Department of Plastic and Hand Surgery, University Hospital of Erlangen, Friedrich-Alexander Universität Erlangen-Nürnberg (FAU), 91054 Erlangen, Germany.
Department of Materials Science and Engineering for Metals, Friedrich-Alexander Universität Erlangen-Nürnberg (FAU), 91058 Erlangen, Germany.
Cells. 2023 Jan 9;12(2):261. doi: 10.3390/cells12020261.
Due to the limitations of current in vivo experimental designs, our comprehensive knowledge of vascular development and its implications for the development of large-scale engineered tissue constructs is very limited. Therefore, the purpose of this study was to develop unique in vivo imaging chambers that allow the live visualization of cellular processes in the arteriovenous (AV) loop model in rats. We have developed two different types of chambers. Chamber A is installed in the skin using the purse sting fixing method, while chamber B is installed subcutaneously under the skin. Both chambers are filled with modified gelatin hydrogel as a matrix. Intravital microscopy (IVM) was performed after the injection of fluorescein isothiocyanate (FITC)-labeled dextran and rhodamine 6G dye. The AV loop was functional for two weeks in chamber A and allowed visualization of the leukocyte trafficking. In chamber B, microvascular development in the AV loop could be examined for 21 days. Quantification of the microvascular outgrowth was performed using Fiji-ImageJ. Overall, by combining these two IVM chambers, we can comprehensively understand vascular development in the AV loop tissue engineering model¯.
由于目前体内实验设计的局限性，我们对血管发育及其对大规模工程组织构建发展的影响的全面了解非常有限。因此，本研究的目的是开发独特的体内成像室，允许在大鼠动静脉（AV）环模型中实时可视化细胞过程。我们已经开发了两种不同类型的腔室。腔室 A 采用荷包缝合固定法安装在皮肤上，而腔室 B 则安装在皮下。两个腔室均充满改良明胶水凝胶作为基质。在注射异硫氰酸荧光素（FITC）标记的葡聚糖和罗丹明 6G 染料后进行活体显微镜检查（IVM）。腔室 A 中的 AV 环在 2 周内具有功能，允许观察白细胞迁移。在腔室 B 中，可以在 21 天内检查 AV 环中的微血管发育。使用 Fiji-ImageJ 对微血管生长进行定量。总的来说，通过结合这两种 IVM 腔室，我们可以全面了解 AV 环组织工程模型中的血管发育。
