Significant Bone Growth Research Articles

Intermittent mechanical loading on mouse tibia accelerates longitudinal bone growth by inducing PTHrP expression in the female tibial growth plate.

It is not clear as to whether weight bearing and ambulation may affect bone growth. Our goal was to study the role of mechanical loading (one of the components of ambulation) on endochondral ossification and longitudinal bone growth. Thus, we applied cyclical, biologically relevant strains for a prolonged time period (4 weeks) to one tibia of juvenile mice, while using the contralateral one as an internal control. By the end of the 4-week loading period, the mean tibial growth of the loaded tibiae was significantly greater than that of the unloaded tibiae. The mean height and the mean area of the loaded tibial growth plates were greater than those of the unloaded tibiae. In addition, in female mice we found a greater expression of PTHrP in the loaded tibial growth plates than in the unloaded ones. Lastly, microCT analysis revealed no difference between loaded and unloaded tibiae with respect to the fraction of bone volume relative to the total volume of the region of interest or the tibial trabecular bone volume. Thus, our findings suggest that intermittent compressive forces applied on tibiae at mild-moderate strain magnitude induce a significant and persistent longitudinal bone growth. PTHrP expressed in the growth plate appears to be one growth factor responsible for stimulating endochondral ossification and bone growth in female mice.

Mechanical Properties, biodegradable Behavior and biocompatibility of Zn-0.5Ti alloy membranes produced by powder metallurgy for Guided bone regeneration

The Zn-0.5Ti alloy bone regeneration membranes, prepared via powder metallurgy, hot extrusion, and hot rolling, exhibit robust mechanical properties (tensile strength: 122.5 ± 1.8 MPa, elongation: 18.3 ± 0.7 %), appropriate degradation rate (0.127 mm/year in SBF solution), and good biocompatibility (cytotoxicity grade 0–1). After 4 and 8 weeks of implantation, the alloy showed significant bone growth (bone volume ratios (BV/TV): 11.3 ± 1.8 % and 23.1 ± 3.5 %, respectively, measured by Micro-CT), indicating promising potential as a GBR membrane material.

Multifunctional Design of Triply Periodic Minimal Surface Structures for Temporary Pediatric Fixation Devices

Fracture fixation and limb deformity correction in pediatric orthopedics often use temporary metallic fixation devices. These devices’ higher stiffness compared to cortical bone leads to stress shielding, causing significant bone density reduction, periprosthetic loosening, and bone growth interference. The 3D‐printed triply periodic minimal surface (TPMS) structures present a promising engineering solution to match bone stiffness while ensuring reliable implant strength. In this study, finite‐element modeling and experimental testing are employed to identify optimal multifunctional TPMS‐based lattices that meet the required design constraints of 1) stiffness in the range of cortical bone, 2) strength in the range of cortical bone, 3) minimum osteointegration to facilitate the implant removal after healing, and 4) manufacturability with limited defect sensitivity. Six different types of TPMS structures in Ti–6Al–4V material manufactured via laser powder bed fusion are evaluated for their ability to target the lower and upper bounds of pediatric cortical bone stiffness. Lattices based on the Primitive unit cell design are superior, demonstrating the highest strength/stiffness ratio, best manufacturability, and potentially reduced osteointegration due to larger pore size, smaller surface area, and smallest negative Gaussian curvature compared to other investigated TPMS types.

Evaluation of safety and efficacy of the bone marrow mesenchymal stem cell and gelatin-nano-hydroxyapatite combination in canine femoral defect repair.

Femoral shaft fracture is a common bone trauma in dogs. The limitation of mesenchymal stem cells in bone defect applications is that the cell suspension cannot be fixed to the bone defect site. In the study, our objective was to substantiate the combined application of canine bone marrow mesenchymal stem cells (cBMSCs) and gelatin-nano-hydroxyapatite (Gel-nHAP) and evaluate its therapeutic effect on bone defect diseases in dogs. Experiments were performed to evaluate the following: (1) the porosity of Gel-nHAP; (2) the adhesion of cBMSCs to Gel-nHAP; and (3) the effect of Gel-nHAP on cBMSC proliferation. The efficacy and safety of the combination of cBMSC and Gel-nHAP in the repair of femoral shaft defects were evaluated in animal experiments. The results showed that Gel-nHAP supported the attachment of cBMSCs and exhibited good biocompatibility. In the animal bone defect repair experiment, significant cortical bone growth was observed in the Gel-nHAP group at week 8 (p < 0.05) and in the cBMSCs-Gel-nHAP group at week 4 (p < 0.01). We demonstrated that Gel-nHAP could promote the repair of bone defects, and the effect of cBMSC-Gel-nHAP on the repair of bone defects was profound.

