Journal of Orthopaedic Research®最新文献

Heterotopic ossification (HO) in Achilles tendon often arises due to endochondral ossification during the healing process following trauma. Retinoic acid receptor γ (RARγ) plays a critical role in this phenomenon. This study aims to elucidate the therapeutic effects of CD1530, an RARγ selective agonist, along with the contributing cells, in Achilles tendon healing, utilizing a cell lineage tracing system. Local injection of CD1530 facilitated histological tendon healing by inhibiting chondrification in a mouse Achilles rupture model. Resident Scleraxis (Scx)⁺ cells in Achilles tendon were not found to be actively involved in HO or　tendon healing following injury. Instead, these processes were primarily driven by tendon stem/progenitor cells (TSPC)-like cells. Furthermore, an in vitro assay revealed that CD1530 attenuated inflammation in injured Achilles tendon-derived tendon fibroblasts (iATF) and inhibited the chondrogenesis of iATF. This dual effect suggests the potential of CD1530 in effectively modulating the healing environment during tendon healing. Together, the present study demonstrated that the local administration of CD1530 accelerated tendon healing by modulating the healing environment, including reducing chondrification via targeting TSPC-like cells in a mouse Achilles tendon rupture model. These results suggest that CD1530 may have the potential to be a novel tendon therapy that offers benefits via the inhibition of chondrogenesis.

Distraction osteogenesis (DO) is widely utilized for treating limb length discrepancy, nonunion, bone deformities and defects. This study sought to develop a 4D time-lapse morphometry method to quantify bone formation and resorption in mouse femur during DO based on image registration of longitudinal in vivo micro-CT scans. Female C57BL/6 mice (n = 7) underwent osteotomy, followed by 5 days of latency, 10 days of distraction and 35 days of consolidation. The mice were scanned with micro-CT at Days 5, 15, 25, 35, 45, and 50. Histological sectioning and Movat Pentachrome straining were performed at Day 50. After registration of two consecutive micro-CT images of the same bone (day x and day y), the spatially- and temporally-linked sequences of formation, resorption and quiescent bones at the distraction gap were identified and bone formation and resorption rates (BFR_dayx-y and BRR_dayx-y) were calculated. The overall percentage error of the registration method was 2.98% ± 0.89% and there was a strong correlation between histologically-measured bone area fraction and micro-CT-determined bone volume fraction at Day 50 (r = 0.89, p < 0.05). The 4D time-lapse morphometry indicated a rapid bone formation during the first 10 days of the consolidation phase (BFR_day15–25 = 0.14 ± 0.05 mm³/day), followed by callus reshaping via equivalent bone formation and resorption rates. The 4D time-lapse morphometry method developed in this study allows for a continuous quantitative monitoring of the dynamic process of bone formation and resorption following distraction, which may offer a better understanding of the mechanism for mechano-regulated bone regeneration and aid for development of new treatment strategies of DO.

Orthopaedic researchers need new strategies for engaging underrepresented minority (URM) students. Our field has demonstrated noticeable gaps in racial, ethnic, and gender diversity, which inhibit our ability to innovate and combat the severe socioeconomic burden of musculoskeletal disorders. Towards this goal, we designed, implemented, and evaluated Learning on a Limb (LoaL), an orthopaedic research outreach module to teach URM high school students about orthopaedic research. During the 4-h module, students completed hands-on activities to learn how biomechanical testing, microcomputed tomography, cell culture, and histology are used in orthopaedic research. Over 3 years, we recruited 32 high school students from the Greater Philadelphia Area to participate in LoaL. Most participants identified as racial/ethnic or gender minorities in orthopaedic research. Using pre/post-tests, we found that students experienced significant learning gains of 51 percentage points from completing LoaL. In addition to teaching students about orthopaedic research, post-survey data demonstrated that participating in LoaL strongly influenced students' interest in orthopaedic research and scientific confidence. Several students acted on this interest by completing summer research experiences in the McKay Orthopaedic Research Laboratory at the University of Pennsylvania. LoaL instructors also benefited by having the opportunity to “pay it forward” to the next generation of students and build community within their department. Empowering institutions to host modules like LoaL would synergistically inspire URM high school students and strengthen community within orthopaedic departments to ultimately enhance orthopaedic research innovations.

