Differences in bone microarchitecture between genetic and secondary iron-overload mouse models suggest a role for hepcidin deficiency in iron-related osteoporosis.

Iron overload is one of the secondary osteoporosis etiologies. Cellular and molecular mechanisms involved in iron-related osteoporosis are not fully understood. AIM: The aim of the study was to investigate the respective roles of iron excess and hepcidin, the systemic iron regulator, in the development of iron-related osteoporosis. MATERIAL AND METHODS: We used mice models with genetic iron overload (GIO) related to hepcidin deficiency (Hfe-/- and Bmp6-/- ) and secondary iron overload (SIO) exhibiting a hepcidin increase secondary to iron excess. Iron concentration and transferrin saturation levels were evaluated in serum and hepatic, spleen, and bone iron concentrations were assessed by ICP-MS and Perl's staining. Gene expression was evaluated by quantitative RT-PCR. Bone micro-architecture was evaluated by micro-CT. The osteoblastic MC3T3 murine cells that are able to mineralize were exposed to iron and/or hepcidin. RESULTS: Despite an increase of bone iron concentration in all overloaded mice models, bone volume/total volume (BV/TV) and trabecular thickness (Tb.Th) only decreased significantly in GIO, at 12 months for Hfe-/- and from 6 months for Bmp6-/- . Alterations in bone microarchitecture in the Bmp6-/- model were positively correlated with hepcidin levels (BV/TV (ρ = +.481, p < .05) and Tb.Th (ρ = +.690, p < .05). Iron deposits were detected in the bone trabeculae of Hfe-/- and Bmp6-/- mice, while iron deposits were mainly visible in bone marrow macrophages in secondary iron overload. In cell cultures, ferric ammonium citrate exposure abolished the mineralization process for concentrations above 5 µM, with a parallel decrease in osteocalcin, collagen 1, and alkaline phosphatase mRNA levels. Hepcidin supplementation of cells had a rescue effect on the collagen 1 and alkaline phosphatase expression level decrease. CONCLUSION: Together, these data suggest that iron in excess alone is not sufficient to induce osteoporosis and that low hepcidin levels also contribute to the development of osteoporosis.

(-)-Epigallocatechin gallate (EGCG) pharmacokinetics and molecular interactions towards amelioration of hyperglycemia, hyperlipidemia associated hepatorenal oxidative injury in alloxan induced diabetic mice.

Diabetes mellitus has become a serious problem associated with health complications, such as metabolism disorders and liver-kidney dysfunction. The inadequacies associated with conventional medicines have led to a determined search for alternative natural therapeutic agents. The present study was conducted to evaluate the hypoglycemic, antilipidemic, and antioxidant effects of EGCG in surviving diabetic mice. Alloxan diabetic mice were treated with EGCG. Their bloods were collected and submitted to various biochemical measurements, including blood glucose, cholesterol, triglycerides, urea, creatinine, and transaminases. Their livers and kidneys were isolated to assess oxidative damage and to perform histological analysis. Both EGCG and insulin treatment of diabetic mice resulted in a significant reduction in fasting blood glucose levels. EGCG supplementation also ameliorated hepatic as well as renal toxicity indices. Moreover, diabetic mice injected with EGCG exhibited significant changes in antioxidant enzyme activities in the liver and kidney. Histological analyses also showed that it exerted an ameliorative action on these organs and efficiently protected the liver-kidney functions of diabetic mice. EGCG was found to bind α-amylase, PTP1B, and α-glucosidase with good affinities ranging from -6.1 to -8.4 kcal/mol. The findings revealed that EGCG administration induced attractive curative effects on diabetic mice, particularly in terms of liver-kidney function. EGCG can, therefore, be considered as a potential strong candidate for future applications to treat and alleviate diabetic burden. Its pharmacokinetics, high affinities, and molecular interactions with the targeted receptors satisfactory explain the in vivo findings.

