The use of photobiomodulation therapy or LED and mineral trioxide aggregate improves the repair of complete tibial fractures treated with wire osteosynthesis in rodents.

The repair of large bone defects is lengthy and complex. Both biomaterials and phototherapy have been used to improve bone repair. We aimed to describe histologically the repair of tibial fractures treated by wiring (W), irradiated or not, with laser (λ780 nm, 70 mW, CW, spot area of 0.5 cm2, 20.4 J/cm2 (4 × 5.1 J/cm2, Twin Flex Evolution®, MM Optics, Sao Carlos, SP, Brazil) per session, 300 s, 142.8 J/cm2 per treatment) or LED (λ850 ± 10 nm, 150 mW, spot area of 0.5 cm2, 20.4 J/cm2 per session, 64 s, 142.8 J/cm2 per treatment, Fisioled®, MM Optics, Sao Carlos, Sao Paulo, Brazil) and associated or not to the use of mineral trioxide aggregate (MTA, Angelus®, Londrina, PR, Brazil). Inflammation was discrete on groups W and W + LEDPT and absent on the others. Phototherapy protocols started immediately before suturing and repeated at every other day for 15 days. Collagen deposition intense on groups W + LEDPT, W + BIO-MTA + LaserPT and W + BIO-MTA + LEDPT and discrete or moderate on the other groups. Reabsorption was discrete on groups W and W + LEDPT and absent on the other groups. Neoformation varied greatly between groups. Most groups were partial and moderately filed with new-formed bone (W, W + LaserPT, W + LEDPT, W + BIO-MTA + LEDPT). On groups W + BIO-MTA and W + BIO-MTA + LaserPT bone, neoformation was intense and complete. Our results are indicative that the association of MTA and PBMT (λ = 780 nm) improves the repair of complete tibial fracture treated with wire osteosynthesis in a rodent model more efficiently than LED (λ = 850 ± 10 nm).

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.

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.

The efficacy of the use of IR laser phototherapy associated to biphasic ceramic graft and guided bone regeneration on surgical fractures treated with miniplates: a histological and histomorphometric study on rabbits.

The aim of the present study was to assess, by light microscopy and histomorphometry, the repair of surgical fractures fixed with internal rigid fixation (IRF) treated or not with IR laser (λ780 nm, 50 mW, 4 × 4 J/cm(2) = 16 J/cm(2), ϕ = 0.5 cm(2), CW) associated or not to the use of hydroxyapatite and guided bone regeneration. Surgical tibial fractures were created under general anesthesia on 15 rabbits that were divided into 5 groups, maintained on individual cages, at day/night cycle, fed with solid laboratory pelted diet, and had water ad libidum. The fractures in groups II, III, IV, and V were fixed with miniplates. Animals in groups III and V were grafted with hydroxyapatite and GBR technique used. Animals in groups IV and V were irradiated at every other day during two weeks (4 × 4 J/cm(2), 16 J/cm(2) = 112 J/cm(2)). Observation time was that of 30 days. After animal death, specimens were taken, routinely processed to wax, cut and stained with HA and Sirius red, and used for histological assessment. The results of both analyses showed a better bone repair on all irradiated subjects especially when the biomaterial and GBR were used. In conclusion, the results of the present investigation are important clinically as they are suggestive that the association of hydroxyapatite, and laser light resulted in a positive and significant repair of complete tibial fractures treated with miniplates.

Low-level laser therapy enhances the expression of osteogenic factors during bone repair in rats.

