Controlled Stiffness of Direct-Write, Near-Field Electrospun Gelatin Fibers Generates Differences in Tenocyte Morphology and Gene Expression.

Tendinopathy is a leading cause of mobility issues. Currently, the cell-matrix interactions involved in the development of tendinopathy are not fully understood. In vitro tendon models provide a unique tool for addressing this knowledge gap as they permit fine control over biochemical, micromechanical, and structural aspects of the local environment to explore cell-matrix interactions. In this study, direct-write, near-field electrospinning of gelatin solution was implemented to fabricate micron-scale fibrous scaffolds that mimic native collagen fiber size and orientation. The stiffness of these fibrous scaffolds was found to be controllable between 1 MPa and 8 MPa using different crosslinking methods (EDC, DHT, DHT+EDC) or through altering the duration of crosslinking with EDC (1 h to 24 h). EDC crosslinking provided the greatest fiber stability, surviving up to 3 weeks in vitro. Differences in stiffness resulted in phenotypic changes for equine tenocytes with low stiffness fibers (∼1 MPa) promoting an elongated nuclear aspect ratio while those on high stiffness fibers (∼8 MPa) were rounded. High stiffness fibers resulted in the upregulation of matrix metalloproteinase (MMPs) and proteoglycans (possible indicators for tendinopathy) relative to low stiffness fibers. These results demonstrate the feasibility of direct-written gelatin scaffolds as tendon in vitro models and provide evidence that matrix mechanical properties may be crucial factors in cell-matrix interactions during tendinopathy formation.

A Systems Approach to Biomechanics, Mechanobiology, and Biotransport.

The human body represents a collection of interacting systems that range in scale from nanometers to meters. Investigations from a systems perspective focus on how the parts work together to enact changes across spatial scales, and further our understanding of how systems function and fail. Here, we highlight systems approaches presented at the 2022 Summer Biomechanics, Bio-engineering, and Biotransport Conference in the areas of solid mechanics; fluid mechanics; tissue and cellular engineering; biotransport; and design, dynamics, and rehabilitation; and biomechanics education. Systems approaches are yielding new insights into human biology by leveraging state-of-the-art tools, which could ultimately lead to more informed design of therapies and medical devices for preventing and treating disease as well as rehabilitating patients using strategies that are uniquely optimized for each patient. Educational approaches can also be designed to foster a foundation of systems-level thinking.

Neo-Natal Castration Leads to Subtle Differences in Porcine Anterior Cruciate Ligament Morphology and Function in Adolescence.

Female adolescent athletes are at a higher risk of tearing their anterior cruciate ligament (ACL) than male counterparts. While most work related to hormones has focused on the effects of estrogen to understand the increased risk of ACL injury, there are other understudied factors, including testosterone. The purpose of this study was to determine how surgical castration in the male porcine model influences ACL size and function across skeletal growth. Thirty-six male Yorkshire crossbreed pigs were raised to 3 (juvenile), 4.5 (early adolescent), and 6 months (adolescent) of age. Animals were either castrated (barrows) within 2 weeks after birth or were left intact (boars). Posteuthanasia, joint and ACL size were assessed via MRI, and biomechanics were assessed via a robotic testing system. Joint size increased throughout age, yet barrows had smaller joints than boars. ACL cross-sectional area (CSA), length, volume, and in situ stiffness increased with age, as did the percent contribution of the ACL anteromedial (AM) bundle to resisting loads. Boar ACL, AM bundle, and PL bundle volumes were 19%, 25%, and 15% larger than barrows across ages. However, ACL CSA, in situ stiffness, and bundle contribution were similar between boars and barrows. The barrows had smaller temporal increases in AM bundle function than boars, but these data were highly variable. Early and sustained loss in testosterone leads to subtle differences in ACL morphology but may not influence measures associated with increased injury risk, such as CSA or bundle forces in response to applied loads.

Preclinical tendon and ligament models: Beyond the 3Rs (replacement, reduction, and refinement) to 5W1H (why, who, what, where, when, how).

