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Medial proximal sesamoid bones (PSBs) from Thoroughbred racehorses that did (Case) or did not (Control) experience unilateral biaxial PSB fracture were evaluated for bone volume fraction (BVF), apparent mineral density (AMD), tissue mineral density (TMD), and microdamage in Case fractured, Case contralateral limb intact, and Control bones. A majority of Case bones had a subchondral lesion with high microdamage density, and low BVF, AMD, and TMD. Lesion microdamage and densitometric measures were associated with training history by robust linear regression. Exercise intensity was negatively related to BVF (0.07 ≤ R2  ≤ 0.12) and positively related to microcrack areal density (0.21 ≤ R2  ≤ 0.29) in the lesion; however, in an undamaged site, the relationships were opposite in direction. Regardless of location, TMD decreased with event frequency for both Case and Control, suggesting increased bone remodeling with exercise. Measures of how often animals were removed from active training (layups) predicted a decrease in TMD, AMD, BVF, and microdamage at regions away from the lesion site. A steady-state compartment model was used to organize the differences in the correlations between variables within the data set. The overall conclusions are that at the osteopenic lesion site, repair of microdamage by remodeling was not successful (e.g., lower bone mass, increased damage, and lower mineralization) but that in regions away from the lesion remodeling successfully controlled damage (e.g., higher bone mass, less microdamage, and lower mineralization).

ObjectiveTo evaluate the biomechanical properties of the mandibles of cats with experimentally created osteotomies simulating oblique ramus fractures, which were stabilized with malleable L-miniplates with either locking screws [locking construct (LC)] or nonlocking screws [nonlocking construct (NLC)], compared with those for intact mandibles.Samples20 mandibles from 10 adult cat cadavers.ProceduresA block study design was adopted to allocate the mandibles of each cadaver to 2 of the 3 test groups (LC, NLC, or intact mandible). Mandibles within each cadaver were allocated systematically to a test group. For mandibles assigned to an LC and an NLC, a complete oblique osteotomy was performed from the mid rostral aspect of the ramus in a caudoventral direction. All mandibles were loaded in a single-load-to-failure test through cantilever bending. Load and actuator displacement were recorded simultaneously. Mode of failure and radiographic evidence of damage to tooth roots and the mandibular canal were evaluated. Biomechanical properties were compared among the groups.ResultsNo iatrogenic tooth root damage was evident, but all mandibles with an LC and an NLC had evidence of screw invasion into the mandibular canal. Plated mandibles had significantly less stiffness and bending moment than intact mandibles. Stiffness was not significantly different between the LC and the NLC; the NLC had a greater bending moment at failure than the LC. The pre-yield stiffness of plated mandibles decreased when the number of screw holes overlapping the mandibular canal increased.Clinical relevanceThe use of a malleable L-miniplate in a caudal mandibular fracture model is feasible. Both the LC and the NLC were inferior mechanically to intact mandibles. Type of construct used did not affect the construct stiffness significantly in tested mandibles.

The most prevalent causes of death in racehorses are musculoskeletal injuries, causing ~83% of deaths within the racing industry in California and elsewhere. The vast majority of these injuries have preexisting lesions that predispose to fatal injury. A 4-y-old Thoroughbred colt suffered an acute suspensory apparatus failure, including biaxial proximal sesamoid bone fractures of the right front fetlock, causing loss of support of the fetlock joint and consequent fall with fractures of the cervical and sacral spine. Cervical fracture caused spinal cord damage that resulted in sudden death. A preexisting lesion in the medial proximal sesamoid bone likely predisposed to complete fracture of this bone and fetlock breakdown. Interestingly, a comparable osteopenic lesion was present in the intact medial proximal sesamoid bone of the left forelimb, which is consistent with bilateral repetitive overuse injury in racehorses. The morphologic features of the cervical and sacral spine fractures were compatible with acute injury; no evidence of preexisting lesions was seen. Most likely, these acute vertebral fractures occurred as a result of the horse falling. This case emphasizes the importance of performing a detailed autopsy in horses that suffer an appendicular musculoskeletal injury, particularly in fatal cases when the horse dies following a leg injury.

OBJECTIVE:To evaluate and quantify the kinematic behavior of canine mandibles before and after bilateral rostral or unilateral segmental mandibulectomy as well as after mandibular reconstruction with a locking reconstruction plate in ex vivo conditions. SAMPLE:Head specimens from cadavers of 16 dogs (range in body weight, 30 to 35 kg). PROCEDURE:Specimens were assigned to undergo unilateral segmental (n = 8) or bilateral rostral (8) mandibulectomy and then mandibular reconstruction by internal fixation with locking plates. Kinematic markers were attached to each specimen in a custom-built load frame. Markers were tracked in 3-D space during standardized loading conditions, and mandibular motions were quantified. Differences in mandibular range of motion among 3 experimental conditions (before mandibulectomy [ie, with mandibles intact], after mandibulectomy, and after reconstruction) were assessed by means of repeated-measures ANOVA. RESULTS:Both unilateral segmental and bilateral rostral mandibulectomy resulted in significantly greater mandibular motion and instability, compared with results for intact mandibles. No significant differences in motion were detected between mandibles reconstructed after unilateral segmental mandibulectomy and intact mandibles. Similarly, the motion of mandibles reconstructed after rostral mandibulectomy was no different from that of intact mandibles, except in the lateral direction. CONCLUSIONS AND CLINICAL RELEVANCE:Mandibular kinematics in head specimens from canine cadavers were significantly altered after unilateral segmental and bilateral rostral mandibulectomy. These alterations were corrected after mandibular reconstruction with locking reconstruction plates. Findings reinforced the clinical observations of the beneficial effect of reconstruction on mandibular function and the need for reconstructive surgery after mandibulectomy in dogs.

