Shi Lijuan, Xu Yiwen, Li Jingying, He Li, Li Kaiyu, Yin Shigang, Nie Minhai, Liu Xuqian
Department of Periodontics & Oral Mucosal Diseases, The Affiliated Stomatology Hospital of Southwest Medical University, Luzhou, Sichuan, China.
Oral & Maxillofacial Reconstruction and Regeneration of Luzhou Key Laboratory, Luzhou, Sichuan, China.
J Tissue Eng. 2024 Sep 18;15:20417314241268912. doi: 10.1177/20417314241268912. eCollection 2024 Jan-Dec.
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.
我们之前的研究有效地构建了组织工程化血管化口腔黏膜替代物（VOME）；然而，遇到了诸如重复性和稳定性低以及无法准确复制真实血管复杂性等挑战。因此，本研究旨在通过与激光捕获显微切割和人血管生成抗体阵列技术相关的串联质量标签蛋白质组学筛选VOME和天然口腔黏膜血管稳态表型。然后，构建慢病毒并将其稳定转染至血管内皮样细胞（VELC）以检测血管生成能力。使用苏木精-伊红染色、EdU Apollo示踪剂染色、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来分析血管化稳态的特征并监测血管化稳态表型口腔黏膜替代物（VHPOME）中的功能再生。结果表明，磷酸甘油酸变位酶1（PGAM1）、Ⅴ型胶原α1链（COL5A1）、血管生成素（ANG）和核糖核酸酶H1（RNH1）是潜在的特定血管生成表型。PGAM1、COL5A1和ANG高表达和/或RNH1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组中PGAM1、COL5A1和ANG的表达高于对照组，而RNH1的表达低于对照组。COL5A1/ANG可显著改善力学性能。ANG/shRNH1组的血流信号最为显著。PGAM1、COL5A1、ANG、shRNH1、PGAM1/ANG、COL5A1/ANG、PGAM1/shRNH1、PGAM1/shRNH1、COL5A1/shRNH1和ANG/shRNH1可能是建立口腔黏膜替代物血管化稳态和功能再生的基因（组）靶点，且ANG/shRNH1的作用最为显著。
