High glucose inhibits proliferation, migration, and osteogenic differentiation of human placenta-derived mesenchymal...

Chronic hyperglycemia is recognized as an important contributor to chronic inflammation, oxidative stress, and organ dysfunction that causes serious complications in diabetes and aging. This study investigates the effects of elevated glucose levels on human placenta-derived mesenchymal stem cells (hP-MSCs), especially since these cells hold a promise for tissue engineering and regenerative medicine due to their multipotency and self-renewal capabilities. hP-MSCs were treated with 10-40 mM D-glucose to study the effects of high glucose on hP-MSCs functions. hP-MSCs viability and proliferation were determined by using thiazolyl blue tetrazolium bromide (MTT), cell cycle analysis, and senescence assays. The migration and osteogenic differentiation capacity were also determined by migration assay, alkaline phosphatase (ALP) activity assay, and Alizarin Red S staining. Quantitative real-time RT-PCR, Western blot, and Nanostring® nCounter assay were performed to study the effect of high glucose on the expression levels of genes involved in various aspects of hP-MSCs functions. The results demonstrated that high glucose significantly inhibited proliferation and cell cycle progression of hP-MSCs at the G1/S phase and induced replicative senescence in hP-MSCs possibly by decreasing the expression of proliferation-promoting genes, CCND1 and LMNB1, and increasing the expression of several senescence-associated proteins, p16, p21, and p53. Furthermore, high glucose also inhibited the migration and osteogenic differentiation of hP-MSCs, possibly by suppressing the expression of SDF1, CXCR4, RUNX2, OSX, OCN, and COL1A. The additional Nanostring analysis also showed that high glucose significantly affects multiple genes involved in inflammation, DNA repair, autophagy, and oxidative stress response in hP-MSCs. This study provides significant insights into the wide-ranging effects of high glucose on the expression of the hP-MSCs genes that affect various aspects of its function, including proliferation, viability, senescence, oxidative stress response, and DNA repair, highlighting its implications for regenerative medicine in the context of diabetes and metabolic disorders.