The aim of this study was to evaluate the effects of low-level laser therapy (LLLT) on bone formation, immunoexpression of osteogenic factors, and biomechanical properties in a tibial bone defect model in rats. Sixty male Wistar rats were distributed into bone defect control group (CG) and laser irradiated group (LG). Animals were euthanized on days 15, 30, and 45 post-injury. The histological and morphometric analysis showed that the treated animals presented no inflammatory infiltrate and a better tissue organization at 15 and 30 days postsurgery. Also, a higher amount of newly formed bone was observed at 15 days postsurgery. No statistically significant difference was observed in cyclooxygenase-2 immunoexpression among the groups at 15, 30, and 45 days in the immunohistochemical analysis. Considering RUNX-2, the immunoexpression was statistically higher in the LG compared to the CG at 45 days. BMP-9 immunoexpression was significantly higher in the LG in comparison to CG at day 30. However, there was no expressivity for this immunomarker, both in the CG and LG, at the day 45 postsurgery. No statistically significant difference was observed in the receptor activator of nuclear factor kappa-B ligand immunoexpression among the groups in all periods evaluated. No statistically significant difference among the groups was observed in the maximal load in any period of time. Our findings indicate that laser therapy improved bone healing by accelerating the development of newly formed bone and activating the osteogenic factors on tibial defects, but the biomechanical properties in LG were not improved.

The efficacy of the use of IR laser phototherapy associated to biphasic ceramic graft and guided bone regeneration on surgical fractures treated with miniplates: a Raman spectral study on rabbits.

The aim of the present study was to assess, by Raman spectroscopy, the repair of surgical fractures fixed with internal rigid fixation (IRF) treated or not with IR laser (λ780 nm, 50 mW, 4 × 4 J/cm(2) = 16 J/cm(2), ϕ = 0.5 cm(2), CW) associated or not to the use of hydroxyapatite and guided bone regeneration (GBR). Surgical tibial fractures were created under general anesthesia on 15 rabbits that were divided into five groups, maintained on individual cages, at day/night cycle, fed with solid laboratory pelted diet and had water ad libitum. The fractures in groups II, III, IV and V were fixed with miniplates. Animals in groups III and V were grafted with hydroxyapatite and GBR technique used. Animals in groups IV and V were irradiated at every other day during 2 weeks (4 × 4 J/cm(2), 16 J/cm(2) = 112 J/cm(2)). Observation time was that of 30 days. After animal death, specimens were taken and kept in liquid nitrogen and used for Raman spectroscopy. Raman spectroscopy showed significant differences between groups (p < 0.001). Basal readings showed mean value of 1,234 ± 220.1. Group internal rigid fixation + biomaterial + laser showed higher readings (3,521 ± 2,670) and group internal rigid fixation + biomaterial the lowest (212.2 ± 119.8). In conclusion, the results of the present investigation are important clinically as spectral analysis of bone component evidenced increased levels of CHA on fractured sites by using the association of laser light to a ceramic graft.

The efficacy of the use of IR laser phototherapy associated to biphasic ceramic graft and guided bone regeneration on surgical fractures treated with wire osteosynthesis: a comparative laser fluorescence and Raman spectral study on rabbits.

The aim of the present study was to assess, by Raman spectroscopy and laser fluorescence, the repair of surgical fractures fixed with wire osteosynthesis treated or not with infrared laser (λ780 nm, 50 mW, 4 × 4 J/cm(2) =16 J/cm(2), ϕ=0.5 cm(2), CW) associated or not to the use of hydroxyapatite and guided bone regeneration. Surgical tibial fractures were created under general anesthesia on 15 rabbits that were divided into five groups, maintained on individual cages, at day/night cycle, fed with solid laboratory pelted diet, and had water ad libitum. The fractures in groups II, III, IV, and V were fixed with wires. Animals in groups III and V were grafted with hydroxyapatite (HA) and guided bone regeneration (GBR) technique used. Animals in groups IV and V were irradiated at every other day during 2 weeks (4 × 4 J/cm(2), 16 J/cm(2) =112 J/cm(2)). Observation time was that of 30 days. After animal death, specimens were taken and kept in liquid nitrogen and used for Raman spectroscopy. The Raman results showed basal readings of 1,234.38 ± 220. Groups WO+B+L showed higher readings (1,680.22 ± 822) and group WO+B the lowest (501.425 ± 328). Fluorescence data showed basal readings of 5.83333 ± 0.7. Groups WO showed higher readings (6.91667 ± 0.9) and group WO+B+L the lowest (1.66667 ± 0.5). There were significant differences between groups on both cases (p<0.05). Pearson correlation was negative and significant (R (2) = -0.60; p<0.001), and it was indicative that, when the Raman peaks of calcium hydroxyapatite (CHA) are increased, the level of fluorescence is reduced. It is concluded that the use of near-infrared lasertherapy associated to HA graft and GBR was effective in improving bone healing on fractured bones as a result of the increasing deposition of CHA measured by Raman spectroscopy and decrease of the organic components as shown by the fluorescence readings.

