Bone Repair In Rats Research Articles

Biomimetic composite hydrogel promotes new bone formation in rat bone defects through regulation of miR-19b-3p/WWP1 axis by loaded extracellular vesicles

ObjectiveThis study aims to investigate the mechanism by which biomimetic composite hydrogels loaded with bone marrow mesenchymal stem cells (BMSCs) derived microRNA-19b-3p/WWP1 axis through extracellular vesicles (EVs) affect the new bone formation in rat bone defects.MethodsFirst, synthesize the bionic composite hydrogel Gel-OCS/MBGN. Characterize it through field-emission scanning electron microscopy (FE-SEM), X-ray diffraction (XRD), and FTIR. Then, conduct performance tests such as rheology, dynamic mechanical analysis, in vitro mineralization, and degradation. Rat BMSCs were selected for in vitro cell experiments, and EVs derived from BMSCs were obtained by differential centrifugation. The EVs were loaded onto Gel-OCS/MBGN to obtain Gel-OCS/MBGN@EVs hydrogel. Cell viability and proliferation were detected by live/dead cell staining and CCK-8 assay, respectively. ALP and ARS staining was used to evaluate the osteogenic differentiation of BMSCs. Differential gene expression analysis of osteogenic differentiation was performed using high-throughput sequencing. TargetScan database predicted the binding site between miR-19b-3p and WWP1, and a dual-luciferase reporter assay was performed to confirm the targeting binding site. A rat bone defect model was established, and new bone formation was evaluated by Micro-CT, H&E staining, and Masson's trichrome staining. Immunofluorescence staining and immunohistochemistry were used to detect the expression levels of osteogenic-related factors in rat BMSCs. RT-qPCR and Western blot were used to detect the expression levels of genes and proteins in tissues and cells.ResultGel-OCS/MBGN was successfully constructed and loaded with EVs, resulting in Gel-OCS/MBGN@EVs. The in vitro drug release experiment results show that Gel-OCS/MBGN could sustainably release EVs. Further experiments have shown that Gel-OCS/MBGN@EVs could significantly promote the differentiation of BMSCs into osteoblasts. Experiments have shown that WWP1 is a key factor in osteogenic differentiation and is regulated by miR-19b-3p. EVs promote osteogenic differentiation by suppressing WWP1 expression through the transmission of miR-19b-3p. In vivo animal experiments have demonstrated that Gel-OCS/MBGN@EVs significantly promote bone repair in rats with bone defects by regulating the miR-19b-3p/WWP1 signaling axis.ConclusionFunctional Gel-OCS/MBGN@EVs were obtained by constructing Gel-OCS/MBGN and loading EVs onto it. EVs could deliver miR-19b-3p to BMSCs, inhibit the expression of WWP1, and promote the osteogenic differentiation of BMSCs, ultimately promoting bone regeneration in rats with bone defects.Graphical

Single intraoperative infrared laser optimized bone repair in rat femoral osteotomies with experimentally induced osteoporosis.

This study aimed to evaluate the effect of infrared laser (IRL) on bone repair in ovariectomized rats subjected to femoral osteotomies. Of 32 rats, half underwent bilateral ovariectomy (OVX) and the other half underwent sham ovariectomy (SHAM). A period of 3months was defined to observe the presence of osteoporosis. The rats were subjected to osteotomies in the femurs and then fixed with a miniplate and 1.5-mm system screws. Thereafter, half of the rats from both SHAM and OVX groups were not irradiated, and the other half were irradiated by IRL using the following parameters: wavelength, 808nm; power, 100 mW; 60s for each point; 6J/point; and a total of 5 points of bone gap. All animals were euthanized 60days after surgery. The femur gap was scanned using micro-computed tomography (micro-CT). The samples were then examined under a confocal laser microscope to determine the amounts of calcein and alizarin red. The slides were stained with alizarin red and Stevenel's blue for histometric analysis. In the micro-CT analysis, the OVX groups had the lowest bone volume (P < 0.05). When the laser was applied to the OVX groups, bone turnover increased (P < 0.05). New bone formation (NBF) was comparable between SHAM and OVX/IR (P > 0.05) groups; however, it was less in the OVX groups (P < 0.05). In conclusion, the results encourage the use of IRL intraoperatively as it optimizes bone repair, mainly in animals with low bone mineral density.

