Influence of zoledronic acid and low-intensity laser on collagen fibers during the bone repair process.

PURPOSE: To evaluate collagen fibers during the bone repair process in critical defects created in the tibias of rats, treated with zoledronic acid (AZ) associated with low-level laser therapy (LLLT). METHODS: Ten rats were distributed according to treatment: group 1) saline solution; group 2) LLLT; group 3) AZ; group 4) AZ and LLLT. AZ was administered at the dose of 0.035 mg/kg at fortnightly intervals over eight weeks. Next, 2-mm bone defects were created in the tibias of all animals. The bone defects in groups 2 and 4 were irradiated LLLT in the immediate postoperative period. After periods 14 and 28 of application, the animals were euthanized, and birefringence analysis was performed. RESULTS: Approximately 90% of the total area was occupied by collagen fibers within the red color spectrum, this area being statistically larger in relation to the area occupied by collagen fibers within the green and yellow spectrum, in the four groups. Over the 14-day period, there was no statistically significant difference between the groups. In the 28-day period, group 2 (14.02 ± 15.9%) was superior in quantifying green birefringent fibers compared to group 1 (3.06 ± 3.24%), with p = 0.009. CONCLUSIONS: LLLT associated with ZA is effective in stimulating the neoformation of collagen fibers. The LLLT group without the association with ZA showed a greater amount of immature and less organized matrix over a period of 28 days.

Effects of green light emitting diode light during incubation and dietary organic macro and trace minerals during rearing on tibia characteristics of broiler chickens at slaughter age.

This study was designed to evaluate the effects of green light emitting diode (LED) light during incubation and dietary organic macro and trace minerals during rearing on tibia morphological, biophysical, and mechanical characteristics of broiler chickens at slaughter age. The experiment was setup as a 2 × 2 × 2 factorial arrangement, with the following treatments: 1) light during incubation (green LED light or darkness), 2) macro mineral source during rearing (organic or inorganic Ca and P), and 3) trace mineral source during rearing (organic or inorganic Fe, Cu, Mn, Zn, and Se). A total of 2,400 eggs (Ross 308) were either incubated under green LED light (16L:8D) or in complete darkness. After hatch, a total of 864 male broiler chickens were reared until slaughter age (day 42) and provided with 1 of 4 diets, differing in macro and/or trace mineral source. During rearing, the experiment had a complete randomized block design with 9 replicate pens per treatment and 12 chickens per pen. At slaughter age (day 42), 2 chickens per replicate were randomly selected and tibia bones were obtained. Tibia weight, length, thickness, osseous volume, pore volume, total volume, mineral content, mineral density, ultimate strength, and stiffness were determined. Green LED light during incubation did not affect any of the tibia characteristics. Dietary organic macro minerals positively affected most of the tibia morphological, biophysical, and mechanical characteristics compared to the inorganic macro minerals, whereas trace mineral sources did not affect tibia characteristics. It can be concluded that dietary organic macro minerals Ca and P stimulated tibia characteristics, whereas green LED light during incubation and dietary trace minerals during rearing did not affect tibia characteristics, locomotion, or leg disorders.

Altered mechanical behavior of demineralized bone following therapeutic radiation.

Post-radiotherapy (RTx) bone fragility fractures are a late-onset complication occurring in bone within or underlying the radiation field. These fractures are difficult to predict, as patients do not present with local osteopenia. Using a murine hindlimb RTx model, we previously documented decreased mineralized bone strength and fracture toughness, but alterations in material properties of the organic bone matrix are largely unknown. In this study, 4 days of fractionated hindlimb irradiation (4 × 5 Gy) or Sham irradiation was administered in a mouse model (BALB/cJ, end points: 0, 4, 8, and 12 weeks, n = 15/group/end point). Following demineralization, the viscoelastic stress relaxation, and monotonic tensile mechanical properties of tibiae were determined. Irradiated tibiae demonstrated an immediate (day after last radiation fraction) and sustained (4, 8, 12 weeks) increase in stress relaxation compared to the Sham group, with a 4.4% decrease in equilibrium stress (p < .017). While tensile strength was not different between groups, irradiated tibiae had a lower elastic modulus (-5%, p = .027) and energy to failure (-12.2%, p = .012) with monotonic loading. Gel electrophoresis showed that therapeutic irradiation (4 × 5 Gy) does not result in collagen fragmentation, while irradiation at a common sterilization dose (25 kGy) extensively fragmented collagen. These results suggest that altered collagen mechanical behavior has a role in postirradiation bone fragility, but this can occur without detectable collagen fragmentation. Statement of Clinical Significance: Therapeutic irradiation alters bone organic matrix mechanics and which contribute to diminished fatigue strength, but this does not occur via collagen fragmentation.

