that meet the recommendations of CONSORT to prove the effectiveness and safety of this intervention and support its recommendation in clinical practice.

Photobiomodulation therapy (PBMT) in bone repair: A systematic review.

BACKGROUND: Photobiomodulation therapy (PBMT) using low-level laser influences the release of several growth factors involved in the formation of epithelial cells, fibroblasts, collagen and vascular proliferation, besides accelerating the synthesis of bone matrix due to the increased vascularization and lower inflammatory response, with significant increase of osteocytes in the irradiated bone. Considering its properties, beneficial effects and clinical relevance, the aim of this review was to analyze the scientific literature regarding the use of PBMT in the process of bone defect repair. METHODS: Electronic search was carried out in PubMed/MEDLINEⓇ and Web of Science databases with combination of the descriptors low-level laser therapy AND bone repair, considering the period of publication until the year 2018. RESULTS: The literature search identified 254 references in PubMed/MEDLINE and 204 in Web of Science, of which 33 and 4 were selected, respectively, in accordance with the eligibility requirements. The analysis of researches showed articles using PBMT in several places of experimentation in the subjects, different types of associated biomaterials, stimulatory effects on cell proliferation, besides variations in the parameters of use of laser therapy, mainly in relation to the wavelength and density of energy. Only four articles reported that the laser did not improve the osteogenic properties of a biomaterial. CONCLUSIONS: Many studies have shown that PBMT has positive photobiostimulatory effects on bone regeneration, accelerating its process regardless of parameters and the use of biomaterials. However, standardization of its use is still imperfect and should be better studied to allow correct application concerning the utilization protocols.

Optimal currents for electrical stimulation of bone fracture repair: A computational analysis including variations in frequency, tissue properties, and fracture morphology.

Fracture healing happens naturally in most bone break cases. Occasionally prolongation of restoration period or non-union of the fracture may occur, where electrical stimulation has been shown to facilitate bone restoration by stimulating osteoblasts. Despite clinical use, a comprehensive computational model linking the applied currents to the stimulating field in the fracture has been missing. In this paper, we investigate the input current needed to stimulate osteoblasts in a fracture in the human forearm. Optimal current is computed for various fracture configurations, and sensitivity to frequency and inter/intrapersonal variance in dielectric properties are analyzed. Stimulation thresholds at the fracture site are based on detailed review of experimental studies. Our results show that for a 1 mm thick 30° fracture with a 15 Hz sinusoidal field, the input current amounts to a maximum of 3.77 µA. Minimum and maximum required current levels are plotted versus fracture parameters, all of which comply with the ICNIRP standard. Simulation results are supported by several experimental reports. Our model is useful for understanding the effects of various geometrical and electrical factors on clinical outcome, and serves as a theoretical aid in the design of more efficient systems. Bioelectromagnetics. 40:128-135, 2019. © 2019 Bioelectromagnetics Society.

Evaluation of the Effects of Photobiomodulation on Partial Osteotomy in Streptozotocin-Induced Diabetes in Rats.

