RESUMO
Herein we report a new library of 2,3-pyrrolidinedione analogues that expands on our previous report on the antimicrobial studies of this heterocyclic scaffold. The novel 2,3-pyrrolidinediones reported herein have been evaluated against S. aureus and methicillin-resistant S. aureus (MRSA) biofilms, and this work constitutes our first report on the antibiofilm properties of this class of compounds. The antibiofilm activity of these 2,3-pyrrolidinediones has been assessed through minimum biofilm eradication concentration (MBEC) and minimum biofilm inhibition concentration (MBIC) assays. The compounds displayed antibiofilm properties and represent intriguing scaffolds for further optimization and development.
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Antibacterianos , Staphylococcus aureus Resistente à Meticilina , Antibacterianos/farmacologia , Staphylococcus aureus , Resistência a Meticilina , Biofilmes , Testes de Sensibilidade MicrobianaRESUMO
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 (â¼1 MPa) promoting an elongated nuclear aspect ratio while those on high stiffness fibers (â¼8 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.
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Gelatina , Tenócitos , Alicerces Teciduais , Gelatina/química , Animais , Cavalos , Tenócitos/citologia , Tenócitos/metabolismo , Alicerces Teciduais/química , Fenômenos Mecânicos , Regulação da Expressão Gênica , Forma Celular , Fenômenos BiomecânicosRESUMO
OBJECTIVE: The objective of the study was to determine the effect of gentamicin on CD3+ T-lymphocyte proliferation and cell viability using an in vitro cell culture model as a means of investigating the mechanism of action of low-dose intravitreal gentamicin injection. ANIMALS STUDIED: Three adult horses with no evidence of ophthalmic or systemic disease. PROCEDURE: Peripheral blood lymphocytes were treated with gentamicin at concentrations 37.5 µg/mL, 112.5 µg/mL, 187 µg/mL, 375 µg/mL, or 750 µg/mL then stimulated to proliferate with concanavalin A (ConA). 4',6-diamidino-2-phenylindole (DAPI) and carboxyfluoroscein succinimidyl ester (CSFE) were used as markers of cell viability and cell proliferation, respectively. Following 5-day culture, live cell counts and CSFE fluorescent intensity data were collected via automated cell count and flow cytometry. The experimental design was duplicated using preservative-free gentamicin and a proprietary brand formulation. Statistical analysis was performed using two-way ANOVA with Tukey's multiple comparison test. RESULTS: No statistically significant comparisons in CD3+ T-lymphocyte live cell counts and geometric mean fluorescent intensity of CSFE were identified between gentamicin concentrations or formulations. CONCLUSIONS: Gentamicin had no effect on equine peripheral blood CD3+ T-lymphocyte cell viability and proliferation in concentrations ranging from "safe" to "retinotoxic" in relation to intravitreal injection volumes. Low-dose intravitreal gentamicin may not suppress the Th1- and Th17-mediated immune response.
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Doenças dos Cavalos , Uveíte , Animais , Cavalos , Gentamicinas/farmacologia , Gentamicinas/uso terapêutico , Projetos de Pesquisa , Uveíte/tratamento farmacológico , Uveíte/veterinária , Linfócitos T , Proliferação de Células , Doenças dos Cavalos/tratamento farmacológicoRESUMO
Limited information exists regarding associations between distal interphalangeal joint (DIPJ) abnormalities and synovial invagination changes in the distal sesamoid (navicular) bone. This retrospective, analytical study aimed to measure specific characteristics of the synovial invaginations of the navicular bone to determine whether any single characteristic was associated with abnormalities in the DIPJ or navicular apparatus (NA) using high field MRI and a sample of 200 horses' feet. The DIPJ and NA were graded independently by three scorers. The grades were averaged, creating a global pathology score for the DIPJ, NA, and synovial invaginations. Higher global scores represented more severe pathology. The number of invaginations, depth of penetration, invagination shape, and cross-sectional area (CSA) of the largest invagination were recorded. Interobserver agreement was measured using Cohen's Kappa. Associations of global scores of the DIPJ and NA with individual invagination characteristics were assessed using linear mixed modeling. A significant relationship was found between the number of invaginations and global DIPJ score, with higher invagination numbers associated with higher DIPJ scores. For invagination depth and CSA, a significant relationship was noted with global scores of both the DIPJ and NA. Reliable relationships between the shape of synovial invaginations and global scores of DIPJ and NA were not found, likely due to poor interobserver scoring (0.305). These findings suggest that primary DIPJ disease and NA pathology should be considered when noticing alterations to navicular synovial invaginations on MRI. This contrasts traditional views that synovial invagination abnormalities are indicative solely of NA pathology.