An agent-based model of prostate Cancer bone metastasis progression and response to Radium223.

BACKGROUND: Bone metastasis is the most frequent complication in prostate cancer patients and associated outcome remains fatal. Radium223 (Rad223), a bone targeting radioisotope improves overall survival in patients (3.6 months vs. placebo). However, clinical response is often followed by relapse and disease progression, and associated mechanisms of efficacy and resistance are poorly understood. Research efforts to overcome this gap require a substantial investment of time and resources. Computational models, integrated with experimental data, can overcome this limitation and drive research in a more effective fashion. METHODS: Accordingly, we developed a predictive agent-based model of prostate cancer bone metastasis progression and response to Rad223 as an agile platform to maximize its efficacy. The driving coefficients were calibrated on ad hoc experimental observations retrieved from intravital microscopy and the outcome further validated, in vivo. RESULTS: In this work we offered a detailed description of our data-integrated computational infrastructure, tested its accuracy and robustness, quantified the uncertainty of its driving coefficients, and showed the role of tumor size and distance from bone on Rad223 efficacy. In silico tumor growth, which is strongly driven by its mitotic character as identified by sensitivity analysis, matched in vivo trend with 98.3% confidence. Tumor size determined efficacy of Rad223, with larger lesions insensitive to therapy, while medium- and micro-sized tumors displayed up to 5.02 and 152.28-fold size decrease compared to control-treated tumors, respectively. Eradication events occurred in 65 ± 2% of cases in micro-tumors only. In addition, Rad223 lost any therapeutic effect, also on micro-tumors, for distances bigger than 400 µm from the bone interface. CONCLUSIONS: This model has the potential to be further developed to test additional bone targeting agents such as other radiopharmaceuticals or bisphosphonates.

The effect of hyperbaric oxygen therapy on bone macroscopy, composition and biomechanical properties after ionizing radiation injury.

BACKGROUND: Radiotherapy used in tumor treatment compromises vascularization of bone tissue. Hyperbaric oxygenation (HBO) increases oxygen availability and improves vascularization, minimizing the deleterious effects of ionizing radiation (IR). Therefore, the aim of this study was to evaluate HBO therapy effect on bone macroscopy, composition and biomechanical properties after IR damage. METHODS: Twenty male Wistar rats weighing 300 ± 20 g (10 weeks of age) were submitted to IR (30 Gy) to the left leg, where the right leg was not irradiated. After 30 days, ten animals were submitted to HBO therapy, which was performed daily for 1 week at 250 kPa for 90-min sessions. All animals were euthanized 37 days after irradiation and the tibia were separated into four groups (n = 10): from animals without HBO - right tibia Non-irradiated (noIRnoHBO) and left tibia Irradiated (IRnoHBO); and from animals with HBO - right tibiae Non-irradiated (noIRHBO) and left tibia Irradiated (IRHBO). The length (proximal-distal) and thickness (anteroposterior and mediolateral) of the tibiae were measured. Biomechanical analysis evaluated flexural strength and stiffness. Attenuated Total Reflectance Fourier Transform Infrared Spectroscopy (ATR-FTIR) was used to calculate the amide I ratio, crystallinity index, and matrix to mineral ratios. RESULTS: In the macroscopic and ATR-FTIR analysis, the IRnoHBO showed lower values of length, thickness and amide I ratio, crystallinity index and matrix to mineral ratios compared to noIRnoHBO (p < 0.03). IRnoHBO showed no statistical difference compared to IRHBO for these analyses (p > 0.05). Biomechanics analysis showed that the IRnoHBO group had lower values of flexural strength and stiffness compared to noIRnoHBO and IRHBO groups (p < 0.04). In addition, the noIRHBO group showed higher value of flexural strength when compared to noIRnoHBO and IRHBO groups (p < 0.02). CONCLUSIONS: The present study concluded that IR arrests bone development, decreases the collagen maturation and mineral deposition process, thus reducing the flexural strength and stiffness bone mechanical parameters. Moreover, HBO therapy minimizes deleterious effects of irradiation on flexural strength and the bone stiffness analysis.

