Distal forelimb radiographic bone morphology in Thoroughbred foals during the first 10 months post-partum. Part 1: Carpus.

BACKGROUND: The risk of carpal injury in racehorses may be related to the morphology, yet whether carpal morphologies are set from birth or change through growth remains unclear. OBJECTIVE: To quantify carpal bone changes through growth. METHOD: Twenty privately owned Thoroughbred foals born between January 2022 and May 2023 were radiographed bimonthly from birth to 10 months of age. Imprint training was used to take radiographs safely without chemical restraints. Fifteen individual and 11 relative angular carpal parameters were measured using ImageJ on dorsopalmar radiographs of the carpus at zero degrees of vertical and horizontal rotation. Associations with age (growth), sex and the differences between left and right limbs were analysed separately using a linear mixed effects model. RESULTS: Six individual carpal parameters changed with age (radial carpal joint [RCJ], Prx.dor. radial carpal [Cr], Prx.Cu, Dis.dor. third carpal [C3], Dis.pal.C3 and Dis.pal. intermediate carpal), and one was influenced by side, that is higher in the left carpus (Dis.pal.Cr). Seven relative parameters changed with age, and one relative parameter was influenced by side, that is higher in the left (Ra.met-RCJ). The proximo-dorsal bone surface angle of Cr and disto-dorsal bone surface angle of C3 became flatter over time, which may be associated with the re-direction of the load towards the sagittal carpal plane. Sex did not influence any of the carpal parameters, nor did the combined effect of age, side of the limb and sex. CONCLUSION: Specific individual and relative angular carpal parameters changed significantly over time and some differed between the left and right limb, whereas other parameters did not change. The steeper carpal bone angles achieved proximally with the parameters that did change may improve stability by redirecting the load more medially through the carpus and the proximal and distal bones.

Finite element analysis of patient-specific additive-manufactured implants.

Introduction: Bone tumors, characterized by diverse locations and shapes, often necessitate surgical excision followed by custom implant placement to facilitate targeted bone reconstruction. Leveraging additive manufacturing, patient-specific implants can be precisely tailored with complex geometries and desired stiffness, enhancing their suitability for bone ingrowth. Methods: In this work, a finite element model is employed to assess patient-specific lattice implants in femur bones. Our model is validated using experimental data obtained from an animal study (n = 9). Results: The results demonstrate the accuracy of the proposed finite element model in predicting the implant mechanical behavior. The model was used to investigate the influence of reducing the elastic modulus of a solid Ti6Al4V implant by tenfold, revealing that such a reduction had no significant impact on bone behavior under maximum compression and torsion loading. This finding suggests a potential avenue for reducing the endoprosthesis modulus without compromising bone integrity. Discussion: Our research suggests that employing fully lattice implants not only facilitates bone ingrowth but also has the potential to reduce overall implant stiffness. This reduction is crucial in preventing significant bone remodeling associated with stress shielding, a challenge often associated with the high stiffness of fully solid implants. The study highlights the mechanical benefits of utilizing lattice structures in implant design for enhanced patient outcomes.

Reducing the prosthesis modulus by inclusion of an open space lattice improves osteogenic response in a sheep model of extraarticular defect.

Introduction: Stress shielding is a common complication following endoprosthetic reconstruction surgery. The resulting periprosthetic osteopenia often manifests as catastrophic fractures and can significantly limit future treatment options. It has been long known that bone plates with lower elastic moduli are key to reducing the risk of stress shielding in orthopedics. Inclusion of open space lattices in metal endoprostheses is believed to reduce the prosthesis modulus potentially improving stress shielding. However, no in vivo data is currently available to support this assumption in long bone reconstruction. This manuscript aims to address this hypothesis using a sheep model of extraarticular bone defect. Methods: Initially, CT was used to create a virtual resection plan of the distal femoral metaphyses and to custom design endoprostheses specific to each femur. The endoprostheses comprised additively manufactured Ti6Al4V-ELI modules that either had a solid core with a modulus of ∼120 GPa (solid implant group) or an open space lattice core with unit cells that had a modulus of 3-6 GPa (lattice implant group). Osteotomies were performed using computer-assisted navigation followed by implantations. The periprosthetic, interfacial and interstitial regions of interest were evaluated by a combination of micro-CT, back-scattered scanning electron microscopy (BSEM), as well as epifluorescence and brightfield microscopy. Results: In the periprosthetic region, mean pixel intensity (a proxy for tissue mineral density in BSEM) in the caudal cortex was found to be higher in the lattice implant group. This was complemented by BSEM derived porosity being lower in the lattice implant group in both caudal and cranial cortices. In the interfacial and interstitial regions, most pronounced differences were observed in the axial interfacial perimeter where the solid implant group had greater bone coverage. In contrast, the lattice group had a greater coverage in the cranial interfacial region. Conclusion: Our findings suggest that reducing the prosthesis modulus by inclusion of an open-space lattice in its design has a positive effect on bone material and morphological parameters particularly within the periprosthetic regions. Improved mechanics appears to also have a measurable effect on the interfacial osteogenic response and osteointegration.