Several tendon and ligament animal models were presented at the 2022 Orthopaedic Research Society Tendon Section Conference held at the University of Pennsylvania, May 5 to 7, 2022. A key objective of the breakout sessions at this meeting was to develop guidelines for the field, including for preclinical tendon and ligament animal models. This review summarizes the perspectives of experts for eight surgical small and large animal models of rotator cuff tear, flexor tendon transection, anterior cruciate ligament tear, and Achilles tendon injury using the framework: "Why, Who, What, Where, When, and How" (5W1H). A notable conclusion is that the perfect tendon model does not exist; there is no single gold standard animal model that represents the totality of tendon and ligament disease. Each model has advantages and disadvantages and should be carefully considered in light of the specific research question. There are also circumstances when an animal model is not the best approach. The wide variety of tendon and ligament pathologies necessitates choices between small and large animal models, different anatomic sites, and a range of factors associated with each model during the planning phase. Attendees agreed on some guiding principles including: providing clear justification for the model selected, providing animal model details at publication, encouraging sharing of protocols and expertise, improving training of research personnel, and considering greater collaboration with veterinarians. A clear path for translating from animal models to clinical practice was also considered as a critical next step for accelerating progress in the tendon and ligament field.

Neo-natal castration leads to subtle differences in porcine anterior cruciate ligament morphology and function in adolescence.

Female adolescent athletes are at a higher risk of tearing their anterior cruciate ligament (ACL) than male counterparts. While most work related to hormones has focused on the effects of estrogen to understand the increased risk of ACL injury, there are other understudied factors, including testosterone. The purpose of this study was to determine how surgical castration in the male porcine model influences ACL size and function across skeletal growth. Thirty-six male Yorkshire crossbreed pigs were raised to 3 (juvenile), 4.5 (early adolescent), and 6 months (adolescent) of age. Animals were either castrated (barrows) within 1-2 weeks after birth or were left intact (boars). Post-euthanasia, joint and ACL size were assessed via MRI, and biomechanics were assessed via a robotic testing system. Joint size increased throughout age, yet barrows had smaller joints than boars (p<0.001 for all measures). ACL cross-sectional area (CSA), length, volume, and stiffness increased with age (p<0.0001), as did ACL anteromedial (AM) bundle percent contribution to resisting loads (p=0.012). Boar ACL, AM bundle, and PL bundle volumes were 19% (p=0.002), 25% (p=0.003), and 15% (p=0.04) larger than barrows across ages. However, CSA, stiffness, and bundle contribution were similar between boars and barrows (p>0.05). The barrows had smaller temporal increases in AM bundle percent function than boars, but these data were highly variable. Thus, early and sustained loss in testosterone leads to subtle differences in ACL morphology, but may not influence measures associated with increased injury risk, such as CSA or bundle forces in response to applied loads.

Towards a standardized multi-tissue decellularization protocol for the derivation of extracellular matrix materials.

The goal of tissue decellularization is to efficiently remove unwanted cellular components, such as DNA and cellular debris, while retaining the complex structural and molecular milieu within the extracellular matrix (ECM). Decellularization protocols to date are centered on customized tissue-specific and lab-specific protocols that involve consecutive manual steps which results in variable and protocol-specific ECM material. The differences that result from the inconsistent protocols between decellularized ECMs affect consistency across batches, limit comparisons between results obtained from different laboratories, and could limit the transferability of the material for consistent laboratory or clinical use. The present study is the first proof-of-concept towards the development of a standardized protocol that can be used to derive multiple ECM biomaterials (powders and hydrogels) via a previously established automated system. The automated decellularization method developed by our group was used due to its short decellularization time (4 hours) and its ability to reduce batch-to-batch variability. The ECM obtained using this first iteration of a unified protocol was able to produce ECM hydrogels from skin, lung, muscle, tendons, cartilage, and laryngeal tissues. All hydrogels formed in this study were cytocompatible and showed gelation and rheological properties consistent with previous ECM hydrogels. The ECMs also showed unique proteomic composition. The present study represents the first step towards developing standardized protocols that can be used on multiple tissues in a fast, scalable, and reproducible manner.

