Department of Plastic and Hand Surgery, University Hospital of Erlangen, Friedrich-Alexander-University Erlangen-Nürnberg (FAU), Erlangen, 91054, Germany.
Laboratory for Tissue Engineering and Regenerative Medicine, University Hospital of Erlangen, Friedrich-Alexander-University Erlangen-Nürnberg (FAU), Erlangen, 91054, Germany.
Biomed Mater. 2021 Sep 6;16(6). doi: 10.1088/1748-605X/ac1e9d.
tissue engineering is an emerging field aiming at the generation of ready-to-use three-dimensional tissues. One solution to supply a proper vascularization of larger tissues to provide oxygen and nutrients is the arteriovenous loop (AVL) model. However, for this model, suitable scaffold materials are needed that are biocompatible/non-immunogenic, slowly degradable, and allow vascularization. Here, we investigate the suitability of the known gelatin methacryloyl (GelMA)-based hydrogel fortissue engineering utilizing the AVL model. Rat AVLs are embedded by two layers of GelMA hydrogel in an inert PTFE chamber and implanted in the groin. Constructs were explanted after 2 or 4 weeks and analyzed. For this purpose, gross morphological, histological, and multiphoton microscopic analysis were performed. Immune response was analyzed based on anti-CD68 and anti-CD163 staining of immune cells. The occurrence of matrix degradation was assayed by anti-MMP3 staining. Vascularization was analyzed by anti--smooth muscle actin staining, multiphoton microscopy, as well as expression analysis of 53 angiogenesis-related proteins utilizing a proteome profiler angiogenesis array kit. Here we show that GelMA hydrogels are stable for at least 4 weeks in the rat AVL model. Furthermore, our data indicate that GelMA hydrogels are biocompatible. Finally, we provide evidence that GelMA hydrogels in the AVL model allow connective tissue formation, as well as vascularization, introducing multiphoton microscopy as a new methodology to visualize neovessel formation originating from the AVL. GelMA is a suitable material forandTE in the AVL model.
组织工程学是一个旨在生成即用型三维组织的新兴领域。为了给较大的组织提供适当的血管化以提供氧气和营养，可以使用动静脉环（AVL）模型。然而，对于这种模型，需要使用生物相容性/非免疫原性、缓慢降解且允许血管生成的合适支架材料。在这里，我们研究了已知的明胶甲基丙烯酰（GelMA）基水凝胶在利用 AVL 模型进行组织工程中的适用性。将大鼠 AVL 嵌入两层惰性 PTFE 腔室中的 GelMA 水凝胶中，并植入腹股沟。在 2 或 4 周后取出构建体并进行分析。为此，进行了大体形态学、组织学和多光子显微镜分析。根据免疫细胞的抗 CD68 和抗 CD163 染色分析免疫反应。通过抗 MMP3 染色检测基质降解的发生。通过抗平滑肌肌动蛋白染色、多光子显微镜以及利用蛋白质组分析试剂盒进行的 53 种与血管生成相关的蛋白质表达分析来分析血管生成。在这里，我们表明 GelMA 水凝胶在大鼠 AVL 模型中至少稳定 4 周。此外，我们的数据表明 GelMA 水凝胶具有生物相容性。最后，我们提供了证据表明 GelMA 水凝胶在 AVL 模型中允许结缔组织形成和血管生成，引入多光子显微镜作为一种新的方法来可视化源自 AVL 的新血管形成。GelMA 是 AVL 模型中组织工程学的一种合适材料。
