Mesoscopic structural analysis via deep learning processing, with a special reference to in vitro alteration in collagen fibre induced by a gap junction inhibitor
Mesoscopic structural analysis via deep learning processing, with a special reference to in vitro alteration in collagen fibre induced by a gap junction inhibitor
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通过深度学习处理进行细观结构分析，特别参考间隙连接抑制剂诱导的胶原纤维的体外变化
DOI：
10.1093/jmicro/dfac044
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发表时间：
2022
期刊：
影响因子：
1.8
通讯作者：
Nakamura Kei-ichiro
中科院分区：
文献类型：
--
作者：
Hirashima Shingo;Ohta Keisuke;Togo Akinobu;Tsuneyoshi Risa;Kusukawa Jingo;Nakamura Kei-ichiro
致密结缔组织，包括韧带、肌腱、筋膜和角膜，由成纤维细胞（Fbs）合成的规则排列的胶原纤维形成。确定纤维取向的机制仍不清楚。牙周膜韧带Fbs始终通过间隙连接（GJ）与周围环境沟通，从而形成广泛的细胞网络。Schaferet等人先前报道了培养Hb合成的胶原纤维的方法。（“培养的人成纤维细胞中的抗坏血酸缺乏症”，细胞生物学杂志，34：83-95，1967）。该方法已被应用于调查的能力和活动的Fb胶原蛋白的合成/吞噬作用，使用传统的电子显微镜（EM）。然而，胶原纤维的三维介观结构和GJ抑制剂对体外胶原纤维形成的影响知之甚少。在这项研究中，三维介观分析被用来阐明定向纤维形成的机制。我们研究了GJ抑制剂对牙周膜Fbin体外驱动胶原形成的影响，组织形态计量学，以及在中尺度上定量的体外胶原纤维的结构特性，使用相关光和EM优化的苦天狼星红染色和聚焦离子束扫描EM断层扫描。我们的研究结果表明，在培养条件下，在GJ抑制剂的存在下，胶原纤维的取向变得比对照组更无序。这表明，GJ可能参与决定胶原纤维形成过程中的纤维取向。阐明这一机制可能有助于开发新的治疗策略结缔组织定向障碍。图形摘要
Dense connective tissue, including the ligament, tendon, fascia and cornea, is formed by regularly arranged collagen fibres synthesized by fibroblasts (Fbs). The mechanism by which fibre orientation is determined remains unclear. Periodontal ligament Fbs consistently communicate with their surroundings via gap junctions (GJs), leading to the formation of a wide cellular network. A method to culture Fb–synthesized collagen fibres was previously reported by Schaferet al.(‘Ascorbic acid deficiency in cultured human fibroblasts’.J. Cell Biol.34: 83–95, 1967). This method has been applied to investigate the ability and activity of Fb collagen synthesis/phagocytosis using conventional electron microscopy (EM). However, the three-dimensional mesoscopic architecture of collagen fibres and the influence of GJ inhibitors on collagen fibre formationin vitroare poorly understood. In this study, three-dimensional mesoscopic analysis was used to elucidate the mechanism of directional fibre formation. We investigated the influence of GJ inhibitors on collagen formation driven by periodontal ligament Fbsin vitro, histomorphometrically, and the structural properties ofin vitrocollagen fibre on a mesoscale quantitatively, using correlative light and EM optimized for picrosirius red staining and focused ion beam-scanning EM tomography. Our results indicate that under culture conditions, in the presence of a GJ inhibitor, the orientation of collagen fibres becomes more disordered than that in the control group. This suggests that the GJ might be involved in determining fibre orientation during collagen fibre formation. Elucidation of this mechanism may help develop novel treatment strategies for connective tissue orientation disorders.Graphical Abstract