Regeneration Potential of Bone Marrow-derived Mesenchymal Stromal Cells Lysate for H₂O₂ (In-Vitro) Injured Cells

Abstract

Background: Mesenchymal stromal cells (MSCs) and their paracrine factors make them a suitable cell-free-therapeutic candidate. Cellular lysate usage could be an effective treatment strategy that circumvents the possible associated drawbacks of stem cell therapies. Objective: Thus, this research aims to examine the injury regeneration potential of MSCs cellular lysate derived from bone marrow by studying its anti-apoptotic, proliferative, and anti-oxidative effects. Methods: Hydrogen peroxide (H2O2) was used to induce cellular injury. MTT assay, trypan blue, and crystal violet assays were used to assess bone marrow-derived mesenchymal stromal cell (BMSCs) lysate treated cells' viabilities. Next, to investigate the BMSCs lysate anti-oxidative potential anti-oxidants, ascorbate peroxidase (APX), glutathione reductase (GR), and superoxide dismutase (SOD) assays were performed. Simultaneously, the proliferative and anti-apoptotic potential was measured via vascular endothelial growth factor (VEGF A) and p53 expression analysis through immunostaining and ELISA. Results: It was observed that BMSCs lysate enhances the viability of H2O2 injured cells. APX, GR, and SOD's results indicated that after H2O2 injury, the anti-oxidant status decreased significantly and was uplifted by BMSCs lysate treatment. Additionally, the results of p53, BAX, and caspase-3 expression revealed that BMSCs lysate inhibits apoptosis by downregulating their expression in treated cells. The VEGF protein expression findings demonstrated that BMSCs lysate upregulates the downregulated expression of VEGF in H2O2 injured cells. The expression of proliferative markers (TOP2A, PCNA, and Ki-67) was also elevated in BMSCs treated cells. Conclusion: To conclude this study's findings, it was observed that BMSCs lysate could decrease H2O2 injury and possibly regenerate the injured cells by enhancing their viability and proliferation, improving anti-oxidants levels, and alleviating apoptosis.