Tsai Ching-Wen, Kao Yu-Ting, Chiang I-Ni, Wang Jyh-Horng, Young Tai-Horng
Institute of Biomedical Engineering, College of Medicine and College of Engineering, National Taiwan University, Taipei 100, Taiwan, No.1, Sec. 1, Jen - Ai Rd., Taipei 100, Taiwan.
Institute of Biomedical Engineering, College of Medicine and College of Engineering, National Taiwan University, Taipei 100, Taiwan, No.1, Sec. 1, Jen - Ai Rd., Taipei 100, Taiwan; Department of Urology, National Taiwan University Hospital, Taipei 100, Taiwan, No.7, Chung-Shan S. Rd., Taipei 100, Taiwan.
PLoS One. 2015 Oct 14;10(10):e0140747. doi: 10.1371/journal.pone.0140747. eCollection 2015.
Fibroblasts have been extensively used as a model to study cellular senescence. The purpose of this study was to investigate whether the human foreskin fibroblast aging process could be regulated by using the biomaterial chitosan. Fibroblasts cultured on commercial tissue culture polystyrene (TCPS) entered senescence after 55-60 population doublings (PDs), and were accompanied by larger cell shape, higher senescence-associated β-galactosidase (SA β-gal) activity, lower proliferation capacity, and upregulation of senescence-associated molecular markers p21, p53, retinoblastoma (pRB), and p16. Before senescence was reached, PD48 cells were collected from TCPS and seeded on chitosan for three days (PD48-Cd3) to form multicellular spheroids. The protein expression of senescence-associated secretory phenotypes (SASPs) and senescence-associated molecular markers of these cells in PD48-Cd3 spheroids were downregulated significantly. Following chitosan treatment, fibroblasts reseeded on TCPS showed lower SA β-gal activity, increased cellular motility, and a higher proliferation ability of 70-75 PDs. These phenotypic changes were not accompanied by colonies forming in soft agar and a continuous decrease in the senescence-associated proteins p53 and pRB which act as a barrier to tumorigenesis. These results demonstrate that chitosan treatment could delay the induction of senescence which may be useful and safe for future tissue engineering applications.
成纤维细胞已被广泛用作研究细胞衰老的模型。本研究的目的是调查使用生物材料壳聚糖是否可以调节人包皮成纤维细胞的衰老过程。在商业组织培养聚苯乙烯(TCPS)上培养的成纤维细胞在经历55 - 60次群体倍增(PDs)后进入衰老状态,同时伴随着细胞形态变大、衰老相关β - 半乳糖苷酶(SA β - gal)活性升高、增殖能力降低以及衰老相关分子标志物p21、p53、视网膜母细胞瘤(pRB)和p16的上调。在达到衰老之前,从TCPS收集PD48细胞并接种到壳聚糖上三天(PD48 - Cd3)以形成多细胞球体。PD48 - Cd3球体中这些细胞的衰老相关分泌表型(SASPs)和衰老相关分子标志物的蛋白质表达显著下调。经过壳聚糖处理后,重新接种到TCPS上的成纤维细胞显示出较低的SA β - gal活性、增加的细胞运动性以及70 - 75次PDs的更高增殖能力。这些表型变化并未伴随着在软琼脂中形成集落以及作为肿瘤发生屏障的衰老相关蛋白p53和pRB的持续减少。这些结果表明,壳聚糖处理可以延迟衰老的诱导,这对于未来的组织工程应用可能是有用且安全的。