大分子拥挤会增强基质蛋白在组织工程血管移植物中的沉积。

IF 3.5 3区 医学 Q3 CELL & TISSUE ENGINEERING
Tissue Engineering Part A Pub Date : 2024-08-01 Epub Date: 2024-03-01 DOI:10.1089/ten.TEA.2023.0290
Qing Liu, Jiang Liu, Xu-Heng Sun, Jian-Yi Xu, Cong Xiao, Hong-Jing Jiang, Yin-Di Wu, Zhan-Yi Lin
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引用次数: 0

摘要

小直径组织工程血管移植物(TEVG)的成功体外培养需要血管平滑肌细胞(VSMC)分泌的生物大分子在聚乙醇酸(PGA)网状支架的三维(3D)多孔环境中快速沉积。然而,与体内组织液相比,普通介质的拥挤度较低。此外,在构建的早期阶段,细胞分泌到培养基中的大部分生物大分子都会流失,这对 TEVG 的培养过程产生了负面影响。在本研究中,我们提出了利用大分子拥挤(MMC)来增强培养基的拥挤度,以提高 TEVG 培养早期主要生物大分子的沉积和自组装效率。加入卡拉胶(CR)可显著提高培养基中的大分子拥挤度,而不会影响细胞活力、增殖和代谢活性。蛋白质分析表明,在添加大分子拥挤剂后,二维和三维平滑肌细胞培养物的细胞层中 I 型胶原蛋白、III 型胶原蛋白和纤连蛋白的沉积明显增加。培养基中的 I 型胶原蛋白与未添加大分子拥挤剂的培养基中的 I 型胶原蛋白相比明显减少。扫描电子显微镜显示,在三维培养的早期阶段,大分子拥挤剂大大促进了基质蛋白结构的形成。因此,MMC 改变了培养基的拥挤度,从而快速形成了大量基质蛋白和纤维结构。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Macromolecular Crowding Enhances Matrix Protein Deposition in Tissue-Engineered Vascular Grafts.

Successful in vitro culture of small-diameter tissue-engineered vascular grafts (TEVGs) requires rapid deposition of biomacromolecules secreted by vascular smooth muscle cells in a polyglycolic acid mesh scaffold's three-dimensional (3D) porous environment. However, common media have lower crowding conditions than in vivo tissue fluids. In addition, during the early stages of construction, most of the biomolecules secreted by the cells into the medium are lost, which negatively affects the TEVG culture process. In this study, we propose the use of macromolecular crowding (MMC) to enhance medium crowding to improve the deposition and self-assembly efficiency of major biomolecules in the early stages of TEVG culture. The addition of carrageenan significantly increased the degree of MMC in the culture medium without affecting cell viability, proliferation, and metabolic activity. Protein analysis demonstrated that the deposition of collagen types I and III and fibronectin increased significantly in the cell layers of two-dimensional and 3D smooth muscle cell cultures after the addition of a MMC agent. Collagen type I in the culture medium decreased significantly compared with that in the medium without a MMC agent. Scanning electron microscopy demonstrated that MMC agents considerably enhanced the formation of matrix protein structures during the early stages of 3D culture. Hence, MMC modifies the crowding degree of the culture medium, resulting in the rapid formation of numerous matrix proteins and fiber structures. Impact Statement Small-diameter tissue-engineered vascular grafts (TEVGs) are one of the most promising means of treating cardiovascular diseases; however, the in vitro construction of TEVGs has some limitations, such as slow deposition of extracellular matrix (ECM), long culture period, and poor mechanical properties. We hypothesized that macromolecular crowding can increase the crowding of the culture medium to construct a more bionic microenvironment, which enhances ECM deposition in the medium to the cell layer and reduces collagen loss, accelerating and enhancing TEVG culture and construction in vitro.

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来源期刊
Tissue Engineering Part A
Tissue Engineering Part A Chemical Engineering-Bioengineering
CiteScore
9.20
自引率
2.40%
发文量
163
审稿时长
3 months
期刊介绍: Tissue Engineering is the preeminent, biomedical journal advancing the field with cutting-edge research and applications that repair or regenerate portions or whole tissues. This multidisciplinary journal brings together the principles of engineering and life sciences in the creation of artificial tissues and regenerative medicine. Tissue Engineering is divided into three parts, providing a central forum for groundbreaking scientific research and developments of clinical applications from leading experts in the field that will enable the functional replacement of tissues.
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