Knee meniscus tearing is a common orthopaedic injury that can heal poorly if left untreated, increasing the risk of post-traumatic Osteoarthritis. Intraarticular injection of human cartilage-derived progenitor cells (CPCs) has been shown to promote meniscus healing after injury. However, the mechanism by which CPCs stimulated this effect was unclear. The purpose of this study was to determine the paracrine effects that CPC-derived extracellular vesicles (EVs) have on native meniscal cells during healing. EVs from human CPCs and marrow-derived stromal cells were isolated via ultracentrifugation. EVs produced by each cell type were quantified, and their sizes were determined via NanoSight. EV protein expression was characterized via western blot. Meniscal fibrochondrocyte cellular metabolic activity (as an indicator of cell viability and proliferation) following treatment with EVs, was quantified using MTT and ATP assays. A 2D wound healing assay was used to determine the effects of treating inner meniscal fibrochondrocytes with EVs in a dose-dependent manner. Gene expression analysis for chondrogenesis genes was performed via RT-qPCR on inner meniscal fibrochondrocytes following treatment with EVs. Our results showed that CPCs produced a wide size range of EVs expressing CD9, CD81, and HSP70. Treatment of inner meniscal fibrochondrocytes with CPC-EVs improved 2D wound healing, in comparison to EVs isolated from marrow-derived stromal cell controls. CPC-EV treatment increased Type II Collagen mRNA expression in inner meniscal fibrochondrocytes. These findings demonstrate that CPC-EVs stimulate chondrogenic matrix production and wound healing in meniscal cells at the optimal dose of 1.0 × 10⁷ particles/mL, significantly outperforming the effects of marrow stromal cell-derived EVs.

Fracture site motion is thought to play an important role in the healing of complex fractures of the distal femur via mechanotransduction. Measuring this motion in vivo is challenging, and this has led researchers to turn to finite element modeling approaches to gain insights into the mechanical environment at the fracture site. Developing a systematic understanding of the effect of different model choices for distal femur fractures may allow more accurate prediction of fracture site motion from these types of simulations. In this study, we aim to assess the effect of four different modeling choices and parameters. We looked at the effect of using bone specific density distributions vs generic values, employing landmark-based geometry generation, varying fracture alignment within clinically relevant ranges, and determining whether direct apposition of the fracture to the plate was achieved. For validation, five cadaveric femurs had fractures created and repaired with plated constructs, and these were then loaded and fracture site motion was directly measured. We found that using landmark based bone geometry and patient-specific bone density distributions had a minimal effect on the overall model predictions. Changing the alignment, particularly into varus and procurvatum could have a large (>50%) effect on predicted shear motion, as could direct apposition of the bone to the plate. These findings demonstrate that modeling choices can play an important role in simulating distal femur fracture mechanics, and it is particularly critical that patient customized models attempt to accurately represent alignment of the bone fragments and lateral plate apposition.

Osteoarthritis (OA) is a debilitating disease that impacts millions of individuals and has limited therapeutic options. A significant hindrance to therapeutic discovery is the lack of in vitro OA models that translate reliably to in vivo preclinical animal models. An alternative to traditional inflammatory cytokine models is the matrikine stimulation model, in which fragments of matrix proteins naturally found in OA tissues and synovial fluid, are used to stimulate cells of the joint. The objective of this study was to determine if matrikine stimulation of equine synovial fibroblasts and chondrocytes with fibronectin fragments (FN7-10) would result in an OA phenotype. We hypothesized that FN7-10 stimulation of equine articular cells would result in an OA phenotype with gene and protein expression changes similar to those previously described for human chondrocytes stimulated with FN7-10. Synovial fibroblasts and chondrocytes isolated from four horses were stimulated in monolayer culture for 6 or 18 h with 1 µM purified recombinant 42 kD FN7-10 in serum-free media. At the conclusion of stimulation, RNA was collected for targeted gene expression analysis and media for targeted protein production analysis. Consistent with our hypothesis, FN7-10 stimulation resulted in significant alterations to many important genes that are involved in OA pathogenesis including increased expression of IL-1β, IL-4, IL-6, CCL2/MCP-1, CCL5/RANTES, CXCL6/GCP-2, MMP-1, MMP-3, and MMP13. The results of this study suggest that the equine matrikine stimulation model of OA may prove useful for in vitro experiments leading up to preclinical trials.

Post-traumatic osteoarthritis develops following an inciting injury to a joint and results in cartilage degeneration. Mechanical loading, including articulation, drives anabolic responses in cartilage clinically, in vivo, and in vitro. Tribological articulation, or sliding of cartilage on a glass counterface, has long been used as an in vitro tool to study cartilage tissue behavior. However, it is unclear if tribological articulation affects chondrocyte fate following injury, and if the timing of articulation impacts the resultant effect. The goal of this study was to investigate the effect of tribological articulation on injured cartilage tissue at two time points: (i) performed immediately after injury and (ii) 24 h after injury. Neonatal bovine femoral cartilage explants were injured using a rapid spring-loaded impactor and subsequently subjected to tribological articulation. Cell death due to impact injury was highest near the articular surface, suggesting a strain-dependent mechanism. Immediate articulation following injury mitigated cell death compared to injury alone or delayed articulation; markers for both general cell death and early-stage apoptosis were markedly decreased in the explants that were immediately slid. Interestingly, mitigation of cell death due to sliding was most predominant at the cartilage surface. Tribological articulation is known to create fluid flow within the tissue, predominantly at the articular surface, which could drive the protective response seen here. Altogether, this work shows that perturbations to the cellular environment immediately following cartilage injury significantly impact chondrocyte fate.

