Walawalkar Sonal, Almelkar Shahdab
Division of Tissue Engineering & Cell Science (TECS), HEAL BIOLABS, Shree Hospital & Research Institute (SHRI), Maharashtra, India.
J Biomater Appl. 2021 Jul;36(1):165-178. doi: 10.1177/0885328220968388. Epub 2020 Nov 1.
There is a significant pitfall in clinical translation of large-sized tissue-engineered grafts - a lack of vascularization. This study was carried out to find the answer in a plant leaf, as plants and animals share structural similarities.
We fabricated a scaffold using leaves (10%SDS) and expanded the endothelial cells onto them. The vascularity was demarcated by angiography. The thermal decomposition confirmed that the oxidation resistance of the scaffold is parallel to the natural leaf. The acellularity of the scaffold as well as the presence of cellular establishment after culture on the scaffold was confirmed by histology, scanning electron microscopy, periodic acid-Schiff, and DNA quantification. Further, we estimated various biochemical markers like MDA, catalase, total proteins, and total nitric oxide for confirming their metabolic activities. Cell-specific markers like vWF, lectin established their phenotype. Cytotoxicity and live-dead assay showed the viability of cells.
Our findings proved that the decellularized leaf scaffold preserves vascularity, exhibits non-toxicity, maintains the cell identity, and supports mammalian cells for their metabolic activities. The study gives a futuristic hope in combating the ever-growing issues of clinical applicability of large-sized grafts.
大型组织工程移植物的临床转化存在一个重大缺陷——缺乏血管化。由于植物和动物在结构上有相似之处，本研究旨在从植物叶片中寻找答案。
我们使用叶片（10%十二烷基硫酸钠）制作了一个支架，并在其上扩增内皮细胞。通过血管造影确定血管分布。热分解证实该支架的抗氧化性与天然叶片相当。通过组织学、扫描电子显微镜、高碘酸希夫反应和DNA定量分析，证实了支架的脱细胞状态以及在支架上培养后细胞的生长情况。此外，我们评估了丙二醛、过氧化氢酶、总蛋白和总一氧化氮等多种生化标志物，以确认它们的代谢活性。血管性血友病因子、凝集素等细胞特异性标志物确定了细胞的表型。细胞毒性和活死检测显示了细胞的活力。
我们的研究结果证明，脱细胞叶片支架保留了血管分布，表现出无毒性，维持了细胞特性，并支持哺乳动物细胞进行代谢活动。该研究为解决大型移植物临床应用中日益突出的问题带来了未来的希望。