Osteoporosis has been associated with pulmonary silicosis in California horses exposed to soils rich in cytotoxic silica dioxide crystals, a syndrome termed silicate associated osteoporosis (SAO). The causal mechanism for the development of osteoporosis is unknown. Osteoporotic lesions are primarily located in bone marrow-rich sites such as ribs, scapula and pelvis. Gene transcription patterns within bone marrow and pulmonary lymph nodes of affected horses may offer clues to disease pathobiology. Bone marrow core and tracheobronchial lymph node tissue samples harvested postmortem from affected and unaffected horses were examined histologically and subjected to RNA sequencing (RNA-seq). Sequenced data were analyzed for differential gene expression and gene ontology. Metatranscriptomic and metagenomic assays evaluated samples for infectious agents. Thirteen of 17 differentially expressed transcripts in bone marrow were linked to bone and cartilage formation such as integrin binding bone sialoprotein (log2FC = 3.39, PFDR = 0.013) and chondroadherin (log2FC = 4.48, PFDR = 0.031). Equus caballus solute carrier family 9, subfamily A2 (log2FC = 3.77, PFDR = 0.0034) was one of the four differentially expressed transcripts linked to osteoclast activity. Osteoblasts were hyperplastic and hypertrophic in bone marrow from affected horses. Biological pathways associated with skeletal morphogenesis were significantly enriched in affected horses. The 30 differentially expressed genes in affected lymph nodes were associated with inflammatory responses. Evidence of infectious agents was not found. The SAO affected bone marrow molecular signature demonstrated increased transcription and heightened activation of osteoblasts. Increased osteoblastic activity could be part of the pathological mechanism for osteoporosis or a compensatory response to the accelerated osteolysis. Transcriptome data offer gene targets for inquiries into the role of osteocytes and osteoblasts in SAO pathogenesis. Viral or bacterial infectious etiology in SAO is less likely based on metatranscriptomic and metagenomic data but cannot be completely ruled out.

OBJECTIVE To evaluate biomechanical properties of intact feline mandibles, compared with those for mandibles with an experimentally created osteotomy that was stabilized with 1 of 2 internal fixation configurations. SAMPLE 20 mandibles from 10 adult feline cadavers. PROCEDURES An incomplete block study design was used to assign the mandibles of each cadaver to 2 of 3 groups (locking plate with locking screws [locking construct], locking plate with nonlocking screws [nonlocking construct], or intact). Within each cadaver, mandibles were randomly assigned to the assigned treatments. For mandibles assigned to the locking and nonlocking constructs, a simple transverse osteotomy was created caudal to the mandibular first molar tooth after plate application. All mandibles were loaded in cantilever bending in a single-load-to-failure test while simultaneously recording load and actuator displacement. Mode of failure (bone or plate failure) was recorded, and radiographic evidence of tooth root and mandibular canal damage was evaluated. Mechanical properties were compared among the 3 groups. RESULTS Stiffness, bending moments, and most post-yield energies for mandibles with the locking and nonlocking constructs were significantly lower than those for intact mandibles. Peak bending moment and stiffness for mandibles with the locking construct were significantly greater than those for mandibles with the nonlocking construct. Mode of failure and frequency of screw damage to tooth roots and the mandibular canal did not differ between the locking and nonlocking constructs. CONCLUSIONS AND CLINICAL RELEVANCE Results indicated that both fixation constructs were mechanically inferior to intact mandibles. The locking construct was mechanically stronger than the nonlocking construct.

ObjectiveTo determine if an interlocking bolt would limit subsidence of the biological fixation universal hip (BFX(®)) femoral stem under cyclic loading and enhance construct stiffness, yield, and failure properties.Study designEx vivo biomechanical study.AnimalsCadaveric canine femora (10 pairs).MethodsPaired femora implanted with a traditional stem or an interlocking stem (constructs) were cyclically loaded at walk, trot, and gallop loads while implant and bone motions were captured using kinematic markers and high-speed video. Constructs were then loaded to failure to evaluate failure mechanical properties.ResultsImplant subsidence was greater (P = .037) for the traditional implant (4.19 mm) than the interlocking implant (0.78 mm) only after gallop cyclic loading, and cumulatively after walk, trot, and gallop cyclic loads (5.20 mm vs. 1.28 mm, P = .038). Yield and failure loads were greater (P = .029 and .002, respectively) for the interlocking stem construct (1155 N and 2337 N) than the traditional stem construct (816 N and 1405 N). Version angle change after cyclic loading was greater (P = .020) for the traditional implant (3.89 degrees) than for the interlocking implant (0.16 degrees), whereas stem varus displacement at failure was greater (P = .008) for the interlocking implant (1.5 degrees) than the traditional implant (0.17 degrees).ConclusionAddition of a stabilizing bolt enhanced construct stability and limited subsidence of a BFX(®) femoral stem. Use of the interlocking implant may decrease postoperative subsidence. However, in vivo effects of the interlocking bolt on osseointegration, bone remodeling, and stress shielding are unknown.

