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The purpose of this study was to evaluate the effect of single and repeat injection of amniotic suspension allograft (ASA) on pain, function, and cytokine levels using a destabilization of the medial meniscus (DMM) rat model of OA. Post\u2010DMM surgery, animals were treated with a single injection of either ASA, vehicle, or triamcinolone, or repeated injection of either ASA or vehicle. Behavioral testing including knee swelling, pain threshold, dynamic weight bearing (DWB), and gait analysis were evaluated during the in\u2010life phase. Postsacrifice, histopathology and serum and synovial fluid analyses were evaluated. Significant improvements in both DWB differentials and pain threshold were seen in response to repeated injection of ASA, while a single injection of ASA and triamcinolone resulted in significant improvements in pain threshold. Histopathology analysis found no significant differences regardless of treatment compared to vehicle, except for an increase in synovitis following repeated injection of ASA. A single injection of ASA and triamcinolone resulted in increased anti\u2010inflammatory cytokines; repeated ASA injection resulted in significant increases in several immune\u2010modulating factors relevant to OA. When comparing the impact of single and repeat ASA treatments on behavioral testing, repeated injection provided significant additional improvements in both pain and function. This study provides evidence demonstrating the impact of a second injection while also providing additional data for evaluating the use of ASA as a nonsurgical treatment for knee OA.<\/jats:p>","DOI":"10.1002\/jor.25872","type":"journal-article","created":{"date-parts":[[2024,5,23]],"date-time":"2024-05-23T10:35:00Z","timestamp":1716460500000},"page":"2159-2171","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":1,"title":["Effect of amniotic suspension allograft in a rat destabilization of medial meniscus osteoarthritis model"],"prefix":"10.1002","volume":"42","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-2167-3816","authenticated-orcid":false,"given":"Katrina A.","family":"Harmon","sequence":"first","affiliation":[{"name":"Department of Research and Development Organogenesis Birmingham Alabama 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