Gene Expression of TGF-β1 , Matrix Metalloproteinases, and Collagen in Keloid Patients and Their Growth Patterns in 2D and 3D Cell Cultures: An In Vitro Study

Abstract

Background/Aim: Keloids result from abnormal wound healing and are characterized by excess fibroblast activity, extracellular matrix (ECM) accumulation, and elevated collagen types. The contemporary research often relies on 2D cell cultures, which do not fully replicate keloid pathophysiology. Spheroid models offer a promising alternative. This study analyzed gene expression differences in keloid and normal skin tissues, evaluated mRNA expression from 2D cultures, and assessed spheroid viability as proof of concept. Materials and Methods: Keloid and normal skin samples were collected from patients with auricular keloids. The tissues underwent histological and immunohistochemical analyses, RNA extraction for RT-qPCR, and 2D and 3D cell culture. Spheroids were cultivated for 21 days and analyzed for morphology, viability, and histological staining. Results: Histological analysis revealed mixed results, with higher mRNA levels of COL1A1, COL3A1, TGF-β1, and TGF-βR1 in the keloids. In the 2D cultures, normal skin showed elevated MMP-1, MMP-3, and TGF-β1 expression. The keloid spheroids demonstrated comparable morphology, but decreased viability after 21 days. Conclusion: Keloid gene and protein expression patterns varied among native tissue, 2D cultures, and 3D cultures. Keloid-derived spheroids displayed distinct histomorphological features compared with normal skin spheroids, suggesting that they may serve as more physiologically relevant models for future keloid research.