Helium-neon laser improves bone repair in rabbits: comparison at two anatomic sites.

The purpose of this study was to evaluate the influence of helium-neon laser on bone repair of femur and tibia in rabbits. For this purpose, 15 New Zealand rabbits underwent bilateral bone damage (tibia and femur) using a spherical bur. Helium-neon laser light, at a fluency of 6 J∕cm(2) and wavelength of 632.8 nm was applied on the left legs (laser group). The right tibia or femur lesions (control group) served as negative control. All sections were histopathologically analyzed using HE sections and the morphometric data from bone tissue and hyaline cartilage were achieved. Histopathological analysis showed regular bone trabeculae covered by osteoblastic cells after 1 week in the group exposed to laser therapy from femur and tibia indistinctly. After 3 weeks, the laser group showed new bone formation coming from the bony walls in the femur and tibia as well. On the 5th week, well-defined trabecula undergoing remodeling process was detected for the most intense pattern in tibia only. Morphometric analysis revealed significant statistical differences (p < 0.05) in the bone tissue for the laser-exposed group on 1st and 3rd weeks. After 5th week, bone formation was increased to tibia only. Taken together, such findings suggest that helium-neon laser is able to improve bone repair in rabbits being the most pronounced effect in tibia.

The effects of low-level laser therapy, 670 nm, on epiphyseal growth in rats.

The longitudinal growth of long bones is attributed to epiphyseal growth. However, the effects of low-level laser therapy (LLLT) in such structures has still not been studied extensively in the literature. Therefore, the aim of this study was to evaluate the use of LLLT, 670 nm, at three different doses on the epiphyseal growth of the right tibia of rats. Twenty-one Wistar rats, aged four weeks, were subjected to the application of LLLT, with dosage according to the group (G4: were submitted to the application of 4 J/cm(2); G8: were submitted to the application of 8 J/cm(2); G16: were submitted to the application of 16 J/cm(2)). After completion of protocol they were kept until they were 14 weeks of age and then submitted to a radiological examination (evaluation of limb length) and euthanised. The histological analysis of the growth plates (total thickness and hypertrophic and proliferative zones) was then performed. Comparisons were made with the untreated left tibia. No differences were observed in any of the reviews (radiological and histological), when comparing the right sides (treated) to the left (untreated). It was concluded that the treatment with LLLT within the parameters used caused changes neither in areas of the epiphyseal cartilage nor in the final length of limbs.

Comparative study of the effects of low-intensity pulsed ultrasound and low-level laser therapy on bone defects in tibias of rats.

The aim of this study was to investigate and to compare the effects of low intensity ultra-sound (LIPUS) and low-level laser therapy (LLLT) during the process of bone healing by means of histopathological and morphometric analysis. The animals were randomly distributed into three groups of 30 animals each: the control group (bone defect without treatment); the laser-treated group: (bone defect treated with laser), and the LIPUS-treated (bone defect treated with ultrasound). Each group was further divided into three different subgroups (n = 10) and on days 7, 13, and 25 post-injury, rats were killed with an intra-peritoneal injection of general anesthetic. The rats were treated with a 30-mW/cm(2) low-intensity pulsed ultrasound and a 830-nm laser at 50 J/cm(2). The results showed intense new bone formation surrounded by highly vascularized connective tissue presenting a slight osteogenic activity, with primary bone deposition being observed in the group exposed to laser in the intermediary (13 days) and late stages of repair (25 days). This was confirmed by morphometric analysis in which significant statistical differences (p < 0.05) were noticed when compared to the control. No remarkable differences were noticed in the specimens treated with ultrasound with regard to the amount of newly formed bone in comparison to the control group. Taken together, our results indicate that laser therapy improves bone repair in rats as depicted by histopathological and morphometric analysis, mainly at the late stages of recovery. Moreover, it seems that this therapy was more effective than US to accelerate bone healing.

