title: Intra-articular injection of synthetic microRNA-210 accelerates avascular meniscal healing in rat medial meniscal injured model.
abstract: BACKGROUND
  The important functions of the meniscus are shock absorption, passive stabilization and load transmission of the knee. Because of the avascularity of two-thirds of the meniscal center region, the treatment of tears in this area is hard. Recently, microRNAs have been proven to play an important role in the pathogenesis of diseases. We focused on microRNA (miR)-210, which plays a wide spectrum of roles comprising mitochondrial metabolism, angiogenesis, DNA repair and cell survival. This study aimed to investigate the effect of intra-articular injection of synthetic miR-210 on the injured meniscus in the avascular zone.
  
  
  METHODS
  The middle segments of the medial meniscus of Spraque Dawley rats were incised longitudinally with a scalpel. An intra-articular injection of double-stranded (ds) miR-210 (for control group using control dsRNA) with atelocollagen was administered immediately after injury. Four weeks and 12 weeks after the injection, we conducted a histologic evaluation, immunohistochemical evaluation and Real-time PCR analysis. In vitro, the inner meniscus and synovial cells were isolated from rat knee joint, and were transfected with ds miR-210 or control dsRNA. Real-time PCR and immunohistochemical evaluations were performed.
  
  
  RESULTS
  Twenty-four hours after the injection, FAM (Fluorescein amidite) labeled miR-210 was observed in the cells around the injured site. Four weeks after the injection, the injured site of the miR-210 group was filled with repaired tissue while that of the control was not repaired. In gene expression analysis of the meniscus, the expression of miR-210, Collagen type 2 alpha 1 (Col2a1), Vascular endothelial growth factor (VEGF), and Fibroblast growth factor-2 (FGF2) in the miR-210 group was significantly higher than that in the control. At 12 weeks, the intra-articular injection of miR-210 had healed the injured site of the meniscus and had prevented articular cartilage degeneration. In vitro, miR-210 upregulated Col2a1 expression in the meniscus cells and VEGF and FGF2 expression in the synovial cells.
  
  
  CONCLUSIONS
  An intra-articular injection of ds miR-210 was effective in the healing of the damaged white zone meniscus through promotion of the collagen type 2 production from meniscus cells and through upregulated of VEGF and FGF2 from synovial cells.
authors.forename: Yoshitaka
authors.initials: Y
authors.lastname: Kawanishi
authors.forename: Tomoyuki
authors.initials: T
authors.lastname: Nakasa
authors.forename: Takeshi
authors.initials: T
authors.lastname: Shoji
authors.forename: Michio
authors.initials: M
authors.lastname: Hamanishi
authors.forename: Ryo
authors.initials: R
authors.lastname: Shimizu
authors.forename: Naosuke
authors.initials: N
authors.lastname: Kamei
authors.forename: Muhammad Andry
authors.initials: MA
authors.lastname: Usman
authors.forename: Mitsuo
authors.initials: M
authors.lastname: Ochi
mesh_terms: D000818:Animals; D004195:Disease Models, Animal; D007270:Injections, Intra-Articular; D008297:Male; D008592:Menisci, Tibial / Q000187:drug effects* / Q000473:pathology; D035683:MicroRNAs / Q000008:administration & dosage* / Q000138:chemical synthesis*; D051381:Rats; D017207:Rats, Sprague-Dawley; D000070600:Tibial Meniscus Injuries; D014945:Wound Healing / Q000187:drug effects* / Q000502:physiology
chemical_list: C546388:MIRN210 microRNA, rat; D035683:MicroRNAs