Guided Bone Regeneration in a Periodontally Compromised Individual with Autogenous Tooth Bone Graft: A Radiomics Analysis.

Autogenous tooth bone graft material (AutoBT) has been advocated as a bone substitute when conducting alveolar ridge preservation. This study is aimed at using a radiomics approach in order to evaluate and testify whether AutoBT can stimulate bone growth during socket preservation in severe periodontal cases. For this study, 25 cases with severe periodontal diseases were selected. The patients' AutoBTs were inserted into the extraction sockets and covered with Bio-Gide® collagen membranes. 3D CBCT scans and 2D X-rays were taken of the patients before surgery and after 6 months post-surgery. For the retrospective radiomics analysis, the maxillary and mandibular images were compared in different groups. Maxillary bone height was analyzed at the buccal, middle, and palatal crest sites, while the mandibular bone height was compared at the buccal, center, and lingual crest sites. In the maxilla, the alveolar height was increased by -2.15 ± 2.90 mm at the buccal crest; -2.45 ± 2.36 mm at the center of the socket, and -1.62 ± 3.19 mm at the palatal crest, while the height of the buccal crest was increased by 0.19 ± 3.52 mm, and the height at the center of the socket was increased by -0.70 ± 2.71 mm in the mandible. The three-dimensional radiomics analysis demonstrated significant bone growth in the local alveolar height and high density. Based on clinical radiomics analysis, AutoBT could be used as an alternative bone material in socket preservation after tooth extraction in patients with severe periodontitis.

Degradability and in vivo biocompatibility of micro-alloyed Mg-Ca-La alloys as orthopedic implants

This study describes the development of Mg-Ca-La alloys with trace La addition for orthopedic application, which aims to control the biosafety and degradation simultaneously. The Mg-0.2Ca-0.2La alloy exhibited slow and uniform degradation with only 5.9% reduction in implant volume 12 weeks post-implantation. Cytocompatibility study, using MC3T3-E1, showed significantly higher cell proliferation and osteogenic differentiation on Mg-0.2Ca-0.2La alloy surface. When implanted in rat femur, significant bone growth with increased trabecular thickness and number was found around Mg-0.2Ca-0.2La rod. Moreover, no local or systematic adverse effect was observed during the implantation period.

Functionally graded membrane: A novel approach in the treatment of gingival recession defects.

Background:Guided tissue regeneration has recently been advocated in re-constructing soft-tissue dimensions in recession defects. Advancement in nanotechnology has led to increased zest for approaches such as electrospinning of biologically active; nanofibrous functionally graded regenerative membranes for periodontal tissue engineering. A functionally graded membrane (FGM) had been tailored by incorporating chitosan and nano-hydroxyapatite over Amnion membrane and used in gingival recession defects.Study Design:It was single-blind, randomized controlled study. Split-mouth study was conducted in nine patients and 22 sites with recession defects were selected. Sites were divided into Group A (Amnion membrane with coronal advanced flap) and Group B (FGM with coronal advanced flap).Materials and Methods:Sites were assessed clinically by recording plaque index (PI), gingival index (GI), vertical recession defect depth (VRDD), relative clinical attachment level (CAL), and width of keratinized tissue at baseline, 3–6 months; and radiographically by recording linear bone growth by dentascan at baseline and 6 months.Result:Both groups showed statistically significant reduction in PI, GI and VRDD, and CAL and nonsignificant reduction in width of keratinized tissue at 3 and 6 months postoperatively. Group A showed statistically significant linear bone growth at 6 months. Group B also showed gain in linear bone growth at 6 months; however, result was statistically nonsignificant.Conclusion:FGM had shown favorable results by enhancing bone growth while preventing the gingival tissue downgrowth.