OBJECTIVE: We examined the effects of photobiomodulation (PBM) on stereological parameters, and gene expression of Runt-related transcription factor 2 (RUNX2), osteocalcin, and receptor activator of nuclear factor kappa-B ligand (RANKL) in repairing tissue of tibial bone defect in streptozotocin (STZ)-induced type 1 diabetes mellitus (TIDM) in rats during catabolic response of fracture healing. BACKGROUND DATA: There were conflicting results regarding the efficacy of PBM on bone healing process in healthy and diabetic animals. MATERIALS AND METHODS: Forty-eight rats have been distributed into four groups: group 1 (healthy control, no TIDM and no PBM), group 2 (healthy test, no TIDM and PBM), group 3 (diabetic control, TIDM and no PBM), and group 4 (diabetic test, no TIDM and PBM). TIDM was induced in the groups 3 and 4. A partial bone defect in tibia was made in all groups. The bone defects of groups second and fourth were irradiated by a laser (890 nm, 80 Hz, 1.5 J/cm2). Thirty days after the surgery, all bone defects were extracted and were submitted to stereological examination and real-time polymerase chain reaction (RT-PCR). RESULTS: PBM significantly increased volumes of total callus, total bone, bone marrow, trabecular bone, and cortical bone, and the numbers of osteocytes and osteoblasts of callus in TIDM rats compared to those of callus in diabetic control. In addition, TIDM increased RUNX2, and osteocalcin in callus of tibial bone defect compared to healthy group. PBM significantly decreased osteocalcin gene expression in TIDM rats. CONCLUSIONS: PBM significantly increased many stereological parameters of bone repair in an STZ-induced TIDM during catabolic response of fracture healing. Further RT-PCR test demonstrated that bone repair was modulated in diabetic rats during catabolic response of fracture healing by significant increase in mRNA expression of RUNX2, and osteocalcin compared to healthy control rats. PBM also decreased osteocalcin mRNA expression in TIDM rats.

Laser/LED phototherapy on the repair of tibial fracture treated with wire osteosynthesis evaluated by Raman spectroscopy.

The aim of the present study was to assess, by means of Raman spectroscopy, the repair of complete surgical tibial fractures fixed with wire osteosynthesis (WO) treated or not with infrared laser (λ780 nm) or infrared light emitting diode (LED) (λ850 ± 10 nm) lights, 142.8 J/cm2 per treatment, associated or not to the use of mineral trioxide aggregate (MTA) cement. Surgical tibial fractures were created on 18 rabbits, and all fractures were fixed with WO and some groups were grafted with MTA. Irradiated groups received lights at every other day during 15 days, and all animals were sacrificed after 30 days, being the tibia removed. The results showed that only irradiation with either laser or LED influenced the peaks of phosphate hydroxyapatite (~ 960 cm-1). Collagen (~ 1450 cm-1) and carbonated hydroxyapatite (~ 1070 cm-1) peaks were influenced by both the use of MTA and the irradiation with either laser or LED. It is concluded that the use of either laser or LED phototherapy associated to MTA cement was efficacious on improving the repair of complete tibial fractures treated with wire osteosynthesis by increasing the synthesis of collagen matrix and creating a scaffold of calcium carbonate (carbonated hydroxyapatite-like) and the subsequent deposition of phosphate hydroxyapatite.

Comparison of effects of LLLT and LIPUS on fracture healing in animal models and patients: A systematic review.

The aim of this paper is to study the in vivo potency of low-level laser therapy (LLLT) and low intensity pulsed ultrasound (LIPUS) alone, accompanied by bone grafts, or accompanied by other factors on fracture healing in animal models and patients. In this paper, we aim to systematically review the published scientific literature regarding the use of LLLT and LIPUS to accelerate fracture healing in animal models and patients. We searched the PubMed database for the terms LLLT or LIPUS and/or bone, and fracture. Our analysis also suggests that both LIPUS and LLLT may be beneficial to fracture healing in patients, and that LIPUS is more effective. These finding are of considerable importance in those treatments with a LIPUS, as a laser device may reduce healing time. The most clinically relevant impact of the LIPUS treatment could be a significant reduction in the proportion of patients who go on to develop a nonunion. If it is confirmed that the therapeutic influence is true and reliable, patients will obtain benefits from LIPUS and LLLT. Further clinical trials of high methodological quality are needed in order to determine the optimal role of LIPUS and LLLT in fracture healing in patients.

Evaluation of the Effect of Phototherapy in Patients with Mandibular Fracture on Mandibular Dynamics, Pain, Edema, and Bite Force: A Pilot Study.