Microarchitecture of titanium cylinders obtained by additive manufacturing does not influence osseointegration in the sheep.

Large bone defects are a challenge for orthopedic surgery. Natural (bone grafts) and synthetic biomaterials have been proposed but several problems arise such as biomechanical resistance or viral/bacterial safety. The use of metallic foams could be a solution to improve mechanical resistance and promote osseointegration of large porous metal devices. Titanium cylinders have been prepared by additive manufacturing (3D printing/rapid prototyping) with a geometric or trabecular microarchitecture. They were implanted in the femoral condyles of aged ewes; the animals were left in stabling for 90 and 270 days. A double calcein labeling was done before sacrifice; bones were analyzed by histomorphometry. Neither bone volume, bone/titanium interface nor mineralization rate were influenced by the cylinder's microarchitecture; the morphometric parameters did not significantly increase over time. Bone anchoring occurred on the margins of the cylinders and some trabeculae extended in the core of the cylinders but the amount of bone inside the cylinders remained low. The rigid titanium cylinders preserved bone cells from strains in the core of the cylinders. Additive manufacturing is an interesting tool to prepare 3D metallic scaffolds, but microarchitecture does not seem as crucial as expected and anchoring seems limited to the first millimeters of the graft.

In vivo osseointegration and erosion of nacre screws in an animal model.

The use of resorbable devices for osteosynthesis has become a subject of interest. Nacre has been proposed as a resorbable and osteoconductive material favoring bone apposition without triggering an inflammatory reaction. We compared the in vivo osseointegration and erosion of nacre screws in an animal model with titanium screws. Implantation of similar nacre and titanium screws was performed in the femoral condyles of adult rats. Animals (n = 41) were randomized in four groups sacrificed at day one, 1, 6, and 12 months. Microcomputed tomography (microCT) allowed 3D morphometry of erosion of nacre. Osseointegration was measured as the volume of trabecular bone bone volume/tissue volume (BV/TV) in a standardized volume of interest around each screw. Undecalcified bone histology was also done. Gross examination revealed a similar clinical osseointegration for titanium and nacre screws. A progressive erosion of nacre screws, but no erosion of titanium screws, was observed in microCT. The volume of nacre screws progressively decreased over time whereas no modification occurred for titanium. For titanium screws, BV/TV remained stable throughout the study. For nacre screws, the BV/TV decrease was not statistically different. A significant difference was found between nacre and titanium screws at 6 months but not at 12 months. The screw heads, outside the bone shaft, were not eroded even after 12 months. Erosion of nacre occurred during the entire study period, only within the bone shaft in direct contact with bone marrow. Bone apposition was observed on nacre surfaces without signs of erosion. Nacre is a promising biomaterial in maxillofacial surgery.

Histochemical identification of wear debris released by alumina-on-alumina hip prostheses in the periprosthetic tissues.

BACKGROUND: Tribological studies have shown that the most used couples for hip prostheses consist of metal-on-polyethylene and alumina-on-alumina prostheses. Over time, wear products accumulate in the joint cavity and in the periprosthetic tissues. Although polyethylene and metal are easily identifiable by microscopy in periprosthetic tissues, alumina particles are very difficult to identify. HYPOTHESIS: The fluorescent azo-dye lumogallion was evaluated as a suitable histochemical stain for alumina particles in periprosthetic tissues. MATERIAL AND METHOD: In 28 patients who had a prosthetic revision of an alumina-on-alumina prosthesis, periprosthetic tissues were removed and embedded in paraffin; sections were stained with HPS (for conventional diagnosis) or with lumogallion. Sections were examined for wear particles in light and fluorescence microscopy. Some sections were counter-stained using DAPI for visualization of cell nuclei. RESULTS: The wear particles of the alumina-alumina prostheses were very difficult to identify on the HPS stained sections; they were clearly evidenced by lumogallion staining with a bright orange fluorescence. The stain revealed large quantities of particles (of the order of several thousand per section). Only two patients had no particles. The staining technique identified numerous particles that were not visible on HPS-stained sections in macrophages, synoviocytes and fibroblasts. CONCLUSION: This staining, which has been validated in neuromuscular pathology for the identification of alumina used as a vaccine adjuvant, gave successful results in the present study. Alumina particles are modified when they are phagocytized by macrophages. lumogallion staining easily shows the presence of thousands of wear particles released by alumina-on-alumina prostheses in periprosthetic tissues. LEVEL OF EVIDENCE: V expert opinion study.

