Tissue Biomechanical Properties Research Articles

Effect of Biomechanical Properties of Perioral Soft Tissues on Lip Response to Simulated Protraction of Upper Front Teeth.

Proclination of front teeth in orthodontics and oral rehabilitation may influence lip protrusion and the overall facial profile. We hypothesized that the degree of profile changes is determined by the biomechanical properties of the lips. (1) to investigate the upper lip changes induced by a simulated protraction of upper front teeth; and (2) to assess the relationship between biomechanical properties of the lips and their response to tooth protraction. Thirty-four participants were recruited at the Faculty of Dentistry, University of Otago, New Zealand. Simulated protraction of upper front teeth was achieved by placing a customised stent covering the buccal surface of the maxillary upper incisors and canines, along with the corresponding buccal mucosa and gingiva. Stereophotogrammetry was used to assess lip changes in three dimensions. Biomechanical properties of the lips, including muscle tone, stiffness and elasticity, were measured using a non-invasive digital palpation device. During the simulated protraction of upper front teeth, the upper lip advanced approximately 50% of the stent thickness, with a considerable inter-individual variation in lip displacement. Soft tissue responses were correlated with the biomechanical properties of the lips with significant correlation coefficients ranging from 0.38 to 0.40. Higher lip tone was associated with increased lip displacement, while greater lip elasticity resulted in reduced lip displacement. Multivariate analyses indicated that upper lip displacement was associated with biomechanical properties and ethnicity, but not with age, sex and body mass index. The response of the lips to protraction of front teeth varies largely between individuals and can be partly explained by variations in the biomechanical properties of perioral muscles.

MR Elastography Using the Gravitational Transducer

MR elastography is a non-invasive imaging technique that provides quantitative maps of tissue biomechanical properties, i.e., elasticity and viscosity. Currently, hepatic MR elastography is deployed in the clinic to assess liver fibrosis in MAFLD patients. In addition, research has demonstrated MR elastography’s ability to non-invasively assess chronic liver disease and to characterize breast cancer lesions and brain tumors. MR elastography requires efficient mechanical wave generation and penetration, motion-sensitized MRI sequences, and MR elastography inversion algorithms to retrieve the biomechanical properties of the tissue. MR elastography promises to enable non-invasive and versatile assessment of tissue, leading to better diagnosis and staging of several clinical conditions.

A Low-Cost Open-Source Uniaxial Tensile System for Soft Tissue Testing

The evaluation of soft tissue biomechanical properties is of paramount importance not only for a comprehensive understanding of human physiology and physiopathology, but also in the research and development of bio-compatible artificial tissues with viscoelastic properties. Contrarily to standard tensile testing devices, a system intended for biomaterials testing should consider low stress and high strain ranges, characteristic of human tissues; moreover, such a system should enable the ex vivo simulation of biological environmental conditions. Commercial solutions address these challenges, although they are expensive for most academic and research institutions. This study presents a low-cost open-source design solution for soft tissue tensile testing, offering an affordable solution, yet without compromising the high quality and precision of the results. The proposed uniaxial tensile system allows for sample testing at room temperature as well as in a temperature-controlled liquid environment. Moreover, custom clamps ensure the fixation of tissue samples without slipping or tearing. System validation is performed using the tensile testing of springs and 3D-printed soft polymeric samples, demonstrating accurate results compared to the available data. The system is suitable for educational, research, and development applications.

Magnetic Resonance Elastography for Breast Cancer Diagnosis Through the Assessment of Tissue Biomechanical Properties.

