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Abstract

OBJECTIVE

To investigate matrix metalloproteinase (MMP) and their inhibitors tissue inhibitor matrix metalloproteinase (TIMP) gene expression and secretion during equine deep digital flexor tendon (DDFT) tenocyte and macrophage (undifferentiated, proinflammatory, and regulatory) co-culture.

SAMPLE

Third passage DDF tenocytes and donor-matched macrophages differentiated from peripheral blood CD14+ monocytes from 5 healthy horses ages 9–11 years, euthanized for reasons unrelated to musculoskeletal conditions.

METHODS

Passage 3 DDT tenocyte aggregate cultures were co-cultured with undifferentiated (control), proinflammatory (granulocyte-macrophage colony-stimulating factor; GM-CSF pretreated and lipopolysaccharide + interferon gamma-primed; LPS+IFN-γ) or regulatory (interleukin-4 and interleukin-10-primed; IL-4 + IL-10) macrophages in direct and transwell co-cultures for 72 hours. MMP-1, -2, -3, -9, -13, and TIMP -1, -2 mRNA were measured via real-time Polymerase Chain Reaction (rtPCR). Co-culture media MMP -3, -9, and TIMP -1, -2 concentrations were quantified via ELISA.

RESULTS

Direct co-culture of DDF tenocytes with proinflammatory macrophages for 72 hours increased MMP-1, -3, and -13 mRNA levels whereas, MMP-9 mRNA levels decreased. Direct and transwell co-culture with proinflammatory and regulatory macrophages resulted in increased MMP-3 and decreased MMP-9 media concentrations. While direct co-culture with regulatory macrophages significantly increased TIMP-1 mRNA, overall, TIMP mRNA and culture media concentrations were largely unchanged.

CLINICAL RELEVANCE

Cell-to-cell contact between DDF tenocytes and macrophages is not essential to induce MMP gene expression and secretion. Co-culture systems offer a viable in vitro platform to screen and evaluate immunomodulatory properties of therapies aimed at improving equine intrasynovial tendon healing.

Open access
in American Journal of Veterinary Research

Abstract

OBJECTIVE

To investigate (1) variables associated with the likelihood of obtaining a positive culture, (2) commonly isolated microorganisms, and (3) antimicrobial resistance patterns of isolates from horses with presumptive synovial sepsis.

SAMPLES

Synovial fluid, synovium, and bone samples from equine cases with presumptive synovial sepsis submitted to the Cornell University Animal Health Diagnostic Center from 2000 to 2020 for microbial culture and antimicrobial sensitivity testing.

PROCEDURES

Univariable and multivariable analyses were performed to determine the effect of variables on the likelihood of positive culture. Frequency distributions for isolated organisms and antimicrobial resistance were generated. Multidrug resistance patterns and associations were assessed with association rule mining.

RESULTS

The positive culture rate for all samples was 37.4%, while the positive culture rate among samples confirmed to be septic by a combination of clinical pathological variables and case details was 43%. Blood culture vial submissions were 1.7 times more likely to yield a positive culture compared to samples submitted in a serum tube. Structure sampled, tissue submitted, and horse age were associated with a positive culture. Staphylococcus spp (23.7%), Streptococcus spp (22.4%), and Enterococcus spp (9.67%) were commonly isolated. Multidrug resistance prevalence decreased from 92% (2000 to 2009) to 76% (2010 to 2020) of gram-negative isolates and 60% (2000 to 2009) to 52% (2010 to 2020) of gram-positive isolates.

CLINICAL RELEVANCE

The positive culture rate from synovial fluid submissions with traditional sampling and culture methods remains low and may be optimized by submitting samples in blood culture vials. Overall, antimicrobial resistance was frequently observed but did not increase from the first to second decade for most genera.

Open access
in American Journal of Veterinary Research