Affiliation:
1. Regenerative Medicine Laboratory, Dr. D. Y. Patil Dental College and Hospital Dr. D. Y. Patil Vidyapeeth, Pimpri Pune India
2. Department of Biotechnology Sinhgad College of Engineering affiliated to Savitribai Phule Pune University Pune India
Abstract
AbstractBackgroundMesenchymal stem cells (MSCs) from type 2 diabetes mellitus (T2DM) individuals exhibit increased adipogenesis and decreased osteogenesis. We investigated the potential of adipose tissue‐derived MSCs (ADMSCs) secretome obtained from healthy individuals in restoring the tumor necrosis factor‐α (TNF‐α) mediated imbalance in the adipo/osteogenic differentiation in the dental pulp‐derived MSCs obtained from T2DM individuals (dDPMSCs).MethodsdDPMSCs were differentiated into adipocytes and osteocytes using a standard cocktail in the presence of (a) induction cocktail, (b) induction cocktail + TNF‐α, and (c) induction cocktail+ TNF‐α + ADMSCs‐secretome (50%) for 15 and 21 days resp. Differentiated adipocytes and osteocytes were stained by oil red O and alizarin red and analyzed by using ImageJ software. Molecular expression of the key genes involved was analyzed by using reverse‐transcription polymerase chain reaction (RT‐PCR).ResultsTreatment of TNF‐α augmented the adipogenesis (9571 ± 765 vs. 19,815 ± 1585 pixel, p < 0.01) and decreased the osteogenesis (15,603 ± 1248 vs. 11,894 ± 951 pixel, p < 0.05) of dDPMSCs as evidenced by the oil red O and alizarin red staining respectively. Interestingly, dDPMSCs differentiated along with TNF‐α and 50% ADMSCs secretome exhibited enhanced osteogenesis (11,894 ± 951 vs. 41,808 ± 3344 pixel, p < 0.01) and decreased adipogenesis (19,815 ± 1585 vs. 4480 ± 358 pixel, p < 0.01). Additionally, dDPMSCs differentiated along with ADMSCs secretome exhibited decreased expression of PPARg (p < 0.01), C/EBPa (p < 0.05), and FAS (p < 0.01) whereas mRNA expression of Runx2 (p < 0.05), Osterix (p < 0.01), and OCN (p < 0.05) was upregulated as revealed by the RT‐PCR analysis.ConclusionADMSCs secretome from healthy individuals restore the TNF‐α influenced differentiation fate of dDPMSCs and therefore can be explored for T2DM clinical management in the future.