Reproducibility and Radiation Effect of High-Resolution In Vivo Micro Computed Tomography Imaging of the Mouse Lumbar Vertebra and Long Bone.

A moderate radiation dose, in vivo µCT scanning protocol was developed and validated for long-term monitoring of multiple skeletal sites (femur, tibia, vertebra) in mice. A customized, 3D printed mouse holder was designed and utilized to minimize error associated with animal repositioning, resulting in good to excellent reproducibility in most cortical and trabecular bone microarchitecture and density parameters except for connectivity density. Repeated in vivo µCT scans of mice were performed at the right distal femur and the 4th lumbar vertebra every 3 weeks until euthanized at 9 weeks after the baseline scan. Comparing to the non-radiated counterparts, no radiation effect was found on trabecular bone volume fraction, osteoblast and osteoblast number/surface, or bone formation rate at any skeletal site. However, trabecular number, thickness, and separation, and structure model index were sensitive to ionizing radiation associated with the µCT scans, resulting in subtle but significant changes over multiple scans. Although the extent of radiation damage on most trabecular bone microarchitecture measures are comparable or far less than the age-related changes during the monitoring period, additional considerations need to be taken to minimize the confounding radiation factors when designing experiments using in vivo µCT imaging for long-term monitoring of mouse bone.

A Mouse Model for Skeletal Structure and Function Changes Caused by Radiation Therapy and Estrogen Deficiency.

Radiation therapy and estrogen deficiency can damage healthy bone and lead to an increased fracture risk. The goal of this study is to develop a mouse model for radiation therapy using a fractionated biologically equivalent dose for cervical cancer treatment in both pre- and postmenopausal women. Thirty-two female C57BL/6 mice 13 weeks of age were divided into four groups: Sham + non-irradiated (SHAM + NR), Sham + irradiated (SHAM + IRR), ovariectomy + non-irradiated (OVX + NR) and ovariectomy + irradiated (OVX + IRR). The irradiated mice received a 6 Gy dose of X-rays to the hindlimbs at Day 2, Day 4 and Day 7 (18 Gy total). Tissues were collected at Day 35. DEXA, microCT analysis and FEA were used to quantify structural and functional changes at the proximal tibia, midshaft femur, proximal femur and L1 vertebra. There was a significant (p < 0.05) decline in proximal tibia trabecular BV/TV from (1) IRR compared to NR mice within Sham (- 46%) and OVX (- 41%); (2) OVX versus Sham within NR mice (- 36%) and IRR mice (- 30%). With homogenous material properties applied to the proximal tibia mesh using FEA, there was (1) an increase in whole bone (trabecular + cortical) structural stiffness from IRR compared to NR mice within Sham (+ 10%) and OVX (+ 15%); (2) a decrease in stiffness from OVX versus Sham within NR mice (- 18%) and IRR mice (- 14%). Fractionated irradiation and ovariectomy both had a negative effect on skeletal microarchitecture. Ovariectomy had a systemic effect, while skeletal radiation damage was largely specific to trabecular bone within the X-ray field.

Thermal damage of osteocytes during pig bone drilling: an in vivo comparative study of currently available and modified drills.

