Imeidopf Gianna, Khaimov Dara, John Sashane, Merna Nick
Fred DeMatteis School of Engineering and Applied Science, Hofstra University, Hempstead, NY 11549, USA.
Cardiothoracic Surgery, Northwell Health, New York, NY 10022, USA.
Int J Mol Sci. 2025 Mar 19;26(6):2752. doi: 10.3390/ijms26062752.
Vascular graft failure rates remain unacceptably high due to thrombosis and poor integration, necessitating innovative solutions. This study optimized plant-derived extracellular matrix scaffolds as a scalable and biocompatible alternative to synthetic grafts and autologous vessels. We refined decellularization protocols to achieve >95% DNA removal while preserving mechanical properties comparable to native vessels, significantly enhancing endothelial cell seeding. Leatherleaf viburnum leaves were decellularized using sodium dodecyl sulfate-based and Trypsin/Tergitol-based treatments, achieved via clearing in bleach and Triton X-100 for 6 to 72 h. To assess the environmental influence on scaffold performance, leaves from multiple collection sites were processed using sodium dodecyl sulfate-based protocols. Scaffold performance was evaluated through tensile testing and histological analysis to assess structural integrity, while DNA quantification and endothelial cell recellularization measured biological compatibility. Sodium dodecyl sulfate-treated scaffolds with shorter clearing durations demonstrated the highest DNA removal (≥95%) while preserving mechanical properties, significantly outperforming Trypsin/Tergitol treatments. Longer clearing times reduced fiber diameter by 60%, compromising scaffold strength. Shorter clearing times preserved extracellular matrix integrity and significantly improved endothelial cell seeding efficiency. Larger leaves supported significantly higher endothelial cell densities than smaller leaves, highlighting the need for standardized material sources. Permeability tests demonstrated minimal leakage at 120 mmHg and structural stability under dynamic flow conditions, suggesting their suitability for vascular applications. These findings establish a reliable framework for optimizing plant-derived grafts, improving their reproducibility and performance for tissue engineering applications.
由于血栓形成和整合不良，血管移植物的失败率仍然高得令人无法接受，因此需要创新的解决方案。本研究优化了植物源细胞外基质支架，将其作为合成移植物和自体血管的可扩展且生物相容的替代品。我们改进了脱细胞方案，以实现>95%的DNA去除率，同时保持与天然血管相当的机械性能，显著提高内皮细胞的接种效率。使用基于十二烷基硫酸钠和基于胰蛋白酶/特吉托尔的处理方法对雪叶莲叶片进行脱细胞处理，通过在漂白剂和 Triton X-100中清除6至72小时来实现。为了评估环境对支架性能的影响，使用基于十二烷基硫酸钠的方案处理来自多个采集地点的叶片。通过拉伸测试和组织学分析评估支架性能，以评估结构完整性，同时通过DNA定量和内皮细胞再细胞化来测量生物相容性。清除时间较短的十二烷基硫酸钠处理的支架在保持机械性能的同时显示出最高的DNA去除率（≥95%），明显优于胰蛋白酶/特吉托尔处理。较长的清除时间使纤维直径减小了60%，损害了支架强度。较短的清除时间保留了细胞外基质的完整性，并显著提高了内皮细胞的接种效率。较大的叶片比较小的叶片支持显著更高的内皮细胞密度，突出了标准化材料来源的必要性。渗透性测试表明，在120 mmHg压力下渗漏最小，在动态流动条件下结构稳定，表明它们适用于血管应用。这些发现为优化植物源移植物建立了一个可靠的框架，提高了它们在组织工程应用中的可重复性和性能。
