Autologous Bone Marrow-Derived Mesenchymal Stem Cells Modulate Molecular Markers of Inflammation in Dogs with Cruciate Ligament Rupture

Abstract

Mid-substance rupture of the canine cranial cruciate ligament rupture (CR) and associated stifle osteoarthritis (OA) is an important veterinary health problem. CR causes stifle joint instability and contralateral CR often develops. The dog is an important model for human anterior cruciate ligament (ACL) rupture, where rupture of graft repair or the contralateral ACL is also common. This suggests that both genetic and environmental factors may increase ligament rupture risk. We investigated use of bone marrow-derived mesenchymal stem cells (BM-MSCs) to reduce systemic and stifle joint inflammatory responses in dogs with CR. Twelve dogs with unilateral CR and contralateral stable partial CR were enrolled prospectively. BM-MSCs were collected during surgical treatment of the unstable CR stifle and culture-expanded. BM-MSCs were subsequently injected at a dose of 2x106 BM-MSCs/kg intravenously and 5x106 BM-MSCs by intra-articular injection of the partial CR stifle. Blood (entry, 4 and 8 weeks) and stifle synovial fluid (entry and 8 weeks) were obtained after BM-MSC injection. No adverse events after BM-MSC treatment were detected. Circulating CD8+ T lymphocytes were lower after BM-MSC injection. Serum C-reactive protein (CRP) was decreased at 4 weeks and serum CXCL8 was increased at 8 weeks. Synovial CRP in the complete CR stifle was decreased at 8 weeks. Synovial IFNγ was also lower in both stifles after BM-MSC injection. Synovial/serum CRP ratio at diagnosis in the partial CR stifle was significantly correlated with development of a second CR. Systemic and intra-articular injection of autologous BM-MSCs in dogs with partial CR suppresses systemic and stifle joint inflammation, including CRP concentrations. Intra-articular injection of autologous BM-MSCs had profound effects on the correlation and conditional dependencies of cytokines using causal networks. Such treatment effects could ameliorate risk of a second CR by modifying the stifle joint inflammatory response associated with cranial cruciate ligament matrix degeneration or damage.

Mesenchymal stem cell basic research and applications in dog medicine

“…All these in vivo studies support their therapeutic potential, although variably Hall, Rosenkrantz, Hong, Griffin, & Mendelsohn, 2010;Kim et al, 2011;Muir et al, 2016;Sousa et al, 2011). Majority of the studies have not reported any major adverse effect (Black et al, 2007(Black et al, , 2008Cuervo et al, 2014;Pérez-Merino, Usón-Casaús, Zaragoza-Bayle, et al 2015a;Bçrziòð et al, 2018;Villatoro et al, 2015), except for few studies wherein reactions like selfcontrollable localized inflammation and pulmonary parenchymal edema and hemorrhage have been reported (Kang & Park, 2014;Mokbel et al, 2011;Park et al, 2012b).…”

“…Cranial cruciate ligament (CCL) rupture represents one of the most common causes of hindlimb lameness and the most common stifle joint disease in dogs (Johnson, Austin, & Breur, 1994). Although variable but an overall improvement in the repair has been reported with MSCs (Canapp, Leasure, Cox, Ibrahim, & Carr, 2016;Muir et al, 2016). dBM-MSCs implanted intra-articularly engraft cranial cruciate ligament and thus, may have effective role to play (Linon, Spreng, Rytz, & Forterre, 2014).…”

Extracellular Vesicle-Educated Macrophages Promote Early Achilles Tendon Healing

“…Macrophage polarization, both in vitro and in vivo, can be induced by mesenchymal stromal/stem cell (MSC) paracrine activity . In mammalian models of ligament or tendon healing, MSC treatments increased the presence of endogenous M2 macrophages and associated anti‐inflammatory cytokines, which subsequently resulted in improved tendon and ligament healing . We previously reported that MSCs uniquely educate macrophages toward an anti‐inflammatory phenotype (MSC‐educated macrophages [MEMs]) .…”

Radiographic and magnetic resonance imaging predicts severity of cruciate ligament fiber damage and synovitis in dogs with cranial cruciate ligament rupture

“…Notably, eleven dogs had Synovial:Serum CRP above 0.2 in the partial CR stifle, suggesting elevated risk of subsequent contralateral rupture [32]. …”