A customized 3D-printed histological microgrinder for the study of metallic endoprostheses.

Use of histology is the key when evaluation of bone and soft tissue integration of any implanted metallic prosthesis is required. This relies on the ability to prepare very thin sections by grinding down resin embedded samples. Manual grinding has historically been used with variable success, and thus, a number of commercial microgrinders have been previously marketed, however, at a significant cost. The following describes a practical method to 3D print and build a microgrinder construct retrofitted to a metallurgic wheel grinder/polisher previously available. The design files are also supplied, which allow one to implement customized modifications for virtually all types of wheel grinders/polishers circumventing the need to procure highly costly appliances. Recommendations are included on how to safely and reproducibly prepare microscopic sections using the described construct.

Effect of a protease-activated receptor-2 antagonist (GB88) on inflammation-related loss of alveolar bone in periodontal disease.

BACKGROUND AND OBJECTIVE: Protease-activated receptor-2 (PAR2 ), a pro-inflammatory G-protein coupled receptor, has been associated with pathogenesis of periodontitis and the resulting bone loss caused by oral pathogens, including the keystone pathogen Porphyromonas gingivalis (P. gingivalis). We hypothesised that administration of a PAR2 antagonist, GB88, might prevent inflammation and subsequent alveolar bone resorption in a mouse model of periodontal disease. METHODS: Periodontitis was induced in mice by oral inoculations with P. gingivalis for a total of eight times over 24 days. The infected mice were treated with either GB88 or vehicle for the duration of the trial. Following euthanasia on day 56, serum was collected and used for the detection of mast cell tryptase. The right maxillae were defleshed and stained with methylene blue to measure the exposed cementum in molar teeth. The left maxillae were prepared for cryosections followed by staining for tartrate-resistant acid phosphatase to identify osteoclasts or with toluidine blue to identify mast cells. Reverse transcription quantitative PCR (RT-qPCR) was used to quantify the expression of inflammatory cytokines in the gingival tissue. Supernatants of T-lymphocyte cultures isolated from the regional lymph nodes were assayed using a cytometric bead array to measure the Th1/Th2/Th17 cytokine levels. RESULTS: Measurement of the exposed cementum showed that GB88 reduced P. gingivalis-induced alveolar bone loss by up to 69%. GB88 also prevented the increase in osteoclast numbers observed in the infected mice. Serum tryptase levels were significantly elevated in both the infected groups, and not altered by treatment. RT-qPCR showed that GB88 prevented the upregulation of Il1b, Il6, Ifng and Cd11b. In T-lymphocyte supernatants, only IFNγ and IL-17A levels were increased in response to infection, but this was prevented by GB88 treatment. CONCLUSIONS: GB88 significantly reduced osteoclastic alveolar bone loss in mice infected with P. gingivalis, seemingly by preventing the upregulation of several inflammatory cytokines. PAR2 antagonism may be an effective treatment strategy for periodontal disease.

Protease-activated receptor-2 dependent and independent responses of bone cells to prostate cancer cell secretory products.