Breast cancer and normal breast tissue exhibit different degrees of stiffness, indicating distinct biomechanical properties. Study results reveal that breast cancer tissue is several times stiffer than normal breast tissue. These variations can serve as indicative factors for imaging purposes. Depicting markers can significantly enhance the process of breast cancer diagnosis and treatment. This article provides a brief review of the biomechanical properties of breast cancer tissue, highlighting the role of the magnetic resonance elastography (MRE) technique in utilizing these properties for diagnosing breast cancer. In breast MRE, low-frequency shear waves are employed to measure breast stiffness. This method not only offers a quantitative diagnosis but also generates an elastogram, determining the stiffness of each area through its colors. MRE represents a diagnostic technique with heightened sensitivity, based on depicting the viscoelasticity properties of breast tissue and describing tumors in terms of biomechanical properties. Combining tissue biomechanical properties, such as tissue stiffness, with contrast-enhanced breast Magnetic Resonance Imaging (MRI) leads to tumor diagnosis. The value of MRE in oncological imaging aims at the early detection of tumors and evaluating the prognosis of breast cancer. Breast MRE can identify the reduction of interstitial pressure in tumors by detecting changes in tissue stiffness, making it an effective tool for monitoring treatment responses. This technique is safe, repeatable, and highly precise, significantly aiding in patient screening.

Superior Measurement Accuracy of Digital Thickness Gauge Versus Digital Vernier Caliper in Determining Venous Tissue Thickness.

Background In determining mechanical characteristics, the accuracy of the thickness of the specimens can influence the biomechanical behavior, especially in the case of human tissues, where there is an important variability. This study aims to compare the accuracy of two routine measuring instruments, i.e., the digital vernier caliper and the digital thickness gauge, when measuring the thickness of venous specimens multiple times. Methodology In this study, we used 12 tubular vena cava specimens obtained from common breed pigs aged 18-24 weeks at the time of sacrifice from a local slaughterhouse. The measurements were performed using a digital vernier caliper (Multicomp PRO MP012475) for the first four protocols and a digital thickness gauge (Mitutoyo 547-500S) for the fifth protocol. In the first protocol, three measurements were taken on the same side, and their average was recorded as the sample thickness. The second protocol involved taking measurements on two opposite sides, and the average of these measurements was recorded as the sample thickness. In the third protocol, the thickness of each side was measured at its midpoint, and the average of the four measurements was recorded as the sample thickness. In the last protocol using a digital vernier caliper, the thickness of the vernier specimens was calculated as the average of the measurements taken at each corner of the square sample. Finally, for the fifth protocol, three consecutive measurements were taken using the digital thickness gauge, and their average was recorded as the final thickness of the sample. Results In the first protocol, we observed lower values during the first measurement in comparison to the second (0.409 ± 0.063 vs. 0.536 ± 0.064, p < 0.0001) and the third (0.409 ± 0.063 vs. 0.528 ± 0.055, p = 0.0001). Moreover, with the second protocol, we observed lower values during the first two measurements in comparison to the third measurement (p = 0.0279 and p = 0.0054). Regarding protocols three and four, we recorded higher values for the second and third measurements than the first one, with higher values for the third measurement than the second one. In the fifth protocol, there were no significant statistical differences between the three consecutive measurements (p = 0.953, p = 0.742, and p = 0.897). Further, we examined the variations in sample thickness determined using each of the protocols proposed for the digital vernier caliper, as well as the values obtained with the digital thickness gauge protocol. As a result, during the first and second measurements, we observed lower thickness values for the venous wall samples using the first four protocols compared to the fifth protocol (for all p < 0.05). However, no differences were noted between the five protocols during the third measurement. Conclusions The digital thickness gauge Mitutoyo 547-500S provided superior accuracy with no difference between three successive measurements of venous wall thickness, regardless of the examiner's experience. Accurately determining the thickness of venous specimens is crucial for calculating the tissue's biomechanical properties.

Use of atomic force microscopy to assess the biomechanical properties of 3D tumor cell models

Multicellular spheroids are a unique object-model for toxicological studies. Cells in a 3D cluster have microenvironment, intercellular communication, which allows spheroids to be used as more realistic model compared to traditional cell cultures. It is known that tumor microregions consist of heterogeneous populations of cancer cells, in which cell growth and response to antitumor drugs depend on their 3D architecture, intercellular contacts and interaction with the microenvironment. In addition, tumor growth and progression are also strongly influenced by mechanical cues Currently, 3D cell culture models are a powerful tool for studying the toxicity of drug compounds and nanomaterials of different composition and morphology. This review presents data on the use of various techniques, in particular atomic force microscopy, to investigate changes in the mechanical properties of cells in spheroids. In particular, the use of atomic force microscopy as a tool to reveal physicochemical parameters of cells during pathophysiological processes or drug exposure is considered. The relevance of this review is related to the increasing interest in the role of biomechanical properties of tissues, cells and subcellular structures as markers of pathophysiological conditions.

