Assessment of specific mRNA levels in cartilage regions in a lapine model of osteoarthritis.
Osteoarthritis (OA) is the most common form of arthritis and patients with meniscal and ligament injuries of the knee are at high risk to develop the disease. The purpose of this study was to evaluate molecular and structural changes occurring in four articular cartilage (AC) regions from the knees of anterior cruciate ligament (ACL)-transected rabbits at 3 and 8 weeks post-surgery. Rabbit AC from the lateral and medial femoral condyles (LFC and MFC) as well as from the medial and lateral tibial plateau (MTP and LTP) were processed for histology and for semi-quantitative reverse transcriptase-polymerase chain reaction (RT-PCR) analysis for a subset of relevant molecules (collagen II, aggrecan. biglycan, decorin, fibromodulin, MMP-1, -3, -13, and TIMP-1). While the most severe histological changes were observed in the MTP starting as early as 3 weeks post-ACL transection based on Mankin scores, histological examination demonstrated a progression of osteoarthritic changes in the MFC from 3 to 8 weeks post-surgery. In contrast, very few changes were observed within both the LFC and LTP, and these changes did not worsen with increasing time after surgery. The water content increased significantly in the MFC at 8 weeks post-ACL transection and at both 3 and 8 weeks post-ACL transection in the MTP. Significant decreases in DNA content were observed for the MFC, LTP and MTP at 8 weeks post-ACL transection. Total RNA yields from the MFC and MTP were significantly elevated at 8 weeks post-ACL transection, while in the lateral compartment total RNA was unchanged following ACL transection. Analysis of mRNA levels for a subset of matrix molecules, proteinases and proteinase inhibitors, by RT-PCR demonstrated significant region-specific changes at the mRNA level following ACL transection. These results show that following ACL transection, complex molecular, as well as structural changes occur early in cartilage and that the observed changes are both region-specific and time-dependent.
Le Graverand
Marie-Pierre Hellio
MP
Faculty of Medicine, McCaig Center for Joint Injury and Arthritis Research, University of Calgary HSC, Alberta, Canada.
Eggerer
Jonna
J
Vignon
Eric
E
Otterness
Ivan G
IG
Barclay
Leona
L
Hart
David A
DA
D000818:Animals; D000070598:Anterior Cruciate Ligament Injuries; D001834:Body Water / Q000378:metabolism; D002358:Cartilage, Articular / Q000378:metabolism* / Q000473:pathology; D004247:DNA / Q000378:metabolism; D010450:Endopeptidases / Q000235:genetics; D005109:Extracellular Matrix / Q000378:metabolism; D046911:Macromolecular Substances; D009994:Osmolar Concentration; D010003:Osteoarthritis / Q000209:etiology / Q000378:metabolism* / Q000473:pathology; D012313:RNA / Q000378:metabolism; D012333:RNA, Messenger / Q000378:metabolism*; D011817:Rabbits; D020133:Reverse Transcriptase Polymerase Chain Reaction; D019715:Tissue Inhibitor of Metalloproteinase-1 / Q000235:genetics; D014950:Wounds, Penetrating / Q000150:complications
D046911:Macromolecular Substances; D012333:RNA, Messenger; D019715:Tissue Inhibitor of Metalloproteinase-1; D012313:RNA; D004247:DNA; D010450:Endopeptidases