OBJECTIVE: The aim of this study was to evaluate the effect of phototherapy on bite force, facial swelling, mandibular movements, and pain in patients having undergone surgical treatment for mandibular fractures. BACKGROUND: These are among the predominant types of facial fractures, and treatment involving surgical fixation with titanium plates is one of the most common procedures in oral-maxillofacial surgery. Phototherapy has been used to accelerate the muscle healing process and significantly improves muscle regeneration by inducing the formation of new muscle fibers. METHODS: The patients were divided into two groups: Group 1-active phototherapy, and Group 2-sham phototherapy. Both groups underwent the surgical procedure by the same surgeon using the same surgical technique. Dosimetric parameters are wavelength, 660 nm; power, 108 mW; radiant energy, 21.6 J; fluency, 21.6 J/cm2; radiance, 38197 mW/cm2; exposure time, 200 sec per point, 10 points bilaterally. Photobiomodulation was performed in 15 sessions. RESULTS: The primary variable was bite force measured with a gnathodynamometer and the secondary variables were facial swelling, mandibular movements (measured with digital calipers), and pain. The Student's t-test was used to determine intergroup differences. CONCLUSIONS: The findings suggest improvements in the laser group in comparison with the sham group with regard to mandibular dynamics, a reduction in postoperative facial swelling, a reduction in pain, and an increase in bite force.

Evaluation of the Effects of Photobiomodulation on Bone Healing in Healthy and Streptozotocin-Induced Diabetes in Rats.

OBJECTIVE: This study aimed to assess the effects of Photobiomodulation (PBM) with pulsed wave laser on Hounsfield unit (HU) and bone strength at a catabolic response (bone resorption) of a callus bone defect in healthy and streptozotocin (STZ)- induced type I diabetes mellitus (TI DM) in rats. BACKGROUND DATA: Conflicting results exist regarding the effect of PBM on bone healing in healthy and diabetic animals. MATERIALS AND METHODS: We randomly divided 20 adult female rats into the following groups: (1) control, no TI DM, and no PBM; (2) no TI DM and PBM; (3) TI DM and no PBM; and (4) TI DM and PBM. TI DM was induced by STZ. All rats underwent partial transversal standardized osteotomies in their right tibias. The rats received PBM (890 nm, 80 Hz, 1.5 J/cm2) thrice per week during 30 days. At 4 weeks after the surgery, the rats were sacrificed and their tibias submitted to computed tomography scanning to measure HU. The samples underwent a three-point bending test to evaluate bone strength. RESULTS: Analysis of variance (ANOVA) (p = 0.013) results showed that treatment by PBM significantly increased the biomechanical property (stress high load) of the callus defect from the partial tibia osteotomy in healthy rats compared to the control groups. However, we observed no significant increase in the biomechanical properties of the laser-treated diabetic bone defect compared to the control diabetic group. The ANOVA for the HU of callus density produced a p value of 0.000. A significant increase existed in the mean callus density in the healthy groups compared to the diabetic groups. CONCLUSIONS: The 80-Hz laser did not significantly enhance bone repair from an osteotomy of the tibia in an experimental model of TI DM rats.

Effects of local vibration and pulsed electromagnetic field on bone fracture: A comparative study.

The effectiveness of various therapeutic methods on bone fracture has been demonstrated in several studies. In the present study, we tried to evaluate the effect of local low-magnitude, high-frequency vibration (LMHFV) on rat tibia fracture in comparison with pulsed electromagnetic fields (PEMF) during the healing process. Mid-diaphysis tibiae fractures were induced in 30 Sprague-Dawley rats. The rats were assigned into groups such as control (CONT), LMHFV (15 min/day, 7 days/week), and PEMF (3.5 h/day, 7 days/week) for a three-week treatment. Nothing was applied to control group. Radiographs, serum osteocalcin levels, and stereological bone analyses of the three groups were compared. The X-rays of tibiae were taken 21 days after the end of the healing process. PEMF and LMHFV groups had more callus formation when compared to CONT group; however, the difference was not statistically significant (P = 0.375). Serum osteocalcin levels were elevated in the experimental groups compared to CONT (P ≤ 0.001). Stereological tests also showed higher osteogenic results in experimental groups, especially in LMHFV group. The results of the present study suggest that application of direct local LMHFV on fracture has promoted bone formation, showing great potential in improving fracture outcome. Bioelectromagnetics. 38:339-348, 2017. © 2017 Wiley Periodicals, Inc.