Effects of oral contrast agent on the viscoelastic properties of the terminal ileum investigated using magnetic resonance elastography.

While standard clinical magnetic resonance (MR) enterography can detect inflammatory bowel disease, it is of limited value in deciding between medical versus surgical treatment. Alternatively, intestinal MR elastography has the potential to contribute additional information to therapeutic decision-making; however, the influence of bowel distension by oral contrast agent on viscoelastic tissue properties remains elusive. Therefore, we aimed to investigate the influence of oral contrast agent-induced bowel distension on the viscoelastic properties of the terminal ileum in healthy volunteers. In this prospective pilot study, 20 healthy volunteers (33.2±8.2 years; 10 men, 10 women) underwent multifrequency MR elastography using a single-shot spin-echo echo planar imaging sequence at 1.5 Tesla and drive frequencies of 40, 50, 60 and 70 Hz. Maps of shear wave speed (c in ms-1) and loss angle (φ in rad), representing stiffness and viscous properties, respectively, were generated using tomoelastography data processing. The volunteers were scanned before and after ingestion of 1,000 mL of 2% mannitol solution as oral contrast agent. There was no significant difference in terminal ileum biomechanical properties before vs. after ingestion of an oral contrast agent (mean c: 1.47±0.24 vs. 1.40±0.25 ms-1 with P=0.37; mean φ: 0.70±0.12 rad vs. 0.68±0.12 rad with P=0.61). Moreover, there was no statistically significant correlation between MR elastography parameters before and after the ingestion of oral contrast (c: r=0.22, P=0.36; φ: r=0.24, P=0.30). The results of this study suggest that bowel distension for intestinal MR elastography has no systematic effect on the biomechanical tissue properties of the terminal ileum determined by MR elastography. Therefore, future study protocols appear feasible with or without oral contrast agents.

Transplantation of Mesenchymal Stem Cells Derived from Old Rats Improves Healing and Biomechanical Properties of Vaginal Tissue Following Surgical Incision in Aged Rats.

Pelvic floor dysfunction encompasses a group of disorders that negatively affect the quality of women's lives. These include pelvic organ prolapse (POP), urinary incontinence, and sexual dysfunction. The greatest risk factors for prolapse are increased parity and older age, with the largest group requiring surgical intervention being post-menopausal women over 65. Prolapse recurrence rates following surgery were reported to be as high as 30%. This may be attributed to ineffective healing in the elderly. Autologous stem cell transplantation during surgery may improve surgical results. In our previous studies, we showed that the transplantation of bone marrow-derived mesenchymal stem cells (MSCs) from young donor rats improved the healing of full-thickness vaginal surgical incision in the vaginal wall of old rats, demonstrated by both histological and functional analysis. In order to translate these results into the clinical reality of autologous MSC transplantation in elderly women, we sought to study whether stem cells derived from old donor animals would provide the same effect. In this study, we demonstrate that MSC transplantation attenuated the inflammatory response, increased angiogenesis, and exhibited a time-dependent impact on MMP9 localization. Most importantly, transplantation improved the restoration of the biomechanical properties of the vagina, resulting in stronger healed vaginal tissue. These results may pave the way for further translational studies focusing on the potential clinical autologous adjuvant transplantation of MSCs for POP repair for the improvement of surgical outcomes.

