Evaluation of the effects of extracellular matrix on cellular aging using a cell culture-based model

Human diploid fibroblasts have the capacity to complete a finite number of cell divisions before entering a replicative senescence state, characterized by growth arrest, morphological changes, and altered gene expression. During this process, the interaction with the extracellular matrix (ECM) plays a significant role in the onset of cellular aging and senescence. Understanding the biological processes underlying cellular aging is of critical importance for tissue engineering and therapeutic strategies aimed at combating aging. However, the impact of environmental factors and, particularly, the role of the cellular microenvironment in this process has yet to be fully elucidated. In this context, it has been suggested that the interaction with the ECM derived from young dermal tissue can reverse the senescence process and restore aged cells to a seemingly youthful state. Additionally, the effect of aged ECM on young cells has also been examined. In this study, the aging process of fibroblast cells was investigated using a decellularized ECM (dECM) model produced by three-dimensional bioprinting technology. To observe the aging process of fibroblasts in a more natural cellular microenvironment, a three-dimensional culture model was employed. This model enabled a more realistic investigation of aging mechanisms in an in vivo-like environment. dECM derived from both young and aged rat skin was used in conjunction with aged and young cells to investigate the impact of ECM's biochemical properties on aging. In this context, the chemical composition of dECM was verified by Fourier-transform infrared spectroscopy (FTIR) and DNA quantification. The expression levels of genes involved in aging, such as SIRT1, p53, GLB1, COL1A1, NF-κB, and MMP1, were analyzed using real-time polymerase chain reaction (qPCR) on samples taken on days 0, 3, 7, and 10. The results demonstrated that dECM has the capacity to restore some youthful phenotypic features in aged cells and that the youthful properties of ECM can be transferred to aged cells. These findings provide a new perspective on the potential of ECM to slow down or partially reverse aging and highlight the impact of cell-microenvironment interactions on the aging process. Our study, by elucidating the effects of ECM on cellular aging, offers data that can contribute to anti-aging strategies in tissue engineering and regenerative medicine.