Vascularized characteristics and functional regeneration of three-dimensional cell reconstruction of oral mucosa equivalents based on vascular homeostasis phenotypic modification.

IF 6.7 1区 工程技术 Q1 CELL & TISSUE ENGINEERING Journal of Tissue Engineering Pub Date : 2024-09-18 eCollection Date: 2024-01-01 DOI:10.1177/20417314241268912
Lijuan Shi, Yiwen Xu, Jingying Li, Li He, Kaiyu Li, Shigang Yin, Minhai Nie, Xuqian Liu
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Abstract

Our prior research has effectively developed tissue-engineered vascularized oral mucosa equivalents (VOME); however, challenges such as low repeatability and stability, as well as the inability to accurately replicate the complexity of real blood vessels, were encountered. Therefore, this study aimed to screen the VOME and native oral mucosa vascular homeostasis phenotypes by tandem mass tag-tagged proteomics associated with laser capture microdissection and human angiogenesis antibody array technology. Then, lentiviruses were constructed and stably transfected with vascular endothelial-like cells (VELCs) to detect angiogenic capacity. HE, EdU Apollo tracer staining, immunofluorescence staining, scanning electron microscopy, biomechanical testing, and a small animal ultrasound imaging system were used to analyze the characteristics of vascularization homeostasis and monitor functional regeneration of the vascularized homeostatic phenotypic oral mucosal equivalents (VHPOME). The results showed that PGAM1, COL5A1, ANG, and RNH1 are potential specific angiogenesis phenotypes. High expression of PGAM1, COL5A1, and ANG and/or low expression of RNH1 can promote the angiogenesis of VOME. ANG/shRNH1 has the most significant tube-like structure-formation ability. The expression of PGAM1, COL5A1, and ANG in the VHPOME group was higher than that of the control group, and the expression of RNH1 was lower than that of the control group. COL5A1/ANG can significantly improve the mechanical properties. The blood flow signal was most significant in the ANG/shRNH1 group. PGAM1, COL5A1, ANG, shRNH1, PGAM1/ANG, COL5A1/ANG, PGAM1/shRNH1, PGAM1/shRNH1, COL5A1/shRNH1, and ANG/shRNH1 may be the targets for establishing vascularization homeostasis and functional regeneration of oral mucosal equivalent genes (groups), and ANG/shRNH1 has the most significant effect.

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基于血管稳态表型修饰的口腔黏膜等值体三维细胞重建的血管化特征和功能再生。
我们之前的研究有效地开发了组织工程化血管化口腔粘膜等效物(VOME),但也遇到了可重复性和稳定性低以及无法精确复制真实血管的复杂性等挑战。因此,本研究旨在通过串联质量标签标记蛋白质组学,结合激光捕获显微切割和人类血管生成抗体阵列技术,筛选VOME和原生口腔黏膜血管稳态表型。然后,构建慢病毒并稳定转染血管内皮样细胞(VELCs),以检测血管生成能力。利用HE、EdU阿波罗示踪染色、免疫荧光染色、扫描电子显微镜、生物力学测试和小动物超声成像系统分析血管化平衡的特征,并监测血管化平衡表型口腔粘膜等值体(VHPOME)的功能再生。结果显示,PGAM1、COL5A1、ANG和RNH1是潜在的特异性血管生成表型。PGAM1、COL5A1和ANG的高表达和/或RNH1的低表达可促进VOME的血管生成。ANG/shRNH1具有最显著的管状结构形成能力。VHPOME组PGAM1、COL5A1和ANG的表达高于对照组,而RNH1的表达低于对照组。COL5A1/ANG能显著改善机械性能。ANG/shRNH1组的血流信号最明显。PGAM1、COL5A1、ANG、shRNH1、PGAM1/ANG、COL5A1/ANG、PGAM1/shRNH1、PGAM1/shRNH1、COL5A1/shRNH1和ANG/shRNH1可能是建立血管化平衡和口腔粘膜等效基因(组)功能再生的靶点,其中ANG/shRNH1的效果最明显。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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来源期刊
Journal of Tissue Engineering
Journal of Tissue Engineering Engineering-Biomedical Engineering
CiteScore
11.60
自引率
4.90%
发文量
52
审稿时长
12 weeks
期刊介绍: The Journal of Tissue Engineering (JTE) is a peer-reviewed, open-access journal dedicated to scientific research in the field of tissue engineering and its clinical applications. Our journal encompasses a wide range of interests, from the fundamental aspects of stem cells and progenitor cells, including their expansion to viable numbers, to an in-depth understanding of their differentiation processes. Join us in exploring the latest advancements in tissue engineering and its clinical translation.
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