Dapagliflozin and Liraglutide Therapies Rapidly Enhanced Bone Material Properties and Matrix Biomechanics at Bone Formation Site in a Type 2 Diabetic Mouse Model.

The aim of this study is to compare head-to-head the effects of dapagliflozin and liraglutide on bone strength and bone material properties in a pre-clinical model of diabetes-obesity. Combined low-dose streptozotocin and high fat feeding were employed in mice to promote obesity, insulin resistance, and hyperglycaemia. Mice were administered daily for 28 days with saline vehicle, 1 mg/kg dapagliflozin or 25 nmol/kg liraglutide. Bone strength was assessed by three-point bending and nanoindentation. Bone material properties were investigated by Fourier transform infrared microspectroscopy/imaging. Although diabetic controls presented with dramatic reductions in mechanical strength, no deterioration of bone microarchitecture was apparent. At the tissue level, significant alterations in phosphate/amide ratio, carbonate/phosphate ratio, tissue water content, crystal size index, collagen maturity and collagen glycation were observed and linked to alteration of matrix biomechanics. Dapagliflozin and liraglutide failed to improve bone strength by 3-point bending or bone microarchitecture during the 28-day-treatment period. At bone formation site, dapagliflozin enhanced phosphate/amide ratio, mineral maturity, and reduced tissue water content, crystal size index, and collagen glycation. Liraglutide had significant effects on phosphate/amide ratio, tissue water content, crystal size index, mature collagen crosslinks, collagen maturity, and collagen glycation. At bone formation site, both drugs modulated matrix biomechanics. This study highlighted that these two molecules are effective in improving bone material properties and modulating matrix biomechanics at bone formation site. This study also highlighted that the resulting effects on bone material properties are not identical between dapagliflozin and liraglutide and not only mediated by lower blood glucose.

Computational fluid dynamics simulation from microCT stacks of commercial biomaterials usable for bone grafting.

Granules of calcium/phosphate biomaterials are used to fill small bone defects in oral and maxilla-facial surgery. Granules of natural (e.g., trabecular bone, coral) or synthetic biomaterials are provided by industry. Small granules can also form of putty. The 3D geometry of granules creates a macroporosity allowing invasion of vascular and bone cells when pores are larger than 300 µm. We analyzed the 3D-porosity of 11 different stacks of biomaterials: Osteopure®, CopiOs®, Bio-Oss®, TCP Dental HP®, KeraOs®, TCH®, Biocoral®, EthOss® and Nanostim®. For each granular biomaterial, two sizes of granules were analyzed: small and large. Microcomputed tomography (microCT) determined porosity and microarchitectural characteristics of the biomaterials stacks. Computational fluid dynamics (CFD), a simulation method, was used on the stacks of microCT images. Stacks of small granules had a much lower permeation and fluid velocity than large granules and the hydraulic tortuosity was increased. Significant correlations were observed between microarchitecture parameters (porosity, mean pore diameter and specific surface) and fluid dynamic parameters. The two putties were associated with low (or absence of) porosity and permeation study revealed a very low (or absence) of flow rate. Stacks of granules represent 3D scaffolds resembling trabecular bone with an interconnected porous microarchitecture. Small granules create pores less than 300 µm in diameter; this induces a low fluid flow rate. CFD simulates the accessibility of body fluids and progenitor cells and confirms that it is depending on the shape and 3D arrangements of granules within a stack. Large granules must be preferred to putties and small granules.