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Doenças dos Cavalos , Ossos do Tarso , Animais , Cavalos , Estudos Retrospectivos , Articulações , Ossos do Tarso/diagnóstico por imagem , Imageamento por Ressonância Magnética/veterinária , PéRESUMO
Increasing antimicrobial resistance in veterinary practice has driven the investigation of novel therapeutic strategies including regenerative and biologic therapies to treat bacterial infection. Integration of biological approaches such as platelet lysate and mesenchymal stromal cell (MSC) therapy may represent adjunctive treatment strategies for bacterial infections that minimize systemic side effects and local tissue toxicity associated with traditional antibiotics and that are not subject to antibiotic resistance. In this review, we will discuss mechanisms by which biological therapies exert antimicrobial effects, as well as potential applications and challenges in clinical implementation in equine practice.
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Doenças dos Cavalos , Células-Tronco Mesenquimais , Cavalos , Animais , Doenças dos Cavalos/terapia , Plaquetas , AntibacterianosRESUMO
While MRI is the modality of choice for the diagnosis of longitudinal tears (LTs) of the deep digital flexor tendon (DDFT) of horses, differentiating between various grades of tears based on imaging characteristics is challenging due to overlapping imaging features. In this retrospective, exploratory, diagnostic accuracy study, a machine learning (ML) scheme was applied to link quantitative features and qualitative descriptors to leverage MRI characteristics of different grades of tearing of the DDFT of horses. A qualitative MRI characteristic scheme, combining tendon morphologic features, altered signal intensity, and synovial sheath distention, was used for LT classification with an excellent diagnostic accuracy of the high-grade tears but more limited accuracy for the detection of low-grade tears. A quantitative ML approach was followed to measure the contribution of 30 quantitative phenotypic features for characterizing and classifying tendinous tears. Among the 30 imaging features, boundary curvature represented by the standard deviation and maximum had the most significant discriminatory power (P < 0.05) between normal and abnormal tendons and could be used as an aid for classifying the different grades of LTs of DDFTs. Imaging analysis-based 3D interactive surface plot supports qualitative characterization of different grades of LTs of the DDFT through clearer visualization of the tendon in three dimensions and simple integration of two perspectives features (i.e., margin/distribution and intensity/distribution). A systematic approach combining quantitative features with qualitative analyses using ML was diagnostically beneficial in MRI characterization and in discriminating between different grades of LTs of the DDFT of horses.
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Doenças dos Cavalos , Animais , Doenças dos Cavalos/diagnóstico , Cavalos , Aprendizado de Máquina , Imageamento por Ressonância Magnética/veterinária , Estudos Retrospectivos , Tendões/diagnóstico por imagemRESUMO
The failure of frontline antibiotics in the clinic is one of the most serious threats to human health and requires a multitude of novel therapeutics and innovative approaches to treatment so as to curtail the growing crisis. In addition to traditional resistance mechanisms resulting in the lack of efficacy of many antibiotics, most chronic and recurring infections are further made tolerant to antibiotic action by the presence of biofilms. Herein, we report an expanded set of 5-benzylidene-4-oxazolidinones that are able to inhibit the formation of Staphylococcus aureus biofilms, disperse preformed biofilms, and, in combination with common antibiotics, are able to significantly reduce the bacterial load in a robust collagen-matrix model of biofilm infection.
