The Role of Ecto-5'-Nucleotidase (CD73) in Regeneration and Immunomodulation of Human Bone Marrow Mesenchymal Stem Cells

Abstract

Introduction: Ecto-5’-nucleotidase (CD73) regulates inflammatory responses by balancing extracellular ATP and adenosine, while simultaneously influencing the regenerative and migratory potential of bone marrow mesenchymal stem cells (BM-MSCs), as well as their interactions within the bone marrow. Revealing the role of CD73 in BM-MSCs will provide a better understanding of the BM-MSCs’ behavior, which will be important for cell therapy and advances in regenerative medicine.

 Aim: Investigation of short-term CD73 inhibition with APCP on the biological functions of BM-MSCs, including proliferation, viability, differentiation, cytokine secretion and protein detection, and migration.

 Materials and Methods: BM-MSCs were isolated from the iliac bone of patients, and treated with adenosine 5’-(α,β-methylene) diphosphate (APCP) to inhibit CD73 signalling. Viability was evaluated by MTT assay, while cytokine secretion was analysed by microbead-based flow cytometry. Migratory potential was assessed by scratch assay, while cytoskeletal components were evaluated by immunofluorescence. Images were captured by fluorescence microscopy and quantified with ImageJ.

 Results: We demonstrated that short-term CD73 inhibition does not compromise cell viability, but it significantly influences osteogenesis. Furthermore, CD73 inhibition modulated cytokine secretion and reduced the BM-MSCs’ migratory capacity. Additionally, APCP decreased the expression of cytoskeletal elements, including vimentin, α-SMA, and actin, further supporting these findings and confirming the role of CD73 in regulating MSC migratory potential.

 Conclusion: Short-term CD73 inhibition reduces BM-MSCs’ migratory capacity and cytoskeletal protein expression, while altering their differentiation potential and cytokine profile. These changes highlight CD73 as an important regulator of differentiation, migration, and immunomodulation that should be thoroughly understood and exploited for potential therapeutic purposes.