The Effect of Combined Cellular Therapy on Sperm in Experimental Diabetic Damage

Abstract Diabetes (DB)-related sperm damage is one of the most significant issues contributing to infertility. Mesenchymal Stem Cells (MSCs) are an important product that has recently gained clinical relevance. The medium composed of MSC secretions after 48 h of culture is called Conditioned Medium (CM), and its effectiveness has been demonstrated in experimental studies. Platelet-Rich Plasma (PRP) and Stromal Vascular Fraction (SVF) are products commonly used in clinical settings with proven efficacy. However, there is limited research on the combined use of these products in treatment. This study examined the effectiveness of combined therapy (CT) compared to individual treatments in sperm damage induced by DB damage (DBD) simulated using a high glucose medium (HGM) in culture. Sperm obtained from male Wistar Albino rats were placed in 6-well flasks and divided into 6 different groups: Control (CG), DBD with high glucose (HG), HG + MSC, HG + CM, HG + PRP, HG + SVF, and HG + MSC + CM (CT). After one hour of treatment, the samples were evaluated for viability, motility, and morphology. Slides prepared from these samples were analyzed immunohistochemically for oxidative stress using endothelial Nitreous Acide Syntease (eNOS) and for apoptosis using Caspase 3. The damage to sperm caused by HG led to significant increases in eNOS and Caspase 3, as well as significant reductions in viability, motility, and morphological abnormalities. Although all single treatments reversed all parameters, the most effective treatment was found to be the combined application of MSC and CM. The DB in vitro model demonstrated that sperm damage can occur and that this damage can potentially be prevented with cell therapy treatments, highlighting the importance of exploring other treatment possibilities. The combination of MSC and CM was shown to be quite beneficial by reducing oxidative stress and apoptosis. Cellular therapy, a cost-effective and newly developed regenerative method, could offer hope to families wishing to have children, especially in the context of the high prevalence and increasing incidence of male infertility. Validation of these findings through studies in experimental animals and humans, as well as clarification of the underlying mechanisms, will facilitate their clinical application.