Chinese Journal of Tissue Engineering Research ›› 2026, Vol. 30 ›› Issue (31): 8100-8107.doi: 10.12307/2026.848

Previous Articles     Next Articles

Modulation of nasal mucosa-derived ectomesenchymal stem cell senescence by mechanical modulus

Zhou Jinlong1, 2, Wang Juan2, Bian Lu2, 3, Lyu Long2, Que Yunduan1, Qian Wenwu1   

  1. 1Department of Orthopedics, Nanjing Gaochun People’s Hospital Affiliated to Jiangsu Health Vocational College, Nanjing 211300, Jiangsu Province, China; 2Central Laboratory, Gaochun Hospital Affiliated to Jiangsu University, Nanjing 211300, Jiangsu Province, China; 3Wuxi Medical School, Jiangnan University, Wuxi 214026, Jiangsu Province, China
  • Received:2025-09-26 Accepted:2026-02-13 Online:2026-11-08 Published:2026-05-23
  • Contact: Qian Wenwu, Master’s supervisor, Associate chief physician, Department of Orthopedics, Nanjing Gaochun People’s Hospital Affiliated to Jiangsu Health Vocational College, Nanjing 211300, Jiangsu Province, China
  • About author:Zhou Jinlong, MS candidate, Attending physician, Department of Orthopedics, Nanjing Gaochun People’s Hospital Affiliated to Jiangsu Health Vocational College, Nanjing 211300, Jiangsu Province, China; Central Laboratory, Gaochun Hospital Affiliated to Jiangsu University, Nanjing 211300, Jiangsu Province, China
  • Supported by:
    Jiangsu Health Vocational College Institutional Research Project, No. YJXTG202309 (to ZJL); Nanjing Municipal Health Science and Technology Development Special Fund Project (Key Project), No. ZKX21063 (to QYD)

PDF

8

Abstract: BACKGROUND: The mechanical properties of the cellular microenvironment play a pivotal role in regulating the fate of nasal mucosa–derived ectomesenchymal stem cells.
OBJECTIVE: To investigate the regulatory effects of mechanical modulus on the senescence of ectomesenchymal stem cells and the potential molecular mechanisms.
METHODS: Gelatin methacryloyl hydrogels with 50, 100, and 150 g/L mass concentrations were fabricated, and low-, medium-, and high-mechanical moduli were established, with standard culture plates serving as controls. Ectomesenchymal stem cells of SD rats were isolated, and identified. Passage 5 cells were seeded onto hydrogels of different moduli and cultured to passage 7. Cells were further treated with the Piezo1 activator Yoda1 or the transglutaminase 2 inhibitor KCC009. RT-qPCR, western blotting, and immunofluorescence were performed to assess the expression of senescence markers (p16 and p21), proliferation-associated genes (Mki67 and Pcna), osteogenic markers (Spp1, Sp7, type I collagen), Piezo1, calcium ions, transglutaminase 2, and fibrotic proteins (α-smooth muscle actin and type I collagen). β-Galactosidase staining was used to quantify the proportion of senescent cells. Alkaline phosphatase staining was used to measure alkaline phosphatase activity. Alizarin Red S staining was applied to assess mineralized nodule formation.
RESULTS AND CONCLUSION: Low-modulus hydrogels significantly downregulated p16 and p21 expression, reduced β-galactosidase–positive cells, alleviated fibrosis, and enhanced osteogenic differentiation, whereas high-modulus hydrogels induced the opposite effects. Further mechanistic analysis revealed that ectomesenchymal stem cells cultured on high-modulus substrates exhibited increased Piezo1 expression, enhanced Ca²⁺ influx, and elevated transglutaminase 2 levels. Inhibition of transglutaminase 2 effectively attenuated high-modulus–induced ectomesenchymal stem cell senescence. These findings indicate that a low-modulus microenvironment can reverse ectomesenchymal stem cell senescence, and the Piezo1/transglutaminase 2 signaling pathway serves as a key regulator in this process.

Key words: ">nasal mucosa–derived ectomesenchymal stem cells, mechanical modulus, senescence, piezo1, transglutaminase 2, gelatin methacryloyl, osteogenic differentiation, fibrosis

CLC Number: