Engraftment of autologous bone marrow cells into the injured cranial cruciate ligament in dogs

Abstract

Current research indicates that exogenous stem cells may accelerate reparative processes in joint disease but, no previous studies have evaluated whether bone marrow cells (BMCs) target the injured cranial cruciate ligament (CCL) in dogs. The objective of this study was to investigate engraftment of BMCs following intra-articular injection in dogs with spontaneous CCL injury. Autologous PKH26-labelled BMCs were injected into the stifle joint of eight client-owned dogs with CCL rupture. The effects of PKH26 staining on cell viability and PKH26 fluorescence intensity were analysed in vitro using a MTT assay and flow cytometry. Labelled BMCs in injured CCL tissue were identified using fluorescence microscopy of biopsies harvested 3 and 13 days after intra-articular BMC injection. The intensity of PKH26 fluorescence declines with cell division but was still detectable after 16 days. Labelling with PKH26 had no detectable effect on cell viability or proliferation. Only rare PKH26-positive cells were present in biopsies of the injured CCL in 3/7 dogs and in synovial fluid in 1/7 dogs. No differences in transforming growth factor-β1, and interleukin-6 before and after BMC treatment were found and no clinical complications were noted during a 1 year follow-up period. In conclusion, BMCs were shown to engraft to the injured CCL in dogs when injected into the articular cavity. Intra-articular application of PKH26-labelled cultured mesenchymal stem cells is likely to result in higher numbers of engrafted cells that can be tracked using this method in a clinical setting.

Manufacturing Mesenchymal Stromal Cells for the Treatment of Osteoarthritis in Canine Patients: Challenges and Recommendations

“…The clinical application of MSC-based therapy in veterinary medicine comprises of multiple target diseases, with the vast majority being focused on musculoskeletal conditions such as osteoarthritis (8,(25)(26)(27)(28)(29)(30)(31)(32)(33)(34), cruciate ligament disease (9,(35)(36)(37)(38), hip dysplasia (39), bone fractures and lesions (40,41), muscle tears (42,43) and tendinopathies (44,45). Canine clinical studies have also shown promising results for the treatment of diseases of the gastro-intestinal tract (46)(47)(48), central nervous system (49)(50)(51)(52)(53), cardiovascular system (54,55), cutaneous (56)(57)(58)(59) and ocular conditions (60)(61)(62)(63), resulting in encouraging safety and efficacy data (Figure 2).…”

Stem Cells in Veterinary Medicine—Current State and Treatment Options

“…It was demonstrated that the level of post-operative lameness and pain after single intra-articular injection of allogeneic BMMSC could be a valuable alternative to 1 month course of oral administration of non-steroidal anti-inflammatory drugs (NSAIDs) in dogs requiring tibial plateau leveling osteotomy (TPLO) (178). It was shown that intraarticularly injected autologous BMMSCs engraft to the site of the injured cranial crucial ligament (179) and have an anti-inflammatory effect. Post-operatively intraarticular or IV injection of autologous MSC in dogs with the same condition resulted in a decreased level of CD8+ Tcells, decreased serum and synovia CRP, and decreased synovial IFN-γ levels that persisted over 8 weeks after BMMSC injection (180).…”

High-content analysis reveals senescence of cultured canine adipose-derived mesenchymal stromal cells to be reversible with a novel immortalization approach

“…Even though insight into the mechanisms of senescence in human medicine has evolved dramatically 18 , knowledge about replicative senescence of canine ( Canis lupus familiaris ) MSCs is still very scarce 19–21 . Therapeutic use of stem cells in veterinary medicine has not yet progressed beyond pioneering work 22,23 . Correspondingly, there is a great interest in this field 24 , both for the importance of dogs as a model organism for human diseases in pre-clinical studies and for the relevance of dogs as a companion animal.…”