Effect of Energy Level of Photobiomodulation Therapy on Bone Repair in Rats

The aim of this experimental study was to determine the effect of photobiomodulation therapy on bone repair in a rat tibia osteotomy model at 15 and 30 days. The sample consisted of 36 male Holtzman rats that were randomized into 6 equal groups. Groups A1 and A2: osteotomy + 1 J laser energy. Groups B1 and B2: osteotomy + 3 J laser energy. Groups C1 and C2 (controls): osteotomy only. The bone repair was analyzed by histological evaluation of osteoblasts and osteocytes both at 15 days (groups A1, B1, and C1) and at 30 days (groups A2, B2, and C2). Within the results, in all groups a greater number of osteoblasts was found at 15 days vs 30 days (p&lt;0.05), and a greater number of osteocytes in B1 and C2 vs B2 and C1, respectively (p&lt;0.05). When evaluating the 3 groups worked up to 15 days, more osteoblasts were found in A1 and C1 vs B1 (p&lt;0.001); and osteocytes predominated in A1 and B1 vs C1 (p&lt;0.001). At 30 days there was a greater quantity of osteoblasts in C2 vs A2 and B2 (p&lt;0.05) and of osteocytes in C2 vs B2 (p&lt;0.05). It is concluded that 1 J photobiomodulation therapy improved bone repair at 15 days; however, this improvement was not observed at 30 days because there were no differences between the irradiated groups and the control.

Does resveratrol favor peri-implant bone repair in rats with ovariectomy-induced osteoporosis? Gene expression, counter-torque and micro-CT analysis

This study investigated the influence of resveratrol on peri-implant repair and its effects on bone-related markers in ovariectomy-induced osteoporosis in rats. Animals were divided into: OVX+PLAC (n = 10): ovariectomized animals treated with placebo; OVX+RESV (n = 10): OVX treated with resveratrol; OVX+PLAC+ZOL (n = 10): OVX treated with PLAC and zoledronate; OVX+RESV+ZOL (n = 10): OVX treated with RESV and ZOL; and SHOVX+PLAC (n = 10): sham ovariectomy treated with PLAC. RESV and PLAC were administrated after ovariectomy and ZOL after six weeks after OVX, until the end of experiment. One implant was inserted in each tibiae of animals 18 weeks after ovariectomy. After 4 weeks, one implant was removed for counter-torque, and peri-implant tissue was collected for mRNA quantification of several osteogenic markers by PCR. The other tibia was submitted to micro-computed tomography analysis. Reduced counter-torque values, bone-implant contact (BIC) and bone volume fraction (BV/TV), and higher bone porosity (BP) were detected in OVX+PLAC group when compared to SHOVX+PLAC (p < 0.05). OVX+RESV rats presented lower BIC, BV/TV, and trabecular number (Tb.N), and augmented BP and trabecular spacing (Tb.Sp) when compared to SHOVX+PLAC (p < 0.05). Higher Tb.N and connectivity density (Conn.Dn) and reduced Tb.Sp were observed in OVX rats treated with ZOL, independently of RESV, when compared to OVX+PLAC and OVX+RESV groups (p < 0.05), whereas the combination ZOL+RESV promoted lower BP when compared to OVT+PLAC and OVX+RESV (p < 0.05). Gene expression was not influenced by RESV (p > 0.05), whereas ZOL promoted up-regulation of BMP-2 (p<0.05). RESV did not improve peri-implant bone repair in rats with ovariectomy-induced osteoporosis.