Laser 904 nm action on bone repair in rats with osteoporosis.

SUMMARY: The aim of the present study was to determine the action of AsGA laser irradiation on bone repair in the tibia of osteopenic rats. The animals were randomly divided into eight experimental groups according to the presence of ovarian hormone (sham group) or the absence of the hormone (OVX group), as well as being irradiated or non-irradiated. Low-level 904-nm laser (50 mJ/cm(2)) accelerated the repair process of osteopenic fractures, especially in the initial phase of bone regeneration. INTRODUCTION: The development of new techniques to speed the process of bone repair has provided significant advances in the treatment of fractures. Some attention recently focused on the effects of biostimulation on bone. METHODS: Forty-eight adult rats were randomly divided into eight experimental groups (six animals in each group) according to the presence of ovarian hormone (sham group) or absence of the hormone (ovariectomized (OVX) group) as well as being irradiated or non-irradiated. For the application of low-level laser therapy, the animals were anesthetized with one third of the dose sufficient to immobilize the animal and irradiated with AsGa laser (904 nm, 50 mJ/cm(2) for 2 s, point form and in contact). The control animals received the same type of manipulation as the irradiated animals, but with the laser turned off. Half of the animals were killed 7 days following the confection of the bone defect, and the other half were killed 21 days after the surgery. After complete demineralization, the tibias were cut cross-sectionally in the central region of the bone defect and embedded in paraffin blocks. The blocks were then cut in semi-seriated slices and stained with hematoxylin and eosin. RESULTS: There was new bone formation in the animals in the OVX group with laser treatment killed after 7 days (p < 0.001). The lowest percentage of bone formation was observed in the OVX without laser killed after 7 days (p > 0.05). All animals killed after 21 days exhibited linear closure of the lesion. CONCLUSION: Low-level 904-nm laser (50 mJ/cm(2)) accelerated the repair process of osteopenic fractures, especially in the initial phase of bone regeneration.

The effects of helium-neon light therapy on healing of partial osteotomy of the tibia in streptozotocin induced diabetic rats.

OBJECTIVE: The effect of light therapy (LT) on surgically created partial osteotomy in streptozotocin (STZ)-induced diabetic rats was examined. BACKGROUND DATA: LT has been shown to enhance bone repair in healthy human and animal models. MATERIALS AND METHODS: Forty male rats were divided into groups 1 to 5. Diabetes was induced in rats of groups 1, 2, and 3 using an intraperitoneal injection of STZ. All diabetic rats were maintained for 30 days after STZ injection. Under general anesthesia and sterile conditions, a partial transversal standardized osteotomy was made in the mid-portion of the right tibia. The defects in groups 2, 3, and 5 were treated using a helium-neon (He-Ne) laser (632.8 nm, 10 mW, circular beam shape). Groups 1 and 4 were diabetic placebo and normal placebo groups, respectively. A dose of 369.4 J/cm2 for groups 2 and 5 and a dose of 66.8 J/cm2 for group 3 were applied three times a week. Six weeks after surgery, the right tibia was collected. The specimen was subjected to a three-point bending test. RESULTS: LT with 369.4 J/cm2 energy density resulted in significantly greater bending stiffness in group 5 (41.8+/-5.2) than in groups 1 (18.5+/-4.1), 2 (17.7+/-1.6), and 3 (11.5+/-4) (least significant difference (LSD) test, p<0.01, p<0.001, and p<0.001, respectively). LT with 369.4 J/cm2 energy density resulted in a significantly higher stress load in group 5 (10+/-0.4) than in groups 1 (4.9+/-1.5), 2 (5.7+/-0.52), and 3 (3.9+/-1.1) (LSD test, p<0.01, p<0.01, p<0.001, respectively). CONCLUSION: LT with a He-Ne laser in STZ-induced diabetic rats did not enhance bone repair of a partial transversal standardized osteotomy.

The effects of low-level laser therapy on bone in diabetic and nondiabetic rats.

