Bone substitutes and photobiomodulation in bone regeneration: A systematic review in animal experimental studies.

In general, bone fractures are able of healing by itself. However, in critical situations such as large bone defects, poor blood supply or even infections, the biological capacity of repair can be impaired, resulting in a delay of the consolidation process or even in non-union fractures. Thus, technologies able of improving the process of bone regeneration are of high demand. In this context, ceramic biomaterials-based bone substitutes and photobiomodulation (PBM) have been emerging as promising alternatives. Thus, the present study performed a systematic review targeting to analyze studies in the literature which investigated the effects of the association of ceramic based bone substitutes and PBM in the process of bone healing using animal models of bone defects. The search was conducted from March and April of 2019 in PubMed, Web of Science and Scopus databases. After the eligibility analyses, 16 studies were included in this review. The results showed that the most common material used was hydroxyapatite (HA) followed by Biosilicate associated with infrared PBM. Furthermore, 75% of the studies demonstrated positive effects to stimulate bone regeneration from association of ceramic biomaterials and PBM. All studies used low-level laser therapy (LLLT) device and the most studies used LLLT infrared. The evidence synthesis was moderate for all experimental studies for the variable histological analysis demonstrating the efficacy of techniques on the process of bone repair stimulation. In conclusion, this review demonstrates that the association of ceramic biomaterials and PBM presented positive effects for bone repair in experimental models of bone defects.

Evaluation of photobiomodulation therapy to accelerate bone formation in the mid palatal suture after rapid palatal expansion: a randomized clinical trial.

To evaluate the efficiency of photobiomodulation therapy (PBMT) in the midpalatal suture (MPS) and pain sensation in patients undergoing rapid palatal expansion (RPE). Thirty-four individuals with the diagnosis of skeletal maxillary hypoplasia were divided in two groups: laser (n = 18) and control (n = 16). Treatment plan consisted of the use of the Hyrax expander in all patients. Subjects in the laser group were irradiated with diode laser (980 nm, 0.3 W) in six spots bilaterally distributed along the MPS for 10 s during the active phase of treatment and after overcorrection (passive phase of RPE). Control group received sham irradiations with the laser in standby mode to characterize the placebo effect. Digital occlusal radiographs were performed at different time-points for bone formation evaluation in both groups. The effects of laser irradiation on pain were assessed by the visual analog scale (Wong-Baker Faces Pain Scale). Bone formation between groups was not significantly different (p = 0.2273). At 3 months, bone formation was not yet complete in both groups. Pain sensation was similar between groups (p = 0.3940). However, pain was significantly higher for the first 7 days of treatment compared with the 14th day. PBMT did not accelerate bone regeneration in the MPS and pain sensation was similar.

Photobiomodulation as adjunctive therapy for guided bone regeneration. A microCT study in osteoporotic rat model.

Regeneration of diseased bone is challenging. Guided bone regeneration (GBR) has been applied to favor the bone repair. Photobiomodulation (PBM) is also a recognized therapy able to improve bone repair in healthy and diseased individuals. Thus, with the hypothesis that PBM therapy could improve the GBR of diseased bone, this study evaluated the effect of PBM as adjunctive therapy to GBR in osteoporotic rats. Osteoporosis was induced in rats using the oophorectomy model. Then, 5-mm calvaria bone defects were created and treated according to the experimental groups, as follows: with no further treatment (Control); conventional GBR (Membrane), GBR and PBM applied with 3 s, 4 J/cm2 and 0.12 J per point (PBM-1) and GBR and PBM applied with 10s, 14 J/cm2, 0.4 J per point (PBM-2). PBM therapy (808 nm, 40 mW, 1.42 W/cm2) was applied immediately, 48 and 96 h postoperatively. Four and eight weeks later, the samples were harvested and processed for micro-computerized tomography (Micro CT). Data were statistically compared (p < 0.05). From 4 to 8 weeks mostly significant changes were observed in the PBM groups. The bone volume fraction and number of trabeculae of the PBM groups, especially the PBM-1, were significantly higher than those of Control (p < 0.0001). The values of thickness and separation of the trabeculae and structural model index of the PBM groups were significantly smaller than Control (p < 0.0001). The connectivity density was significantly higher on Membrane and PBM groups than Control (p < 0.0004). The application of PBM as adjunctive therapy to GBR results in enhanced bone formation and maturation in comparison to the conventional GBR in the regeneration of lesions of osteoporotic bone in rats. Overviewing the challenges that face bone regeneration in patients with osteoporosis, our findings open new perspectives on the treatment of bone defects under osteoporotic conditions.