Synthesis of icariin in tobacco leaf by overexpression of a glucosyltransferase gene from Epimedium sagittatum

Epimedium plants are globally known for gardening and medicinal purposes. They are a potential resource of biologically active pharmaceutical compounds that can be used to treat various diseases. Among these compounds, icariin, a glycosylated flavonoid, has significant anti-tumor, bone growth, and neuroprotective effects; however, its biosynthetic pathway remains unknown. In this study, icariin accumulation during the development of leaves of Epimedium sagittatum was investigated. We identified a glycosyltransferase (Epimedium sagittatum glycosyltransferase, EsGT1) gene from this species using transcriptional analysis. Heterogeneously expressed EsGT1 protein transferred icariside II to icariin at the 7-OH site and its catalytic function was validated in vivo by the overexpression of the gene. In the recombinant protein assay, EsGT1 was also active in the glycosylation of kaempferol, an intermediate product of icariin. These results characterized the final step of glycosyltransferase in icariin synthesis and suggested an economical and efficient alternative source of icariin using a tobacco biotechnological method.

A composite hydrogel-3D printed thermoplast osteochondral anchor as example for a zonal approach to cartilage repair: in vivo performance in a long-term equine model

Recent research has been focusing on the generation of living personalized osteochondral constructs for joint repair. Native articular cartilage has a zonal structure, which is not reflected in current constructs and which may be a cause of the frequent failure of these repair attempts. Therefore, we investigated the performance of a composite implant that further reflects the zonal distribution of cellular component both in vitro and in vivo in a long-term equine model. Constructs constituted of a 3D-printed poly(ϵ-caprolactone) (PCL) bone anchor from which reinforcing fibers protruded into the chondral part of the construct over which two layers of a thiol-ene cross-linkable hyaluronic acid/poly(glycidol) hybrid hydrogel (HA-SH/P(AGE-co-G)) were fabricated. The top layer contained Articular Cartilage Progenitor Cells (ACPCs) derived from the superficial layer of native cartilage tissue, the bottom layer contained mesenchymal stromal cells (MSCs). The chondral part of control constructs were homogeneously filled with MSCs. After six months in vivo, microtomography revealed significant bone growth into the anchor. Histologically, there was only limited production of cartilage-like tissue (despite persistency of hydrogel) both in zonal and non-zonal constructs. There were no differences in histological scoring; however, the repair tissue was significantly stiffer in defects repaired with zonal constructs. The sub-optimal quality of the repair tissue may be related to several factors, including early loss of implanted cells, or inappropriate degradation rate of the hydrogel. Nonetheless, this approach may be promising and research into further tailoring of biomaterials and of construct characteristics seems warranted.

Orthodontical Management of Secondary Dental Migration Associated with a Reduced Periodontium: A Case Report

Introduction: Dental migrations following periodontal pathology may lead to significant aesthetic and functional complaints. In this case report, the presence of evident gingival recessions and mandibular crowding pushed us to opt for an ortho-periodontal multidisciplinary treatment plan, using extractions and incisive repositioning. Through this, we ensured tissue gain at the level of the superficial and deep periodontitis, thus avoiding the use of periodontal surgery.Clinical Case: A Moroccan 26-years-old female medical secretary who, being very concerned about her appearance and facial aesthetics, presented with pathological dental migrations following severe periodontitis, an unsatisfactory periodontal status, a clear mandibular crowding of the anterior inter-incisive diastemas, gingival recessions, attachment losses and quite large pocket depths with advanced bone resorptions in different areas. This required a specific multidisciplinary approach aiming at creating a healthy and well-structured periodontal environment through extensive periodontal treatment, combined with regular plaque control, and supplemented by an orthodontic treatment using extractions and incisive repositioning in order to avoid any vestibular movement that may worsen the recessions and will require possible overlapping. After 22 months of combined treatment, a stable occlusion was obtained with class I molar and canine relationship, perfect dental alignment, healthy periodontal architecture, and significant attachment gain and bone growth. In addition, the patient’s facial aesthetics and self-confidence have been significantly improved. Conclusion: The orthodontic treatment supplementing the meticulously planned periodontal therapy that was administered for this patient has helped to improve functionality, facial aesthetics as well as psychological self-confidence. However, it must be gradual with an application of mild forces and constant control of the periodontal status, for optimal tissue response.

Degradability and in vivo biocompatibility of doped magnesium phosphate bioceramic scaffolds

In the present study, degradability and in vivo biocompatibility of doped magnesium phosphate (MgP) scaffolds were evaluated. 0.5 wt% Zn, 0.5 wt% Si, and 2 wt% Sr doped MgP scaffolds were prepared with up to 47% porosity. Zn doped MgP scaffolds showed nearly 13% weight loss when immersed in SBF for 8 weeks whereas least degradation was noticed for Sr doped samples. In vitro cytocompatibility study, using MC3T3-E1, showed significantly higher cell proliferation at day 5 in a composition independent manner. When implanted in rabbit femur, micro-CT (µ-CT) results showed extensive degradation for Zn doped MgP scaffolds after 2 months while significant bone growth was noticed for both Si and Sr doped scaffolds.

