Human primary chronic wound derived fibroblasts demonstrate differential pattern in expression of fibroblast specific markers, cell cycle arrest and reduced proliferation

• Published in: Experimental and molecular pathology Vol. 127; p. 104803
• Main Authors: Monika, P., Chandraprabha, M.N., Murthy, K.N. Chidambara, Rangarajan, Annapoorni, Waiker, P. Veena, Sathish, M.
• Format: Journal Article
• Language: English
• Published: Netherlands Elsevier Inc 01-08-2022
• Subjects: Acute wound fibroblasts; Cell cycle; Chronic wound fibroblasts; Differential expression; Proliferation; Vimentin; α-SMA; Vimentin; α-SMA; Chronic wound fibroblasts; Cell cycle; Proliferation; Differential expression; Acute wound fibroblasts
• ISSN: 0014-4800; 1096-0945
• DOI: 10.1016/j.yexmp.2022.104803

Although wound refers to simple cut in the skin, most wounds don't heal because of the various local and systemic factors that lead to its complexity and chronicity. Thus, prior understanding of the status of the wound is necessary and methods that can differentiate between the healing and non-healing wounds at a much earlier stage is crucial for a successful treatment.
The current study aims at differentiating Acute Wound Fibroblasts (AWFs) and Chronic Wound Fibroblasts (CWFs) based on differential expression of fibroblast specific markers such as Vimentin and Alpha Smooth Muscle Actin (α-SMA) and compare its cell cycle and proliferation.
Immunostaining and western blotting analysis showed that, AWFs and CWFs differentially expressed vimentin and α-SMA, with AWFs and CWFs showing higher expression of vimentin and α-SMA respectively. AWFs showed higher distributions in G0/G1 (67.43% vs. 62.16%), S phase (22.61% vs. 8.51%) compared to CWFs. However, AWFs showed decreased distributions compared to CWFs in G2 + M phase (8.14% vs. 10.6%). Thus, it was observed that CWFs showed cell cycle arrest in the G1/G0 phase and inhibited DNA synthesis, which was further confirmed by reduced proliferation of CWFs. We suggest that, differential expression of the cell specific markers can be attributed to its pathophysiological status and chronicity of the wound and reduced proliferation rate of CWFs is due to lesser expression of vimentin, which is a key protein for in vitro cell proliferation.
Outcome of the study serve as an immunological tool to guide the chronicity of the wound, which helps to understand the wound towards design of personalized care. The findings also represent a promising opportunity to gain insight into how cell cycle arrest can impact on wound healing and clinical outcomes.
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•Immunofluorescence analysis using fibroblast specific markers confirms pure population of human primary fibroblasts isolated from acute and chronic wound samples.•AWFs (normal) showed greater expression for vimentin, while CWFs (pathological condition) showed greater expression for αSMA.•Higher vimentin expression and DNA synthesis in AWFs enhances cell proliferation and can be used as better targets to address chronic wounds.•Cell cycle arrest and inhibition of fibroblast proliferation was observed in CWFs. Differential expression pattern, cell cycle arrest and reduced proliferation together serve as biomarkers and display a versatile method for better identification of chronic wounds.