NIR light-assisted phototherapies for bone-related diseases and bone tissue regeneration: A systematic review.

Recently, the rapid development of biomaterials has induced great interest in the precisely targeted treatment of bone-related diseases, including bone cancers, infections, and inflammation. Realizing noninvasive therapeutic effects, as well as improving bone tissue regeneration, is essential for the success of bone­related disease therapies. In recent years, researchers have focused on the development of stimuli-responsive strategies to treat bone-related diseases and to realize bone regeneration. Among the various external stimuli for targeted therapy, near infrared (NIR) light has attracted considerable interests due to its high tissue penetration capacity, minimal damage toward normal tissues, and easy remote control properties. The main objective of this systematic review was to reveal the current applications of NIR light-assisted phototherapy for bone-related disease treatment and bone tissue regeneration. Database collection was completed by June 1, 2020, and a total of 81 relevant studies were finally included. We outlined the various therapeutic applications of photothermal, photodynamic and photobiomodulation effects under NIR light irradiation for bone­related disease treatment and bone regeneration, based on the retrieved literatures. In addition, the advantages and promising applications of NIR light-responsive drug delivery systems for spatiotemporal-controlled therapy were summarized. These findings have revealed that NIR light-assisted phototherapy plays an important role in bone-related disease treatment and bone tissue regeneration, with significant promise for further biomedical and clinical applications.

Low-level laser therapy (LLLT) in sites grafted with osteoconductive bone substitutes improves osseointegration.

The aim of this study was to evaluate the osseointegration of implants placed in areas grafted with different osteoconductive bone substitutes irradiated with infrared low-level laser therapy (LLLT). Fifty-six rats were randomly allocated into 4 groups: DBB, bone defects filled with deproteinized bovine bone graft (DBB); HA/TCP, bone defects filled with biphasic ceramic made of hydroxyapatite and ß-tricalcium phosphate (HA/TCP); DBB-L, bone defects filled with DBB and treated by LLLT; HA/TCP-L, bone defects filled with HA/TCP and treated by LLLT. Bone defects were performed in the tibia of each animal and filled with the different biomaterials. The grafted areas were treated with LLLT (λ 808 nm, 100 mW, ϕ ∼ 0.60 mm) in 7 sessions with 48 h between the irradiations. After the 60-day period, the implants were placed, and the animals were euthanized after 15 and 45 days. The osseointegration and bone repair in the grafted area were evaluated by biomechanical, microtomographic and histometric analyses, and the expression of some bone biomarkers was evaluated by immunohistochemistry analysis. LLLT induced higher degree of osseointegration, which was associated with the greater expression of BMP2 and OCN. LLLT performed in areas grafted with osteoconductive bone substitutes prior to implant placement improves osseointegration.

Effects of the combination of low-level laser therapy and anionic polymer membranes on bone repair.