Low level laser therapy accelerates bone healing in spinal cord injured rats.

Bone loss occurs rapidly and consistently after the occurrence of a spinal cord injury (SCI), leading to a decrease in bone mineral density (BMD) and a higher risk of fractures. In this context, the stimulatory effects of low level laser therapy (LLLT) also known as photobiomodulation (PBM) have been highlighted, mainly due to its osteogenic potential. The aim of the present study was to evaluate the effects of LLLT on bone healing using an experimental model of tibial bone defect in SCI rats. Twenty-four female Wistar rats were randomly divided into 3 groups: Sham group (SG), SCI control group (SC) and SCI laser treated group (SL). Two weeks after the induction of the SCI, animals were submitted to surgery to induce a tibial bone defect. Treatment was performed 3days a week, for 2weeks, at a single point over the area of the injury, using an 808nm laser (30mW, 100J/cm(2); 0.028cm(2), 1.7W/cm², 2.8J). The results of the histological and morphometric evaluation demonstrated that the SL group showed a larger amount of newly formed bone compared to the SC group. Moreover, a significant immunoexpression of runt-related transcription factor 2 (RUNX2) was observed in the SL group. There was no statistical difference in the biomechanical evaluation. In conclusion, the results suggest that LLLT accelerated the process of bone repair in rats with complete SCI.

Improvement of bone repair in diabetic rats subjected to ƛ780 nm low-level laser therapy.

PURPOSE: To investigate the effect of low-level laser therapy on bone healing in diabetic rats. METHODS: Bone cavities (19 mm diameter) were performed in the femur of 72 alloxan-induced diabetic rats, which were assigned into four groups: CTR (non-diabetic control), DBT (diabetic) CTRL (non-diabetic irradiated) and DBTL (diabetic irradiated). Low-level laser therapy was performed every 48 h for seven days. Animals were euthanized at seven, 18 and 30 days. Alkaline phosphatase serum levels and bone repair were analyzed. RESULTS: Low-level laser therapy significantly increased alkaline phosphatase in at seven and 18 days (p<0.001), and improved bone healing at seven (p<0.01), 18 (p<0.05) and 30 (p<0.01) in diabetic animals. In addition, bone healing in irradiated diabetic group was statistically similar to control group at 30 days (p>0.05). CONCLUSION: Low-level laser therapy increased the serum levels of alkaline phosphatase and improved bone healing in alloxan-induced diabetic rats.

Improvement of bone repair in diabetic rats subjected to ƛ780 nm low-level laser therapy

PURPOSE: To investigate the effect of low-level laser therapy on bone healing in diabetic rats.METHODS: Bone cavities (19 mm diameter) were performed in the femur of 72 alloxan-induced diabetic rats, which were assigned into four groups: CTR (non-diabetic control), DBT (diabetic) CTRL (non-diabetic irradiated) and DBTL (diabetic irradiated). Low-level laser therapy was performed every 48h for seven days. Animals were euthanized at seven, 18 and 30 days. Alkaline phosphatase serum levels and bone repair were analyzed.RESULTS: Low-level laser therapy significantly increased alkaline phosphatase in at seven and 18 days (p<0.001), and improved bone healing at seven (p<0.01), 18 (p<0.05) and 30 (p<0.01) in diabetic animals. In addition, bone healing in irradiated diabetic group was statistically similar to control group at 30 days (p>0.05).CONCLUSION:Low-level laser therapy increased the serum levels of alkaline phosphatase and improved bone healing in alloxan-induced diabetic rats.

Effect of low-level laser therapy on bone repair: a randomized controlled experimental study.

