Seyedi Jasbi Akram Sadat, Aghamollaei Hossein, Jangravi Zohreh, Sari Soyar, Salimi Ali
Department of Cellular and Molecular Biology, Faculty of Advanced Science and Technology, Tehran Medical Sciences, Islamic Azad University, Tehran, Iran.
Chemical Injuries Research Center, Systems Biology and Poisonings Institute, Baqiyatallah University of Medical Sciences, Tehran, Iran.
Sci Rep. 2025 May 24;15(1):18085. doi: 10.1038/s41598-025-02584-w.
This study aims to develop a novel antibacterial hydrogel scaffold composed of gelatin (Gel), amniotic membrane extract (AME), and aminophylline (AMP) for neural regeneration. We investigate its ability to sustain AMP release, inhibit bacterial growth, and promote neural differentiation of human Wharton's jelly mesenchymal stem cells (hWJMSCs) via the CREB pathway, addressing unmet needs in neural tissue engineering. The composite hydrogels were synthesized and characterized using various methods and techniques, including X-ray diffraction (XRD), field emission scanning electron microscopy (FESEM), Fourier transform infrared spectroscopy (FTIR), porosity, contact angle, water uptake, thermogravimetric analysis (TGA), biodegradation, tensile strength, drug release, and antibacterial activity. Biocompatibility tests (MTT assay, AO/EB staining) confirmed > 95% viability of hWJMSCs over six days and their differentiation to the neural cells was analyzed through immunocytochemistry (ICC) staining and real-time reverse transcription-polymerase chain reaction (RT-PCR) at different time points. The results demonstrate the successful synthesis of porous hydrogels with desirable properties, including hydrophilicity, thermal stability, biodegradability, and mechanical strength. The hydrogels support the sustained release of AMP (53.18% over 336 h) and exhibit antibacterial activity against Pseudomonas aeruginosa (90.52 ± 0.26%) and Staphylococcus aureus (93.06 ± 0.34%) due to the presence of penicillin and streptomycin (P-S) antibiotics. The biocompatibility results show that the hydrogels do not have a cytotoxic effect on the viability of human WJMSCs. The neural differentiation of human WJMSCs seeded on surface hydrogels was confirmed by evaluating specific neural markers at both protein and gene levels. In conclusion, the new antibacterial gel-based hydrogel can support the release of AMP and after further evaluation, can be introduced as a new candidate for neural repair applications.
本研究旨在开发一种由明胶（Gel）、羊膜提取物（AME）和氨茶碱（AMP）组成的新型抗菌水凝胶支架，用于神经再生。我们研究了其维持AMP释放、抑制细菌生长以及通过CREB途径促进人脐带华通氏胶间充质干细胞（hWJMSCs）神经分化的能力，以满足神经组织工程中未满足的需求。使用多种方法和技术对复合水凝胶进行了合成和表征，包括X射线衍射（XRD）、场发射扫描电子显微镜（FESEM）、傅里叶变换红外光谱（FTIR）、孔隙率、接触角、吸水率、热重分析（TGA）、生物降解、拉伸强度、药物释放和抗菌活性。生物相容性测试（MTT法、AO/EB染色）证实hWJMSCs在六天内的存活率>95%，并在不同时间点通过免疫细胞化学（ICC）染色和实时逆转录-聚合酶链反应（RT-PCR）分析它们向神经细胞的分化。结果表明成功合成了具有所需特性的多孔水凝胶，包括亲水性、热稳定性、生物降解性和机械强度。水凝胶支持AMP的持续释放（336小时内释放53.18%），并且由于存在青霉素和链霉素（P-S）抗生素，对铜绿假单胞菌（90.52±0.26%）和金黄色葡萄球菌（93.06±0.34%）表现出抗菌活性。生物相容性结果表明，水凝胶对人WJMSCs的活力没有细胞毒性作用。通过在蛋白质和基因水平评估特定的神经标志物，证实了接种在表面水凝胶上的人WJMSCs的神经分化。总之，新型抗菌凝胶基水凝胶可以支持AMP的释放，经过进一步评估后，可以作为神经修复应用的新候选材料引入。