In view of the limitations of bone reconstruction surgeries using autologous grafts as a gold standard, tissue engineering is emerging as an alternative, which permits the fabrication and improvement of scaffolds to stimulate osteogenesis and angiogenesis, processes that are essential for bone repair. Polymers are used to mimic the extracellular bone matrix and support cell growth. In addition, bone neoformation can be induced by external factors such as laser irradiation, which stimulates bone metabolism. The objective of this study was to evaluate the regeneration of bone defects using collagen and elastin membranes derived from intestinal serosa and bovine auricular cartilage combined with low-level laser application. Thirty-six Wistar rats were operated to create a 3-mm defect in the distal metaphysis of the left femur and divided into six groups: G1 (control, no treatment); G2 (laser); G3 (elastin graft), G4 (elastin+laser); G5 (collagen graft); G6 (collagen+laser). The animals were sacrificed 6 weeks after surgery and the femurs were removed for analysis of bone repair. Macroscopic and radiological results showed the absence of an infectious process in the surgical area. This was confirmed by histological analysis, which revealed no inflammatory infiltrate. Histomorphometry showed that the formation of new bone started from the margins of the bone defect and its volume was greater in elastin+laser and collagen+laser. We conclude that newly formed bone in the graft area was higher in the groups that received the biomaterials and laser. The collagen and elastin matrices showed biocompatibility.

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.

Bone Regeneration Effect of Hyperbaric Oxygen Therapy Duration on Calvarial Defects in Irradiated Rats.

OBJECTIVE: In this study, we evaluated changes in bone remodeling in an irradiated rat calvarial defect model according to duration of hyperbaric oxygen therapy. MATERIALS AND METHODS: The 28 rats were divided into four groups. Radiation of 12 Gy was applied to the skull, and 5-mm critical size defects were formed on both sides of the skull. Bone grafts were applied to one side of formed defects. From the day after surgery, HBO was applied for 0, 1, and 3 weeks. At 6 weeks after bone graft, experimental sites were removed and analyzed for radiography, histology, and histomorphometry. RESULTS: Micro-CT analysis showed a significant increase in new bone volume in the HBO-3 group, with or without bone graft. When bone grafting was performed, BV, BS, and BS/TV all significantly increased. Histomorphometric analysis showed significant increases in %NBA and %BVN in the HBO-1 and HBO-3 groups, regardless of bone graft. CONCLUSION: Hyperbaric oxygen therapy was effective for bone regeneration with only 1 week of treatment.

Fibrin Sealant Derived from Human Plasma as a Scaffold for Bone Grafts Associated with Photobiomodulation Therapy.

Fibrin sealants derived from human blood can be used in tissue engineering to assist in the repair of bone defects. The objective of this study was to evaluate the support system formed by a xenograft fibrin sealant associated with photobiomodulation therapy of critical defects in rat calvaria. Thirty-six rats were divided into four groups: BC (n = 8), defect filled with blood clot; FSB (n = 10), filled with fibrin sealant and xenograft; BCPBMT (n = 8), blood clot and photobiomodulation; FSBPBMT (n = 10), fibrin sealant, xenograft, and photobiomodulation. The animals were killed after 14 and 42 days. In the histological and microtomographic analysis, new bone formation was observed in all groups, limited to the defect margins, and without complete wound closure. In the FSB group, bone formation increased between periods (4.3 ± 0.46 to 6.01 ± 0.32), yet with lower volume density when compared to the FSBPBMT (5.6 ± 0.45 to 10.64 ± 0.97) group. It was concluded that the support system formed by the xenograft fibrin sealant associated with the photobiomodulation therapy protocol had a positive effect on the bone repair process.

Histologic Evaluation of Early Bone Regeneration Treated with Simvastatin Associated with Low-Level Laser Therapy.

