Institute of Technical Chemistry, Leibniz University Hannover, Hannover, Germany.
Institute of Quantum Optics, Hannover, Germany.
Tissue Eng Part A. 2021 Oct;27(19-20):1239-1249. doi: 10.1089/ten.TEA.2020.0232. Epub 2021 Feb 24.
In this study, microvascular network structures for tissue engineering were generated on newly developed macroporous polydioxanone (PDO) scaffolds. PDO represents a polymer biodegradable within months and offers optimal material properties such as elasticity and nontoxic degradation products. PDO scaffolds prepared by porogen leaching and cryo-dried to achieve pore sizes of 326 ± 149.67 μm remained stable with equivalent values for Young's modulus after 4 weeks. Scaffolds were coated with fibrin for optimal cell adherence. To exclude interindividual differences, autologous fibrin was prepared out of human plasma-derived fibrinogen and proved a comparable quality to nonautologous commercially available fibrinogen. Fibrin-coated scaffolds were seeded with recombinant human umbilical vein endothelial cells expressing GFP (GFP-HUVECs) in coculture with adipose tissue-derived mesenchymal stem cells (AD-hMSCs) to form vascular networks. The growth factor content in culture media was optimized according its effect on network formation, quantified and assessed by AngioTool. A ratio of 2:3 GFP-HUVECs/AD-hMSCs in medium enriched with 20 ng/mL vascular endothelial growth factor, basic fibroblast growth factor, and hydrocortisone was found to be optimal. Network structures appeared after 2 days of cultivation and stabilized until day 7. The resulting networks were lumenized that could be verified by dextran staining. This new approach might be suitable for microvascular tissue patches as a useful template to be used in diverse vascularized tissue constructs. Impact statement We consider this work as important for the current research in the field of tissue engineering and the development of new and functional tissue. The approach for the production of vascularized tissue patches, consisting of the biodegradable synthetic polymer polydioxanone and of the physiological, autologous, and patient-specific polymer fibrin, and seeded with endothelial cells and mesenchymal stem cells, displayed within this work, could be useful for the sustaining development of diverse and more complex tissue constructs. Therefore, these scaffolds could be used as a cornerstone for future tissue engineering approaches.
在这项研究中，我们在新开发的多孔聚二氧杂环已酮（PDO）支架上生成用于组织工程的微血管网络结构。PDO 是一种可在数月内生物降解的聚合物，具有弹性和无毒降解产物等最佳材料特性。通过致孔剂浸出和冷冻干燥制备的 PDO 支架，孔径达到 326±149.67μm，在 4 周后保持稳定，杨氏模量具有等效值。支架用纤维蛋白包被以实现最佳细胞黏附。为了排除个体间差异，从人血浆衍生的纤维蛋白原制备了自体纤维蛋白，并证明其质量与非自体商业可得的纤维蛋白原相当。用纤维蛋白包被的支架与脂肪组织来源的间充质干细胞（AD-hMSCs）共培养，接种表达 GFP 的重组人脐静脉内皮细胞（GFP-HUVECs），形成血管网络。根据其对网络形成的影响优化培养基中的生长因子含量，并用 AngioTool 定量和评估。在富含 20ng/mL 血管内皮生长因子、碱性成纤维细胞生长因子和氢化可的松的培养基中，发现 2:3 GFP-HUVECs/AD-hMSCs 的比例是最佳的。培养 2 天后出现网络结构，并稳定至第 7 天。通过葡聚糖染色可以验证形成的管腔化网络。这种新方法可能适用于微血管组织贴片，作为在各种血管化组织构建物中使用的有用模板。