Effect of epitendinous suture augmentation to a double Krackow suture pattern for canine gastrocnemius tendon repair.

OBJECTIVE: To evaluate the effect of a double Krackow suture pattern (DK), with and without epitendinous suture augmentation (ES), in a canine gastrocnemius tendon (GT) model. SAMPLE: Paired GTs from 12 adult dog cadavers and 4 control GT. PROCEDURES: GTs were assigned to 2 groups (n = 12/group). Transverse tenotomy was performed and repaired with a DK or DK + ES. Yield, peak, and failure force, stiffness, occurrence of 1-and 3-mm gapping, and failure mode were examined. RESULTS: Yield, peak, and failure loads were greater for DK + ES. Yield force was 48% greater for DK + ES (mean ± SD, 149.56 ± 53.26 N) versus DK (101.27 ± 37.17 N; P = 0.017). Peak force was 45% greater for DK + ES P < 0.001). Failure force was 47% greater for DK + ES (193.752 ± 31.43 N) versus DK (131.54 ± 22.28 N; P < 0.001). Construct stiffness was 36% greater for DK + ES (P = 0.04). All 12 DK and 10 of 12 DK + ES repairs produced a 1-mm gap, with all DK and 4 DK + ES repairs producing a 3-mm gap (P < 0.001). Loads required to create a 3-mm gap were significantly greater for DK + ES (P < 0.013). Suture breakage occurred in all DK repairs, which differed from DK + ES, where suture breakage (7/12) and tissue failure (5/12; P = 0.037) predominated. CLINICAL RELEVANCE: Augmentation of a primary DK repair with an ES significantly improved construct strength in canine GT constructs while increasing loads required to cause 1- and 3-mm gap formation, respectively. ES augmentation is a simple technique modification that can be used to significantly increase construct strength, compared with DK alone.

Elucidating the combinatorial effect of substrate stiffness and surface viscoelasticity on cellular phenotype.

Cells maintain tensional homeostasis by monitoring the mechanics of their microenvironment. In order to understand this mechanotransduction phenomenon, hydrogel materials have been developed with either controllable linear elastic or viscoelastic properties. Native biological tissues, and biomaterials used for medical purposes, often have complex mechanical properties. However, due to the difficulty in completely decoupling the elastic and viscous components of hydrogel materials, the effect of complex composite materials on cellular responses has largely gone unreported. Here, we characterize a novel composite hydrogel system capable of decoupling and individually controlling both the bulk stiffness and surface viscoelasticity of the material by combining polyacrylamide (PA) gels with microgel thin films. By taking advantage of the high degree of control over stiffness offered by PA gels and viscoelasticity, in terms of surface loss tangent, of microgel thin films, it is possible to study the influence that bulk substrate stiffness and surface loss tangent have on complex fibroblast responses, including cellular and nuclear morphology and gene expression. This material system provides a facile method for investigating cellular responses to complex material mechanics with great precision and allows for a greater understanding of cellular mechanotransduction mechanisms than previously possible through current model material platforms.

Differences in Gait Biomechanics Between Adolescents and Young Adults With Anterior Cruciate Ligament Reconstruction.