PURPOSE: This histologic study aimed at assessing bone healing after treatment with simvastatin in association with low-level laser therapy (LLLT). METHODS: Twenty-four male rats (Wistar) were submitted to surgery to create a bone defect of 5 mm in diameter in the parietal bone. These rats were randomly and equally divided into four treatment groups (n = 6): control (C), in which no treatment was performed; simvastatin (SIM), in which rats received daily subcutaneous doses of 2.5 mg/kg of simvastatin; LLLT, which was daily applied to the bone defect; and SIM-LLLT, in which both SIM and LLLT were daily applied. All laser irradiations were carried out with a 830-nm infrared diode laser (GaAlAs) with maximum output of 100 mW and a dose of 4 J, totaling 16 J per session. Rats were euthanized on the 12th postoperative day. Formalin-fixed paraffin-embedded bone samples were obtained and stained with hematoxylin-eosin (HE) and toluidine blue for optical microscope analysis. Degree of inflammation, new vascular formation, tissue repair, and osteoblastic activity were assessed. RESULTS: Categorical analysis of the histologic slides revealed newly formed bone reaching the center of the surgical wound in two animals from the SIM group, two from the LLLT group, and three from the SIM-LLLT group. Greater new bone formation and a lower degree of inflammation were observed in the animals that had bone neoformation at the center of the defect, especially in the LLLT and SIM-LLLT groups. SIM and C groups presented greater angiogenesis than LLLT and SIM-LLLT. SIMLLLT therapy showed a statistically significant reduction in the degree of inflammation when compared to the control group (P < .05). CONCLUSION: Within the limitations of this study, the present results suggest that a combination of simvastatin and low-level laser therapy may stimulate better bone formation.

Biophysical stimulation of bone and cartilage: state of the art and future perspectives.

INTRODUCTION: Biophysical stimulation is a non-invasive therapy used in orthopaedic practice to increase and enhance reparative and anabolic activities of tissue. METHODS: A sistematic web-based search for papers was conducted using the following titles: (1) pulsed electromagnetic field (PEMF), capacitively coupled electrical field (CCEF), low intensity pulsed ultrasound system (LIPUS) and biophysical stimulation; (2) bone cells, bone tissue, fracture, non-union, prosthesis and vertebral fracture; and (3) chondrocyte, synoviocytes, joint chondroprotection, arthroscopy and knee arthroplasty. RESULTS: Pre-clinical studies have shown that the site of interaction of biophysical stimuli is the cell membrane. Its effect on bone tissue is to increase proliferation, synthesis and release of growth factors. On articular cells, it creates a strong A2A and A3 adenosine-agonist effect inducing an anti-inflammatory and chondroprotective result. In treated animals, it has been shown that the mineralisation rate of newly formed bone is almost doubled, the progression of the osteoarthritic cartilage degeneration is inhibited and quality of cartilage is preserved. Biophysical stimulation has been used in the clinical setting to promote the healing of fractures and non-unions. It has been successfully used on joint pathologies for its beneficial effect on improving function in early OA and after knee surgery to limit the inflammation of periarticular tissues. DISCUSSION: The pooled result of the studies in this review revealed the efficacy of biophysical stimulation for bone healing and joint chondroprotection based on proven methodological quality. CONCLUSION: The orthopaedic community has played a central role in the development and understanding of the importance of the physical stimuli. Biophysical stimulation requires care and precision in use if it is to ensure the success expected of it by physicians and patients.

Near-infrared light control of bone regeneration with biodegradable photothermal osteoimplant.

Mild heat stimulation can promote the restoration of bone defects but unfortunately, the delivery of exo-hyperthermy into human body is not efficient enough. In this study, mild heat-induced osteogenesis with high efficacy is demonstrated on an osteoimplant composed of black phosphorus nanosheets and poly(lactic-co-glycolic acid) (BPs@PLGA) with the participation of near-infrared (NIR) light irradiation. BPs@PLGA with only 0.2 wt% BPs show the highly-efficient NIR photothermal response even when being covered by a biological tissue as thick as 7 mm. In addition, this composite is completely biodegradable and the final degradation products are harmless H2O, CO2 and PO43- which can serve as necessary bone ingredient. The BPs@PLGA specimen mediated by low intensity and periodic NIR irradiation can effectively up-regulate the expressions of heat shock proteins and finally promote osteogenesis in vitro and in vivo. Boasting good biodegradability and NIR-mediated osteogenetic performances, the BPs@PLGA implant has great potential in orthopedic applications and this study provides new insights into the design and fabrication of new-style osteoimplants which can be remotely controlled.