Electrical stimulation shifts healing/scarring towards regeneration in a rat limb amputation model

Different species respond differently to severe injury, such as limb loss. In species that regenerate, limb loss is met with complete restoration of the limbs’ form and function, whereas in mammals the amputated limb’s stump heals and scars. In in vitro studies, electrical stimulation (EStim) has been shown to promote cell migration, and osteo- and chondrogenesis. In in vivo studies, after limb amputation, EStim causes significant new bone, cartilage and vessel growth. Here, in a rat model, the stumps of amputated rat limbs were exposed to EStim, and we measured extracellular matrix (ECM) deposition, macrophage distribution, cell proliferation and gene expression changes at early (3 and 7 days) and later stages (28 days). We found that EStim caused differences in ECM deposition, with less condensed collagen fibrils, and modified macrophage response by changing M1 to M2 macrophage ratio. The number of proliferating cells was increased in EStim treated stumps 7 days after amputation, and transcriptome data strongly supported our histological findings, with activated gene pathways known to play key roles in embryonic development and regeneration. In conclusion, our findings support the hypothesis that EStim shifts injury response from healing/scarring towards regeneration. A better understanding of if and how EStim controls these changes, could lead to strategies that replace scarring with regeneration.

Biodegradable polymerized simvastatin stimulates bone formation

Previous research from our labs demonstrated the synthesis of polymerized simvastatin by ring-opening polymerization and slow degradation with controlled release of simvastatin in vitro. The objective of the present study was to evaluate the degradation and intramembranous bone-forming potential of simvastatin-containing polyprodrugs in vivo using a rat calvarial onlay model. Poly(ethylene glycol)-block-poly(simvastatin) and poly(ethylene glycol)-block-poly(simvastatin)-ran-poly(glycolide) were compared with simvastatin conventionally encapsulated in poly(lactic-co-glycolic acid) (PLGA) and pure PLGA. The rate of degradation was higher for PLGA with and without simvastatin relative to the simvastatin polyprodrugs. Significant new bone growth at the circumference of poly(ethylene glycol)-block-poly(simvastatin) disks was observed beginning at 4 weeks, whereas severe bone resorption (4 weeks) and bone loss (8 weeks) were observed for PLGA loaded with simvastatin. No significant systemic effects were observed for serum total cholesterol and body weight. Increased expression of osteogenic (BMP-2, Runx2, and ALP), angiogenic (VEGF), and inflammatory cytokines (IL-6 and NF-ĸB) genes was seen with all polymers at the end of 8 weeks. Poly(ethylene glycol)-block-poly(simvastatin), with slow degradation and drug release, controlled inflammation, and significant osteogenic effect, is a candidate for use in bone regeneration applications. Statement of SignificanceTraditional drug delivery systems, e.g., drug encapsulated in poly(lactic-co-glycolic acid) (PLGA), are typically passive and have limited drug payload. As an alternative, we polymerized the drug simvastatin, which has multiple physiological effects, into macromolecules (“polysimvastatin”) via ring-opening polymerization. We previously demonstrated that the rate of degradation and drug (simvastatin) release can be adjusted by copolymerizing it with other monomers. The present results demonstrate significant new bone growth around polysimvastatin, whereas severe bone loss occurred for PLGA loaded with simvastatin. This degradable biomaterial with biofunctionality integrated into the polymeric backbone is a useful candidate for bone regeneration applications.

Release of CXCL12 From Apoptotic Skeletal Cells Contributes to Bone Growth Defects Following Dexamethasone Therapy in Rats.