Native bubble nuclei for acoustic cavitation in 3D cell cultures

The safety of biomedical ultrasound largely relies on controlling cavitation bubbles in vivo; however, bubble nuclei in biological tissues remain unexplored compared to water. No previous studies have observed where bubble nuclei form in tissues at a microscopic level or how the variation in tissue biomechanical properties can impact the presence of bubble nuclei for acoustic cavitation. In this study, we evaluated whether bubble nuclei form intracellularly or extracellularly in rat epithelial hepatoma (McA-RH7777) and musculoskeletal myoblast (L6) cell lines (n = 5 each), cultured in 3D using Matrigel™ scaffolds. A 3.68 MHz focused ultrasound transducer with f# = 1 was used to induce low-density cavitation using varying pulse lengths (10 to 100 μs) with pressures ranging up to p+ = 29 and p− = 12 MPa. The spatial location of the bubbles was monitored microscopically using high-speed photography at 20 000 fps. Despite significant radiation force on the cell scaffold, preliminary results show that acoustic cavitation bubbles (r = 20 μm approx.) preferentially form extracellularly near the cell membranes. This result suggests that the hydrophobicity in the amphipathic cell membrane may contribute to bubble nuclei formation. Future work includes investigating the distribution of bubble nuclei in healthy versus cancerous cell cultures. [Work supported by NSF CAREER 1943937.]

Profiling native pulmonary basement membrane stiffness using atomic force microscopy.

Mammalian cells sense and react to the mechanics of their immediate microenvironment. Therefore, the characterization of the biomechanical properties of tissues with high spatial resolution provides valuable insights into a broad variety of developmental, homeostatic and pathological processes within living organisms. The biomechanical properties of the basement membrane (BM), an extracellular matrix (ECM) substructure measuring only ∼100-400 nm across, are, among other things, pivotal to tumor progression and metastasis formation. Although the precise assignment of the Young's modulus E of such a thin ECM substructure especially in between two cell layers is still challenging, biomechanical data of the BM can provide information of eminent diagnostic potential. Here we present a detailed protocol to quantify the elastic modulus of the BM in murine and human lung tissue, which is one of the major organs prone to metastasis. This protocol describes a streamlined workflow to determine the Young's modulus E of the BM between the endothelial and epithelial cell layers shaping the alveolar wall in lung tissues using atomic force microscopy (AFM). Our step-by-step protocol provides instructions for murine and human lung tissue extraction, inflation of these tissues with cryogenic cutting medium, freezing and cryosectioning of the tissue samples, and AFM force-map recording. In addition, it guides the reader through a semi-automatic data analysis procedure to identify the pulmonary BM and extract its Young's modulus E using an in-house tailored user-friendly AFM data analysis software, the Center for Applied Tissue Engineering and Regenerative Medicine processing toolbox, which enables automatic loading of the recorded force maps, conversion of the force versus piezo-extension curves to force versus indentation curves, calculation of Young's moduli and generation of Young's modulus maps, where the pulmonary BM can be identified using a semi-automatic spatial filtering tool. The entire protocol takes 1-2 d.

A promising method for reducing the incidence of diabetic foot ulcers: Regulating foot temperature during walking

Diabetic foot ulcers (DFUs) pose a significant challenge in the medical field, particularly with the rising prevalence of diabetes. Consequently, there’s a growing emphasis on devising effective prevention strategies to combat the high recurrence rate of these ulcers. Daily walking, while essential for maintaining a normal lifestyle and social interaction is also the primary source of external mechanical stress applied to the foot. Preserving the integrity of foot tissues during daily walking is critical for preventing foot ulcers. An increase in foot tissue temperature, a significant environmental factor induced by walking, can diminish the foot tissue’s resistance to external mechanical stimuli by affecting tissue metabolism and biomechanical properties (tensile and compressive properties) during walking. This could be a key factor contributing to the high recurrence rate of foot ulcers, despite the advancement in clinical unloading protection technology. We propose a hypothesis that managing foot temperature and preventing a continuous temperature rise caused by walking could help protect the integrity of diabetic foot tissue. Long-term adherence to this approach could potentially reduce the incidence of DFUs. A clinical randomized controlled trial has been designed to evaluate this hypothesis from the aspects of tissue metabolism and biomechanical properties. This study aims to offer a novel method for protecting against DFUs and guide the design of foot aids.