Computer assisted orthopedic surgery is used to improve precision. Electro-magnetic tracking has been shown to improve precision in mono-planar derotational osteotomies. However, studies are lacking to investigate its use in multiplanar osteotomies. For this purpose, 60 complex (derotation and extension) osteotomies were performed in standardized sawbones. Correction amount was randomly planned before the procedures. In 30 bones, the amount of correction was determined intraoperatively using conventional goniometric measurement while in the other 30 bones electro-magnetic tracking was used to guide the amount of correction. CT-scans were done before and after the procedures in all bones and the amount of correction was determined to compare the precision of the two techniques. Electromagnetic tracking resulted in a precision of 2.25° ± 1.77° for derotation and 1.38° ± 1.29° for extension, while precision for the conventional method was significantly lower. There was a significant relationship between goniometer measurement deviation and the absolute angle change for derotation and extension measurements with larger deviations for greater angle changes. For the electro-magnetic tracking, this correlation was observed only for derotation measurement. Electro-magnetic tracking represents an accurate method to control complex, multiplanar corrective osteotomies with superior precision in comparison to conventional goniometric measurement. Further research is needed to investigate the in-vivo accuracy and the effects on clinical outcome.

The success of radial head arthroplasty (RHA) relies on the design of the implant and precision of the surgical technique, with preoperative planning potentially playing a crucial role. The accurate establishment of a patient-specific anatomical coordinate system (ACS) is essential for this planning process. This study tested the hypothesis that an innovative automated method would be an accurate, reliable, and efficient framework to determine the ACS of the proximal radius, which would be a step toward improving the precision of RHA planning. We used advanced computational techniques to analyze 50 forearm CT scans, comparing the accuracy, reproducibility, reliability, and efficiency of the automated method with manually derived ACS using expert observers as benchmarks. The results showed that the automated approach was more accurate in identifying anatomical landmarks, with smaller mean distance discrepancies (0.6 mm) than manual observers (1 mm). Its reproducibility was also superior, with narrower reproducibility limits, particularly for ulnar notch landmarks (0.6 to 0.8 mm compared to manual selection 1.2 to 1.4 mm) (p = .01). In addition, the limits of agreement and the mean absolute rotational and translational differences of the axes were narrower for the automated method, which also reduced the construction time to an average of 46 s compared to 150 s manually (p < .001). These findings suggest that the automated method has the potential to enhance the accuracy and efficiency of preoperative and postoperative computer-assisted procedures for RHA. Further research is needed to fully understand the utility of this automated system for enhancing RHA computer-assisted surgical planning.

This study aimed to investigate the ultrastructural anatomy of the developing ACL tibial enthesis. We hypothesized that enthesis architecture would progressively mature and remodel, eventually resembling that of the adult by the early postnatal stage. Five fresh-frozen human pediatric cadaveric knees aged 1–36 months underwent anatomical dissection to harvest the ACL insertion and underlying tibial chondroepiphysis. The samples were prepared for scanning electron microscopy (SEM) to examine the ultrastructural anatomy of the enthesis and underwent histological staining for circular polarized light (CPL) and light microscopy imaging. SEM analysis of the 1- and 8-month-old samples revealed a shallow interdigitation between the dense fibrous (ligamentous) tissue and unmineralized chondrogenic tissues, with a minimal transition zone. By 11-month, a more complex transition zone was present. By age 19- and 36-month-old, a progressively more complex and defined fibrocartilage zone was observed. CPL analysis revealed distinct collagen fiber continuity, alignment, and organization changes over time. By 19 and 36 months, the samples exhibited complex fiber arrangements and a progression toward uniform fiber orientation. Similarly, histological analysis demonstrated progressive remodeling of the enthesis with increasing age. Our results suggest that the ACL enthesis of the developing knee begins to mimic that of an adult as early as 19 months of age, as a more complex transition between ligamentous and chondro-epiphyseal tissue can be appreciated. We hypothesize that the observed changes are likely due to mechanical loading of the enthesis with the onset of weightbearing. Future investigations of ACL reconstruction and repair will benefit from improved understanding of the chondro-epiphyseal/ACL regions.