Effect of SPARC expression in Mesenchymal Stem Cells and analysis of their potential for wound regeneration.

ANTONELLA LOMBARDI; MIKELE AMONDARAIN; MARCELA N. GARCÍA; ALEJANDRO LA GRECA; JOAQUÍN SMUCLER; GUSTAVO E SEVLEVER; SANTIAGO MIRIUKA; CARLOS LUZZANI

Mar del Plata

Congreso; LXVII Reunión anual de la Sociedad Argentina de Investigación Clínica; 2022

SAIC

Mesenchymal Stem Cells (MSCs) are widely studied for their interesting cell biology and broad-ranging clinical potential. To date, there has been a rapid expansion of both preclinical and clinical studies using the MSC-derived secretome with the advantages of a cell-free system. The potential for therapeutic effects of this secretome could be optimized by pre-conditioning MSC with small molecules, biological agents, and biomaterials, or by genetically modifying them. Wound regeneration is a complex process that requires cell migration, the development of an inflammatory environment, angiogenesis, granulation, tissue formation, re-epithelialization, and extracellular matrix (EM) remodeling. MSCs and their secretome have an active role in this process. SPARC, also known as osteonectin, is one of the many proteins secreted by MSCs. This protein is known to be involved in EM remodeling and assembly as well as in the regulation of cell migration and proliferation processes. In view of this, the aim of this work was to study whether the modification of SPARC expression in MSCs affects their secretome composition and their regenerative capacity. For this purpose, we used a lentiviral system to obtain SPARC knockdown cells (SPARC-KD-MSC). After characterizing them and validating that they kept their identity as MSCs, we used both wild type (WT) and SPARC-KD-MSC conditioned medium (CM) in wound healing in vitro assays with human skin keratinocytes (HaCat). We observed that wound closure was slower when using SPARC-KD-MSC CM than with WT CM. We also analyzed changes in the HaCat cell cycle distribution and no differences between conditions were found. These results suggest that SPARC may be playing a role in the HaCat wound migration process. In the future, we plan to deepen this analysis in order to characterize the role of SPARC in the wound healing process in vitro and in vivo, as well as to test whether its overexpression increases the regenerative capacity of MSCs.