Anterior cruciate ligament (ACL) reconstruction after injury does not prevent post-traumatic osteoarthritis (PTOA). Circulating microRNA (miRNA) and metabolite changes emerging shortly after ACL injury and reconstruction remain insufficiently defined, potentially harbouring early cues contributing to PTOA evolution. Moreover, their differential expression between females and males also may influence PTOA’s natural trajectory. This study aims to determine alterations in plasma miRNA and metabolite levels in the early stages following ACL reconstruction and between females and males. A cohort of 43 ACL reconstruction patients was examined. Plasma was obtained at baseline, 2 weeks, and 6 weeks post-surgery (129 biospecimens in total). High-throughput miRNA sequencing and metabolomics were conducted. Differentially expressed miRNAs and metabolites were identified using negative binomial and linear regression models, respectively. Associations between miRNAs and metabolites were explored using time and sex as co-variants, (pre-surgery versus 2 and 6 weeks post-surgery). Using computational biology, miRNA-metabolite-gene interaction and pathway analyses were performed. Levels of 46 miRNAs were increased at 2 weeks post-surgery compared to pre-surgery (baseline) using miRNA sequencing. Levels of 13 metabolites were significantly increased while levels of 6 metabolites were significantly decreased at 2 weeks compared to baseline using metabolomics. Hsa-miR-145-5p levels were increased in female subjects at both 2 weeks (log-fold-change 0.71, 95%CI 0.22,1.20) and 6 weeks (log-fold-change 0.75, 95%CI 0.07,1.43) post-surgery compared to males. In addition, hsa-miR-497-5p showed increased levels in females at 2 weeks (log-fold-change 0.77, 95%CI 0.06,1.48) and hsa-miR-143-5p at 6 weeks (log-fold-change 0.83, 95%CI 0.07,1.59). Five metabolites were decreased at 2 weeks post-surgery in females compared to males: L-leucine (−1.44, 95%CI −1.75,−1.13), g-guanidinobutyrate (−1.27, 95%CI 1.54,−0.99), creatinine (−1.17, 95%CI −1.44,−0.90), 2-methylbutyrylcarnitine (−1.76, 95%CI −2.17,−1.35), and leu-pro (−1.13, 95%CI −1.44,−0.83). MiRNA-metabolite-gene interaction analysis revealed key signalling pathways based on post-surgical time-point and in females versus males. MiRNA and metabolite profiles were modified by time and by sex early after ACL reconstruction surgery, which could influence surgical response and ultimately risk of developing PTOA.

中文翻译：

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前交叉韧带重建手术后早期 microRNA 和代谢物的变化


损伤后前十字韧带 (ACL) 重建并不能预防创伤后骨关节炎 (PTOA)。 ACL 损伤和重建后不久出现的循环 microRNA (miRNA) 和代谢物变化仍未得到充分定义，可能蕴藏着促进 PTOA 进化的早期线索。此外，它们在女性和男性之间的差异表达也可能影响 PTOA 的自然轨迹。本研究旨在确定 ACL 重建后早期阶段以及女性和男性之间血浆 miRNA 和代谢物水平的变化。对 43 名 ACL 重建患者进行了检查。在基线、术后 2 周和术后 6 周获取血浆（总共 129 个生物样本）。进行了高通量 miRNA 测序和代谢组学。分别使用负二项式和线性回归模型鉴定差异表达的 miRNA 和代谢物。使用时间和性别作为协变量（手术前与手术后 2 周和 6 周）来探索 miRNA 和代谢物之间的关联。利用计算生物学，进行了 miRNA-代谢物-基因相互作用和通路分析。使用 miRNA 测序，与手术前（基线）相比，术后 2 周 46 种 miRNA 的水平有所增加。与代谢组学基线相比，2 周时 13 种代谢物的水平显着增加，而 6 种代谢物的水平显着降低。女性受试者的 Hsa-miR-145-5p 水平在 2 周（对数倍数变化 0.71，95%CI 0.22,1.20）和 6 周（对数倍数变化 0.75，95%CI 0.07,1.43）时均有所增加术后与男性相比。此外，hsa-miR-497-5p 在 2 周时在雌性中表现出水平升高（对数倍数变化 0.77，95% CI 0.06,1）。48) 和 6 周时的 hsa-miR-143-5p (对数倍数变化 0.83, 95% CI 0.07,1.59)。与男性相比，女性术后 2 周有五种代谢物减少：L-亮氨酸 (-1.44, 95%CI -1.75,-1.13)、g-胍丁酸 (-1.27, 95%CI 1.54,-0.99)、肌酐(-1.17, 95%CI -1.44,-0.90)、2-甲基丁酰肉碱 (-1.76, 95%CI -2.17,-1.35) 和 leu-pro (-1.13, 95%CI -1.44,-0.83)。 MiRNA-代谢物-基因相互作用分析揭示了基于术后时间点以及女性与男性的关键信号传导途径。 ACL 重建手术后早期，miRNA 和代谢谱随时间和性别发生变化，这可能会影响手术反应并最终影响发生 PTOA 的风险。