The aim of this study was to investigate the effect of low-level laser therapy (LLLT) on bone repair in femoral fractures. Sixty adult Wistar rats were randomly assigned into one of two groups: group A (ostectomy + LLLT) or group B (ostectomy + sham laser). An experimental model of complete bone fracture was surgically created by removing a 2-mm fragment from the middle third of the femoral shaft. Data were analyzed on days 8, 13, and 18 after the fracture (subgroups 1, 2, and 3). Samples were assessed for changes in inflammatory infiltration; trabecular bone matrix, periosteal, and new bone formations; and changes in the expression of particular osteogenic-related proteins (osteocalcin, osteopontin, and osteonectin). Microscopic analysis revealed a significant decrease in inflammatory infiltration, intense trabecular bone matrix and periosteal formation, and an increase in newly formed bone after laser irradiation. We also found an increase in the expression of bone matrix proteins with LLLT, with a significant difference measured for osteocalcin in the LLLT group at day 8 (p = 0.007). We show that LLLT plays an important role in augmenting bone tissue formation, which is relevant to fracture healing. LLLT may therefore be indicated as an adjunct therapeutic tool in clinical practice for the treatment or recovery of nonunion injuries.

Low power laser stimulation of the bone consolidation in tibial fractures of rats: a radiologic and histopathological analysis.

The objective of this study is to analyze the effectiveness of low power laser irradiation in the bone consolidation of tibial fractures in rats. An experimental, comparative, prospective study with control group was designed. Twenty Wistar rats were grouped into control (n = 10) and experimental groups (n = 10). A tibial fracture, with a mechanical drill, was inflicted in all rats. The experimental group received ten days of low power arsenide-gallium laser irradiation of 850 nm (KLD, Sao Paulo, Brasil)-100 mW, 8 J/cm(2), 64 s. Before and after the laser treatment, a radiologic analysis was carried out in both groups, in which the rats were graded from 0 to IV according the Montoya scale of bone consolidation. Also, we histopathologically analyzed the bone to estimate the proliferation of fibroblasts, bone matrix, and angiogénesis with a microscopy, which were graded as I (thin layer of fibroblasts and osteoid matrix), II (thick layer of fibroblasts and osteoid matrix), or III (thick layer of fibroblasts and osteoid matrix and new blood vessels). Radiologic data showed that the experimental group had a higher bone consolidation of Montoya scale after ten days of laser irradiation compared to control group (P < 0.004). Histopathologic data showed more fibroblasts and angiogenesis presence in the group receiving laser irradiation, compared to control group (P < .002). The low power laser radiation therapy may expedite the bone repair after tibial fractures in rats, according to radiologic and histopathologic analysis.

Avaliação de diferentes protocolos de aplicação do laser de baixa intensidade associado ou não ao osso bovino inorgânico (Bio-Oss®) na cicatrização de defeitos ósseos criados cirurgicamente em calvárias de ratos: estudo histológico e histométrico / Evaluation of different protocols of low-level laser (LLL) application combined or not with inorganic bovine bone (Bio-Oss®) in the healing of surgically created bone defects in rat calvaria: histological and histometric study

