Dynamic analysis of Porphyromonas gingivalis invasion into blood capillaries during the infection process in host tissues using a vascularized three-dimensional human gingival model
Dynamic analysis of Porphyromonas gingivalis invasion into blood capillaries during the infection process in host tissues using a vascularized three-dimensional human gingival model
复制标题
利用血管化三维人体牙龈模型动态分析牙龈卟啉单胞菌在宿主组织感染过程中侵入毛细血管的动态
DOI：
10.1039/d1bm00831e
复制
发表时间：
2021
影响因子：
6.6
通讯作者：
Matsusaki Michiya
中科院分区：
文献类型：
--
作者：
Sasaki Naoko;Takeuchi Hiroki;Kitano Shiro;Irie Shinji;Amano Atsuo;Matsusaki Michiya
牙龈卟啉单胞菌是牙周病的病原体，被认为通过牙龈组织的毛细血管参与了全身的各种疾病。然而，由于缺乏体内和体外的模型，对感染机制的动态分析，特别是对牙龈组织的深层侵袭过程的研究尚未阐明。在这项研究中，我们开发了一个带有上皮屏障的血管化三维(3D)牙龈模型，利用胶原微纤维(CMF)表达细胞与细胞之间的连接，以实现对牙龈假单胞菌侵袭过程的动态分析。牙龈上皮细胞层的脂筏破坏实验表明，牙龈假单胞菌通过细胞间途径而不是细胞内途径迁移到更深的上皮细胞。在两天的培养过程中，在毛细血管内发现了牙龈假单胞菌，这表明它入侵了3D牙龈模型。值得注意的是，细菌的数量在至少两天后大幅增加，而缺乏半胱氨酸蛋白酶的突变牙龈假单胞菌的存活人数明显减少。在感染野生型Pggivalis的两天内，牙周组织中IL-6的分泌减少，而突变体则相反，提示Pggivalis感染在早期阶段干扰了IL-6的分泌。通过首次动态观察牙龈假单胞菌从上皮细胞层向毛细血管的侵袭，该模型将成为开发治疗牙周感染相关疾病的新疗法的有力工具。
Porphyromonas gingivalis, the pathogen of periodontal disease, is thought to be involved in various diseases throughout the body via gingival tissue blood capillaries. However, the dynamic analysis of the infection mechanism, particularly the deep invasion process of the gingival tissue, has not yet been elucidated because of the lack of both in vivo and in vitro models. In this study, we developed a vascularized three-dimensional (3D) gingival model with an epithelial barrier expressing cell–cell junctions using collagen microfibers (CMFs) to enable the dynamic analysis of the P. gingivalis invasion process. Lipid raft disruption experiments in the gingival epithelial cell layer demonstrated that P. gingivalis migrates into the deeper epithelium via the intercellular pathway rather than intracellular routes. P. gingivalis was shown to invade the 3D gingival model, being found inside blood capillaries during two days of culture. Notably, the number of bacteria had increased greatly at least two days later, whereas the mutant P. gingivalis lacking the cysteine proteases, gingipains, showed a significantly lower number of survivors. The secretion of interleukin-6 (IL-6) from the gingival tissue decreased during the two days of infection with the wild type P. gingivalis, but the opposite was found for the mutant suggesting that P. gingivalis infection disturbs IL-6 secretion at an early stage. By allowing the dynamic observation of the P. gingivalis invasion from the epithelial cell layer into the blood capillaries for the first time, this model will be a powerful tool for the development of novel therapeutics against periodontal infection related diseases.