Different Protocols of Combined Application of Photobiomodulation In Vitro and In Vivo Plus Adipose-Derived Stem Cells Improve the Healing of Bones in Critical Size Defects in Rat Models.

Introduction: Long bone segmental deficiencies are challenging complications to treat. Hereby, the effects of the scaffold derived from the human demineralized bone matrix (hDBMS) plus human adipose stem cells (hADSs) plus photobiomodulation (PBM) (in vitro and or in vivo) on the catabolic step of femoral bone repair in rats with critical size femoral defects (CDFDs) were evaluated with stereology and high stress load (HSL) assessment methods. Methods: hADSs were exposed to PBM in vitro; then, the mixed influences of hDBMS+hADS+PBM on CSFDs were evaluated. CSFDs were made on both femurs; then hDBMSs were engrafted into both CSFDs of all rats. There were 6 groups (G)s: G1 was the control; in G2 (hADS), hADSs only were engrafted into hDBMS of CSFD; in G3 (PBM) only PBM therapy for CSFD was provided; in G4 (hADS+PBM in vivo), seeded hADSs on hDBMS of CSFDs were radiated with a laser in vivo; in G5 (hADSs+PBM under in vitro condition), hADSs in a culture system were radiated with a laser, then transferred on hDBMS of CSFDs; and in G6 (hADS+PBM in conditions of in vivo and in vitro), laser-exposed hADSs were transplanted on hDBMS of CSFDs, and then CSFDs were exposed to a laser in vivo. Results: Groups 4, 5, and 6 meaningfully improved HSLs of CSFD in comparison with groups 3, 1, and 2 (all, P=0.001). HSL of G5 was significantly more than G4 and G6 (both, P=0.000). Gs 6 and 4 significantly increased new bone volumes of CSFD compared to Gs 2 (all, P=0.000) and 1 (P=0.001 & P=0.003 respectively). HSL of G 1 was significantly lower than G5 (P=0.026). Conclusion: HSLs of CSFD in rats that received treatments of hDBMS plus hADS plus PBM were significantly higher than treatments with hADS and PBM alone and control groups.

Use of platelet-rich fibrin for bone repair: a systematic review and meta-analysis of preclinical studies.

This systematic review evaluated the potential utility of platelet-rich fibrin (PRF) in bone repair in animals. The question is: can the use of PRF in bone defects in healthy rats induce bone repair compared to clot? This systematic review was conducted in accordance with the Preferred Reporting Items for Systematic Reviews and Meta-Analyses (Prisma). The protocol was registered with Prospero (CRD [42020162319]). The literature search involved nine databases, including grey literature. All studies evaluated the bone defects created in rats filled with PRF and clots (control). Biomaterial evaluation was also performed in this study. The risk of bias was assessed using the Systematic Review Center for Laboratory Animal Experimentation (Syrcle) tool for animal studies. A meta-analysis of quantitative data was performed to estimate the effect of PRF on bone repair in rats. Heterogeneity among the studies was assessed using the I2 statistic. The literature search retrieved 685 studies, 10 of which fulfilled the eligibility criteria, and 4 were included in the quantitative assessment. Analysis of the risk of bias revealed that most studies had a high risk of bias in performance and detection. Meta-analysis yielded divergent results and the absence of a statistically significant effect: PRF with control (standardized mean difference 2.54, 95% confidence interval -0.80-5.89; p = 0.14). In general, study heterogeneity was high (I2 ≥ 75.0%). The quality of the studies that influenced the conclusion of the review was based on the PICO, the sources and form of the search, the study selection criteria, the form of evaluation of publication bias, the evaluation of the quality of the studies, and data extraction by two researchers. PRF did not provide significant benefits for bone repair, resulting in unpredictable effects.