O propósito deste estudo foi avaliar diferentes protocolos de aplicação do laser de baixa intensidade (LBI) associados ou não ao osso bovino inorgânico (Bio-Oss®) na cicatrização de defeitos ósseos de tamanho crítico (DTC) em calvárias de ratos. Foram utilizados 90 ratos machos adultos (Rattus norvegicus, albinus, Wistar). Um defeito de tamanho crítico (DTC) de 5 mm de diâmetro foi criado cirurgicamente na calvária de cada animal. Os animais foram divididos igualmente (n=10) e aleatoriamente em 9 gruposexperimentais: 1) grupo C (controle), 2) grupo LBI (4J) (laser de baixa intensidade GaAlAs, 730 nm, 100 mW, 4J, 140 J/cm2), 3) Grupo LBI (6J) (laser de baixa intensidade GaAlAs, 730 nm, 100 mW, 6J, 210 J/cm2), 4) Grupo BO (osso bovino inorgânico), 5) Grupo BO + LBI (4J) (osso bovino inorgânico + laser de baixa intensidade 4J), 6) Grupo BO + LBI (6J) (osso bovino inorgânico + laser de baixa intensidade 6J), 7) Grupo OA (osso autógeno), 8) Grupo OA + LBI (4J) (osso autógeno + laser de baixa intensidade 4J), 9) Grupo OA + LBI (6J) (osso autógeno + laser de baixa intensidade 6J). Os animais foram submetidos à eutanásia aos 30 dias pós-operatórios. Foram avaliadas a área de osso neoformado (AON), extensão linear de osso (ELO) e área de partículas remanescentes (APR). Os dados foram submetidos ao teste paramétrico ANOVA, seguido pelo teste de Tukey (p<0,05). O grupo BO+LBI (6J) demonstrou maior média (48,57 ± 28,22%) de AON e o grupo C a menor média (9,96 ± 4,49%) de AON. Os grupos LBI (6J), OA+LBI (6J) e BO+LBI (6J) demonstraram diferenças estatisticamente significativas de AON em relação ao grupo C. Em relação a ELO, apenas os grupos BO e BO+LBI (4J) não demonstraram diferenças estatisticamente significativas quando comparados ao grupo C, e a maior diferença entre as médias de ELO foram nas comparações do grupo LBI (6J) (76,55 ± 15,54%) com o grupo C (16,00 ± 9,86%. Maior APR foi observada nos grupos em que BO não foi irradiado pelo LBI. Porém, quando...

The purpose of this study was to evaluate different protocols of low-level laser (LLL) application combined or not with inorganic bovine bone (Bio-Oss®) in the healing process of bone defects of critical size (CSD) in rat calvaria. 90 male adult rats (Rattusnorvegicus, albinus, Wistar) were used. A critical size defect (CSD) of 5 mm in diameter was surgically created in the calvaria of each rat. The rats were then divided equally (n=10) and randomly into 9 experimental groups: 1) Group C (control) 2) Group LBI (4J) (low-level laser - GaAlAs, 730 nm, 100 mW, 4J, 140 J/cm2), 3) Group LBI (6) (low- level laser - GaAlAs, 730 nm, 100 mW, 6J, 210 J/cm2), 4) Group BO (inorganic bovine bone), 5) Group BO + LBI (4J) (inorganic bovine bone + low-level laser 4J), 6) Group BO LBI (6J) (inorganic bovine bone + low-level laser 6J), 7) Group OA (autogenous bone), 8) Group OA + LBI (4J) (autogenous bone + low-level laser 4J), 9 Group OA + LBI (6J) (autogenous bone + low-level laser 6J). The rats utilized were euthanized 30 days post-operation. The areas of new bone formation (ANB), linear extension bone (LEB), and areas of remaining particles (ARP) were evaluated. The data underwent the parametric ANOVA test, followed by the Tukey test (p<0,05). Group BO+LBI (6J) presented the greatest average (48,57 ± 28,22%) of ANB and Group C presented the lowest average (9,96 ± 4,49%) of ANB. The groups LBI (6J), OA+LBI (6J), and BO+LBI (6J) presented statistically significant differences of ANB in comparison to Group C. Regarding the LEB, only the groups BO and BO+LBI (4J) did not present differences statistically significant in comparison to Group C. The largest difference between the averages of LEB were in the comparison of Group LBI (6J) (76,55 ± 15,54%) with Group C (16,00 ± 9,86%). The largest ARP was observed in the groups where the inorganic bovine bone was not irradiated by the LLL. However, when comparing Group BO+LBI (6J) to Group OA+LBI (4J) and...