CONTEXT: Adolescents and adults are treated similarly in rehabilitation and research despite differences in clinical recovery after anterior cruciate ligament reconstruction (ACLR). Aberrant gait is a clinical outcome associated with poor long-term health post-ACLR but has not been compared between adolescents and adults. OBJECTIVE: To compare gait biomechanical waveforms throughout stance between adolescents (<18 years old) and young adults (≥18 years old) post-ACLR. DESIGN: Case-control study. SETTING: Laboratory. PATIENTS OR OTHER PARTICIPANTS: Adolescents (n = 13, girls = 77%, age = 16.7 ± 0.6 years, height = 1.7 ± 0.1 m, weight = 22.2 ± 3.7 kg/m2) were identified from a cross-sectional cohort assessing clinical outcomes 6 to 12 months post-ACLR. Young adults (n = 13, women = 77%, age = 22.3 ± 4.0 years, height = 1.7 ± 0.1 m, weight = 22.9 ± 3.3 kg/m2) were matched based on sex, time since surgery (±2 months), and body mass index (±3 kg/m2). INTERVENTION(S): Participants performed 5 gait trials at their habitual speed. MAIN OUTCOME MEASURE(S): Three-dimensional gait biomechanics and forces were collected. Vertical ground reaction force normalized to body weight (xBW), knee-flexion angle (°), knee-abduction moment (xBW × height), and knee-extension moment (BW × height) waveforms were calculated during the stance phase of gait (0%-100%). Habitual walking speed was compared using independent t tests. We used functional waveforms to compare gait biomechanics throughout stance with and without controlling for habitual walking speed by calculating mean differences between groups with 95% CIs. RESULTS: Adolescents walked with slower habitual speeds compared with adults (adolescents = 1.1 ± 0.1 m/s, adults = 1.3 ± 0.1 m/s, P < .001). When gait speed was not controlled, adolescents walked with less vertical ground reaction force (9%-15% of stance) and knee-abduction moment (12%-25% of stance) during early stance and less knee-extension moment during late stance (80%-99% of stance). Regardless of their habitual walking speed, adolescents walked with greater knee-flexion angle throughout most stances (0%-21% and 29%-100% of stance). CONCLUSIONS: Adolescents and adults demonstrated different gait patterns post-ACLR, suggesting that age may play a role in altered gait biomechanics.

Age- and Sex-Specific Joint Biomechanics in Response to Partial and Complete Anterior Cruciate Ligament Injury in the Porcine Model.

CONTEXT: Pediatric anterior cruciate ligament (ACL) injury rates are increasing and are highest in female adolescents. Complete ACL tears are typically surgically reconstructed, but few guidelines and very limited data exist regarding the need for surgical reconstruction or rehabilitation for partial ACL tears in skeletally immature patients. OBJECTIVE: To evaluate the effects of partial (anteromedial bundle) and complete ACL transection on joint laxity and tissue forces under anterior and rotational loads in male and female stifle joints throughout skeletal growth in the porcine model. DESIGN: Descriptive laboratory study. SETTING: Laboratory. PATIENTS OR OTHER PARTICIPANTS: We studied 60 male and female Yorkshire crossbreed pigs aged 1.5, 3, 4.5, 6, and 18 months (n = 6 pigs per age per sex). MAIN OUTCOME MEASURE(S): Joint laxity was measured in intact, anteromedial bundle-transected, and ACL-transected joints under applied anterior-posterior drawer and varus-valgus torque using a robotic testing system. Loading of the soft tissues in the stifle joint was measured under each condition. RESULTS: Anterior-posterior joint laxity increased by 13% to 50% (P < .05) after anteromedial bundle transection and 75% to 178% (P < .05) after ACL transection. Destabilization after anteromedial bundle transection increased with age (P < .05) and was greater in late female than late male adolescents (P < .05). In anteromedial bundle-transected joints, the posterolateral bundle resisted the anterior load. In ACL-transected joints, the medial collateral ligament (MCL) contribution was largest, followed by the medial meniscus. The MCL contribution was larger and the medial meniscus contribution was smaller in male versus female specimens. CONCLUSIONS: Partial ACL transection resulted in moderate increases in joint laxity, with the remaining bundle performing the primary ACL function. Destabilization due to partial ACL transection (anteromedial bundle) was largest in late adolescent joints, indicating that operative treatment should be considered in active, late-adolescent patients with this injury. Increased forces on the MCL and medial meniscus after ACL transection suggested that rehabilitation protocols may need to focus on protecting these tissues.

Sex-specific biomechanics and morphology of the anterior cruciate ligament during skeletal growth in a porcine model.