Electrospun PLGA scaffold loaded with osteogenic growth peptide accelerates cranial bone repair in rats

To study the feasibility of electrospun poly(D, L-lactide-co-glycolide) (PLGA) scaffold loaded with osteogenic growth peptide (OGP) for bone tissue engineering. PLGA scaffolds were prepared by electrospinning PLGA solution without OGP(control group)or with 0.1%, 0.2%and 0.4%OGP(0.1%OGP@PLGA, 0.2%OGP@PLGA, and 0.4%OGP@PLGA scaffolds, respectively).The microstructure of the scaffolds was observed by scanning electron microscopy(SEM).The scaffolds were soaked in PBS to confirm the release pattern of OGP.The biocompatibility of the scaffolds was evaluated using CCK-8 assay and live/dead staining after a 7-day coculture with rat bone marrow-derived mesenchymal stem cells(BMSCs).ALP assay and ARS staining were used to evaluate osteoinduction capacity of the scaffolds co-cultured with rat BMSCs for 14 days.In a male SD rat model of skull defect(5 mm in diameter), bone defect repair was evaluated 8 weeks after implantation of the scaffolds using Micro-CT, HE and Masson staining. The electrospun scaffolds had a fibrous structure similar to extracellular matrix(ECM)and were capable of sustained release of OGP for at least one month.Co-culture with 0.2%OGP@PLGA and 0.4%OGP@PLGA scaffolds, as compared with pure PLGA scaffold, significantly promoted the growth of rat BMSCs ((P < 0.01).The cells co-cultured with 0.4%OGP@PLGA scaffold showed the highest ALP activity and the greatest number of calcium nodules, indicating its strong osteoinduction ability (P < 0.01).Micro-CT and HE and Masson staining results showed that 0.4%OGP@PLGA scaffold had significantly better ability for promoting bone repair than the other two OGP-loaded scaffolds(P < 0.01). The electrospun PLGA scaffold loaded with OGP effectively mimics the structure of ECM and has a good biocompatibility and osteoinduction ability, suggesting its potential as a new bone tissue engineering scaffold for bone defect repair.

Effect of S53P4 bioactive glass and low-level laser therapy on calvarial bone repair in rats submitted to zoledronic acid therapy.

ABSTRACTPurposeTo evaluate the influence of bioactive glass and photobiomodulation therapy (PBMT) in calvarial bone repair process in rats submitted to zoledronic acid therapy.MethodsTwenty-four rats were selected and treated with the dose of 0.035 mg/kg of zoledronic acid every two weeks, totalizing eight weeks, to induce osteonecrosis. After the drug therapy, surgical procedure was performed to create 5-mm diameter parietal bone defects in the calvarial region. The rats were then randomly assigned to groups according to the following treatments: AZC: control group, treated with blood clot; AZBIO: bone defect filled with bioactive glass; AZL: treated with blood clot and submitted to PBMT; and AZBIOL: treated with bioactive glass S53P4 and submitted to PBMT. Tissue samples were collected and submitted to histomorphometric analysis after 14 and 28 days.ResultsAt 14 days, bone neoformation in the AZBIO (52.15 ± 9.77) and AZBIOL (49.77 ± 13.58) groups presented higher values (p ≤ 0.001) compared to the AZC (23.35 ± 10.15) and AZL groups (23.32 ± 8.75). At 28 days, AZBIO (80.24 ± 5.41)still presented significant higher bone recovery values when compared to AZC (59.59 ± 16.92)and AZL (45.25 ± 5.41) groups (p = 0.048). In the 28-day period, the AZBIOL group didn’t show statistically significant difference with the other groups (71.79 ± 29.38).ConclusionsThe bioactive glass is an effective protocol to stimulate bone neoformation in critical defects surgically created in rats with drug induced osteonecrosis, in the studied periods of 14 and 28 days.