Photobiomodulation Therapy in Bone Repair Associated with Bone Morphogenetic Proteins and Guided Bone Regeneration: A Histomorphometric Study.

OBJECTIVE: To evaluate the efficacy of photobiomodulation for bone repair of critical surgical wounds with implants of bone morphogenetic proteins (BMPs) and bovine biological membranes, using histological and histomorphometric analysis. BACKGROUND: Tissue engineering has been developing rapidly through the use of various biomaterials for the treatment of bone defects, such as mechanical barriers consisting of biological membranes and implants of biomaterials for bone supply. MATERIALS AND METHODS: Thirty-two male rats were divided into four groups (n = 8): group I-C: control; group II-PT: photobiomodulation therapy; group III-PM: Gen-Pro® BMPs+Gen-Derm® membrane; and group IV-PMPT: Gen-Pro® BMPs+Gen-Derm® membrane+photobiomodulation therapy. A 3 mm bone cavity was performed in the upper third of the lateral surface of the right rat femur to obtain a bone defect considered to be critical. The irradiated groups received seven applications of AlGaAs diode laser 830 nm, P = 40 mW, continuous wave (CW) emission mode, f ∼ 0.6 mm, 4 J/cm2 per point (north, south, east, and west) at 48 h intervals, for a total of 16 J/cm2 per session (final dose: 112 J/cm2). Bone repair was evaluated at sacrifice 15 and 30 days after the procedure. The specimens were embedded in paraffin and stained with hematoxylin and eosin and Picrosirius for analysis by light microscopy and by the Leica interactive measurement module software. Statistical analysis was performed (p < 0.05%). RESULTS: Histological analysis confirmed the histomorphometric results, with the experimental groups showing bone neoformation of significantly higher quality and quantity at the end of 30 days compared with the control group. CONCLUSIONS: Photobiomodulation therapy was effective for bone repair mainly when associated with BMPs and a biological membrane. The results of this study are promising and stimulate further scientific and clinical research.

Effect of Laser Bio-Stimulation on Mandibular Distraction Osteogenesis: An Experimental Study.

PURPOSE: The aim of this study was to evaluate the osseous response to laser bio-stimulation clinically and histologically during distraction osteogenesis (DO) induced in the mandibles of mongrel dogs. MATERIALS AND METHODS: Thirty dogs were divided into 3 groups of 10 (5 with and 5 without laser treatment) according to sacrifice periods (2, 4, and 8 weeks after distraction). DO was performed between the mandibular second and third premolars using an internal linear distractor. After a 7-day latency period, the distractor was activated at the rate of 1 mm per day for 10 days followed by a consolidation period during which the right mandibular side was irradiated with a diode laser (wavelength [λ], 970 nm; power, 2 W; spot size, 320 µm; total energy [E], 840 J), whereas the control group was not irradiated, after distraction. Histologic specimens were prepared and histomorphometric analysis of specimens was performed. RESULTS: Clinical examinations showed that the low-intensity laser diode had a pronounced effect on the quality and quantity of newly formed bone in the DO regenerate in the laser groups compared with the control groups. Histopathologic sections from laser groups I, II, and III displayed the bio-stimulatory effect of laser on new bone through an increased rate of osteoblast proliferation and differentiation, an accelerated rate of intramembranous ossification, and increased neoangiogenesis compared with the control groups. Moreover, the histomorphometric results showed that mean bone trabecular size, bone trabecular total area, and bony area fraction of the regenerate were larger and statistically significant (P < .05) especially in laser groups I and II (early stages of bone formation) compared with the control groups. CONCLUSION: The low-level diode laser had a positive role as a potential bio-stimulator and local inducer in enhancing bone formation during DO and resulted in early stability of the bone regenerate, a shorter total treatment time, and improved new bone quality and quantity.