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Antibacterianos/farmacologia , Compostos de Benzilideno/farmacologia , Biofilmes/efeitos dos fármacos , Oxazolidinonas/farmacologia , Staphylococcus aureus/fisiologia , Acinetobacter baumannii/efeitos dos fármacos , Sinergismo Farmacológico , Staphylococcus aureus Resistente à Meticilina/efeitos dos fármacos , Testes de Sensibilidade Microbiana , Relação Estrutura-AtividadeRESUMO
Equine immune-mediated keratitis (IMMK) leads to increased corneal opacity and inflammation secondary to an alteration of the local immune system. Bone marrow-derived mesenchymal stem cells (BM-MSC) have been shown to modulate the immune system by downregulating inflammation. Four horses with unilateral IMMK poorly responsive to traditional medical treatments underwent novel, autologous subconjunctival BM-MSC therapy. Bone marrow was harvested and processed as previously described for equine orthopedic disease. Horses received autologous subconjunctival BM-MSC injections approximately every 3-4 weeks for 1-5 treatments total. Horses were maintained on their current medical treatment regimen throughout the BM-MSC treatment period. Three horses had a positive response to therapy as demonstrated by an increase in corneal clarity, a decrease in neovascularization and a reduction in surface irregularity. One horse was nonresponsive to therapy. These experimental results demonstrate the safety and potential efficacy of an innovative solution for IMMK.
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Doenças dos Cavalos/terapia , Ceratite/veterinária , Transplante de Células-Tronco Mesenquimais/veterinária , Animais , Feminino , Cavalos , Ceratite/terapia , Masculino , Resultado do TratamentoRESUMO
OBJECTIVE: To characterize the growth factors contained in equine amniotic membrane allograft (eAM; StemWrap scaffold and StemWrap+ injection) and to evaluate the effect of eAM on equine distal limb wound healing. STUDY DESIGN: Prospective experimental controlled study. SAMPLE POPULATION: Eight adult horses. METHODS: Transforming growth factor (TGF)-ß1, vascular endothelial growth factor (VEGF), epidermal growth factor, platelet-derived growth factor-BB, and prostaglandin E2 (PGE2 ) concentrations in StemWrap+ were assessed with enzyme-linked immunosorbent assay. Two full-thickness 6.25-cm2 skin wounds were created on each metacarpus. On one forelimb, one wound was treated with eAM, and the other was left untreated (eAM control). On the contralateral limb, one wound was treated with a silicone dressing, and the other served as negative control. Three-dimensional images were obtained to determine wound circumference and surface area analyses at each bandage change until healed. Excessive granulation tissue was debrided once weekly for 4 weeks. Biopsy samples were taken to evaluate quality of wound healing via histologic and immunohistochemistry assays. RESULTS: StemWrap+ contained moderate concentrations of TGF-ß1 (494.10 pg/mL), VEGF (212.52 pg/mL), and PGE2 (1811.61 pg/mL). Treatment of wounds with eAM did not affect time to healing or histologic quality of the healing compared with other groups but was associated with increased granulation tissue production early in the study, particularly on day 7. CONCLUSION: Application of eAM resulted in increased granulation tissue production while maintaining appropriate healing of experimental wounds. CLINICAL SIGNIFICANCE: Use of eAM is likely most beneficial for substantial wounds in which expedient production of large amounts of granulation tissue is desirable.
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Âmnio , Doenças dos Cavalos/terapia , Cavalos/lesões , Cicatrização/fisiologia , Ferimentos e Lesões/veterinária , Aloenxertos , Animais , Bandagens/veterinária , Tecido de Granulação/fisiologia , Estudos Prospectivos , Pele/lesões , Pele/patologia , Ferimentos e Lesões/terapiaRESUMO
OBJECTIVES: Mesenchymal stem cells (MSCs) are frequently delivered via needle injection for treatment of musculoskeletal injuries. The purpose of this study was to evaluate the effect of needle diameter on the viability of MSCs. METHODS: Equine bone marrow-derived MSCs from 5 horses were suspended in PBS, and held at room temperature for 7 hours to mimic shipping conditions. Two replicate samples for each needle size (20, 22, 23, or 25-gauge [ga]) were aspirated into a 3 mL syringe and re-injected into the holding vial 3 times, to reproduce the resuspension of cells prior to injection in clinical cases. Cells were stained with fluorescein diacetate and propidium iodide to measure viability. Flow cytometry (FC) was performed to compare cell debris and intact cells between groups. RESULTS: MSC viability was higher when cells were passed through a 20-ga rather than a 25-ga needle. Cell suspensions passed through a 20-ga needle contained a larger percentage of intact cells, compared to 25-ga samples. The percentage of debris present in cell suspensions tended to increase with decreasing needle diameter. Neither horse nor passage had a significant effect on viability. CONCLUSIONS: Cell damage is more likely when MSCs are passed through 25-ga rather than 20-ga needles. CLINICAL RELEVANCE: Use of needles larger than 25-ga is recommended to maintain the viability of MSCs injected in horses.
