Wound Healing-Promoting Effects of Apelin-13 in a CCD-1072Sk Fibroblast-Based In Vitro Model
DOI:
https://doi.org/10.58600/eurjther2780Keywords:
Apelin-13, CCD-1072Sk, fibroblast, fibrosis, scratch assay, wound healingAbstract
Objective: As an intricate process, wound healing involves contributions from numerous cell types. Fibroblasts critically support wound healing via sustaining the integrity of the extracellular matrix, promoting collagen synthesis, and driving contraction during proliferation and remodeling. A balanced inflammatory response is essential for this repair. The apelinergic system, comprising Apelin and APJ (apelin receptor)—abundantly found in the skin and scar fibroblasts—plays a significant regulatory role. This is among the first to reveal the role of exogenous Apelin-13 in wound healing using a scratch model in CCD-1072Sk fibroblast cell line.
Methods: The doses of 2 and 5-µg/ml Apelin-13 were chosen based on the cell viability assay result. After seeding cells in 24-well plates, the wound scratch model was applied once they reached approximately 90% confluency. Samples of the control (CTL), A-2, and A-5 groups were collected and analyzed at 0 h (hour) (baseline), 24 h, 48 h, and 72 h. After the wound scratch model, the wound gap area was checked at the relevant time points. ELISA analysis was performed for levels of TGF-β1, TNF-α, and IL-10. Apoptosis was analyzed with a fluorescent microscope using the Annexin V-FITC/PI method.
Results: In the viability assay, 2 and 5 µg/ml Apelin-13 doses were used. Wound closure decreased significantly at all time points in both Apelin groups vs. 0 h (p<0.001). Apoptosis increased significantly in A-2-48 and A-5-48 groups (p<0.001). TNF-α decreased in A-2 vs. CTL (p<0.001) and A-5-72 (p<0.001). TGF-β1 increased in A-5-72 vs. A-2-72 (p<0.05). IL-10 increased in all Apelin groups vs. CTL (p<0.001).
Conclusion: Apelin-13 promoted wound healing by balancing inflammation, enhancing fibroblast proliferation, migration, and reducing apoptosis. 2 µg/ml Apelin-13 showed notable effects, while 5 µg/ml was superior at certain time points. Apelin-13 appears to be a promising wound-healing agent.
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Istanbul Üniversitesi-Cerrahpasa
Grant numbers TSA-2023-37359