Evaluation of breast skin and tissue stiffness using a non-invasive aspiration device and impact of clinical predictors.

A personalized 3D breast model could present a real benefit for preoperative discussion with patients, surgical planning, and guidance. Breast tissue biomechanical properties have been poorly studied in vivo, although they are important for breast deformation simulation. The main objective of our study was to determine breast skin thickness and breast skin and adipose/fibroglandular tissue stiffness. The secondary objective was to assess clinical predictors of elasticity and thickness: age, smoking status, body mass index, contraception, pregnancies, breastfeeding, menopausal status, history of radiotherapy or breast surgery. Participants were included at the Montpellier University Breast Surgery Department from March to May 2022. Breast skin thickness was measured by ultrasonography, breast skin and adipose/fibroglandular tissue stiffnesses were determined with a VLASTIC non-invasive aspiration device at three different sites (breast segments I-III). Multivariable linear models were used to assess clinical predictors of elasticity and thickness. In this cohort of 196 women, the mean breast skin and adipose/fibroglandular tissue stiffness values were 39 and 3 kPa, respectively. The mean breast skin thickness was 1.83 mm. Only menopausal status was significantly correlated with breast skin thickness and adipose/fibroglandular tissue stiffness. The next step will be to implement these stiffness and thickness values in a biomechanical breast model and to evaluate its capacity to predict breast tissue deformations.

Biologically Enhanced Patch in the Healing and Mechanical Stability of Rotator Cuff Tears.

Biological patches have emerged as promising adjuncts in the surgical management of rotator cuff tears, aiming to enhance tissue healing and biomechanical properties of repaired tendons. These patches, derived from human or animal sources such as dermis or small intestinal submucosa, undergo mechanical and pathological changes within the rotator cuff environment post-implantation. These patches provide structural reinforcement to the repair site, distributing forces more evenly across the tendon and promoting a gradual load transfer during the healing process. This redistribution of forces helps alleviate tension on the repaired tendon and surrounding tissues, potentially reducing the risk of re-tears and improving overall repair integrity. Moreover, biological patches serve as scaffolds for cellular infiltration and tissue ingrowth, facilitating the recruitment of cells and promoting collagen synthesis. The integration of these patches into the host tissue involves a cascade of cellular events, including inflammation, angiogenesis, and matrix remodeling. Inflammatory responses triggered by patch implantation contribute to the recruitment of immune cells and the release of cytokines and growth factors, fostering a microenvironment conducive to tissue repair. However, despite their potential benefits, the long-term efficacy and durability of biological patches in rotator cuff repair remain areas of ongoing research and debate. Further studies are needed to elucidate the optimal patch characteristics, surgical techniques, and rehabilitation protocols to maximize clinical outcomes and minimize complications in rotator cuff surgery.

In vivo application of a glutaraldehyde-free, UVA/riboflavin cross-linked bovine pericardium confirms suitability for cardiovascular substitutes

Glutaraldehyde (GA)-treated bovine pericardium is still the gold standard for the fabrication of bioprostheses needed for the surgical treatment of valvular malfunction. Although excellent stability and low immunogenicity are accomplished, the application of GA is considered to be causal for structural valve deterioration, diminishing the long-term durability of bioprosthetic tissue. The novel GA-free SULEEI-treatment of bovine pericardium combines decellularization, riboflavin/UVA-cross-linking, and low-energy electron beam irradiation. In the present study, we initiated an in vivo application. We used a subcutaneous rat model to compare the immune and tissue responses, calcification propensity, and biomechanical properties of the alternatively prepared SULEEI bovine pericardial tissue with standard glutaraldehyde-fixed and industrially produced bovine pericardial patch material. SULEEI pericardium evokes a similar immune reaction and tissue response as the control standard bovine patch material. The calcification propensity of SULEEI tissue was low, and biomechanical analysis revealed a heterogeneous but similar pattern in tissue stiffness compared to the control patch. The results of this study highlight the potential of SULEEI-treated bovine pericardial tissue as a candidate for cutting-edge cardiovascular and valvular biomaterials in reconstructive surgery.