Enteroendocrine K Cells Exert Complementary Effects to Control Bone Quality and Mass in Mice.

The involvement of a gut-bone axis in controlling bone physiology has been long suspected, although the exact mechanisms are unclear. We explored whether glucose-dependent insulinotropic polypeptide (GIP)-producing enteroendocrine K cells were involved in this process. The bone phenotype of transgenic mouse models lacking GIP secretion (GIP-GFP-KI) or enteroendocrine K cells (GIP-DT) was investigated. Mice deficient in GIP secretion exhibited lower bone strength, trabecular bone mass, trabecular number, and cortical thickness, notably due to higher bone resorption. Alterations of microstructure, modifications of bone compositional parameters, represented by lower collagen cross-linking, were also apparent. None of these alterations were observed in GIP-DT mice lacking enteroendocrine K cells, suggesting that another K-cell secretory product acts to counteract GIP action. To assess this, stable analogues of the known K-cell peptide hormones, xenin and GIP, were administered to mature NIH Swiss male mice. Both were capable of modulating bone strength mostly by altering bone microstructure, bone gene expression, and bone compositional parameters. However, the two molecules exhibited opposite actions on bone physiology, with evidence that xenin effects are mediated indirectly, possibly via neural networks. Our data highlight a previously unknown interaction between GIP and xenin, which both moderate gut-bone connectivity. © 2020 American Society for Bone and Mineral Research.

Sclerostin-Antibody Treatment Decreases Fracture Rates in Axial Skeleton and Improves the Skeletal Phenotype in Growing oim/oim Mice.

In osteogenesis imperfecta (OI), vertebrae brittleness causes thorax deformations and leads to cardiopulmonary failure. As sclerostin-neutralizing antibodies increase bone mass and strength in animal models of osteoporosis, their administration in two murine models of severe OI enhanced the strength of vertebrae in growing female Crtap-/- mice but not in growing male Col1a1Jrt/+ mice. However, these two studies ignored the impact of antibodies on spine growth, fracture rates, and compressive mechanical properties. Here, we conducted a randomized controlled trial in oim/oim mice, an established model of human severe OI type III due to a mutation in Col1a2. Five-week-old female WT and oim/oim mice received either PBS or sclerostin antibody (Scl-Ab) for 9 weeks. Analyses included radiography, histomorphometry, pQCT, microcomputed tomography, and biomechanical testing. Though it did not modify vertebral axial growth, Scl-Ab treatment markedly reduced the fracture prevalence in the pelvis and caudal vertebrae, enhanced osteoblast activity (L4), increased cervico-sacral spine BMD, and improved the lumbosacral spine bone cross-sectional area. Scl-Ab did not impact vertebral height and body size but enhanced the cortical thickness and trabecular bone volume significantly in the two Scl-Ab groups. At lumbar vertebrae and tibial metaphysis, the absolute increase in cortical and trabecular bone mass was higher in Scl-Ab WT than in Scl-Ab oim/oim. The effects on trabecular bone mass were mainly due to changes in trabecular number at vertebrae and in trabecular thickness at metaphyses. Additionally, Scl-Ab did not restore a standard trabecular network, but improved bone compressive ultimate load with more robust effects at vertebrae than at metaphysis. Overall, Scl-Ab treatment may be beneficial for reducing vertebral fractures and spine deformities in patients with severe OI.

Mandibular bone effects of botulinum toxin injections in masticatory muscles in adult.

