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Background <p>Biotechnological advances in tissue engineering increasingly exploit naturally derived scaffolds as bioactive delivery systems. Acellular fish skin (AFS) provides a collagen-rich extracellular matrix favorable for tissue regeneration but lacks intrinsic antimicrobial activity. Propolis, a polyphenol-rich natural resin, exhibits antimicrobial, antioxidant, and immunomodulatory properties, yet its direct application is limited by burst release and local cytotoxicity. In this study, we aimed to develop and characterize a propolis-functionalized AFS scaffold as a multifunctional biomaterial capable of combining sustained antimicrobial activity with regenerative support for wound-healing applications.</p> Results <p>Lipid-preserved <i>Tilapia</i>-derived AFS was biofunctionalized with ethanolic propolis extract (EPE) through a vacuum-assisted impregnation process to obtain an EPE-AFS hybrid scaffold. The biotechnological modification preserved the collagenous architecture and porosity of the matrix, enhanced surface hydrophilicity (contact angle reduced from 91° ± 2° to 67° ± 3°), and maintained mechanical integrity (~ 13&#xa0;MPa). Spectroscopic and microscopic analyses (FTIR, FE-SEM) confirmed successful deposition of polyphenolic components. The release profile demonstrated sustained polyphenol diffusion over 96&#xa0;h (&gt; 90% cumulative release). Functionally, cellular assays confirmed enhanced fibroblast adhesion and migration on propolis-functionalized scaffolds. Gene expression analysis revealed transcriptional upregulation of extracellular matrix remodeling and angiogenesis-associated markers (COL1A1, COL3A1, VEGF, TGF-β1), alongside downregulation of inflammation-related transcripts (IL-6, MMP-9). Antibacterial evaluation based on turbidity measurements indicated effective growth inhibition against <i>Staphylococcus aureus</i> and <i>Escherichia coli</i>.</p> Conclusions <p>This study demonstrates a biotechnological strategy for functionalizing natural collagen scaffolds with propolis-derived polyphenols, yielding a dual-action biomaterial that combines sustained antibacterial growth suppression with enhanced regenerative-associated signaling in vitro. The EPE–AFS construct represents a promising nature-derived platform for further preclinical investigation in wound-healing applications. While these findings reveal a pro-regenerative and anti-inflammatory transcriptional profile in fibroblasts, additional validation—including endothelial and immune cell functional assays and in vivo wound models—is required to confirm angiogenic, immunomodulatory, and therapeutic efficacy under physiologically relevant conditions.</p>

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Propolis-functionalized acellular fish skin scaffolds as biotechnological platforms for antimicrobial activity and regenerative wound healing

  • 期刊论文

  • 出版模式：

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  • 发表日期：

    2026年4月21日

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  • Sima Bakhtazad,
  • Sepehr Mehdizadeh,
  • Nosratollah Zarghami,
  • Ali Akbar Shabani,
  • Mehdi Dadashpour,
  • Younes Pilehvar

摘要

Background

Biotechnological advances in tissue engineering increasingly exploit naturally derived scaffolds as bioactive delivery systems. Acellular fish skin (AFS) provides a collagen-rich extracellular matrix favorable for tissue regeneration but lacks intrinsic antimicrobial activity. Propolis, a polyphenol-rich natural resin, exhibits antimicrobial, antioxidant, and immunomodulatory properties, yet its direct application is limited by burst release and local cytotoxicity. In this study, we aimed to develop and characterize a propolis-functionalized AFS scaffold as a multifunctional biomaterial capable of combining sustained antimicrobial activity with regenerative support for wound-healing applications.

Results

Lipid-preserved Tilapia-derived AFS was biofunctionalized with ethanolic propolis extract (EPE) through a vacuum-assisted impregnation process to obtain an EPE-AFS hybrid scaffold. The biotechnological modification preserved the collagenous architecture and porosity of the matrix, enhanced surface hydrophilicity (contact angle reduced from 91° ± 2° to 67° ± 3°), and maintained mechanical integrity (~ 13 MPa). Spectroscopic and microscopic analyses (FTIR, FE-SEM) confirmed successful deposition of polyphenolic components. The release profile demonstrated sustained polyphenol diffusion over 96 h (> 90% cumulative release). Functionally, cellular assays confirmed enhanced fibroblast adhesion and migration on propolis-functionalized scaffolds. Gene expression analysis revealed transcriptional upregulation of extracellular matrix remodeling and angiogenesis-associated markers (COL1A1, COL3A1, VEGF, TGF-β1), alongside downregulation of inflammation-related transcripts (IL-6, MMP-9). Antibacterial evaluation based on turbidity measurements indicated effective growth inhibition against Staphylococcus aureus and Escherichia coli.

Conclusions

This study demonstrates a biotechnological strategy for functionalizing natural collagen scaffolds with propolis-derived polyphenols, yielding a dual-action biomaterial that combines sustained antibacterial growth suppression with enhanced regenerative-associated signaling in vitro. The EPE–AFS construct represents a promising nature-derived platform for further preclinical investigation in wound-healing applications. While these findings reveal a pro-regenerative and anti-inflammatory transcriptional profile in fibroblasts, additional validation—including endothelial and immune cell functional assays and in vivo wound models—is required to confirm angiogenic, immunomodulatory, and therapeutic efficacy under physiologically relevant conditions.