Effect of Plasma Gel on Viability, Proliferation and Migration of Dermal Human Fibroblasts In Vitro

This study evaluated plasma gel preparation protocols with potential to yield more biocompatible materials for facial aesthetic applications. Human dermal fibroblasts were cultured in medium supplemented with 20% plasma gel obtained using four protocols: G1, centrifugation at 580 g for 8 min in tubes with anticoagulant followed by activation with 10% calcium chloride; G2, centrifugation at 580 g for 8 min in tubes with anticoagulant without activation; G3, centrifugation at 210 g for 8 min in tubes without anticoagulant with activation; and G4, centrifugation at 210 g for 8 min in tubes without anticoagulant and without activation. All protocols involved heating plasma at 76 °C for 12 min to produce gels. Cell viability was assessed using MTT and Crystal Violet assays over 72 h. Cell morphology and fibrin formation were analyzed by hematoxylin/eosin staining. Cell migration was evaluated using a wound healing assay. RT-qPCR analysis of COL1A1 and FN1 was performed after 24, 48, and 72 h. Fibroblasts cultured with plasma gel from G3 and G4 showed significantly higher viability compared with the control group. G1 and G2 exhibited increased fibrin formation and reduced cell density. Slower wound closure was observed in G2, which was consistent with increased late-stage expression of COL1A1 and FN1. In the conditions evaluated, low-speed centrifugation and the use of tubes without anticoagulant improved plasma gel biocompatibility. Lack of activation after high-speed centrifugation was associated with slower wound closure.