Bone "stress-whitens," becoming visibly white during mechanical loading, immediately prior to failure. Stress-whitening is known to make materials tougher by dissipating mechanical energy. A greater understanding of stress-whitening, both an optical and mechanical phenomenon, may help explain age-related increases in fracture risk that occur without changes in bone mineralization. In this work, we directly measure the optical properties of demineralized bone as a function of deformation and immersing fluid (with different hydrogen-bonding potentials, water, and ethanol). The change in refractive index of demineralized bone was linear: with deformation and not applied force. Changes in refractive index were likely due to pushing low-refractive-index fluid out of specimens and secondarily due to changes in the refractive index of the collagenous phase. Results were consistent with stress-whitening of demineralized bone previously observed. In ethanol, the refractive index values were lower and less sensitive to deformation compared with deionized water, corroborating the sensitivity to fluid hydration. Differences in refractive index were consistent with structural changes in the collagenous phase such as densification that may also occur under mechanical loading. Understanding bone quality, particularly stress-whitening investigated here, may lead to new therapeutic targets and noninvasive methods to assess bone quality.

There is a need for improved biomaterials for use in treating non-healing bone defects. A number of natural and synthetic biomaterials have been used for the regeneration of bone tissue with mixed results. One approach is to modify native tissue via decellularization or other treatment for use as natural scaffolding for tissue repair. In this study, our goal was to improve on our previously published alternating solution immersion (ASI) method to fabricate a robust, biocompatible, and mechanically competent biomaterial from natural demineralized bone matrix (DBM). The improved method includes an antigen removal (AR) treatment step which improves mineralization and stiffness while removing unwanted proteins. The chemistry of the mineral in the remineralized bone matrix (RBM) was consistent with dicalcium phosphate dihydrate (brushite), a material used clinically in bone healing applications. Mass spectrometry identified proteins removed from the matrix with AR treatment to include α-2 HS-glycoprotein and osteopontin, noncollagenous proteins (NCPs) and known inhibitors of biomineralization. Additionally, the RBM supported the survival, proliferation, and differentiation of human mesenchymal stromal cells (MSCs) in vitro as well or better than other widely used biomaterials including DBM and PLG scaffolds. DNA content increased more than 10-fold on RBM compared to DBM and PLG; likewise, osteogenic gene expression was significantly increased after 1 and 2 weeks. We demonstrated that ASI remineralization has the capacity to fabricate mechanically stiff and biocompatible RBM, a suitable biomaterial for cell culture applications.

BackgroundRacetrack surface is a risk factor for racehorse injuries and fatalities. Current research indicates that race surface mechanical properties may be influenced by material composition, moisture content, temperature, and maintenance. Race surface mechanical testing in a controlled laboratory setting would allow for objective evaluation of dynamic properties of surface and factors that affect surface behavior.ObjectiveTo develop a method for reconstruction of race surfaces in the laboratory and validate the method by comparison with racetrack measurements of dynamic surface properties.MethodsTrack-testing device (TTD) impact tests were conducted to simulate equine hoof impact on dirt and synthetic race surfaces; tests were performed both in situ (racetrack) and using laboratory reconstructions of harvested surface materials. Clegg Hammer in situ measurements were used to guide surface reconstruction in the laboratory. Dynamic surface properties were compared between in situ and laboratory settings. Relationships between racetrack TTD and Clegg Hammer measurements were analyzed using stepwise multiple linear regression.ResultsMost dynamic surface property setting differences (racetrack-laboratory) were small relative to surface material type differences (dirt-synthetic). Clegg Hammer measurements were more strongly correlated with TTD measurements on the synthetic surface than the dirt surface. On the dirt surface, Clegg Hammer decelerations were negatively correlated with TTD forces.ConclusionsLaboratory reconstruction of racetrack surfaces guided by Clegg Hammer measurements yielded TTD impact measurements similar to in situ values. The negative correlation between TTD and Clegg Hammer measurements confirms the importance of instrument mass when drawing conclusions from testing results. Lighter impact devices may be less appropriate for assessing dynamic surface properties compared to testing equipment designed to simulate hoof impact (TTD).Potential relevanceDynamic impact properties of race surfaces can be evaluated in a laboratory setting, allowing for further study of factors affecting surface behavior under controlled conditions.