Sandra J. Shefelbine

Flexural and angular limb deformities (LD) are an important cause of early-life morbidity and mortality/euthanasia in Thoroughbred foals. The majority are congenital in origin but, to date, their precise aetiology is poorly understood. We hypothesized that maternal-and pregnancy-level factors, particularly those with potential to influence in-utero growth and development, could play an important role. The aim of this study was therefore to investigate associations between such factors and early-life LD in Thoroughbred foals. A birth cohort was established on seven farms across the United Kingdom and Ireland and details of veterinary interventions for LD in foals in the first six months of life prospectively recorded. Details of dams' signalment, breeding history and reproductive and veterinary history in the breeding season(s) of interest were retrieved retrospectively from stud farm and veterinary records. Associations between mare-and pregnancy-level factors and LD in offspring were assessed using multivariable logistic regression. Records were available for 275 pregnancies in 235 mares, over two breeding seasons. Pregnancies resulted in the birth of 272 live foals, 21% of which (n = 57/272, 95% CI, 16-26) required veterinary intervention for LD in the first six months of life. Odds of LD decreased by 4% per day increase in gestation length between 314 and 381 days (OR 0.96, 95% CI, 0.93-0.99, P = .01). Longer gestation length appears to reduce the odds of early-life LD, including within the normal range of gestation length for Thoroughbred foals. Further work is required to elucidate biological mechanisms behind this association.

Collagen is a ubiquitous protein with remarkable mechanical properties. It is highly elastic, shows large fracture strength and enables substantial energy dissipation during deformation. Most of the connective tissue in humans consists of collagen fibrils composed of a staggered array of tropocollagen molecules, which are connected by intermolecular cross-links. In this study, we report a three-dimensional coarse-grained model of collagen and analyze the influence of enzymatic cross-links on the mechanics of collagen fibrils. Two representatives immature and mature cross-links are implemented in the mesoscale model using a bottom-up approach. By varying the number, type and mechanical properties of cross-links in the fibrils and performing tensile test on the models, we systematically investigate the deformation mechanisms of cross-linked collagen fibrils. We find that cross-linked fibrils exhibit a three phase behavior, which agrees closer with experimental results than what was obtained using previous models. The fibril mechanical response is characterized by: (i) an initial elastic deformation corresponding to the collagen molecule uncoiling, (ii) a linear regime dominated by molecule sliding and (iii) the second stiffer elastic regime related to the stretching of the backbone of the tropocollagen molecules until the fibril ruptures. Our results suggest that both cross-link density and type dictate the stiffness of large deformation regime by increasing the number of interconnected molecules while cross-links mechanical properties determine the failure strain and strength of the fibril. These findings reveal that cross-links play an essential role in creating an interconnected fibrillar material of tunable toughness and strength.

The cartilage-to-bone transition is an essential process in healthy bone development and repair. Our previous work has shown that when the cells found within the human periosteum (the membrane surrounding the bone) are cultured in human serum (HS) as opposed to the standard animal serum (FBS), these cells have greater bone-forming capacity as assessed in an ectopic assay in nude mice. What is not understood is the molecular interactions that permitted this enhanced biological potency. Herein, virtual networks are created to identify the key proteins driving increased bone formation from these cells. Key signalling factors were identified through a network analysis, where FGFR3 was pinpointed as a major differential regulator between cells grown in HS and cells grown in FBS. This analysis was validated through an analysis of human-derived periosteal progenitor cells (PDCs) containing a constitutively active (ca) FGFR3. Following removal and analysis, we found that the FGFR3-ca cells that were implanted on bone void filler scaffolds in mice had an abundance of bone and cartilage that were present compared to the scaffold containing normal/healthy cells. This suggests that these cells were undergoing enhanced cartilage-to-bone transitions and that this protein may be a potentially novel therapeutic target for diseases where the cartilage-to-bone transition is affected such as during poor fracture healing.