Pediatric anterior cruciate ligament (ACL) injuries are on the rise, and females experience higher ACL injury risk than males during adolescence. Studies in skeletally immature patients indicate differences in ACL size and joint laxity between males and females after the onset of adolescence. However, functional data regarding the ACL and its anteromedial and posterolateral bundles in the pediatric population remain rare. Therefore, this study uses a porcine model to investigate the sex-specific morphology and biomechanics of the ACL and its bundles throughout skeletal growth. Hind limbs from male and female Yorkshire pigs aged early youth to late adolescence were imaged using magnetic resonance imaging to measure the size and orientation of the ACL and its bundles, then biomechanically tested under anterior-posterior drawer using a robotic testing system. Joint laxity decreased (p < 0.001) while joint stiffness increased (p < 0.001) throughout skeletal growth in both sexes. The ACL was the primary stabilizer against anterior tibial loading, while the functional role of the anteromedial bundle increased with age (p < 0.001), with an earlier increase in males. ACL and posterolateral bundle cross-sectional area and ACL and anteromedial bundle length were larger in males than females during adolescence (p < 0.01 for all), while ACL and bundle sagittal angle remained similar between sexes. Additionally, in situ ACL stiffness versus cross-sectional area regressions were significant across skeletal growth (r2 = 0.75, p < 0.001 in males and r2 = 0.64, p < 0.001 in females), but not within age groups. This study has implications for age and sex-specific surgical intervention strategies and suggests the need for human studies.

Age- and sex-specific differences in ACL and ACL bundle size during adolescent growth.

Anterior cruciate ligament (ACL) injuries are increasingly common in adolescents, and injuries in this age-group are associated with many unique challenges. Recent large animal studies suggest that the size and function of the major bundles of the ACL change differently throughout skeletal growth. To better aid clinical treatment of pediatric partial ACL tears and better predict outcomes from age-specific treatments, there is a need to measure changes in ACL bundle size in humans during growth. As such, the objective of this study was to compare changes in the length and cross-sectional area (CSA) of the ACL and its primary bundles in adolescent human subjects. Magnetic resonance imaging (MRI) scans were analyzed to determine the visibility and integrity of the ACL and its anteromedial and posterolateral bundles. MRI scans were considered from a retrospective database of subjects ranging from 10 to 18 years of age. The ACL and its anteromedial and posterolateral bundles were segmented and reconstructed into 3D models, and length and CSA were calculated. Total ACL length and CSA were greater in males compared with females, with a statistically significant interaction between age and sex for CSA. Sex had a significant effect on the CSA of both bundles. These sex-dependent differences emerge with moderate to large effect sizes (range: d = 0.50 to d = 1.23) beginning around 13 years of age. Along with ACL bundle structure-function relationships previously established in preclinical animal models, these findings may point toward biomechanical changes in the adolescent human ACL.

A Direct Comparison of Node and Element-Based Finite Element Modeling Approaches to Study Tissue Growth.

Finite element analysis is a useful tool to model growth of biological tissues and predict how growth can be impacted by stimuli. Previous work has simulated growth using node-based or element-based approaches, and this implementation choice may influence predicted growth, irrespective of the applied growth model. This study directly compared node-based and element-based approaches to understand the isolated impact of implementation method on growth predictions by simulating growth of a bone rudiment geometry, and determined what conditions produce similar results between the approaches. We used a previously reported node-based approach implemented via thermal expansion and an element-based approach implemented via osmotic swelling, and we derived a mathematical relationship to relate the growth resulting from these approaches. We found that material properties (modulus) affected growth in the element-based approach, with growth completely restricted for high modulus values relative to the growth stimulus, and no restriction for low modulus values. The node-based approach was unaffected by modulus. Node- and element-based approaches matched marginally better when the conversion coefficient to relate the approaches was optimized based on the results of initial simulations, rather than using the theoretically predicted conversion coefficient (median difference in node position 0.042 cm versus 0.052 cm, respectively). In summary, we illustrate here the importance of the choice of implementation approach for modeling growth, provide a framework for converting models between implementation approaches, and highlight important considerations for comparing results in prior work and developing new models of tissue growth.