In Vivo Evaluation of Bovine Xenograft Associated With Oxygen Therapy in Alveolar Bone Repair

To preserve alveolar bone thickness and width after extraction, clinical strategies have been adopted to reduce or eliminate the need for future surgical interventions to increase the alveolar ridge. The use of xenogeneic biomaterials has been increasing for such application. The association of bone substitutes with active oxygen-based materials, which is essential in the wound-healing process, could accelerate bone repair, optimizing the maintenance of alveolar architecture after extraction. However, the truth of this hypothesis is not clear. The present study aimed to compare the biological response to inorganic bovine bone graft Bonefill (BF), associated or not with active oxygen-based oral gel Bluem (BF+BM), in alveolar bone repair. Twenty female Wistar rats were randomly allocated. The left upper central incisor was extracted, and the dental sockets were filled with BF in the control group (n = 10) and with BF+BM in the experimental group (n = 10). The animals were euthanized at 7 and 42 days after implantation (n = 5), and the samples were processed for descriptive histological and histomorphometric evaluations. The results showed no significant difference between the groups (P > .05). Both groups presented a time-dependent increase in newly formed bone and biosorption biomaterial (P = .0001). The association between active oxygen-based gel and inorganic bovine bone graft did not interfere with or improve bone repair during the experimental periods of alveolar bone repair in rats.

Peri‐implant bone repair in rats systemically modified by antineoplastic agents and nicotine

Peri‐implant bone repair in rats systemically modified by antineoplastic agents and nicotine

Bone repair in craniofacial defects treated with different doses of alendronate: a histological, histomorphometric, and immunohistochemical study.

The objective of the study is to evaluate bone repair in rats treated with different alendronate doses. Sixty female rats ovariectomized were randomly divided in three groups: group C (control group), group A1 (ALN/1mg/kg), and A2 (ALN/ 3mg/kg). Each animal received subcutaneous applications of sodium alendronate at a dose correspondent to group A1 or A2 three times a week, while the control group received 0.9% saline solution. After 4weeks of application, a critical defect was created in the calvaria of animals of all groups. The defect was filled by particulate autogenous bone. The applications were maintained until euthanasia, which occurred 15 and 60days after the surgical procedure. The pieces were sent for histological, histomorphometric and immunohistochemical analysis. The data were submitted to statistical analysis with significance level of 0.05. The descriptive histological analysis demonstrated an increase in bone neoformation in both groups treated with alendronate when compared to the control group. The histomorphometric analysis showed an increase in the amount of neoformed bone in A1 and A2 groups when compared to group C, both at 15days (p = 0.0002) and at 60days (p = 0.001). In the immunohistochemical analysis, it was possible to observe a difference in immunolabeling just for Mmp2 at the time of 60days in A1 (p = 0.001) and A2 (p = 0.023) when compared to the control group. Systemic delivery of alendronate, regardless of the dose, increased the amount of bone neoformation. Prescription of sodium alendronate at 1mg/kg for improvement of bone neoformation in bone graft procedures.

Controlled release of FGF-2 and BMP-2 in tissue engineered periosteum promotes bone repair in rats

The aim of this study was to prepare chitosan–collagen (CS/COL) scaffolds that could release fibroblast growth factor-2 (FGF-2) and bone morphogenetic protein 2 (BMP-2), and to study the effect of this scaffold on bone repair. By improving the double emulsion/solvent evaporation technique, BMP-2 was encapsulated in poly(lactic acid)–poly(ethylene glycol)–poly(lactic acid) (PELA) microcapsules, to the surface of which FGF-2 was attached. The CS/COL scaffold carrying the microcapsules was prepared by freeze-drying. Periosteum derived cells (PDCs) were extracted and cultured on the scaffolds to study their proliferation and differentiation on the scaffolds. In addition, the effects of the scaffolds were investigated on rats with skull defects by micro-computed tomography and histology. We successfully prepared PELA microcapsules with external adherence to FGF-2 and encapsulated with BMP-2. The CS/COL scaffolds were porous and PDCs adhered, proliferated and underwent osteogenic differentiation on the scaffolds. The sequential release of FGF-2/BMP-2 had better osteogenic efficacy than other groups. Our results suggest that CS/COL scaffolds that bind FGF-2 and BMP-2 in combination with PDCs could be a promising new strategy for tissue engineering periosteum.