OBJECTIVES: The Gekkou-drill® is an industrial drill that is highly efficient due to reduced cutting resistance resulting from its characteristic drill point shape. In this experiment, we compared the degree of thermal damage to bone tissue caused by conventional medical drills and these same drills with Gekkou modifications. METHODS: Holes were created in the tibias of living pigs using two different 3.2-mm diameter drills and their modified versions. Regarding the drilling parameters, the thrust force was 10 N and the drilling speeds were 800 revolutions per minute (rpm) and 1500 rpm. We compared the original and modified drills in terms of the bone temperature around the drill bit and the total time necessary to create each hole, the latter calculated using imaging data captured during drilling. In histopathological examination, the percentages of empty lacunae in osteocytes of the cortical bone beneath the periosteum were evaluated at 400 × magnification with an optical microscope. RESULTS: Compared to the original drills, the modified drills required significantly less time to create each hole and caused a significantly lower temperature rise during bone drilling. With the modified drills, the percentages of empty lacunae around the drilling holes were about 1/2-1/3 of those with the original drills, and were significantly lower for both drilling speeds. CONCLUSIONS: Gekkou-modified medical drills shortened drilling times despite low thrust force, and histopathological assessment demonstrated a significant reduction in osteocyte damage.

Effect of low-level laser therapy and zoledronic acid on bone repair process.

This study aimed to evaluate, through histomorphometric analysis, the bone repair process in the tibia of rats treated with zoledronic acid and submitted to 808-nm low-level laser therapy (LLLT) by using arsenide aluminum gallium laser. For this purpose, 20 rats were used and distributed according to treatment: group 1-saline administration; group 2-treated with LLLT; group 3-treated with zoledronic acid; and group 4-treated with zoledronic acid and LLLT. The zoledronic acid was administered at a dose of 0.035 mg/kg every 2 weeks for 8 weeks. Subsequently, bone defects of 2 mm were prepared in the tibias of all groups. The bone defects in groups 2 and 4 were irradiated with LLLT in the immediate post-operative period. After 14 and 28 days of application, the animals were submitted and euthanized for histomorphometric analysis. The results were submitted to statistical analysis (α = 5%), and the intragroup comparison was performed using the t test. On the other hand, for intergroup comparison, the ANOVA test was performed, and to the groups presenting statistically significant difference, the Student-Newman-Keuls test was used. In intergroup comparison, group 1 (mean ± SD= 45.2 ± 18.56%) showed a lower bone formation compared with groups 2 (64.13 ± 3.51%) (p = 0.358) and 4 (15.2 ± 78.22%) (p = 0.049), at the 14-day period. Group 3 (20.99 ± 7.42%) also presented a lower amount of neoformed bone tissue, with statistically significant difference when compared with groups 1 (p = 0.002), 2, and 4 (p ≤ 0,001). After 28 days, group 1 presented a lower amount of neoformed bone tissue compared with the other groups, with p = 0.020. Thus, it was concluded that LLLT associated with zoledronic acid is effective for stimulating bone formation in surgically created defects in rats, at the periods studied.

The risks following the exposure to radiation associated with the surgical correction of limb deformities in children are minimal.

AIMS: The aim of this study was to quantify the risk of developing cancer from the exposure to radiation associated with surgery to correct limb deformities in children. PATIENTS AND METHODS: A total of 35 children were studied. There were 19 girls and 16 boys. Their mean age was 11.9 years (2 to 18) at the time of surgery. Details of the radiological examinations were recorded during gradual correction using a Taylor Spatial Frame. The dose area product for each radiograph was obtained from the Computerised Radiology Information System database. The effective dose in millisieverts (mSv) was calculated using conversion coefficients for the anatomical area. The lifetime risk of developing cancer was calculated using government-approved Health Protection Agency reports, accounting for the age and gender of the child. RESULTS: Correction was undertaken in five femurs, 18 tibiae, and 12 feet. The median duration of treatment was 45 months (11 to 118). The mean effective dose was 0.31 mSv (0.05 to 0.64) for the femur, 0.29 mSv (0.01 to 0.97) for the tibia, and 0.027 mSv (0.001 to 0.161) for the foot. The cumulative exposure gave 'negligible' risk in 26 children and 'minimal' risk in nine children, according to Public Health England categories. These results are below the mean annual background radiation in the United Kingdom. CONCLUSION: The lifetime attributable risk of developing cancer from repeated exposure to radiation was negligible or minimal in all children. This is the first study to quantify the exposure to radiation from serial radiographs in children with limb deformities who are treated surgically using circular external fixation, linking this to the risk of developing cancer. Cite this article: Bone Joint J 2019;101-B:241-245.

Cell Phone Radiation Effect on Bone-to-Implant Osseointegration: A Preliminary Histologic Evaluation in Rabbits.