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Células da Medula Óssea/fisiologia , Cavalos , Células-Tronco Mesenquimais/fisiologia , Agulhas , Manejo de Espécimes/veterinária , Animais , Técnicas de Cultura de Células , Manejo de Espécimes/instrumentaçãoRESUMO
OBJECTIVE: To report a transnasal, endoscopically guided ventral surgical approach for accessing the cranial and caudal segments of the sphenopalatine sinus for mass removal in a horse. STUDY DESIGN: Case report. ANIMAL: Adult horse with acute onset blindness referable to a soft tissue mass within the sphenopalatine sinus. CLINICAL REPORT: A 7-year-old Warmblood gelding presented with a history of running into a fence and falling. No neurologic signs were identified at initial examination but acute blindness was noted 3 weeks later. On computed tomography (CT) the sphenopalatine sinus was filled with a large homogeneous mass with poor contrast enhancement that extended dorsally with thinning to the dorsal cortex of the sphenoid bone, just rostral to the entrance of the optic canals into the cranial cavity. Surgical access to the sphenopalatine sinus was achieved using a transnasal, endoscopically guided ventral pharyngotomy approach and the mass lesion was removed. A presumptive diagnosis of chondroma was made based on histopathology. The horse recovered well from surgery, and although it has not regained vision as of 6.5 years postoperatively, the disease has not progressed. CONCLUSION: Transnasal, endoscopically-guided ventral surgical access to the sphenopalatine sinus is possible in horses and may improve access in horses with disease extending caudally beyond the palatine portion of the sinus. Use of smaller diameter or specialized instruments, such as various endoscopic bone cutting instruments, and CT image guidance may improve sinus access by this route.
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Condroma/veterinária , Doenças dos Cavalos/cirurgia , Faringectomia/veterinária , Crânio/cirurgia , Animais , Condroma/diagnóstico , Condroma/cirurgia , Endoscopia/veterinária , Doenças dos Cavalos/diagnóstico , Cavalos , Masculino , Faringectomia/métodosRESUMO
OBJECTIVES: To evaluate the veterinary student learning outcome of 2 methods of equine rectal examination training. STUDY DESIGN: Randomized prospective study. SAMPLE POPULATION: Veterinary students (3rd and 4th year; n = 40) and practicing equine veterinarians (n = 10). METHODS: Year 1: Group 1 (n = 11) and Group 2 students (n = 10) received skilled instructor (SI) and laparoscopic-guidance (LG), respectively, during rectal exam instruction. All students were tested on rectal identification of 4 abdominal organs. Year 2: One group of students (n = 19) was trained and subsequently tested using each technique, first SI, followed by LG. Subjective evaluation of laparoscopy as a teaching tool was achieved with veterinary students and equine practitioners. RESULTS: A significantly greater percentage of students having LG compared to SI were able to correctly identify the left kidney (Year 1) and the spleen, cecum, and right ovary (Year 2). A significantly greater proportion of LG trained students in years 1 and 2 (100% and 95%, respectively) were also able to identify 75% of organs compared with SI (27% and 21%, respectively). Both students and veterinarians uniformly provided favorable feedback for LG in teaching rectal palpation skills. CONCLUSION: The LG method of equine rectal examination instruction resulted in improved learning for identification of several key abdominal organs compared with SI.
