期刊
JOURNAL OF PERIODONTAL RESEARCH
卷 56, 期 1, 页码 108-120出版社
WILEY
DOI: 10.1111/jre.12799
关键词
ageing; extracellular matrix; fibroblasts; second harmonic generation imaging; senescence
资金
- National University of Singapore (Faculty of Dentistry)
- Ministry of Education, Singapore [R-221-001-118-720, R-221-000-118-133]
- UROP Project [17-09]
- Agency for Science, Technology and Research (A*STAR): Core funds [H1701a0004]
- Agency for Science, Technology and Research (A*STAR): HBMS IAF-PP Project [H1701a0004]
- National Research Foundation Singapore [NRF2017_ SISFP10]
Ageing of oral fibroblasts impairs extracellular matrix production and organization, particularly affecting the structure of ECM fibers which may impact wound healing in the elderly. Aged oral fibroblasts exhibit slower proliferation rate and a senescence phenotype, leading to altered production and organization of ECM proteins such as collagen and fibronectin.
Background and Objectives Ageing is associated with an impaired cellular function that can affect tissue architecture and wound healing in gingival and periodontal tissues. However, the impact of oral fibroblast ageing on the structural organization of the extracellular matrix (ECM) proteins is poorly understood. Hence, in this study, we investigated the impact of cellular ageing of oral fibroblasts on the production and structural organization of collagen and other ECM proteins. Methods Oral fibroblasts were serially subcultured, and replicative cellular senescence was assessed using population doubling time, Ki67 counts and expression of P21(WAFI). The production and structural organization of ECM proteins were assessed at early (young-oFB) and late (aged-oFB) passages. The thickness and pattern of collagen produced by live cultures of young- and aged-oFB were assessed using a label-free and non-invasive second harmonic generation (SHG)-based multiphoton imaging. Expression of other ECM proteins (fibronectin, fibrillin, collagen-IV and laminins) was evaluated using immunocytochemistry and confocal microscopy-based depth profile analysis. Results Aged-oFB displayed a higher population doubling time, lower Ki67+ cells and higher expression of P21(WAFI)indicative of slower proliferation rate and senescence phenotype. SHG imaging demonstrated that young-oFB produced a thick, interwoven network of collagen fibres, while the aged-oFB produced thin and linearly organized collagen fibres. Similarly, analysis of immunostained cultures showed that young-oFB produced a rich, interwoven mesh of fibronectin, fibrillin and collagen-IV fibres. In contrast, the aged-oFB produced linearly organized fibronectin, fibrillin and collagen-IV fibres. Lastly, there was no observable difference in production and organization of laminins among the young- and aged-oFB. Conclusion Our results suggest that oral fibroblast ageing impairs ECM production and more importantly the organization of ECM fibres, which could potentially impair wound healing in the elderly.
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