BACKGROUND AND PURPOSE: The increased use of cell phones has raised many questions as to whether their use is safe for patients with dental implants. This study aimed to assess the consequences of cell phone-emitted radiation on bone-to-implant osseointegration during the healing phase. MATERIALS AND METHODS: Twelve rabbits were grouped into three groups of four. Group 1 (control) was not exposed to electromagnetic radiation; group 2 (test) was exposed for 8 hours/day in speech mode and 16 hours/day in standby mode; and group 3 (test) was exposed for 24 hours continuously in standby mode for 3 months. Forty-eight implants were placed in tibia and femur bone of rabbits, and after 90 days the rabbits were sacrificed and bone surrounding the implant was retrieved. Histopathologic evaluations of the specimens were done using transmitted light microscope. The differences among the three groups were statistically analyzed with analysis of variance (ANOVA) and pairwise comparisons via Fisher's exact test. RESULTS: Significantly less bone-to-implant contact and bone area surrounding implant threads were found in the test groups compared to the control group. There was a significant difference in regular bone formation (P < .001) among the three groups. CONCLUSION: Implants exposed to cell phone radiation showed more inflammatory reaction when compared to the nonexposed implants, thus indicating that cellular phone overuse could affect the maturation of bone and thus delay osseointegration.

Effects of low-level laser therapy on the organization of articular cartilage in an experimental microcrystalline arthritis model.

The aim of this study was to evaluate the effects of low-level laser therapy using the gallium arsenide laser (λ = 830 nm) on the articular cartilage (AC) organization from knee joint in an experimental model of microcrystalline arthritis in adult male Wistar rats. Seventy-two animals were divided into three groups: A (control), B (induced arthritis), and C (induced arthritis + laser therapy). The arthritis was induced in the right knee using 2 mg of Na4P2O7 in 0.5 mL of saline solution. The treatments were daily applied in the patellar region of the right knee after 48 h of induction. On the 7th, 14th, and 21st days of treatment, the animals were euthanized and their right knees were removed and processed for structural and biochemical analysis of the AC. The chondrocytes positively labeled for the TUNEL reaction were lower in C than in B on the 14th and 21st days. The content of glycosaminoglycans and hydroxyproline in A and C was higher than B on the 21st day. The amount of tibial TNF-α in B and C was lower than in A. The amount of tibial BMP-7 in B and C was higher than in A. The femoral MMP-13 was lower in B and C than for A. The tibial TGF-ß for C was higher than the others. The femoral ADAMT-S4 content of A and C presented similar and inferior data to B on the 21st day. The AsGa-830 nm therapy preserved the content of glycosaminoglycans, reduced the cellular changes and the inflammatory process compared to the untreated group.

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.

Endothelial progenitor cells improve the therapeutic effect of mesenchymal stem cell sheets on irradiated bone defect repair in a rat model.

BACKGROUND: The reconstruction of bone defects is often impaired by radiotherapy since bone quality is compromised by radiation. This study aims to investigate the therapeutic efficacy of the composite cell sheets-bone marrow mesenchymal stem cell (BMSC) sheets cocultured with endothelial progenitor cells (EPCs)-in the healing of irradiated bone defects and the biological effects of EPCs on the osteogenic properties of BMSC sheets. METHODS: BMSCs and EPCs were isolated from rat bone marrow. BMSCs were used to form cell sheets by the vitamin C inducing method. EPCs were seeded on BMSC sheets to make EPCs-BMSC sheets. Osteogenesis of EPCs-BMSC sheets and BMSC sheets were tested. In vitro osteogenesis tests included ALP, Alizarin Red S, Sirius Red staining, qRT-PCR and Western blot analysis after 3 and 7 days of osteogenic incubation. Subcutaneous osteogenesis was tested by H&E staining and immunohistochemical staining 8 weeks after transplantation. EPCs-BMSC sheets and BMSC sheets were used in the 3 mm defects of non-irradiated and irradiated rat tibias. Micro-CT and histological analysis were used to test the healing of bone defects 4 and 8 weeks after transplantation. RESULTS: EPCs-BMSC sheets showed enhanced osteogenic differentiation in vitro with increased expression of osteoblastic markers and osteogenesis related staining compared with BMSC sheets. In subcutaneous osteogenesis test, EPCs-BMSC sheets formed larger areas of new bone and blood vessels. The EPCs-BMSC group had the highest volume of newly formed bone in the defect area of irradiated tibias. CONCLUSIONS: EPCs improved the osteogenic differentiation of BMSC Sheets and enhanced the ectopic bone formation. EPCs-BMSC sheets promoted bone healing in irradiated rat tibias. EPCs-BMSC sheets are potentially useful in the reconstruction of bone defect after radiotherapy.

