Insulin-like growth factor 1 promotes the extension of Tracheal Epithelium in an in Vitro Tracheal organ culture model
Insulin-like growth factor 1 promotes the extension of Tracheal Epithelium in an in Vitro Tracheal organ culture model
复制标题
胰岛素样生长因子 1 在体外气管器官培养模型中促进气管上皮的延伸
DOI：
10.1016/j.anl.2020.09.017
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发表时间：
2021
影响因子：
1.7
通讯作者：
Yamamoto Norio
中科院分区：
文献类型：
--
作者：
Kishimoto Ippei;Ohnishi Hiroe;Yamahara Kohei;Nakagawa Takayuki;Yamashita Masaru;Omori Koichi;Yamamoto Norio
快速上皮化对于维持气管通畅和防止因气管浸润或气管狭窄而进行的气管切除术后气管重建中潜在的移植失败至关重要。胰岛素样生长因子1是一种肝脏分泌的内分泌分子，控制细胞增殖、分化和凋亡，并已报道可促进几种器官的上皮形成。在这里，我们利用小鼠气管器官培养物来研究胰岛素样生长因子1对气管epithelialization.MethodsThe气管的影响，从13周龄的雌性ICR小鼠切除，并切成小板状的气管部分。首先，胰岛素样生长因子1受体的表达通过免疫组织化学进行评估。第二，取气管组织切片在培养液中培养7 d，通过免疫组化、苏木精-伊红染色和扫描电镜观察观察气管组织的形态学变化。将气管切片与不同浓度的胰岛素样生长因子1共同培养48 h（0、0.1、1和10 µg/mL），并测量培养过程中气管上皮的延伸长度，以评估局部IGF 1对气管上皮形成的影响。类生长因子1受体表达于气管上皮。免疫组化、苏木精-伊红染色和扫描电镜显示气管器官培养物至少稳定7天，无明显形态学损伤。在添加或不添加胰岛素样生长因子1的培养基中培养48 h的平板状气管切片中检测胰岛素样生长因子1对气管上皮形成的影响。在含胰岛素样生长因子1的培养液中，气管板状切片的上皮边缘沿气管表面沿着更远。结论在小鼠气管器官体外培养模型中，局部应用胰岛素样生长因子1可促进气管上皮的延伸，提示胰岛素样生长因子1在帮助需要使用组织工程气管进行气管重建的患者快速气管上皮形成中的潜在效用。
ObjectiveRapid epithelialization is crucial to maintain tracheal patency and prevent potential graft failure in tracheal reconstruction after tracheal resection for cancer with tracheal infiltration or tracheal stenosis. Insulin-like growth factor 1 is a liver-secreted endocrine molecule that controls cell proliferation, differentiation, and apoptosis and has been reported to promote epithelialization in several organs. Here, we utilized mouse tracheal organ cultures to examine the effect of insulin-like growth factor 1 on tracheal epithelialization.MethodsThe trachea was resected from thirteen-week-old female ICR mice, and cut into small plate-shaped tracheal sections. First, the expression of insulin-like growth factor 1 receptor was assessed by immunohistochemistry. Secondly, the tracheal sections were cultured for seven days in the culture medium, and the morphological change during the seven-day culture was assessed by immunohistochemistry, hematoxylin and eosin staining, and scanning electron microscopy. Moreover, the tracheal sections were cultured for 48 h with different concentration of insulin-like growth factor 1 (0, 0.1, 1 and 10 µg/mL) in the culture medium, and the extension length of the tracheal epithelium during culture was measured in order to assess the effect of topical IGF1 on tracheal epithelialization.ResultsImmunohistochemistry showed that insulin-like growth factor 1 receptor was expressed in tracheal epithelium. Immunohistochemistry, hematoxylin and eosin staining, and scanning electron microscopy showed that the tracheal organ cultures were stable for at least seven days without apparent morphological damage. The effect of insulin-like growth factor 1 on tracheal epithelialization was examined in plate-shaped tracheal sections cultured in medium supplemented with or without insulin-like growth factor 1 for 48 h. We also found that the epithelial edge of plate-shaped tracheal sections extended further along the surface of the tracheal section in culture medium containing insulin-like growth factor 1 compared with that in culture medium without insulin-like growth factor 1.ConclusionThe current study using anin vitromouse tracheal organ culture model demonstrated that topical insulin-like growth factor 1 treatment promoted the extension of tracheal epithelium, suggesting the potential utility of insulin-like growth factor 1 in aiding rapid tracheal epithelialization in patients requiring tracheal reconstruction using tissue-engineered tracheas.