Extracellular Matrix Hydrogels Promote Expression of Muscle-Tendon Junction Proteins.

Muscle and tendon injuries are prevalent and range from minor sprains and strains to traumatic, debilitating injuries. However, the interactions between these tissues during injury and recovery remain unclear. Three-dimensional tissue models that incorporate both tissues and a physiologically relevant junction between muscle and tendon may help understand how the two tissues interact. Here, we use tissue specific extracellular matrix (ECM) derived from muscle and tendon to determine how cells of each tissue interact with the microenvironment of the opposite tissue, resulting in junction-specific features. The ECM materials were derived from the Achilles tendon and gastrocnemius muscle, decellularized, and processed to form tissue-specific pre-hydrogel digests. The ECM materials were unique in respect to protein composition and included many types of ECM proteins, not just collagens. After digestion and gelation, ECM hydrogels had similar complex viscosities that were less than type I collagen hydrogels at the same concentration. C2C12 myoblasts and tendon fibroblasts were cultured in tissue-specific ECM conditioned media or encapsulated in tissue-specific ECM hydrogels to determine cell-matrix interactions and the effects on a muscle-tendon junction marker, paxillin. The ECM conditioned media had only a minor effect on the upregulation of paxillin in cells cultured in monolayer. However, cells cultured within ECM hydrogels had 50-70% higher paxillin expression than cells cultured in type I collagen hydrogels. Contraction of the ECM hydrogels varied by the type of ECM used. Subsequent experiments with a varying density of type I collagen (and thus contraction) showed no correlation between paxillin expression and the amount of gel contraction, suggesting that a constituent of the ECM was the driver of paxillin expression in the ECM hydrogels. In addition, another junction marker, type XXII collagen, had similar expression patterns as paxillin, with smaller effect sizes. Using tissue-specific ECM allowed for the de-construction of the cell-matrix interactions similar to muscle-tendon junctions to study the expression of myotendinous junction-specific proteins. Impact statement The muscle-tendon junction is an important feature of muscle-tendon units; however, despite crosstalk between the two tissue types, the junction is often overlooked in current research. Deconstructing the cell-matrix interactions will provide the opportunity to study significant junction-specific features and markers that should be included in tissue models of the muscle-tendon unit, while gaining a deeper understanding of the natural junction. This research aims at informing future methods to engineer a more relevant multi-tissue platform to study the muscle-tendon unit.

Fibrin-Based Biomaterial Systems to Enhance Anterior Cruciate Ligament Healing.

Anterior cruciate ligament (ACL) tears are a common and potentially career-ending injury, particularly for athletes and soldiers. Partial and complete ruptures of this ligament cause instability in the knee, and the ACL does not have the capacity for healing due, in part, to its position within the highly thrombolytic synovial fluid environment of the knee joint. Traditional methods of ACL reconstruction, such as graft replacement with attached bone anchors for bone integration, restore stability, but do not prevent the development of post-traumatic osteoarthritis. To enhance therapeutic treatment options, novel fibrin-based technologies and repair techniques have been recently explored and show promise for improved patient outcomes. Through modification of existing surgical methods, such as the use of fibrin glues incorporating growth factors and cells and the implementation of scaffolds containing platelet-rich plasma, platelet-rich fibrin, and other blood derivatives, surgeons are attempting to overcome the shortcomings of traditional treatments. This mini-review will detail current efforts using fibrin-based treatments and discuss opportunities to further enhance ACL healing.

Ex vivo evaluation of novel core tenorrhaphy patterns in dogs.