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.

[Extracorporeal irradiation : Reimplantation of bone segments in the treatment of malignant bone tumours]. / Die extrakorporale Bestrahlung : Replantation von Knochensegmenten bei malignen Knochentumoren.

BACKGROUND: Malignant bone tumors themselves and the wide resection required because of them may cause huge bone defects in the bone segment involved. Autologous bone grafts are a reliable option to cover these defects in many cases but their availability is limited. Besides common alternative reconstruction methods, including the use of allografts and/or prostheses, especially extracoroporeal irradiation (ECI) and reimplantation of the bone segment involved is attracting increasingly more attention nowadays. DISCUSSION: In the following, we report on indications/contraindications, details of the operative technique, as well as the recommended rehabilitation regime of ECI. Furthermore, we compare our own results with those published in the recent literature. Especially the advantages and disadvantages of this method, the risks and the complications are illustrated and critically discussed. CONCLUSION: Extracorporeal irradiation of a tumor bearing bone segment is a valuable alternative reconstruction technique following tumor resections of the pelvis, femur and tibia, with encouraging results with respect to local control, complication risks and functional outcome.

The effects of low-level laser therapy on the healing of bone defects in streptozotocin-induced diabetic rats: A histological and morphometric evaluation.

BACKGROUND: The aim of the present study was to evaluate the effects of low-level laser therapy (LLLT) on the healing of bone defects in rats with streptozotocin (STZ)-induced DM. METHODS: 28 male Sprague-Dawley rats were used in this study. 14 animals received a single dose of STZ intraperitoneally (65 mg/kg) to induce Type I DM, whereas others were injected only with sterile saline solution. Four weeks later, standard bone defects were created in the tibiae of rats. Surgical wounds in one group from each of the diabetic and non-diabetic animals were irradiated with diode laser for every other day for 4 weeks and they were described as DM + LLLT and CONT + LLLT groups, respectively. Remaining two groups received no laser treatment. New bone formation, osteoblast and blood vessel counts were calculated in histologic sections. RESULTS: DM group had significantly smaller bone area and lower blood vessel count when compared to DM + LLLT, CONT and CONT + LLLT groups (p < 0.05 for each). CONT and CONT + LLLT groups had significantly larger bone area than DM + LLLT group (p < 0.05 for both). CONCLUSIONS: LLLT application promoted vascularization and new bone formation in animals with DM to a limited extent, since it was unable to support the healing process up to the level of non-diabetic animals.

Effect of low-intensity whole-body vibration on bone defect repair and associated vascularization in mice.

Low-intensity whole-body vibration (LIWBV) may stimulate bone healing, but the involvement of vascular ingrowth, which is essential for bone regeneration, has not been well examined. We thus investigated the LIWBV effect on vascularization during early-stage bone healing. Mice aged 13 weeks were subjected to cortical drilling on tibial bone. Two days after surgery (day 0), mice were exposed daily to sine-wave LIWBV at 30 Hz and 0.1 g peak-to-peak acceleration for 20 min/day (Vib) or were sham-treated (sham). Following vascular casting with a zirconium-based contrast agent on days 6, 9, or 12 and sacrifice, vascular and bone images were obtained by K-edge subtraction micro-CT using synchrotron lights. Bone regeneration advanced more in the Vib group from days 9 to 12. The vascular volume fraction decreased from days 6 to 9 in both groups; however, from days 9 to 12, it was increased in shams, while it stabilized in the Vib group. The vascular volume fraction tended to be or was smaller in the Vib group on days 6 and 12. The vessel number density was higher on day 9 but lower on day 12 in the Vib group. These results suggest that the LIWBV-promoted bone repair is associated with the modulation of vascularization, but additional studies are needed to determine the causality of this association.