Surface-Wave-Induced Photoacoustic Elastography in the Feasibility Study of Tissue Biomechanical Properties

Surface-Wave-Induced Photoacoustic Elastography in the Feasibility Study of Tissue Biomechanical Properties

Accuracy of an Air-Puff Dynamic Tonometry Biomarker to Discriminate the Corneal Biomechanical Response in Patients With Keratoconus.

The aim of this study was to assess accuracy of the mean corneal stiffness ( kc , N/m) parameter to discriminate between patients with keratoconus and age-matched healthy subjects. Dynamic Scheimpflug imaging tonometry was performed with Corvis ST (Oculus Optikgeräte GmbH, Germany) in patients with keratoconus (n = 24; study group) and age-matched healthy subjects (n = 32; control). An image processing algorithm was developed to analyze the video sequence of the Corvis ST air-puff event and to determine the geometric and temporal parameters that correlated with the corneal tissue biomechanical properties. A modified 3-element viscoelastic model was used to derive the kc parameter, which represented the corneal tissue resistance to deformation under load. Receiver operating characteristic curves were used to assess the overall diagnostic performance for determining the area under the curve, sensitivity, and specificity of the kc in assessing the corneal tissue deformation to the Corvis ST air-puff event in keratoconus and control eyes. The Corvis Biomechanical Index ( CBI ) was analyzed for external validation. The kc parameter was significantly different between keratoconus and controls ( P < 0.001), ranging from 24.9 ±3.0 to 34.2 ±3.5 N/m, respectively. It was highly correlated with CBI (r = -0.69; P < 0.001); however, the kc parameter had greater specificity (94%) than CBI (75%), whereas the 2 biomarkers had similar area under the curve (0.98 vs. 0.94) and sensitivity (96% vs. 92%) in predicting the occurrence of keratoconus. The kc parameter extracted by video processing analysis of dynamic Scheimpflug tonometry data was highly accurate in discriminating patients with clinically manifest keratoconus compared with controls.

The use of brain tissue mechanics for time since death estimations

Time since death estimation is a vital part of forensic pathology. Despite the known tissue degradation after death, the efficacy of using biomechanical tissue properties to estimate time since death remains unexplored. Here, eight brain tissue localizations were sampled from the frontal lobe, parietal lobe, anterior and posterior deep brain, superior colliculi, pons, medulla, and cerebellum of 30 sheep; were then stored at 20 °C; and subsequently subjected to rheometry tests on days zero to four after death. Overall, the measured tissue storage modulus, loss modulus, and complex shear modulus decreased after death for all of the tested regions in a site-specific manner. Day zero to day one changes were the only 24-h interval, for which statistically significant differences in tissue mechanical moduli were observed for some of the tested brain regions. Based on receiver operator characteristic analyses between day zero and the pooled data of days one to four, a post mortem interval of at least 1 day can be determined with a sensitivity of 90%, a specificity of 92%, and a positive likelihood ratio of 10.8 using a complex shear modulus cut-off value of 1461 Pa for cerebellar samples. In summary, biomechanical properties of brain tissue can discriminate between fresh and at least 1-day-old samples stored at 20 °C with high diagnostic accuracy. This supports the possible value of biomechanical analyses for forensic time since death estimations. A striking advantage over established methods to estimate the time since death is its usability in cases of disintegrated bodies, e.g. when just the head is found.

Mechanotransduction and Biomechanical Properties of Bone Tissue as a Basis for the Use of Intraosseous Osteopathic Techniques

The development and maintenance of bone mass is critical to movement, health, and quality of life. Bone mass is regulated by various factors, activation of biologically active substances, enzymes, including changes in mechanical load. The osteocyte network provides a wide-ranging load monitoring "network" that penetrates every cubic millimeter of bone tissue. Deformities or trauma create a limitation that can cause asymmetric bone growth, such as plagiocephaly. A deep understanding of the cellular mechanisms responsible for bone remodeling as well as bone mechanotransduction is essential for the development of effective exercise, osteopathic techniques, and pharmaceutical strategies to increase and/or prevent bone loss. This review summarizes current data on the main molecular mechanisms involved in bone mechanotransduction, which provides insight into bone tissue, methods of working with it, and points of application for correcting various dysfunctions to promote bone growth in length. This process is carried out by increasing the proliferation of chondrocytes and the activity of growth plates. This contributes to the correction of such consequences of intraosseous dysfunctions as the development of anatomical differences in the size of the lower extremities in children.

