Designing, fabrication, and characterization of a bilayer wound dressing made from decellularized amniotic membrane coated with emulsion based PCU Silk electerospun for Drug delivery and cornea tissue engineering
Jila Majidi1 , Seyed-Hashem Daryabari2 *, Mazaher Gholipourmalekabadi3
- Department of Tissue Engineering & Regenerative Medicine, Faculty of Advanced Technologies in Medicine, Iran University of Medical Sciences, Tehran, Iran
- Vision Health Research Center, Semnan University of Medical Sciences, Semnan, Iran
- Department of Medical Biotechnology, Faculty of Allied Medicine, Iran University of Medical Sciences, Tehran, Iran
Abstract: Effective communication between immune and corneal epithelial-forming cells is crucial for successfully healing chemical corneal defects. In this study, for the first time, we fabricated an emulsion-based (EM) electrospun core-shell mat using Betamethasone Dipropionate (BD) loaded polycarbonate urethane-Silk (EM) as a system for inducing M1/M2 polarization in macrophages and promoting corneal epithelial regeneration, both in vitro and in vivo.
Methods: In this study, BD-loaded EM solution was electrospun directly onto a decellularized human amniotic membrane (AM) to engineer an ultrathin bilayer membrane (AM/EM) and conducted a comprehensive characterization of its biomechanical properties, and biocompatibility. In vitro, we examined the influence of the BD-loaded AM/EM on the polarization of macrophages through nitric oxide assays, phagocytosis tests, and real-time polymerase chain reaction (PCR). For in vivo assessment, we utilized histological evaluations, and real-time PCR to determine the impact of the BD-loaded AM/EM on cornea regeneration in a chemical corneal burn defect model.
Results: The culture of macrophages on the BD-loaded AM/EM led to increased production of IL-10, upregulation of CD206, Arg1 gene expression, and enhanced phagocytic capacity for apoptotic thymocytes. Concurrently, it reduced the secretion of TNF-α and nitric oxide (NO), downregulated the expression of CD86, and NOS2 genes, and decreased macrophage phagocytosis of yeast. These results indicated the polarization of macrophages toward an M2-like phenotype. In vivo, the presence of the BD-loaded AM/EM resulted in enhanced cornea recapitalization at the defect site, as compared to the control defect.
Additionally, various inflammatory factors, including IL-1β, IL-10, TNF-alpha, MM9, VEGF, and COL1A2 were investigated through quantitative real-time RT-PCR. AM/EM 6%BD group showed histologically alleviated signs of corneal inflammation and regeneration compared with another group, which decreased IL-1β, TNF-alpha, and MMP9 expression. Notably, AM/EM 6%BD treatment significantly suppressed neovascularization and enhanced the anti-inflammatory function of BD during the acute phase of ocular inflammation.
Conclusion: In conclusion, this immunomodulatory effect, coupled with its biocompatibility and biomechanical properties resembling natural cornea, positions the BD-loaded AM/EM as an attractive candidate for further exploration in the field of cornea tissue engineering and regenerative medicine.
