RESUMEN
Reconstruction surgery for acute proximal anterior cruciate ligament (ACL) tears remains controversial. Recently, ACL primary repair has received increasing attention in ACL treatment. This study aimed to explore the histological characteristics of ACL healing in primary repair and compare its therapeutic and prognostic results with the reconstruction of acute proximal ACL tears. Histological experiments using rabbits and a prospective clinical trial were conducted. We established a rabbit model of ACL primary repair, and histological changes were observed using haematoxylin and eosin (HE) and toluidine blue staining. We performed immunohistochemical analysis of CD34 and S-100 and measured the expression of collagen I and II using qRT-PCR, Western blotting, and immunohistochemistry. The prospective clinical trial involved performing ACL primary repair and reconstruction in patients with acute proximal ACL tears to detect proprioception and evaluate the function of joints. We discovered that primary repair promoted cell proliferation in the tendon-bone transition and ligament portions, reduced osteoarthritis-like pathological changes, and maintained blood vessels and proprioceptors within the ACL. In the clinical trial, primary repair achieved similar therapeutic outcomes, including recovery of knee function and proprioception, in the follow-up period as ACL reconstruction. However, the primary repair had a significantly shorter operative time and lower cost than reconstruction. Therefore, doctors should consider the benefit of primary repair in treating acute proximal ACL tears.
RESUMEN
BACKGROUND/AIMS: Chondrocyte apoptosis is the most common pathological feature in cartilage in osteoarthritis (OA). Transient receptor potential channel vanilloid 5 (TRPV5) is important in regulating calcium ion (Ca2+) influx. Accumulating evidences suggest that Ca2+ is a major intracellular second messenger that can trigger cell apoptosis. Therefore, we investigate the potential role of TRPV5 in mediating Ca2+ influx to promote chondrocyte apoptosis in OA. METHODS: The monoiodoacetic acid (MIA)-induced rat OA model was assessed by macroscopic and radiographic analyses. Calmodulin protein immunolocalization was detected by immunohistochemistry. The mRNA and protein level of TRPV5, calmodulin and cleaved caspase-8 in articular cartilage were assessed by real time polymerase chain reaction and western blotting. Primary chondrocytes were isolated and cultured in vitro. TRPV5 small interfering RNA was used to silence TRPV5 in chondrocytes. Then, calmodulin and cleaved caspase-8 were immunolocalized by immunofluorescence in chondrocyte. Fluo-4AM staining was used to assess intracellular Ca2+ to reflect TRPV5 function of mediation Ca2+ influx. Annexin V-fluorescein isothiocyanatepropidium iodide flow cytometric analysis was performed to determine chondrocytes apoptosis. Western blotting techniques were used to measure the apoptosis-related proteins in chondrocyte level. RESULTS: Here, we reported TRPV5 was up-regulated in MIA-induced OA articular cartilage. Ruthenium red (a TRPV5 inhibitor) can relieve progression of joint destruction in vivo which promoted us to demonstrate the effect of TRPV5 in OA. We found that TRPV5 had a specific role in mediating extracellular Ca2+ influx leading to chondrocytes apoptosis in vitro. The apoptotic effect was inhibited even reversed by silencing TRPV5. Furthermore, we found that the increase Ca2+ influx triggered apoptosis by up-regulating the protein of death-associated protein, FAS-associated death domain, cleaved caspase-8, cleaved caspase-3, cleaved caspase-6, and cleaved caspase-7, and the up-regulated proteins were abolished by silencing TRPV5 or 1, 2-bis-(o-Aminophenoxy)-ethane-N,N,N',N'-tetraacetic acid, tetraacetoxymethyl ester (a Ca2+ chelating agent). CONCLUSION: The up-regulated TRPV5 could used be as an initiating factor that induces extrinsic chondrocyte apoptosis via the mediation of Ca2+ influx. These findings suggested TRPV5 could be an intriguing mediator for drug target in OA.