Acid ceramidase treatment enhances the outcome of autologous chondrocyte implantation in a rat osteochondral defect model

Abstract

Objective The overall aim of this study was to evaluate how supplementation of chondrocyte media with recombinant acid ceramidase (rhAC) influenced cartilage repair in a rat osteochondral defect model. Methods Primary chondrocytes were grown as monolayers in polystyrene culture dishes with and without rhAC (added once at the time of cell plating) for 7 days, and then seeded onto Bio-Gide® collagen scaffolds and grown for an additional 3 days. The scaffolds were then introduced into osteochondroal defects created in Sprague-Dawley rat trochlea by a micordrilling procedure. Analysis was performed 6 weeks post-surgery macroscopically, by micro-CT, histologically, and by immunohistochemistry. Results Treatment with rhAC led to increased cell numbers and glycosaminoglycan production (~2 and 3-fold, respectively) following 7 days of expansion in vitro. Gene expression of collagen 2, aggrecan and Sox-9 also was significantly elevated. After seeding onto Bio-Gide®, more rhAC treated cells were evident within 4 hours. At 6 weeks post-surgery, defects containing rhAC-treated cells exhibited more soft tissue formation at the articular surface, as evidenced by microCT, as well as histological evidence of enhanced cartilage repair. Notably, collagen 2 immunostaining revealed greater surface expression in animals receiving rhAC treated cells as well. Collagen 10 staining was not enhanced. Conclusion The results further demonstrate the positive effects of rhAC treatment on chondrocyte growth and phenotype in vitro, and reveal for the first time the in vivo effects of the treated cells on cartilage repair.

Enzyme replacement therapy for Farber disease: Proof-of-concept studies in cells and mice

“…Addition of rhAC to the culture media could correct this phenotype, further indicating the bioactivity of the enzyme and the fact that it can be taken up by this important cell type. We and others have shown that there is an important and unique relationship between sphingolipids and cartilage integrity [32] , [33] , and in the future it will be interesting to examine this further in the context of Farber disease.…”

Acid Ceramidase Protects Against Hepatic Ischemia/Reperfusion Injury by Modulating Sphingolipid Metabolism and Reducing Inflammation and Oxidative Stress

“…In addition to Farber disease, the therapeutic potential of recombinant AC also has been investigated in other diseases where ceramide pathology has been demonstrated, for example to reduce inflammation/infection in Cystic Fibrosis and to assist in cartilage repair after joint injury ( Pewzner-Jung et al, 2014 ; Xia et al, 2015 ; Frohbergh et al, 2016 ; He et al, 2017 ; Becker et al, 2018 ). In the current work we have focused on investigating the potential protective effects of recombinant AC on hepatic IR injury, and further understanding the underlying mechanism(s) contributing to these effects.…”

Elusive Roles of the Different Ceramidases in Human Health, Pathophysiology, and Tissue Regeneration

“…In vivo studies have also demonstrated a positive effect of ACDase in chondrocyte differentiation. In cartilage replacement therapy, pre-treatment with ACDase induces chondrocyte proliferation, the production of glycosaminoglycan, the expression of COL2 , the adhesion of chondrocytes to a scaffold, a reduced resorption after implantation, and an improved differentiation to cartilage [ 116 ]. Furthermore, a variation of FRBRL characterized by peripheral osteolysis not associated with MMP-2 and MMP-14 was found, suggesting the involvement of ASAH1 in bone remodeling [ 117 ].…”