Publication

Cellular ageing of oral fibroblasts differentially modulates extracellular matrix organization

Atkuru, Srividya
Muniraj, Giridharan
Sudhaharan, Thankiah
Chiam, Keng-Hwee
Wright, Graham Daniel
Sriram, Gopu
Citations
Altmetric:
Alternative Title
Abstract
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 P21WAFI. 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 P21WAFI 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.
Keywords
Science & Technology, Life Sciences & Biomedicine, Dentistry, Oral Surgery & Medicine, ageing, extracellular matrix, fibroblasts, second harmonic generation imaging, senescence, HUMAN GINGIVAL FIBROBLASTS, PHENOTYPIC DIFFERENCES, SKIN, EXPRESSION, AGE, COLLAGEN, FIBRONECTIN, FIBRILLIN, LAMININ, GENES
Source Title
JOURNAL OF PERIODONTAL RESEARCH
Publisher
WILEY
Series/Report No.
Organizational Units
Organizational Unit
Rights
Date
2020-09-23
DOI
10.1111/jre.12799
Type
Article
Additional Links
Related Datasets
Related Publications