Changes of articular cartilage and subchondral bone after extracorporeal shockwave therapy in osteoarthritis of the knee.

We assessed the pathological changes of articular cartilage and subchondral bone on different locations of the knee after extracorporeal shockwave therapy (ESWT) in early osteoarthritis (OA). Rat knees under OA model by anterior cruciate ligament transaction (ACLT) and medial meniscectomy (MM) to induce OA changes. Among ESWT groups, ESWT were applied to medial (M) femur (F) and tibia (T) condyles was better than medial tibia condyle, medial femur condyle as well as medial and lateral (L) tibia condyles in gross osteoarthritic areas (p<0.05), osteophyte formation and subchondral sclerotic bone (p<0.05). Using sectional cartilage area, modified Mankin scoring system as well as thickness of calcified and un-calcified cartilage analysis, the results showed that articular cartilage damage was ameliorated and T+F(M) group had the most protection as compared with other locations (p<0.05). Detectable cartilage surface damage and proteoglycan loss were measured and T+F(M) group showed the smallest lesion score among other groups (p<0.05). Micro-CT revealed significantly improved in subchondral bone repair in all ESWT groups compared to OA group (p<0.05). There were no significantly differences in bone remodeling after ESWT groups except F(M) group. In the immunohistochemical analysis, T+F(M) group significant reduced TUNEL activity, promoted cartilage proliferation by observation of PCNA marker and reduced vascular invasion through observation of CD31 marker for angiogenesis compared to OA group (P<0.001). Overall the data suggested that the order of the effective site of ESWT was T+F(M) ≧ T(M) > T(M+L) > F(M) in OA rat knees.

Apoptosis in bone defect of diabetic rats treated with low intensity laser: radiological and immunohistochemical approach / Apoptosis en ratones diabéticos con defecto oseo tratados con láser de baja intensidad: evaluación radiológica e inmunohistoquímica

The aim of this study is evaluate the efficacy of 904 nm laser diode in bone regeneration in the bone defect in diabetic rats. Six groups of 10 male Wistar rats and 2 mm bone defects drilled on the left and right tibia were used. The diabetic animals were treated with streptozotocin (40 mg/kg, i.v.). We compared the diode laser doses of treatment of bone defects 50 w ­ 4 J/cm and 100 w ­ 4J/. The right tibia was used for immunohistochemical analysis with the apoptosis markers XIAP and Caspase-3 and the left tibia was submitted to computer tomography (CT). Caspase-3 marker showed greater amount of apoptosis in all the untreated groups compared to both laser treatments. There was no statistical significance for XIAP marker. CT scan showed improvement of bone defect area and volume in both laser treated groups, control and diabetic. Therefore the low intensity laser therapy was effective in accelerating bone repair in both, control and diabetic groups. It was evidenced in our study that diabetes influences bone repair negatively.

Los objetivos de este estudio fueron evaluar la eficacia del láser diodo de 904 nm en la regeneración ósea del defecto óseo en ratas diabéticas. Se utilizaron seis grupos de 10 ratas Wistar macho y se generó un defecto óseo de 2 mm en las tibias izquierda y derecha de los animales. El animal diabético fue generado con estreptozotocina (40 mg / kg, i.v.). Se compararon las dosis de tratamiento de los defectos óseos con láser de diodo de 50 w - 4 J / cm y 100w - 4 J /. La tibia derecha fue utilizada para el análisis inmunohistoquímico con los marcadores de apoptosis XIAP y Caspasa-3 y la tibia izquierda fue sometida a tomografía computarizada. El marcador caspasa-3 mostró mayor cantidad de apoptosis en todos los grupos no tratados en comparación con ambos tratamientos con láser. No hubo significación estadística para el marcador XIAP. La tomografía computarizada mostró una mejoría del área y el volumen de los defectos óseos en ambos grupos tratados con láser, control y diabéticos. Por lo tanto, la terapia con láser de baja intensidad fue eficaz en la aceleración de la reparación ósea tanto en los grupos control como en los diabéticos. Se evidenció en nuestro estudio que la diabetes afecta negativamente la reparación ósea.