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Competência Clínica , Doenças dos Cavalos/patologia , Laparoscopia/veterinária , Palpação/veterinária , Reto/patologia , Animais , Educação em Veterinária , Cavalos , Humanos , Laparoscopia/métodos , Avaliação de Programas e Projetos de Saúde , Estudos ProspectivosRESUMO
OBJECTIVE: The study objectives were to 1) determine the mesenchymal stem cell (MSC) surface expression of major histocompatibility complex (MHC) class I and transcriptome-wide gene expression changes following IL-1ß + TGF-ß2 dual licensing and 2) evaluate if IL-1ß + TGF-ß2 dual-licensed MSCs had a greater ability to positively modulate tenocyte function compared to naive MSCs. SAMPLE: Equine bone marrow-derived MSCs from 6 donors and equine superficial digital flexor tenocytes from 3 donors. METHODS: Experiments were performed in vitro. Flow cytometry and bulk RNA sequencing were utilized to determine naive and dual-licensed MSC phenotype and transcriptome-wide changes in gene expression. Conditioned media were generated from MSCs and utilized in tenocyte cell culture assays as a method to determine the effect of MSC paracrine factors on tenocyte function. RESULTS: Dual-licensed MSCs have a reduced expression of MHC class I and exhibit enrichment in functional pathways associated with the extracellular matrix, cell signaling, and tissue development. Additionally, dual-licensed MSC-conditioned media significantly improved in vitro tenocyte migration and metabolism to a greater degree than naive MSC-conditioned media. In tenocytes exposed to IL-1ß, dual-licensed conditioned media also positively modulated tenocyte gene expression. CLINICAL RELEVANCE: Our data indicate that conditioned media containing paracrine factors secreted from dual-licensed MSCs significantly modulates in vitro tenocyte function, which may confer benefits in vivo to healing tendons following injury. Additionally, due to reduced MHC class I expression in dual-licensed MSCs, this technique may also provide an avenue to provide an effective "off-the-shelf" allogenic source of MSCs.
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Interleucina-1beta , Células-Tronco Mesenquimais , Tenócitos , Fator de Crescimento Transformador beta2 , Animais , Células-Tronco Mesenquimais/metabolismo , Cavalos , Interleucina-1beta/metabolismo , Interleucina-1beta/genética , Fator de Crescimento Transformador beta2/metabolismo , Fator de Crescimento Transformador beta2/genética , Tenócitos/metabolismo , Movimento Celular/efeitos dos fármacos , Regulação da Expressão Gênica/efeitos dos fármacos , Antígenos de Histocompatibilidade Classe I/metabolismo , Antígenos de Histocompatibilidade Classe I/genética , Células CultivadasRESUMO
Steroid-associated laminitis remains a major concern with use of corticosteroids in horses. Individual case factors such as joint pathology, pre-existing endocrinopathies, or corticosteroid type, dose, and timing influencing steroid-induced laminitis risk have not been investigated. This study aimed to determine if systemic absorption of triamcinolone acetonide (TA) varies between intrasynovial (antebrachiocarpal) and extrasynovial (sacroiliac) injection sites, and to determine the effects of TA absorption on glucose, insulin, cortisol, and adrenocorticotropic hormone (ACTH). Twenty adult horses were randomized into antebrachiocarpal or sacroiliac joint injection groups, and each horse received bilateral injections with a total dose of 18 mg triamcinolone. Blood was collected prior to injection and at 1, 2, 4, 6, 8, 10, 12, 16, 20, 24, 36, 48, 60, and 72 h post-injection. Peak TA absorption occurred at 8 h in both groups, and was significantly higher in the intrasynovial group compared to the extrasynovial group (1.397 ng/mL, 0.672 ng/mL, p < 0.05). Plasma TA levels were significantly higher in the intrasynovial group from 8 to 36 h post-injection (p < 0.05). There was no difference in glucose, insulin, cortisol, or ACTH between groups at any time point. Insulin and glucose were significantly increased from baseline at all timepoints from 10-72 h and 1-72 h post-injection, respectively. Horses with elevated baseline insulin values (>20 µU/mL) from both groups experienced a more marked hyperinsulinemia, reaching a mean peak insulin of 197.5 µU/mL as compared to 90.06 µU/mL in those with normal baseline insulin. Cortisol and ACTH were significantly decreased from baseline at timepoints from 4-72 h post-injection in both groups. This study is the first to evaluate drug absorption from the sacroiliac site and demonstrates that drug absorption varies between intrasynovial and extrasynovial injection sites. TA absorption causes metabolic derangements, most notably a marked hyperinsulinemia that is more severe in horses with elevated baseline insulin values. The influence of baseline endocrinopathies on response to corticosteroid administration as well as the effect of corticosteroid-induced metabolic derangements warrant further investigation as risk factors for corticosteroid-associated laminitis.
