IN VITRO ASSESSMENT OF TGFβ3/TGFβ1 RATIO VARIATION PATTERN FOLLOWING ANTISEPTIC TREATMENT ON ADIPOSE-DERIVED STEM CELLS AND FIBROBLASTS IN 2D VS. 3D CULTURE SYSTEMS
Abstract
Wound management, especially chronic wounds and burns, remain a challenge despite advances regarding topical antimicrobial agents and emerging stem-cell based strategies. Moreover, the current available data is controversial regarding antiseptics drawbacks on wound healing and minimal on the effects on stem cells, although it seems they modulate cytokines, including TGFβ. The aim of the study was to evaluate in vitro the effects of commonly used antiseptics on the TGFβ3/TGFβ1 ratio in cell cultures. Material and methods: Four cell constructs were developed with different cell ratios using adipose-derived stem cells and normal human dermal fibroblasts in both 2D and 3D mono and co-cultures. Following treatment with either 1% povidone-iodine, 0.05% chlorhexidine digluconate or silver-coated dressing, TGFβ1 and TGBβ3 levels were evaluated using ELISA method. Subsequently, TGFβ3/TGFβ1 ratio was calculated, and statistical analysis performed. Results: TGFβ3/TGFβ1 ratio for untreated stem cells and co-cultures increase pattern follows the stem cell ratio in both 2D and 3D culture systems. Following povidone-iodine treatment for all cell constructs in 2D systems, TGFβ3/TGFβ1 ratio increases significantly, whereas in 3D culture systems increased only in stem cell monoculture and co-culture with fibroblast as a majority cell fraction. Moreover, there are significant differences in TGFβ3/TGFβ1 ratio when 2D and 3D culture systems are compared, especially after chlorhexidine digluconate and silver-coated dressing exposure for fibroblast monocultures and co-cultures. Conclusions: TGFβ3/TGFβ1 ratio pattern modulation in vitro is dependent on antiseptic, cell type, cell ratio and culture system.
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