Dexamethasone (Dex) is known to cause significant bone growth impairment in childhood. Although previous studies have suggested roles of osteocyte apoptosis in the enhanced osteoclastic recruitment and local bone loss, whether it is so in the growing bone following Dex treatment requires to be established. The current study addressed the potential roles of chemokine CXCL12 in chondroclast/osteoclast recruitment and bone defects following Dex treatment. Significant apoptosis was observed in cultured mature ATDC5 chondrocytes and IDG-SW3 osteocytes after 48 hours of 10-6 M Dex treatment, and CXCL12 was identified to exhibit the most prominent induction in Dex-treated cells. Conditioned medium from the treated chondrocytes/osteocytes enhanced migration of RAW264.7 osteoclast precursor cells, which was significantly inhibited by the presence of the anti-CXCL12 neutralizing antibody. To investigate the roles of the induced CXCL12 in bone defects caused by Dex treatment, young rats were orally gavaged daily with saline or Dex at 1 mg/kg/day for 2 weeks, and received an intraperitoneal injection of anti-CXCL12 antibody or control IgG (1 mg/kg, three times per week). Aside from oxidative stress induction systemically, Dex treatment caused reductions in growth plate thickness, primary spongiosa height, and metaphysis trabecular bone volume, which are associated with induced chondrocyte/osteocyte apoptosis and enhanced chondroclast/osteoclast recruitment and osteoclastogenic differentiation potential. CXCL12 was induced in apoptotic growth plate chondrocytes and metaphyseal bone osteocytes. Anti-CXCL12 antibody supplementation considerably attenuated Dex-induced chondroclast/osteoclast recruitment and loss of growth plate cartilage and trabecular bone. CXCL12 neutralization did not affect bone marrow osteogenic potential, adiposity, and microvasculature. Thus, CXCL12 was identified as a potential molecular linker between Dex-induced skeletal cell apoptosis and chondroclastic/osteoclastic recruitment, as well as growth plate cartilage/bone loss, revealing a therapeutic potential of CXCL12 functional blockade in preventing bone growth defects during/after Dex treatment. © 2018 American Society for Bone and Mineral Research.

Degradation of Bioresorbable Mg-4Zn-1Sr Intramedullary Pins and Associated Biological Responses in Vitro and in Vivo.

This article reports the degradation and biological properties of as-drawn Mg-4Zn-1Sr (designated as ZSr41) and pure Mg (P-Mg) wires as bioresorbable intramedullary pins for bone repair. Specifically, their cytocompatibility with bone marrow derived mesenchymal stem cells (BMSCs) and degradation in vitro, and their biological effects on peri-implant tissues and in vivo degradation in rat tibiae were studied. The as-drawn ZSr41 pins showed a significantly faster degradation than P-Mg in vitro and in vivo. The in vivo average daily degradation rates of both ZSr41 and P-Mg intramedullary pins were significantly greater than their respective in vitro degradation rates, likely because the intramedullary site of implantation is highly vascularized for removal of degradation products. Importantly, the concentrations of Mg2+, Zn2+, and Sr2+ ions in the BMSC culture in vitro and their concentrations in rat blood in vivo were all lower than their respective therapeutic dosages, i.e., in a safe range. Despite of rapid degradation with a complete resorption time of 8 weeks in vivo, the ZSr41 intramedullary pins showed a significant net bone growth because of stimulatory effects of the metallic ions released. However, proportionally released OH- ions and hydrogen gas caused adverse effects on bone marrow cells and resulted in cavities in surrounding bone. Thus, properly engineering the degradation properties of Mg-based implants is critical for harvesting the bioactivities of beneficial metallic ions, while controlling adverse reactions associated with the release of OH- ions and hydrogen gas. It is necessary to further optimize the alloy processing conditions and/or modify the surfaces, for example, applying coatings onto the surface, to reduce the degradation rate of ZSr41 wires for skeletal implant applications.

Vertical Bone Growth after Autotransplantation of Mature Third Molars: 2 Case Reports with Long-term Follow-up

Tooth autotransplantation is a viable treatment option for tooth replacement when a suitable donor tooth is available. These case reports described significant vertical bone growth after autotransplantation of a mature third molar. The left mandible third molars (n = 2) were transplanted to the missing tooth in the left mandible. The patient follow-up period was 10 years after transplantation. Clinical examination revealed no mobility of the transplanted tooth. Radiographic examination indicated that bone regeneration occurred around the transplanted tooth. Vertical bone growth was observed in the cervical area of the root surface and the recipient bone. In autotransplantation of mature teeth, long-term follow-up results indicate that vertical bone growth can be expected if viability of the periodontal ligament cells is maintained.