OBJECTIVE: The aim of the present study was to examine the effects of low-level laser therapy (LLLT) on the tibia of streptozotocin-induced diabetic (STZ-D) rats. BACKGROUND DATA: LLLT has been found to accelerate fracture healing in animals. Diabetes mellitus decreases bone volume and its biomechanical parameters. MATERIALS AND METHODS: Twenty rats were divided randomly into four groups. Rats in the first two groups were administrated a single injection of STZ to induce diabetes, while animals in groups 3 and 4 were given a sham injection of distilled water. The right tibia in groups 1 and 2 was treated with a He-Ne laser (632.8 nm, 10 mW) of 28.6 and 382.2 J/cm(2), respectively. LLLT was performed daily for 14 consecutive days. The right tibia of rats in group 3 was treated with LLLT the same as group 2. The right tibia of rats in group 4 was used for based line studies. After 14 d, right tibiae and left tibiae (control bone) were extracted and subjected to the three-point bending test and histological study. RESULTS: Maximum force (N) was significantly greater in laser-treated bones of groups 2 and 3 compared with their relevant control groups (paired Student t test, p = 0.05 and p = 0.007, respectively). Density of the bone lamella meshwork of compact bone in group 2 was significantly greater in comparison with its control group (paired Student t test, p = 0.005). CONCLUSION: LLLT on tibia of STZ-D rats increased the bone lamella meshwork density of compact bone and also increased its strength.

Low-level laser therapy modulates cyclo-oxygenase-2 expression during bone repair in rats.

The goal of this study was to analyze the role of cyclo-oxygenase-2 following bone repair in rats submitted to low-level laser therapy. A total of 48 rats underwent surgery to inflict bone defects in their tibias having been randomly distributed into two groups: negative control and laser exposed group, i.e., the animals were treated with low-level laser therapy by means of gallium arsenide laser at 16 J/cm(2). The animals were killed after 48 h, 7 days, 14 days, or 21 days. The tibias were removed for morphological, morphometric, and immunohistochemistry analysis for cyclo-oxygenase-2. Statistical significant differences (P < 0.05) were observed in the quality of bone repair and quantity of formed bone between groups 14 days after surgery in the laser exposed group. In the same way, cyclo-oxygenase-2 immunoreactivity was more intense in bone cells for intermediate periods evaluated in this group. Taken together, such results suggest that low-level laser therapy is able to improve bone repair in the tibia of rats after 14 days of surgery as a result of an up-regulation for cyclo-oxygenase-2 expression in bone cells.

Effect of lower-level laser therapy on rabbit tibial fracture.

OBJECTIVE: The purpose of the study was to demonstrate the biological effects of low-level laser therapy (LLLT) on tibial fractures using radiographic, histological, and bone density examinations. METHODS: Fourteen New Zealand white rabbits with surgically induced mid-tibial osteotomies were included in the study. Seven were assigned to a group receiving LLLT (LLLT-A) and the remaining seven served as a sham-treated control group (LLLT-C). A low-energy laser apparatus with a wavelength of 830 nm, and a sham laser (a similar design without laser diodes) were used for the study. Continuous outflow irradiation with a total energy density of 40 J/cm(2) and a power level of 200 mW/cm(2) was directly delivered to the skin for 50 seconds at four points along the tibial fracture site. Treatment commenced immediately postsurgery and continued once daily for 4 weeks. RESULTS: Radiographic findings revealed no statistically significant fracture callus thickness difference between the LLLT-A and LLLT-C groups (p > 0.05). However, the fractures in the LLLT-A group showed less callus thickness than those in LLLT-C group 3 weeks after treatment. The average tibial volume was 14.5 mL in the LLLT-A group, and 11.25 mL in the LLLT-C group. The average contralateral normal tibial volume was 7.1 mL. Microscopic changes at 4 weeks revealed an average grade of 5.5 and 5.0 for the LLLT-A group and the LLLT-C group, respectively. The bone mineral density (BMD) as ascertained using a grey scale (graded from 0 to 256) showed darker coloration in the LLLT-A group (138) than in the LLLT-C group (125). CONCLUSION: The study suggests that LLLT may accelerate the process of fracture repair or cause increases in callus volume and BMD, especially in the early stages of absorbing the hematoma and bone remodeling. Further study is necessary to quantify these findings.