Electrospun Poly(3-hydroxybutyrate-co-4-hydroxybutyrate)/Graphene Oxide Scaffold: Enhanced Properties and Promoted in Vivo Bone Repair in Rats.

Bone tissue engineering emerges as an advantageous technique to achieve tissue regeneration. Its scaffolds must present excellent biomechanical properties, where bare polymers poorly perform. Development of new biomaterials with high osteogenic capacity is urgently pursued. In this study, an electrospun poly(3-hydroxybutyrate-co-4-hydroxybutyrate)/graphene oxide (P34HB/GO) nanofibrous scaffold is successfully fabricated and characterized. The effects of GO amount on scaffold morphology, biomechanical properties, and cellular behaviors are investigated. GO reduces the fiber diameter and enhances porosity, hydrophilicity, mechanical properties, cellular performance, and osteogenic differentiation of scaffolds. P34HB/GO triumphs over P34HB in in vivo bone regeneration in critical-sized calvarial defect of rats. We believe that this study is the first to evaluate the capability of in vivo bone repair of electrospun P34HB/GO scaffold. With facile fabrication process, favorable porous structures, enhanced biomechanical properties, and fast osteogenic capability, P34HB/GO scaffold holds practical potentials for bone tissue engineering application.

Impact of resveratrol on bone repair in rats exposed to cigarette smoke inhalation: histomorphometric and bone-related gene expression analysis

This study investigated the effect of resveratrol on bone healing and its influence on the gene expression of bone-related markers in rats exposed to cigarette smoke. Two calvarial defects were created in each of 60 rats, which were assigned equally (n=20) to three groups: (1) resveratrol (10mg/kg)+smoke exposure (SMK+RESV); (2) placebo+smoke exposure (SMK+PLA); or (3) placebo+no smoke exposure (NS+PLA). Substances were administered daily for 30days following surgery. Smoke inhalation was started 7days before surgery and continued for 30days after surgery. One defect was processed for histomorphometric analysis and the other was used for mRNA quantification of bone-related gene expression by qPCR. The remaining defect was smaller in the SMK+RESV (2.27±0.61mm, P=0.0003) and NS+PLA (2.17±0.74mm, P=0.0005) groups than in the SMK+PLA group (3.12±0.47mm). Higher levels of Runx2 were observed in the NS+PLA group than in the smoke exposure groups (vs. SMK+PLA, P=0002; vs. SMK+RESV, P=0.052); levels of Lrp-5 were also higher in the no smoke exposure group (vs. SMK+RESV, P=0.009; vs. SMK+PLA, P=0.003). Resveratrol therapy decreased RANKL/OPG expression when compared to placebo (SMK+RESV vs. SMK+PLA, P=0.017). Dkk1 levels were decreased in the SMK+RESV group when compared to the SMK+PLA (P=0.006) and NS+PLA groups (P=0.005). In conclusion, resveratrol optimizes the repair of critical-sized bone defects, up-regulating the gene expression of important bone remodelling markers in rats exposed to cigarette smoke inhalation.

Calcium-phosphate ceramics and polysaccharide-based hydrogel scaffolds combined with mesenchymal stem cell differently support bone repair in rats