OBJECTIVE: Botulinum toxin (BTX) is injected into masticatory muscles to treat various conditions. Animal studies have demonstrated bone loss at the condylar and alveolar regions of the mandible after BTX injection into masticatory muscles. The aim of the present study was to investigate mandibular bone changes in patients who received BTX injections in masticatory muscles. STUDY DESIGN: Twelve adult patients who received BTX injections into masticatory muscles were included in this study. Cone beam computed tomography (CBCT) was performed before and 12 months after the injection. The condylar and alveolar regions of the mandible were analyzed by using texture analysis of the CBCT images with the run length method. Condylar cortical thickness was measured, and 3-dimensional analysis of the mandible was also performed. Six patients who did not receive BTX injections were used as controls. RESULTS: A run length parameter (gray level nonuniformity) was found to be increased in condylar and alveolar bones. A significant cortical thinning was found at the anterior portion of the right condyle. Three-dimensional analysis showed significant changes in the condylar bone and at the digastric fossa. No changes in mandibular angles were found. CONCLUSIONS: This study identified mandibular bone changes in adult patients who received BTX injection into masticatory muscles.

GIP analogues augment bone strength by modulating bone composition in diet-induced obesity in mice.

Receptors to glucose-dependent insulinotropic polypeptide (GIP), have been identified on bone and GIP receptor (GIPr) knockout mice exhibit reduced bone strength and quality. Despite this, little is known on the potential beneficial bone effects of exogenous GIP on bone physiology. The aim of the present study was to assess whether stable GIP analogues were capable of ameliorating bone strength in mice with diet-induced obesity. The stable GIP analogue (D-Ala²)-GIP, and (D-Ala²)-GIP-Tag, a specific GIP analogue homing exclusively to bone, were employed. In vitro studies were used to assess effects of (D-Ala²)-GIP and (D-Ala²)-GIP-Tag on bone mineralization, lysyl oxidase activity, collagen maturity as well as osteoclast formation and activity. Subsequent in vivo studies employed obese-prediabetic Swiss NIH mice subjected to a 42-day period of daily administration of saline, (D-Ala²)-GIP or (D-Ala²)-GIP-Tag. In vitro studies confirmed that (D-Ala²)-GIP and (D-Ala²)-GIP-Tag had similar beneficial biological effects on bone cells. Administration of (D-Ala²)-GIP and (D-Ala²)-GIP-Tag resulted in lower blood glucose levels without any effects on body weight. Both GIP analogues augmented bone strength to a similar extent. Trabecular or cortical bone microarchitecture were not changed over the time course of the study. However, (D-Ala²)-GIP and (D-Ala²)-GIP-Tag augmented enzymatic collagen crosslinking as well as the heterogeneity of enzymatic collagen crosslinking, mineral-to-matrix ratio and significantly reduced the heterogeneity in mineral bone crystallite size. This study demonstrates that activation of skeletal GIPr by stable GIP analogues enhance bone strength in prediabetes and suggest that these analogues may be beneficial in the treatment of bone disease.

Osteocyte staining with rhodamine in osteonecrosis and osteoarthritis of the femoral head.

Death of osteocytes is synonymous of bone death. Aseptic osteonecrosis of the femoral head is a lesion characterized by the death of osteocytes occurring after major vascular changes. The evolution may lead to hip osteoarthritis, which requires total hip arthroplasty in most cases. Evolution of aseptic osteonecrosis in four radiological stages is well known. We analyzed 24 femoral heads from patients with osteonecrosis or osteoarthritis, retrieved at the time of surgery for a hip arthroplasty. The aim of the study was to clearly identify the necrotic bone from the living bone in the histological samples. The femoral heads were sawed, and a large sample was harvested in the superior zone; it was stained en-bloc with rhodamine dissolved in formalin to make the osteocytes fluorescent under UV light microscopy. Undecalcified sections, 7 µm thick, were obtained on a heavy-duty microtome. A micrographic analysis using two UV excitation wavelengths visualized the living osteocytes (in green) and the bone matrix (in blue). A simple method to prepare combined images is described. In addition, the blocks can be analyzed by confocal microscopy to visualize more details. It is possible to identify at low magnification the osteocytes within the bone matrix and the osteonecrotic areas where osteocytes have disappeared. Identification of osteocytes showed that newly formed bone packets are laid on dead trabeculae in patients with aseptic osteonecrosis or with osteoarthritis. In the osteosclerotic areas, the enlarged trabeculae have a dead central core surrounded by recently apposed bone structure units.

