Specific Calcium Signal Responses in Human Keloid-Derived Fibroblasts During Cyclical Stretching: Basic Research

人类瘢痕疙瘩来源成纤维细胞在周期性拉伸过程中的特异性钙信号反应:基础研究

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Abstract

BACKGROUND: Keloids most commonly develop in the regions where the skin is constantly stretched. Although some keloid-derived fibroblasts exhibit higher single calcium spikes than normal dermal fibroblasts during short-time cyclical stretching, the calcium signal responses to long-time stretching remain unclear. METHODS: This study compared the intracellular Ca(2+) dynamics induced by cyclical stretching stimuli between the control group (normal dermal fibroblasts) and the keloid group (keloid-derived fibroblasts). Each group was cyclically exposed to a two-dimensional stretch (10% strain). A confocal laser microscope was used to examine intracellular Ca(2+) for 30 min fluorescently. The fluorescence intensity ratio (Fluo-8H/calcein red-orange) was used to evaluate intracellular Ca(2+) concentration every 0.5 s. A calcium spike was a transient ratio increase of ≥ 20%. Receiver operating characteristic analysis was performed to determine the cutoff value of a normal calcium spike. RESULTS: No significant difference was observed between the keloid and control groups in the calcium signal response-positive rates (26.9% vs. 25.0%; p = 0.9). However, the calcium spike amplitudes were significantly higher in the keloid group than in the control group (1.66 vs. 1.41; p = 0.02). The cutoff value was 2.12, and 9.6% of keloid-derived fibroblasts exhibited multiple hypercalcium spikes. DISCUSSION: We are conducting further research based on the hypothesis that this keloid-specific subpopulation triggers the pathogenesis of keloid formation, that is, collagen overproduction, accelerated angiogenesis, and chronic inflammation.

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