Avaliação de diferentes protocolos de aplicação do laser de baixa intensidade associado ou não ao osso bovino inorgânico (Bio-Oss®) na cicatrização de defeitos ósseos criados cirurgicamente em calvárias de ratos: estudo histológico e histométrico / Evaluation of different protocols of low-level laser (LLL) application combined or not with inorganic bovine bone (Bio-Oss®) in the healing of surgically created bone defects in rat calvaria: histological and histometric study

O propósito deste estudo foi avaliar diferentes protocolos de aplicação do laser de baixa intensidade (LBI) associados ou não ao osso bovino inorgânico (Bio-Oss®) na cicatrização de defeitos ósseos de tamanho crítico (DTC) em calvárias de ratos. Foram utilizados 90 ratos machos adultos (Rattus norvegicus, albinus, Wistar). Um defeito de tamanho crítico (DTC) de 5 mm de diâmetro foi criado cirurgicamente na calvária de cada animal. Os animais foram divididos igualmente (n=10) e aleatoriamente em 9 gruposexperimentais: 1) grupo C (controle), 2) grupo LBI (4J) (laser de baixa intensidade GaAlAs, 730 nm, 100 mW, 4J, 140 J/cm2), 3) Grupo LBI (6J) (laser de baixa intensidade GaAlAs, 730 nm, 100 mW, 6J, 210 J/cm2), 4) Grupo BO (osso bovino inorgânico), 5) Grupo BO + LBI (4J) (osso bovino inorgânico + laser de baixa intensidade 4J), 6) Grupo BO + LBI (6J) (osso bovino inorgânico + laser de baixa intensidade 6J), 7) Grupo OA (osso autógeno), 8) Grupo OA + LBI (4J) (osso autógeno + laser de baixa intensidade 4J), 9) Grupo OA + LBI (6J) (osso autógeno + laser de baixa intensidade 6J). Os animais foram submetidos à eutanásia aos 30 dias pós-operatórios. Foram avaliadas a área de osso neoformado (AON), extensão linear de osso (ELO) e área de partículas remanescentes (APR). Os dados foram submetidos ao teste paramétrico ANOVA, seguido pelo teste de Tukey (p<0,05). O grupo BO+LBI (6J) demonstrou maior média (48,57 ± 28,22%) de AON e o grupo C a menor média (9,96 ± 4,49%) de AON. Os grupos LBI (6J), OA+LBI (6J) e BO+LBI (6J) demonstraram diferenças estatisticamente significativas de AON em relação ao grupo C. Em relação a ELO, apenas os grupos BO e BO+LBI (4J) não demonstraram diferenças estatisticamente significativas quando comparados ao grupo C, e a maior diferença entre as médias de ELO foram nas comparações do grupo LBI (6J) (76,55 ± 15,54%) com o grupo C (16,00 ± 9,86%. Maior APR foi observada nos grupos em que BO não foi irradiado pelo LBI. Porém, quando...

The purpose of this study was to evaluate different protocols of low-level laser (LLL) application combined or not with inorganic bovine bone (Bio-Oss®) in the healing process of bone defects of critical size (CSD) in rat calvaria. 90 male adult rats (Rattusnorvegicus, albinus, Wistar) were used. A critical size defect (CSD) of 5 mm in diameter was surgically created in the calvaria of each rat. The rats were then divided equally (n=10) and randomly into 9 experimental groups: 1) Group C (control) 2) Group LBI (4J) (low-level laser - GaAlAs, 730 nm, 100 mW, 4J, 140 J/cm2), 3) Group LBI (6) (low- level laser - GaAlAs, 730 nm, 100 mW, 6J, 210 J/cm2), 4) Group BO (inorganic bovine bone), 5) Group BO + LBI (4J) (inorganic bovine bone + low-level laser 4J), 6) Group BO LBI (6J) (inorganic bovine bone + low-level laser 6J), 7) Group OA (autogenous bone), 8) Group OA + LBI (4J) (autogenous bone + low-level laser 4J), 9 Group OA + LBI (6J) (autogenous bone + low-level laser 6J). The rats utilized were euthanized 30 days post-operation. The areas of new bone formation (ANB), linear extension bone (LEB), and areas of remaining particles (ARP) were evaluated. The data underwent the parametric ANOVA test, followed by the Tukey test (p<0,05). Group BO+LBI (6J) presented the greatest average (48,57 ± 28,22%) of ANB and Group C presented the lowest average (9,96 ± 4,49%) of ANB. The groups LBI (6J), OA+LBI (6J), and BO+LBI (6J) presented statistically significant differences of ANB in comparison to Group C. Regarding the LEB, only the groups BO and BO+LBI (4J) did not present differences statistically significant in comparison to Group C. The largest difference between the averages of LEB were in the comparison of Group LBI (6J) (76,55 ± 15,54%) with Group C (16,00 ± 9,86%). The largest ARP was observed in the groups where the inorganic bovine bone was not irradiated by the LLL. However, when comparing Group BO+LBI (6J) to Group OA+LBI (4J) and...