Evaluation of photobiomodulation therapy associated with guided bone regeneration in critical size defects. In vivo study.

The repair of bone defects raises the interest of investigators in several health specialties. Grafting techniques with bone substitutes and laser therapies have been investigated to replace autogenous bone and accelerate the bone healing process. Objective To evaluate the effect of photobiomodulation therapy (PBMT) associated with guided bone regeneration (GBR) in critical size defects. Material and Methods The study was conducted on 80 male rats (Rattus norvegicus albinus, Wistar) submitted to surgical creation of a critical size defect on the calvaria, divided into eight study groups: group C (control - only blood clot); group M (collagen membrane); group PBMT (photobiomodulation therapy); group AB (autogenous bone); group AB+PBMT; group AB+M; group PBMT+M; group AB+PBMT+M. The animals were killed 30 days postoperatively. After tissue processing, bone regeneration was evaluated by histomorphometric analysis and statistical analyses were performed (Tukey test, p<0.05). Results All groups had greater area of newly formed bone compared to group C (9.96±4.49%). The group PBMT+M (achieved the greater quantity of new bone (64.09±7.62%), followed by groups PBMT (47.67±8.66%), M (47.43±15.73%), AB+PBMT (39.15±16.72%) and AB+PBMT+M (35.82±7.68%). After group C, the groups AB (25.10±16.59%) and AB+M (22.72±13.83%) had the smallest quantities of newly formed bone. The area of remaining particles did not have statistically significant difference between groups AB+M (14.93±8.92%) and AB+PBMT+M (14.76±6.58%). Conclusion The PBMT utilization may be effective for bone repair, when associated with bone regeneration techniques.

Effect of Low-Level Laser on the Healing of Bone Defects Filled with Autogenous Bone or Bioactive Glass: In Vivo Study.

PURPOSE: This study evaluated the effect of low-level laser therapy (LLLT) on the healing of bone defects filled with autogenous bone or bioactive glass. MATERIALS AND METHODS: A critical size defect with 5-mm diameter was created on the calvaria of 60 adult male rats divided into 6 groups (n = 10): group C (control), group LLLT (LLLT - GaAlAs, wavelength of 780 nm, power of 100 mW, energy density of 210 J/cm2 per point during 60 seconds/point, in five points, only once, after creation of the surgical defect), group AB (autogenous bone), group AB+LLLT (autogenous bone + LLLT), group BG (bioactive glass), group BG+LLLT (bioactive glass + LLLT). All animals were sacrificed at 30 days after surgery. The areas of newly formed bone (ANFB) and areas of remaining particles (ARP) were calculated in relation to the total area (TA). RESULTS: The highest mean ± SD ANFB was observed for group LLLT (47.67% ± 8.66%), followed by groups AB+LLLT (30.98% ± 16.59%) and BG+LLLT (31.13% ± 16.98%). There was a statistically significant difference in relation to ANFB between group C and the other groups, except for comparison with group BG (Tukey test, P > .05). There was no statistically significant difference in ANFB values between group AB and the other study groups (Tukey test, P > .05), group AB+LLLT and groups BG and BG+LLLT (Tukey test, P > .05), and between groups BG and BG+LLLT (Tukey test, P > .05). The highest mean ± SD ARP was found for group BG (25.15% ± 4.82%), followed by group BG+LLLT (17.06% ± 9.01%), and there was no significant difference between groups (t test, P > .05). CONCLUSION: The LLLT, in the present application protocol, did not increase the area of new bone formation when associated with autogenous bone or bioactive glass.