Research in bone tissue engineering is focused on the development of alternatives to autologous bone grafts for bone reconstruction. Although multiple stem cell-based products and biomaterials are currently being investigated, comparative studies are rarely achieved to evaluate the most appropriate approach in this context. Here, we aimed to compare different clinically relevant bone tissue engineering methods and evaluated the kinetic repair and the bone healing efficiency supported by mesenchymal stem cells and two different biomaterials, a new hydrogel scaffold and a commercial hydroxyapatite/tricalcium phosphate ceramic, alone or in combination.Syngeneic mesenchymal stem cells (5 × 105) and macroporous biphasic calcium phosphate ceramic granules (Calciresorb C35®, Ceraver) or porous pullulan/dextran-based hydrogel scaffold were implanted alone or combined in a drilled-hole bone defect in rats. Using quantitative microtomography measurements and qualitative histological examinations, their osteogenic properties were evaluated 7, 30, and 90 days after implantation. Three months after surgery, only minimal repair was evidenced in control rats while newly mineralized bone was massively observed in animals treated with either hydrogels (bone volume/tissue volume = 20%) or ceramics (bone volume/tissue volume = 26%). Repair mechanism and resorption kinetics were strikingly different: rapidly-resorbed hydrogels induced a dense bone mineralization from the edges of the defect while ceramics triggered newly woven bone formation in close contact with the ceramic surface that remained unresorbed. Delivery of mesenchymal stem cells in combination with these biomaterials enhanced both bone healing (>20%) and neovascularization after 1 month, mainly in hydrogel.Osteogenic and angiogenic properties combined with rapid resorption make hydrogels a promising alternative to ceramics for bone repair by cell therapy.

Degradation of Mg-1.5Sr alloy during bone repair in rats

Objective To explore the effect of degrading process of Mg-Sr alloy (containing 1.5 wt% Sr) on bone growth in rats. Methods Twelve male SD rats of 8 weeks old were randomly divided into 2 even groups (n=6). In the experimental group, a stent made of Mg-1.5Sr alloy was put into the left femoral condyle of each rat while a pure Mg stand was used in the control group. The 2 groups were subjected to Micro-CT scanning at 4, 8, and 12 weeks after surgery. Cross-sectional images were generated for clear observation of the degrading process of the stands and the skeletal growth in the rats. Next, 3D models were made to compare changes in stand volume to calculate their degrading rates. The skeletal growth in the rats were also analyzed by measuring a series of indexes, including bone mineral density (BMD), bone volume fraction (BV/TV), trabecular thickness (Tb.Th) and trabecular number (Tb.N.). Results The cross-sectional images at 4, 8 and 12 weeks showed that the Mg-1.5Sr stands degraded more slowly than the pure Mg ones and more new bone grew out around the Mg-1.5Sr stands than around the pure Mg ones. The degrading rates of Mg-1.5Sr alloy inside the rats at 4, 8 and 12 weeks were respectively 0.34±0.04 mm/y, 0.28±0.09 mm/y and 0.31±0.05 mm/y, significantly lower than those of pure Mg (0.51±0.01 mm/y, 0.37±0.08 mm/y and 0.39±0.06 mm/y)(P<0.05). The BMD, BV/TV, Tb.Th and Tb.N at 12 weeks in the experimental group (0.62±0.08 g/mm3, 53%±11%, 298±50 μm, 0.0027±0.0002/μm) were significantly greater than those in the control group (0.56±0.08 g/mm3, 48%±8%, 220±40 μm, 0.0023±0.0002/μm) (P<0.05). Conclusion Mg-1.5Sr alloy may be a promising orthopedic implant material with great biocompatibility due to its relatively low degrading rate and good bone formation performance. Key words: Magnesium; Strontium; Alloys; Biodegradation; Bone repair

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

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

Stimulating angiogenesis mitigates the unloading-induced reduction in osteogenesis in early-stage bone repair in rats.

