Biological evaluation of gelatin-based hemostatic agents - cytocompatibility and irritation assessment

The effective management of hemostasis is essential in surgical practice. Topical hemostatic agents become indispensable when conventional methods are insufficient or impractical. However, the wide range of commercially available products with similar compositional bases presents clinicians with numerous choices and potentially variable performance profiles. Notably, to the best of the authors' knowledge, no previous study has comprehensively compared the biological functionality of these commercially available materials. Addressing this knowledge gap is essential for improving the precision and effectiveness of hemostatic interventions, ultimately contributing to better patient outcomes and advancing healthcare practice.

This study aimed to conduct a comprehensive evaluation of the biological response to clinically available gelatin-based hemostatic agents, namely Hemospon®, Clinix®, Gelatamp®, and Octocolagen®. The evaluation encompassed in vitro studies to characterize the response of human fibroblasts upon exposure, together with an in vivo irritation assessment using the hen's egg test - chorioallantoic membrane (HET-CAM) assay, thereby providing a comprehensive understanding of their biological effects.

Human fibroblasts were cultured and exposed to the leachables extracted from the hemostatic agents, prepared according to the ISO 10993-12:2021 guidelines, at concentrations of 50%, 25% and 12.5%. The cultures were characterized in terms of cellular viability (the live/dead assay and flow cytometry) and metabolic activity at multiple time points. In addition, irritation potential was evaluated using the HET-CAM assay.

Data analysis revealed that the fibroblast cultures exposed to the leachables from the 4 hemostatic sponges proliferated actively throughout the culture period, with minimal evidence of cell death or significant impairment of metabolic activity. Furthermore, no irritant effects were observed in the HET-CAM assay.

Overall, despite minor variations in the biological response, the comprehensive analysis supports the overall biocompatibility of commercially available gelatin-based hemostatic sponges. These findings advance the current understanding of the biological safety of these materials, further reinforcing their clinical suitability and highlighting their potential to improve surgical healing outcomes.