Statins as Bioactive Molecules to Support Bone Regeneration

Statins are currently used as an effective cholesterol-lowering medication through inhibition of the mevalonate pathway, but recent studies show their potential for bone repair. The bone anabolic effects of statins have been largely attributed to their ability to enhance BMP-2 expression in osteoblast cells. In vitro studies have demonstrated that statins can increase the expression of osteogenic and angiogenic markers such as alkaline phosphatase, vascular endothelial growth factor, and osteocalcin in cells. In vivo, statins have been shown to promote significant new bone growth when injected systemically or locally in combination with a scaffold. The potential anabolic effects of statins on bone make them attractive candidates to support bone regeneration. Since the molecular pathways by which statins induce osteoblast differentiation are still unclear, further investigations are required to elucidate the detailed cellular signaling mechanisms involved to determine the type of statin, optimal dose and mode of delivery to effectively utilize their anabolic effect. This also warrants the development of novel vehicles to locally deliver statins for the desired time periods to support optimal tissue regeneration in vivo.

Research on Biological Materials with Effect of Constant Magnetic Field Intervention on Bone Growth and Development in Rabbits

To investigate the effect of constant magnetic fields on bone deformation, 16 rabbits were randomly divided into the control group and the experimental group. Biological materials with effect of constant magnetic field were attached to the hind legs of rabbits in the experimental group for a period of 4 months. Rabbit bones were periodically measured and compared. Results showed that compared with the control group, rabbits in the experimental group exhibited significant bone growth, indicating that constant magnetic field may significantly activate bone growth in rabbits.

Controlled‐release of bone morphogenetic protein‐2 from a microsphere coating applied to acid‐etched Ti6AL4V implants increases biological bone growth in vivo

A central clinical challenge regarding the surgical treatment of bone and joint conditions is the eventual loosening of an orthopedic implant as a result of insufficient bone ingrowth at the bone-implant interface. We investigated the in vivo effectiveness of a coating containing recombinant human bone morphogenetic protein-2 (rhBMP-2)-loaded microspheres applied to acid-etched Ti6Al4V cylinders for implantation. Three groups of rabbits (24 per group) were used for implantation: (1) acid-etched Ti6Al4V implants coated with a mixture of rhBMP-2-loaded microspheres (125 ng rhBMP-2/mg microspheres) and α-butyl cyanoacrylate; (2) acid-etched, uncoated implants; and (3) bare, smooth uncoated implants. After implantation, 12 rabbits from each group were used for bone ingrowth determination at 4, 5, 6, 7, 8, and 12 weeks (2 rabbits per time point), while the remainder were used for histological analysis and push-out testing at 12 weeks. Scanning electron microscopy showed significant improvement in bone growth of the rhBMP-2 microspheres/α-butyl cyanoacrylate group compared with the other groups (p<0.01). Histological analysis and push-out testing also demonstrated enhanced bone growth of the rhBMP-2 group over that in the other two groups (p<0.01). The rhBMP-2 group showed the most significant bone growth, suggesting that coating acid-etched implants with a mixture of rhBMP-2-loaded microspheres and α-butyl cyanoacrylate may be an effective method to improve the osseointegration of orthopedic implants.

Efeitos do ultra-som terapêutico sobre o crescimento longitudinal do fêmur e da tíbia em ratos

OBJETIVO: Estudar efeitos do ultra-som terapêutico sobre o crescimento do fêmur e da tíbia, em ratos jovens. MÉTODO: Ratus norvegicus com quatro semanas de vida, machos, totalizando 115 animais, divididos em quatro grupos, foram submetidos ao ultra-som terapêutico (0,8 MHz, cabeçote fixo, pulso contínuo, por dez minutos, durante dez dias), na face medial do joelho direito, nas potências de 0,0 W/cm2 (grupo controle), 0,5 W/cm2 (grupo G2), 1,0 W/cm2 (grupo G3), e 1,5 W/cm2 (grupo G4). Lâminas histológicas da epífise, placa de crescimento e metáfise e as medidas dos comprimentos do fêmur e da tíbia foram estudadas na sexta, décima terceira e vigésima sexta semanas de vida. Os dados foram submetidos à análise de variância - fatorial inteiramente aleatorizado. RESULTADO: Nenhuma alteração estatística do crescimento ósseo foi estabelecida entre quaisquer dos três grupos tratados e o grupo controle. Entretanto, alterações sugestivas de diminuição do crescimento do fêmur e da tíbia de G4 em relação a G2 e G3, foram evidenciadas. No grupo G4 alterações histopatológicas como necroses celulares e neoformação óssea, pós-necrose, foram encontradas. CONCLUSÃO: Quando comparados os grupos tratados com o grupo controle, não há evidência estatística de estímulo ou inibição ao crescimento ósseo pela aplicação do ultra-som terapêutico. Nível de Evidência: Nível II, estudo coorte transversal.