OBJECTIVE: To compare the biomechanical properties and gapping characteristics of four novel tenorrhaphy patterns in a canine flexor tendon model. STUDY DESIGN: Ex vivo, randomized, biomechanical study. SAMPLE POPULATION: Superficial digital flexor tendons of 60 forelimbs (30 dogs). METHODS: Each tendon was transected 25 mm distal to its musculotendinous junction prior to tenorrhaphy with 2-0 polypropylene. Repair patterns included the three-loop pulley (3LP, control), exposed double-cross-lock (ExDCrL), embedded double-cross-lock (EmDCrL), triple-circle-lock (TCiL), and Modified-Tang patterns (MTang) were randomly assigned to each experimental group (n = 12/group). Yield, peak, and failure loads, gap formation and failure modes were compared. RESULTS: Tendons repaired with ExDCrL (p < .0001), EmDCrL (p < .0001), and MTang (p < .0001) sustained yield, peak, and failure loads ~2.2x, ~2.0x, and ~1.9x, respectively, greater than those repaired with 3LP. Loads to 1 and 3 mm gapping were also higher for ExDCrL (p < .0001), EmDCrL (p < .0004), and MTang constructs (p < .0017) compared to 3LP. Although TCiL constructs sustained higher loads, their resistance to gap formation did not differ from that of 3LP repairs. Failure mode differed between groups (p < .0001), EmDCrL, ExDCrL, MTang, and TCiL constructs failing predominantly by suture breakage compared to 3LP repairs that failed by suture pull-through. CONCLUSION: Use of novel patterns ExDCrL, EmDCrL, and MTang improved resistance to loads and gap formation and were biomechanically superior compared to 3LP in healthy canine tendon repairs. CLINICAL SIGNIFICANCE: These results justify in vivo evaluation of ExDCrL, EmDCrL, or MTang pattern for tenorrhaphy in dogs.

Effect of epitendinous suture caliber on the tensile strength of repaired canine flexor tendons.

OBJECTIVE: To determine the effect of epitendinous suture (ES) caliber on the tensile strength of flexor tendon repairs in cadaveric specimens from dogs. SAMPLE: 60 cadaveric superficial digital flexor tendons (SDFTs) from 30 skeletally mature dogs. PROCEDURES: Specimens were randomly assigned to 5 suture caliber groups (n = 12 SDFTs/group). After sharp transection, SDFTs were repaired by placement of a simple continuous circumferential ES created with size-0, 2-0, 3-0, 4-0, or 5-0 polypropylene suture. Constructs were preloaded to 2 N and load tested to failure. Loads at yield, peak, and failure and mode of failure were compared among groups by statistical methods. RESULTS: Yield, peak, and failure loads for SDFT repair constructs were positively correlated with ES caliber and did not differ between the size-0 and 2-0 groups on pairwise comparisons. Yield load was significantly greater for size-0, 2-0, and 3-0 groups than for the 4-0 and 5-0 groups. Peak and failure loads were significantly greater for the size-0 and 2-0 groups than for the remaining groups. Most size-0 (12/12), 2-0 (12/12), and 3-0 (10/12) group constructs failed because of ES pull-through; several constructs in the 4-0 group (5/12) and most in the 5-0 group (11/12) failed because of ES breakage. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggested size-0 and 2-0 sutures should be considered when placing an ES for flexor tendon repairs in dogs. However, in vivo studies are needed determine the effects of increasing ES caliber on clinical outcomes for dogs undergoing these procedures.

Biomechanical analysis of accessory tendon graft augmentation for primary gastrocnemius tendon reconstruction in dogs.

OBJECTIVE: To evaluate the effect of accessory tendon graft (ATG) augmentation as an adjunct to a core locking-loop (LL) and epitendinous suture (ES) repair in a gastrocnemius tendon (GT) model. STUDY DESIGN: Randomized, ex vivo, biomechanical. POPULATION: Twenty-two canine GT musculotendinous constructs. METHODS: GT repair constructs were randomly divided into two groups (n = 10/group). After transection, paired GT were repaired with LL + ES alone or with concurrent ATG augmentation. Yield, peak and failure loads, tensile loads required to create 1 and 3 mm gapping, and failure modes were evaluated. Four GT were used as intact controls for validation of testing methodology. ATG constructs were compared to LL + ES and control specimens. RESULTS: Yield (p < .0001), peak (p = .0001) and failure loads (p = .0003) were greater when ATG was used for repair. Greater force was required to cause 1 mm (p = .0001) and 3 mm (p = .0002) gap formation in the ATG group, however, the frequency of gap formation did not differ between groups. All repaired constructs failed exclusively by suture pull-through. CONCLUSION: Autologous ATG augmentation as an adjunct to primary GT repair increased yield, peak and failure forces by approximately 1.6×, 1.9×, 1.8× respectively and required 2.1× greater force to cause 1 and 3 mm formation respectively compared to LL + ES repairs alone. CLINICAL SIGNIFICANCE: ATG augmentation should be considered as an autologous method to support and strengthen the primary GT repair. These results justify studies to determine the effect of ATG on clinical function following graft harvest in dogs.