Evaluation of photobiomodulation therapy associated with guided bone regeneration in critical size defects. In vivo study

Abstract The repair of bone defects raises the interest of investigators in several health specialties. Grafting techniques with bone substitutes and laser therapies have been investigated to replace autogenous bone and accelerate the bone healing process. Objective To evaluate the effect of photobiomodulation therapy (PBMT) associated with guided bone regeneration (GBR) in critical size defects. Material and Methods The study was conducted on 80 male rats (Rattus norvegicus albinus, Wistar) submitted to surgical creation of a critical size defect on the calvaria, divided into eight study groups: group C (control - only blood clot); group M (collagen membrane); group PBMT (photobiomodulation therapy); group AB (autogenous bone); group AB+PBMT; group AB+M; group PBMT+M; group AB+PBMT+M. The animals were killed 30 days postoperatively. After tissue processing, bone regeneration was evaluated by histomorphometric analysis and statistical analyses were performed (Tukey test, p<0.05). Results All groups had greater area of newly formed bone compared to group C (9.96±4.49%). The group PBMT+M (achieved the greater quantity of new bone (64.09±7.62%), followed by groups PBMT (47.67±8.66%), M (47.43±15.73%), AB+PBMT (39.15±16.72%) and AB+PBMT+M (35.82±7.68%). After group C, the groups AB (25.10±16.59%) and AB+M (22.72±13.83%) had the smallest quantities of newly formed bone. The area of remaining particles did not have statistically significant difference between groups AB+M (14.93±8.92%) and AB+PBMT+M (14.76±6.58%). Conclusion The PBMT utilization may be effective for bone repair, when associated with bone regeneration techniques.

Effects of Electrostatic Field on Osteoblast Cells for Bone Regeneration Applications.

Many external stimulations have been shown to promote bone regeneration. The effects of an alternating current (AC) electrostatic field, one of external stimulations, generated from a device with high voltage and low current output on human osteoblastic cell line have been investigated in this study. We investigated how human osteoblasts responded to an AC electrostatic field, and the output parameters were set as 1 kV and 160 µA. Our results showed that, under such condition, the AC electrostatic field had a downregulation effect on the production ability of alkaline phosphatase and type 1 collagen expression. However, the expression of osteocalcin gene was elevated on the end of EFID treatment suggesting that AC electrostatic field might be a potential stimulation for accelerating the differentiation of osteoblastic cells.

Low-level laser therapy (780 nm) combined with collagen sponge scaffold promotes repair of rat cranial critical-size defects and increases TGF-ß, FGF-2, OPG/RANK and osteocalcin expression.

The aim of this study was to evaluate the effect of collagen sponge scaffold (CSS) implantation associated with low-level laser therapy (LLLT) on repairing bone defects. A single 5-mm cranial defect was surgically created in forty Wistar rats, which then received one of the following four interventions (n = 10 per group): no treatment (G0); bone defect implanted with collagen sponge scaffold (CSS) alone (G1); defect treated with low-level laser therapy (LLLT) (wavelength 780 nm; total energy density 120 J/cm2 ; power 50 mW) alone (G2); and CSS associated with LLLT treatment (G3). After surgery, animals in each group were euthanized at 21 days and 30 days (n = 5 per euthanasia time group). Bone formation was monitored by X-ray imaging analysis. Biopsies were collected and processed for histological analysis and immunohistochemical evaluation of transforming growth factor-beta (TGF-ß), fibroblast growth factor-2 (FGF-2), osteoprotegerin (OPG) and receptor activator of nuclear factor ƙ (RANK). Osteocalcin (OCN) was detected by immunofluorescence analysis. Compared to the G0 group, defects in the 30-day G3 group exhibited increased bone formation, both by increase in radiopaque areas (P < 0.01) and by histomorphometric analysis (P < 0.001). The histopathological analysis showed a decreased number of inflammatory cells (P < 0.001). The combined CCS + LLLT (G3) treatment also resulted in the most intense immunostaining for OPG, RANK, FGF-2 and TGF-ß, and the most intense and diffuse OCN immunofluorescent labelling at 30 days postsurgery (G3 vs. G0 group, P < 0.05). Therefore, the use of CCS associated with LLLT could offer a synergistic advantage in improving the healing of bone fractures.

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.