Texture analysis of trabecular bone around RM-Pressfit cementless acetabulum in a series of 46 patients during a 5 year period.

BACKGROUND: Cementless total hip arthroplasty (THA) is a common procedure producing excellent clinical results. Their long-term survival is nevertheless burdened by loosening of the acetabular part caused by changes in the distribution of strains around the cup. In this context the RM-Pressfit® cup has been developed, resulting in a more harmonious distribution of the strains. HYPOTHESIS: Texture analysis of X-ray films can evaluate the evolution of trabecular bone micro-architecture during the five years following THA with a RM-Pressfit® cup. MATERIAL AND METHOD: A monocentric series of 46 hips was reviewed regularly within five years post- surgery. Radiographic evaluation of the operated hip was done on frontal digitized radiographs of the pelvis to follow evolution of bone micro-architecture in the #2 zone of De Lee and Charnley. Texture analysis using fractal algorithms was done at D0, 6 months, 1, 2 and 5 years post-THA. The fractal methods used included the skyscrapers and the dynamic blanket methods with 3 different structuring elements (a cross, a horizontal and a vertical vector). RESULTS: The RM-Pressfit® caused significant changes in the distribution of strains around the acetabulum that preserved the bone volume over a 5-year period post-surgery. This corresponds to an improvement of the trabecular micro-architecture around the acetabular cups. CONCLUSION: A statistically significant increase in the four fractal dimensions considered corresponded to an improved trabecular bone micro-architecture revealed by texture analysis, a non-invasive method that can be used on digitized X-ray images. LEVEL OF EVIDENCE: IIIb, Case control study, retrospective design.

Human macrophages and osteoclasts resorb ß-tricalcium phosphate in vitro but not mouse macrophages.

ß-TCP is a resorbable bony biomaterial but its biodegradation mechanisms in vivo remains unclear. Osteoclast can resorb ß-TCP but a role for macrophages has also been suggested by in vivo studies. However no in vitro study has clearly evidenced the action of macrophages in the resorption process. We prepared flat ß-TCP tablets with a smooth surface to investigate the in vitro capability of murine (RAW 264.7) and human macrophage cells (PBMCs) to resorb the biomaterial. In parallel, these cells were differentiated into multinucleated osteoclasts with M-CSF and RANK-L. The action of these cells was evaluated by scanning electron microscopy and Raman microspectroscopy after a 21 day culture on the tablets. Human macrophages and osteoclasts derived from PBMCs appeared able to resorb ß-TCP by forming resorption pits at the surface of the flat tablets. RAW macrophages were unable to resorb ß-TCP but they exhibited this possibility when they have been differentiated into osteoclasts. These cells can engulf ß-TCP grains in their cytoplasm as evidenced by light and TEM microscopy with production of carbonic anhydrase (revealed by the immunogold technique in TEM). The resorbed areas were characterized by severe degradation of the grains showing speckled and stick-like aspects indicating a chemical corrosion. The effect was maximal at the grain boundaries which have a slightly different chemical composition. Changes in the Raman spectrum were observed between the resorbed and un-resorbed ß-TCP suggesting crystal modifications. In contrast, un-differentiated murine macrophages were not able to chemically attack ß-TCP and no resorption pit was observed. RAW cell is not a representative model of the macrophage-biomaterial interactions that occur in human. This in vitro study evidences that both human osteoclasts and macrophages represent active cell populations capable to resorb ß-TCP.

Maxillary sinus floor elevation using Beta-Tricalcium-Phosphate (beta-TCP) or natural bone: same inflammatory response.