Biomechanical evaluation of a novel barbed suture pattern with epitendinous suture augmentation in a canine flexor tendon model.

OBJECTIVE: To determine the effect of a novel barbed suture pattern (NBSP) compared to a three-loop-pulley (3LP) with and without epitendinous suture (ES) augmentation on the biomechanical strength and gap formation of repaired canine tendons. STUDY DESIGN: Ex vivo, cadaveric, randomized, experimental study. SAMPLE POPULATION: Forty, adult superficial digital flexor tendons (SDFT). METHODS: SDFT were randomly assigned to one of four groups (n = 10/group). Sharp tenotomy was performed and repaired with 3LP, NBSP, 3LP + ES, and NBSP + ES. Constructs were tested to failure while evaluating yield, peak, and failure loads, loads at 1 and 3 mm gap formation, and failure mode. RESULTS: Constructs augmented with ES sustained 80% greater yield (p < .001), peak (p < .001), and failure (p < .001) loads, with no difference between 3LP + ES and NBSP + ES constructs regarding peak (p = .614), and failure forces (p = .865). Loads resulting in 1 and 3 mm gap formation were greater when constructs were augmented with an ES (p ≤ .003). Failure mode differed between groups (p < .001), occurring predominantly due to suture pull-through in 3LP and NBSP groups compared to tissue failure distant to the repair site in ES augmented constructs. CONCLUSION: Tendons repaired with the NBSP used in this study resisted similar forces as those repaired with 3LP. Augmentation with an ES improved the biomechanical properties of repaired constructs, including resistance to gap formation. CLINICAL RELEVANCE: The NBSP repair tested here may be advantageous over monofilament suture repair as it uses a similar-sized barbed core suture but eliminates the requirement for knot tying.

Processing variables of direct-write, near-field electrospinning impact size and morphology of gelatin fibers.

Several biofabrication methods are being investigated to produce scaffolds that can replicate the structure of the extracellular matrix. Direct-write, near-field electrospinning of polymer solutions and electrowriting of polymer melts are methods which combine fine fiber formation with computer-guided control. Research with such systems has focused primarily on synthetic polymers. To better understand the behavior of biopolymers used for direct-writing, this project investigated changes in fiber morphology, size, and variability caused by varying gelatin and acetic acid concentration, as well as process parameters such as needle gauge and height, stage speed, and interfiber spacing. Increasing gelatin concentration at a constant acetic acid concentration improved fiber morphology from large, planar structures to small, linear fibers with a median of 2.3 µm. Further varying the acetic acid concentration at a constant gelatin concentration did not alter fiber morphology and diameter throughout the range tested. Varying needle gauge and height further improved the median fiber diameter to below 2 µm and variability of the first and third quartiles to within ±1 µm of the median. Additional adjustment of stage speed did not impact the fiber morphology or diameter. Repeatable interfiber spacings down to 250 µm were shown to be capable with the system. In summary, this study illustrates the optimization of processing parameters for direct-writing of gelatin to produce fibers on the scale of collagen fibers. This system is thus capable of replicating the fibrous structure of musculoskeletal tissues with biologically relevant materials which will provide a durable platform for the analysis of single cell-fiber interactions to help better understand the impact scaffold materials and dimensions have on cell behavior.