Accelerating fracture healing during bed rest allows early mobilization and avoids prolonged fracture healing times. We tested the hypothesis that stimulating angiogenesis with deferoxamine (DFO) mitigates the unloading-induced reduction in early-stage bone repair. Rats aged 12 weeks were subjected to cortical drilling on their tibial diaphysis under anesthesia and treated with hindlimb unloading (HU), HU and DFO administration (DFOHU), or weight bearing (WB) for 5 or 10 days (HU5/10, DFOHU5/10, WB5/10; n = 8 per groups) until sacrifice for vascular casting with a zirconium dioxide-based contrast agent. Taking advantage of its absorption discontinuity at the K-absorption edge, vascular and bone images in the drill-hole defects were acquired by synchrotron radiation subtraction CT. Bone repair was reduced in HU rats. The bone volume fraction (B.Vf) was 88% smaller in HU5 and 42% smaller in HU10 than in WB5/10. The bone segment densities (B.Seg) were 97% smaller in HU5 and 141% larger in HU10 than in WB5/10, and bone thickness (B.Th) was 38% smaller in HU10 than in WB10. The vascular volume fraction (V.Vf) was 35% and the mean vessel diameter (V.D) was 13% smaller in HU10 than in WB10. When compared according to categorized vessel sizes, V.Vf in the diameter ranges 20–30, 30–40, and >40 μm were smaller in HU10 than in WB10, and V.Seg in the diameter range >40 μm was smaller in HU10 than in WB10. In contrast, there was no difference in B.Vf between DFOHU5/10 and WB5/10 and in V.Vf between DFOHU10 and WB10, though B.Seg remained 86% smaller in DFOHU5 and 94% larger in DFOHU10 than in WB5/10, and B.Th and V.D were 23% and 14% lower in DFOHU10 than in WB10. Vessel size-specific V.Vf in the diameter ranges 10–20 and 20–30 μm was larger in DFOHU5 than in HU5. In conclusion, the enhanced angiogenic ingrowth mitigates the reduction in bone repair during mechanical unloading.

Histological Study of the Effect of Bioactive Glass on Tibial Bone Repair in Rats

The main goal of this study was to histologically evaluate the healing of surgically created defects on the tibiae of adult rats after implantation of bioactive glass. Twenty adult Wistar rats (body weight of 300g) were divided into two groups: bioglass treated group (n=10) and control group (n=10). Unicortical bone defects with 3-mm diameter were performed in both tibiae of the animals and filled with bioglass particles. The rats were then sacrificed at 14, 30 , 60 and 90 days, and the tissues were prepared for histological processing, sectioning, and staining with hematoxylin and eosin, as well as Mallory trichrome, and analyzed under light microscope. Within 7-14 days, both groups presented connective tissue septa with new bone formation, more intense in bioglass treated group. In the subsequent periods (30, 60 and 90 days), these groups presented more mature bone tissue around the glass particles. Bone trabeculae formed in all experimental periods were juxtaposed to the glass particles. It can be concluded that bioglass materials promoted bone formation over the entire extension of the defect, independently of the size of the granules, thus confirming their biological osteoconductive property.

Cumulative effect of low-level laser therapy and low-intensity pulsed ultrasound on bone repair in rats

Many studies have assessed the effects of either low-level laser therapy (LLLT) or low-intensity pulsed ultrasound (LIPUS) on bone repair; however, an evaluation of the combination of these modalities (LLLT+LIPUS) has not yet been considered. The aim of this study was to demonstrate the effects of LLLT+LIPUS on bone repair. Male Wistar rats (n=128; four groups of 32) were used; the animals underwent a partial tibial bone osteotomy. One group had the osteotomized limb treated with LLLT, the second group with LIPUS, and the third group with the combined treatment protocols of the LLLT and LIPUS groups; the fourth group received no further treatment (control). Each group was divided into two subgroups for assessment at two different time-points, 14 and 21 days. After the completion of treatment rats were sacrificed and the tibias submitted to a three-point bending test or to histomorphometric analysis. Histological evaluation showed increased bone trabeculae, increased vascularization, and decreased inflammation in the LLLT+LIPUS group. Mechanical evaluation revealed increased biomechanical properties including maximum force, maximum stress, and stiffness, in the LLLT+LIPUS group. Combined LLLT+LIPUS treatment enhanced bone healing both histologically and mechanically, shortening the length of the treatment period, when compared to treatment with LLLT or LIPUS alone.