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OBJECTIVE: To compare palpation and ultrasound scores of effusion of the medial femorotibial and femoropatellar joints of horses. ANIMALS: 40 horses (80 stifles) were evaluated over a 12-week period. METHODS: Horses > 1 year of age without history of stifle disease were enrolled from September to December 2022. Palpation of right and left medial femorotibial and femoropatellar joint compartments was performed. Amount of effusion was scored by a board-certified large animal surgeon, a third-year large animal surgery resident, and an equine sports medicine intern. Effusion of right and left medial femorotibial and femoropatellar joints was quantified with ultrasound by a board-certified equine sports medicine and rehabilitation clinician. Amount of effusion on palpation and ultrasound was graded as none-mild (1), moderate (2), or severe (3). A 2-way intraclass correlation coefficient evaluated interrater reliability of palpation scores. The Spearman rank correlation determined association between palpation and ultrasound scores. RESULTS: Interrater reliability for palpation of effusion was poor between all observers for all joint compartments. No significant correlation was identified between palpation and ultrasound scores for any joint compartment for any observer. CLINICAL RELEVANCE: Clinicians often rely on palpation of joint effusion as an indication of stifle pathology. We found interrater reliability to be poor for palpation scores, indicating low agreement for palpation of joint effusion between clinicians within our group. No correlation was found between palpation and ultrasound scores for joint effusion, indicating that clinicians should not rely on palpation alone to quantify joint effusion of the medial femorotibial and femoropatellar joints.
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Líquidos Corporais , Doenças dos Cavalos , Cavalos , Animais , Reprodutibilidade dos Testes , Joelho de Quadrúpedes/patologia , Articulações/patologia , Doenças dos Cavalos/diagnóstico por imagem , Doenças dos Cavalos/patologiaRESUMO
Uncontrolled bleeding after trauma represents a substantial clinical problem. The current standard of care to treat bleeding after trauma is transfusion of blood products including platelets; however, donated platelets have a short shelf life, are in limited supply, and carry immunogenicity and contamination risks. Consequently, there is a critical need to develop hemostatic platelet alternatives. To this end, we developed synthetic platelet-like particles (PLPs), formulated by functionalizing highly deformable microgel particles composed of ultralow cross-linked poly (N-isopropylacrylamide) with fibrin-binding ligands. The fibrin-binding ligand was designed to target to wound sites, and the cross-linking of fibrin polymers was designed to enhance clot formation. The ultralow cross-linking of the microgels allows the particles to undergo large shape changes that mimic platelet shape change after activation; when coupled to fibrin-binding ligands, this shape change facilitates clot retraction, which in turn can enhance clot stability and contribute to healing. Given these features, we hypothesized that synthetic PLPs could enhance clotting in trauma models and promote healing after clotting. We first assessed PLP activity in vitro and found that PLPs selectively bound fibrin and enhanced clot formation. In murine and porcine models of traumatic injury, PLPs reduced bleeding and facilitated healing of injured tissue in both prophylactic and immediate treatment settings. We determined through biodistribution experiments that PLPs were renally cleared, possibly enabled by ultrasoft particle properties. The performance of synthetic PLPs in the preclinical studies shown here supports future translational investigation of these hemostatic therapeutics in a trauma setting.