Sinus elevation is a common procedure to increase bone volume in the atrophic maxilla to allow placement of dental implants. Autogenous bone is the gold standard but is limited in quantity and causes morbidity at the donor site. ß-TCP is a synthetic biomaterial commonly used in that purpose. It appears to induce a poor inflammatory response. This study aimed to evaluate the degree of edema of the sinus mucosa after sinus lift surgery according to the type of biomaterial. Forty sinuses (20 patients) were included retrospectively and divided into 2 groups according to the biomaterial that was used: synthetic biomaterial (BTCP group), natural bone (BONE group). A control group (CTRL group) was constituted by the non-grafted maxillary sinuses. Twelve measurements per sinus were realized on pre- and post-operative computed tomography and averaged to provide the sinus membrane thickness value (SM.Th). SM.Th was thicker post-operatively in the BTCP and BONE groups in comparison with the CTRL group and in comparison with pre-operative measurements. No difference was found post operatively between the BTCP and BONE groups. We found that a synthetic biomaterial (ß-TCP) induced the same degree of edema, and thus of inflammation, as natural bone. It constitutes therefore an interesting alternative to autogenous bone for maxillary sinus lifts.

Giant cells and osteoclasts present in bone grafted with nacre differ by nuclear cytometry evaluated by texture analysis.

Nacre (mother of pearl) is a natural biomaterial used to prepare orthopedic devices. We have implanted screws and plates made with nacre in five sheeps. Bone were harvested after two months and embedded in poly(methyl methacrylate). Blocks were saws and the thick slabs were grinded, polished and surface stained. Sections were photographed at an ×1000 magnification. Giant cells were found in contact with nacre in eroded areas and true osteoclasts were found at distance in the neighboring bone in Howship lacunae. A texture analysis of the nuclei of giant cells and osteoclasts was done using the run-length method of the MaZda freeware. The size of the nuclei was reduced in osteoclast and their mean gray level appeared reduced. Texture analysis revealed that chromatin had a completely different pattern in giant cells when compared to osteoclasts. Giant cells had a fine repartition of the chromatin with large clear areas around prominent nucleoli. On the contrary, osteoclast nuclei had chromatin blocks evenly dispersed in the nuclei. This reflects the different origin of these cells expressing different functions.

Sclerostin antibody reduces long bone fractures in the oim/oim model of osteogenesis imperfecta.

Osteogenesis imperfecta type III (OI) is a serious genetic condition with poor bone quality and a high fracture rate in children. In a previous study, it was shown that a monoclonal antibody neutralizing sclerostin (Scl-Ab) increases strength and vertebral bone mass while reducing the number of axial fractures in oim/oim, a mouse model of OI type III. Here, we analyze the impact of Scl-Ab on long bones in OI mice. After 9 weeks of treatment, Scl-Ab significantly reduced long bone fractures (3.6 ±â€¯0.3 versus 2.1 ±â€¯0.8 per mouse, p < 0.001). In addition, the cortical thickness of the tibial midshaft was increased (+42%, p < 0.001), as well as BMD (+28%, p < 0.001), ultimate load (+86%, p < 0.05), plastic energy (+184%; p < 0.05) and stiffness (+172%; p < 0.01) in OI Scl-Ab mice compared to OI vehicle controls. Similar effects of Scl-Ab were observed in Wild type (Wt) mice. The plastic energy, which reflects the fragility of the tissue, was lower in the OI than in the Wt and significantly improved with the Scl-Ab treatment. At the tissue level by nanoindentation, Scl-Ab slightly increased the elastic modulus in bones of both OI and Wt, while moderately increasing tissue hardness (+13% compared to the vehicle; p < 0.05) in Wt bones, but not in OI bones. Although it did not change the properties of the OI bone matrix material, Scl-Ab reduced the fracture rate of the long bones by improving its bone mass, density, geometry, and biomechanical strength. These results suggest that Scl-Ab can reduce long-bone fractures in patients with OI.