Low-dose X-ray irradiation promotes osteoblast proliferation, differentiation and fracture healing.

Great controversy exists regarding the biologic responses of osteoblasts to X-ray irradiation, and the mechanisms are poorly understood. In this study, the biological effects of low-dose radiation on stimulating osteoblast proliferation, differentiation and fracture healing were identified using in vitro cell culture and in vivo animal studies. First, low-dose (0.5 Gy) X-ray irradiation induced the cell viability and proliferation of MC3T3-E1 cells. However, high-dose (5 Gy) X-ray irradiation inhibited the viability and proliferation of osteoblasts. In addition, dynamic variations in osteoblast differentiation markers, including type I collagen, alkaline phosphatase, Runx2, Osterix and osteocalcin, were observed after both low-dose and high-dose irradiation by Western blot analysis. Second, fracture healing was evaluated via histology and gene expression after single-dose X-ray irradiation, and low-dose X-ray irradiation accelerates fracture healing of closed femoral fractures in rats. In low-dose X-ray irradiated fractures, an increase in proliferating cell nuclear antigen (PCNA)-positive cells, cartilage formation and fracture calluses was observed. In addition, we observed more rapid completion of endochondral and intramembranous ossification, which was accompanied by altered expression of genes involved in bone remodeling and fracture callus mineralization. Although the expression level of several osteoblast differentiation genes was increased in the fracture calluses of high-dose irradiated rats, the callus formation and fracture union were delayed compared with the control and low-dose irradiated fractures. These results reveal beneficial effects of low-dose irradiation, including the stimulation of osteoblast proliferation, differentiation and fracture healing, and highlight its potential translational application in novel therapies against bone-related diseases.

Low-intensity pulsed ultrasound shortens the treatment time in tibial distraction osteogenesis.

PURPOSE: Low-intensity pulsed ultrasound (LIPUS) has been used successfully to accelerate healing of fresh fractures and non-unions. It also improved callus maturation with distraction osteogenesis in animal trials. However, only few clinical studies are available to support its widespread use for the latter indication in humans. METHODS: Twenty-one patients undergoing callus distraction for posttraumatic tibial defects were randomized into two groups: the trial group (12 men; mean age 32 years) which received 20 minutes LIPUS daily during treatment and the control group (six men and three women; mean age 29 years) without LIPUS treatment. The Ilizarov ring fixator was used in all cases. Results were examined clinically and radiologically, analysing callus maturation with a computer-assisted measurement. RESULTS: Patients in the LIPUS group needed a mean of 33 days to consolidate every 1 cm of new bone in comparison to 45 days in the control group. The healing index was therefore shortened by 12 days/cm in the LIPUS group. This means that callus maturation was 27 % faster in the LIPUS group. The fixator time was shortened by 95 days in the LIPUS group. The overall daily increase in radiographic callus density was 33 % more in the LIPUS group than in the control group. CONCLUSIONS: LIPUS treatment is an effective non-invasive adjuvant method to enhance callus maturation in distraction osteogenesis. With the help of this treatment, the healing time and the duration of external fixation can be reliably shortened.