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Hemostáticos , Roedores , Animais , Camundongos , Suínos , Roedores/metabolismo , Distribuição Tecidual , Plaquetas/metabolismo , Hemorragia , Fibrina/química , Fibrina/metabolismoRESUMO
Introduction: One major obstacle in validating drugs for the treatment or prevention of hearing loss is the limited data available on the distribution and concentration of drugs in the human inner ear. Although small animal models offer some insights into inner ear pharmacokinetics, their smaller organ size and different barrier (round window membrane) permeabilities compared to humans can complicate study interpretation. Therefore, developing a reliable large animal model for inner ear drug delivery is crucial. The inner and middle ear anatomy of domestic pigs closely resembles that of humans, making them promising candidates for studying inner ear pharmacokinetics. However, unlike humans, the anatomical orientation and tortuosity of the porcine external ear canal frustrates local drug delivery to the inner ear. Methods: In this study, we developed a surgical technique to access the tympanic membrane of pigs. To assess hearing pre- and post-surgery, auditory brainstem responses to click and pure tones were measured. Additionally, we performed 3D segmentation of the porcine inner ear images and used this data to simulate the diffusion of dexamethasone within the inner ear through fluid simulation software (FluidSim). Results: We have successfully delivered dexamethasone and dexamethasone sodium phosphate to the porcine inner ear via the intratympanic injection. The recorded auditory brainstem measurements revealed no adverse effects on hearing thresholds attributable to the surgery. We have also simulated the diffusion rates for dexamethasone and dexamethasone sodium phosphate into the porcine inner ear and confirmed the accuracy of the simulations using in-vivo data. Discussion: We have developed and characterized a method for conducting pharmacokinetic studies of the inner ear using pigs. This animal model closely mirrors the size of the human cochlea and the thickness of its barriers. The diffusion time and drug concentrations we reported align closely with the limited data available from human studies. Therefore, we have demonstrated the potential of using pigs as a large animal model for studying inner ear pharmacokinetics.
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OBJECTIVES: To (1) evaluate the design and use of a global rating scale assessment instrument in veterinary medical education and; (2) examine the effectiveness of 2 surgical techniques courses for improving the surgical skills of veterinary students. STUDY DESIGN: Instrument development; observational; survey-based. SAMPLE POPULATION: Students (n = 16) registered for 2 elective surgical techniques courses were enrolled on a volunteer basis. METHODS: A 5-point global rating scale instrument was designed for the evaluation of 12 basic surgical skills by faculty evaluators and used to obtain student start and end scores during the courses. Upon conclusion of the courses, students completed a survey from which their opinions on their improvement as well as their desire for feedback were obtained. RESULTS: All authors agreed the instrument was easy to use. As groups, 3rd year students, 4th year students, and all students combined had significantly higher total skill scores at the end of the courses compared to the start of the courses. Individually, 10 students (63%) had significant improvement in surgical skills as a result of their participation in the courses: 4 (100%) 3rd year and 6 (50%) 4th year students. Student survey responses revealed a strong desire for feedback as well as support of formal assessment methods. Only weak agreement was found between student opinions on their improvement and the authors' assessment scores. CONCLUSIONS: Assessment instruments are useful for (1) student evaluation and (2) for providing students with feedback on their surgical skills.
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Competência Clínica/normas , Educação em Veterinária/normas , Estudantes , Procedimentos Cirúrgicos Operatórios/educação , AnimaisRESUMO
Tendon injuries are common in both veterinary and human clinical patients and result in morbidity, pain, and lost athletic performance. Consequently, utilizing naturally occurring injuries in veterinary patients as a comparative model could inform the development of novel therapies and increase translation for the treatment of human tendon injuries. Mesenchymal stem cells (MSCs) have shown considerable efficacy for the treatment of experimental and clinical superficial digital flexor tendon injury in the horse; however, the reinjury rate following treatment can remain high and MSC efficacy in treating other tendons is less well known. Additionally, the translation of MSC therapy to human tendon injury has remained poor. Recent evidence indicates that naïve MSC function can be enhanced through exogenous stimulation or manipulation of their environment. This stimulation or activation, herein termed MSC licensing, markedly alters MSC functions associated with immunomodulation, extracellular matrix remodeling, vascular development, bioactive factor production, and endogenous stromal/progenitor cell support. Additionally, a variety of licensing strategies has proven to influence MSC-secreted factors that have positively influenced outcome parameters in both in vitro and in vivo disease models separate from musculoskeletal tissues. Therefore, identifying the optimal licensing strategy for MSCs could ultimately provide an avenue for reliable and repeatable treatment of a broad range of tendon injuries of both veterinary and human clinical patients. This article details current evidence on the effects of licensed MSCs in both in vitro and in vivo disease models of different species and provides commentary on how those effector functions identified may be translated to the treatment of tendon injuries.