BACKGROUND: Metastatic prostate cancer lesions in the skeleton are frequently characterized by excessive formation of bone. Prostate cancer cells secrete factors, including serine proteases, that are capable of influencing the behavior of surrounding cells. Some of these proteases activate protease-activated receptor-2 (PAR2 ), which is expressed by osteoblasts (bone-forming cells) and precursors of osteoclasts (bone-resorbing cells). The aim of the current study was to investigate a possible role for PAR2 in regulating the behavior of bone cells exposed to metastatic prostate cancer cells. METHODS: The effect of medium conditioned by the PC3, DU145, and MDA-PCa-2b prostate cancer cell lines was investigated in assays of bone cell function using cells isolated from wildtype and PAR2 -null mice. Osteoclast differentiation was assessed by counting tartrate-resistant acid phosphatase-positive multinucleate cells in bone marrow cultured in osteoclastogenic medium. Osteoblasts were isolated from calvariae of neonatal mice, and BrdU incorporation was used to assess their proliferation. Assays of alkaline phosphatase activity and quantitative PCR analysis of osteoblastic gene expression were used to assess osteoblast differentiation. Responses of osteoblasts to medium conditioned by MDA-PCa-2b cells were analyzed by RNAseq. RESULTS: Conditioned medium (CM) from all three cell lines inhibited osteoclast differentiation independently of PAR2 . Media from PC3 and DU145 cells had no effect on assays of osteoblast function. Medium conditioned by MDA-PCa-2b cells stimulated BrdU incorporation in both wildtype and PAR2 -null osteoblasts but increased alkaline phosphatase activity and Runx2 and Col1a1 expression in wildtype but not PAR2 -null cells. Functional enrichment analysis of RNAseq data identified enrichment of multiple gene ontology terms associated with lysosomal function in both wildtype and PAR2 -null cells in response to MDA-PCa-2b-CM. Analysis of individual genes identified osteogenesis-associated genes that were either upregulated by MDA-PCa-2b-CM selectively in wildtype cells or downregulated selectively in PAR2 -null cells. CONCLUSIONS: Factors secreted by prostate cancer cells influence bone cell behavior through both PAR2 -dependent and -independent mechanisms. Both PAR2 -independent suppression of osteoclast differentiation and PAR2 -dependent stimulation of osteogenesis are likely to determine the nature of prostate cancer metastases in bone.

Evaluation of Osteogenic Potentials of Avian Demineralized Bone Matrix in the Healing of Osseous Defects in Pigeons.

OBJECTIVES: To evaluate avian allogeneic demineralized bone matrix (DBM) in the healing of long bone defects as a function of geometry and time in a pigeon model. STUDY DESIGN: Experimental. ANIMALS: Adult rock pigeons (n = 60). METHODS: Midshaft ulnar osseous defects were grafted with 2 geometric forms of DBM (tubular vs. chipped) and stabilized with a hybrid fixator. Autologous chips of sternal keel were used in a third group as control. Outcomes were evaluated by radiography and histology/histomorphometry at 4, 8, 12, and 24 weeks postoperatively. RESULTS: Despite an early rapid healing response, autografts plateaued (histologic score and new bone area) by 8 weeks with no significant improvement afterwards. Conversely, allogeneic DBM implants demonstrated continuous temporal improvement in bone healing, and tubular DBM finally outpaced autograft implants after week 12 with values for metrics achieving statistical significance by week 24. Chip DBM was inferior to tubular DBM and autograft. CONCLUSIONS: Avian DBM is osteogenic, biocompatible, and safe in orthotopic sites with potential usefulness in avian bone grafting. Implant geometry (shape and size) affects such osteogenic potentials.

Intramedullary cement osteosynthesis (IMCO): a pilot study in sheep.

The application of bone substitutes and cements has a long standing history in augmenting fractures as a complement to routine fracture fixation techniques. Nevertheless, such use is almost always in conjunction with definite means of fracture fixation such as intramedullary pins or bone plates. The idea of using biomaterials as the primary fixation bears the possibility of simultaneous fixation and bone enhancement. Intramedullary recruitment of bone cements is suggested in this study to achieve this goal. However, as the method needs primary testings in animal models before human implementation, and since the degree of ambulation is not predictable in animals, this pilot study only evaluates the outcomes regarding the feasibility and safety of this method in the presence of primary bone fixators. A number of two sheep were used in this study. Tibial transverse osteotomies were performed in both animals followed by external skeletal fixation. The medullary canals, which have already been prepared by removing the marrow through proximal and distal drill holes, were then injected with calcium phosphate cement (CPC). The outcomes were evaluated postoperatively by standard survey radiographs, morphology, histology and biomechanical testings. Healing processes appeared uncomplicated until week four where one bone fracture recurred due to external fixator failure. The results showed 56% and 48% cortical thickening, compared to the opposite site, in the fracture site and proximal and distal diaphyses respectively. This bone augmentative effect resulted in 264% increase in bending strength of the fracture site and 148% increase of the same value in the adjacent areas of diaphyses. In conclusion, IMCO, using CPC in tibia of sheep, is safe and biocompatible with bone physiology and healing. It possibly can carry the osteopromotive effect of the CPCs to provide a sustained source of bone augmentation throughout the diaphysis. Although the results must be considered preliminary, this method has possible advantages over conventional methods of bone fixation at least in bones with compromised quality (i.e. osteoporosis and bone cysts), where rigid metal implants may jeopardize eggshell cortices.