Small leucine rich proteoglycans: Biology, function and their therapeutic potential in the ocular surface

Small leucine rich proteoglycans (SLRPs) are the largest family of proteoglycans, with 18 members that are subdivided into five classes. SLRPs are small in size and can be present in tissues as glycosylated and non-glycosylated proteins, and the most studied SLRPs include decorin, biglycan, lumican, keratocan and fibromodulin. SLRPs specifically bind to collagen fibrils, regulating collagen fibrillogenesis and the biomechanical properties of tissues, and are expressed at particularly high levels in fibrous tissues, such as the cornea. However, SLRPs are also very active components of the ECM, interacting with numerous growth factors, cytokines and cell surface receptors. Therefore, SLRPs regulate major cellular processes and have a central role in major fundamental biological processes, such as maintaining corneal homeostasis and transparency and regulating corneal wound healing. Over the years, mutations and/or altered expression of SLRPs have been associated with various corneal diseases, such as congenital stromal corneal dystrophy and cornea plana. Recently, there has been great interest in harnessing the various functions of SLRPs for therapeutic purposes. In this comprehensive review, we describe the structural features and the related functions of SLRPs, and how these affect the therapeutic potential of SLRPs, with special emphasis on the use of SLRPs for treating ocular surface pathologies.

45 Quantifying gait quality changes in fragile foal syndrome carriers using artificial intelligence

The domestic horse has remarkable locomotor polymorphisms valuable to study of genetic musculoskeletal diseases and their effect on gait quality. Here we investigate a polymorphism in the PLOD1 gene, responsible for Fragile Foal Syndrome (FFS), as it impacts the organization of collagen fibers. We hypothesize that the changes in the biomechanical properties of tissues will result in change of locomotor phenotypes. We first addressed the need for a gait assessment tool that can accurately quantify various locomotor parameters. We created a pipeline built around the software package DeepLabCut (DLC), a user-friendly graphical user interface (GUI) written in Python, and a custom gait parameter calculator written in MatLab. DLC uses a pre-trained deep neural network model to apply 22 anatomical landmarks on jogging horses observed in video recordings. A custom GUI allows the user to perform error checks of the labels before the pipeline processes the raw marker coordinate files produced by DLC into geometric gait parameters. We then genotyped 138 horses in training for jumping competition by extracting DNA from the bulbs of pulled tail hair and running a PCR-RFLP test. We confirmed these genotypes via sanger sequencing. Of the 138 horses, 58 were purebred Thoroughbreds, 72 were warmbloods, and the remaining 8 were made up of stock horses. The carrier rate was n = 4 in Thoroughbreds, and n = 3in the warmbloods including an Irish Sport Horse, Sillia Argentina, and Holsteiner. We observed a FFS carrier frequency markedly higher in the Thoroughbred population than those reported in previous studies (6.9%, P = 0.015, Χ2 test). A Quadratic Discriminant Analysis then produced standardized coefficient scores for each of the 15 gait parameters to determine the best predictors of genotype within the model. The change in joint extension of both the hind and forelimb were the strongest predictors of genotype and indicated that FFS carriers had greater extension and swing of all 4 limbs. As a result, FFS carriers likely have an altered hoof flight path throughout the stride cycle. Future work will increase the sample size to capture more FFS carriers and will more precisely define the FFS locomotor phenotype. We will also apply this new phenotyping method to investigate changes in gait associated with other genetic variants for connective tissue disorders like HERDA and JEB. Improved quantification of gait parameters, and our understanding of the physiology of this and other known connective tissue disorders in the horse, will aid in future breeding and selection decisions to reduce injury risk and improve performance.