Unknown,Transcriptomics,Genomics,Proteomics

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The equine carpal osteochondral fragment model

ABSTRACT: Synovial fluid was obtained weekly from nine horses throughout a 70-day study period and subjected to small RNA sequencing. Nine skeletally mature Standardbred trotters (seven mares and two geldings, 2.5-7 years old, weighing 397-528 kg) were included in this study. OA was surgically induced in the left middle carpal joint and the right middle carpal joint underwent sham surgery. Under arthroscopic guidance, an osteochondral fragment was created in the left middle carpal joint by chiselling off a fragment on the dorsodistal surface of the radial carpal bone using an 8 mm curved osteotome. The fragment was left adherent to the joint capsule proximally, and an incongruent defect bed of approximately 15 mm in width was created in the exposed subchondral bone between the fragment and radial carpal bone by drilling with an arthroscopic burr. Debris from the procedure was left inside the joint. Sham surgery (arthroscopy alone) was performed in the right middle carpal joint. All portals were sutured using 2-0 propylene (Surgipro, Medtronic Danmark A/S, Copenhagen, Denmark). Both carpi were bandaged with sterile dressings. Bandages were changed at day 2 and removed at day 5. Sutures were removed 10 days after surgery. From 2 weeks following OA induction and onwards, horses were exercised in 2 min of trot (4.4-5.3 m/sec) , 2 min of fast trot/gallop (9 m/sec) and 2 min of trot (4.4-5.3 m/sec) 5 days a week on a treadmill. The horses were euthanized on day 71 or 72 with an overdose of pentobarbital sodium (140 mg/kg, Euthasol Vet, Dechra Veterinary Products, Uldum, Denmark). One horse, Horse 5, was euthanized according to predefined humane endpoints at day 49, as it became lame at the walk.

INSTRUMENT(S): Illumina HiSeq 4000

ORGANISM(S): Equus caballus

SUBMITTER: Yongxiang Fang

PROVIDER: E-MTAB-11840 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Project description:Objective: To quantify gene expression changes in the synovium of osteoarthritis-affected joints in an equine metacarpophalangeal joint (MCPJ) chip model specifically designed to recapitulate early post-traumatic osteoarthritis (PTOA). Design: Synovial samples were collected arthroscopically from the MCPJ of 11 adult horses before (pre-OA) and after (OA) surgical induction of osteoarthritis and from sham-operated joints. Briefly, in this model, an osteochondral fragment is created in one randomly chosen MCPJ at the proximal dorsomedial aspect of the first phalanx. The fragment is replaced in the fragment bed after creation so that subchondral bone is not exposed to the opposing cartilage surface. The opposite MCPJ is sham-operated. After a two week recover period, the horses are treadmill-exercised for 14 weeks and then the osteochondral fragment is removed (16 weeks after creation). Synovial tissue samples were collected arthroscopically from the MCPJ of eleven adult horses before (pre-OA) and 16 weeks after (OA) experimental induction of OA as well as from the sham-operated joints (sham). After sequencing of synovial RNA, Salmon was used to quasi-map reads to the reference genome and quantify transcript abundances. Differential expression analysis was performed with the limma-treat method using a fold-change cutoff of 1.1. Functional annotation was performed with PANTHER and Reactome at FDR < 0.05. Results: RNA was successfully extracted from 28 samples (6 pre-OA, 11 OA, 11 sham). Sequencing yielded 15.7-29.4 million paired-end reads per sample. “Sham” and “pre-OA” were not different and were grouped. 321 genes were upregulated and 351 genes were downregulated in OA synovium compared to unaffected. Gene ontology (GO) terms related to extracellular matrix (ECM) organization and growth factor binding were overrepresented among differentially expressed genes. There were 20 significantly enriched pathways; these included pathways involved in ECM turnover, O-glycosylation of TSR domain-containing proteins, and growth factor signaling. Conclusions: Most enriched pathways and overrepresented GO terms reflect a state of high metabolic activity and tissue turnover in OA-affected tissue, suggesting efforts at healing and restoring homeostasis. Limitations of this study include a small sample size and capture of a single point post-injury. Differentially expressed genes falling within key pathways may represent potential diagnostic markers or therapeutic targets for PTOA.

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The equine carpal osteochondral fragment model

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