Bile Duct Regeneration with an Artificial Bile Duct Made of Gelatin Hydrogel Nonwoven Fabrics.
Bile Duct Regeneration with an Artificial Bile Duct Made of Gelatin Hydrogel Nonwoven Fabrics.
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用明胶水凝胶无纺布制成的人工胆管再生胆管。
DOI：
10.1089/ten.tea.2021.0209
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发表时间：
2022
期刊：
影响因子：
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通讯作者：
Hatano E.
中科院分区：
文献类型：
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作者：
Uemoto Y;Taura K;Nakamura D;Xuefeng L;Nam NH;Kimura Y;Yoshino K;Fuji H;Yoh T;Nishio T;Yamamoto G;Koyama Y;Seo S;Tsuruyama T;Iwaisako K;Uemoto S;Tabata Y;Hatano E.
虽然胆总管空肠吻合术是胆道重建的标准技术，但仍有各种相关问题需要解决，如反流性胆管炎。用人工胆管(ABD)替代切除的胆管，维持了胆汁的生理管道，有望解决这一问题。本研究探讨了明胶水凝胶非织造布(GHNF)制成的ABD的实用性。采用溶液吹塑纺丝法制备了GHNF。在GHNF板上检测小鼠成纤维细胞L929的迁移和活性。L929细胞迁移到GHNF膜中，在那里增殖和合成胶原，提示GHNF是一种有前途的胆管再生支架。将GHNF制成的ABD植入大鼠切除的胆管节段。分别于移植后2、6、12周处死大鼠。对植入部位进行胆管再生的组织学评价。术后2周，ABD小孔内可见移行细胞。植入的ABD在6周时大部分降解，并被胶原纤维取代。移植胆管内可见Ki67阳性的胆管上皮细胞。6周后，腹主动脉中部的神经元数量最多。术后6周，实验组Ki67阳性细胞百分率为31.7 ± 9.1%，假手术组为0.8 ± 0.6%(p<； 0.05)，提示残端成熟的胆管上皮细胞增殖，形成新的胆管上皮。12周时胆管上皮细胞已基本覆盖胆管腔(上皮化率：2周时10.4 ± 6.9%，6周时93.1 ± 5.1%，12周时99.2 ± 1.6%)。再生的胆管上皮细胞角蛋白19阳性。胆管再生伴随着血管生成，CD31阳性血管结构的出现证明了这一点。植入后2周开始诱导毛细血管形成。6周时毛细血管数量最多，12周时降至正常胆管水平。这些结果表明，ABD通过促进细胞外基质合成、血管生成和上皮化所需的细胞迁移，促进了大鼠胆管的成功再生。人工胆管(ABD)的发展使生理性胆管重建成为可能，并可能解决与胆总管空肠吻合相关的临床问题。在本研究中，我们用明胶水凝胶非织造布制作了ABD，并将其植入大鼠切除的胆管中。我们评估了胆管再生和ABD分解的过程，并证明了切除胆管的成功再生，强调了这种新的胆道重建方法取代胆总管空肠吻合术的可能性。
Although choledochojejunostomy is the standard technique for biliary reconstruction, there are various associated problems that need to be solved such as reflux cholangitis. Interposition with an artificial bile duct (ABD) to replace the resected bile duct maintains a physiological conduit for bile and may solve this problem. This study investigated the usefulness of an ABD made of gelatin hydrogel nonwoven fabric (GHNF). GHNF was prepared by the solution blow spinning method. The migration and activity of murine fibroblast L929 cells were examined in GHNF sheets. L929 cells migrated into GHNF sheets, where they proliferated and synthesized collagen, suggesting GHNF is a promising scaffold for bile duct regeneration. ABDs made of GHNF were implanted in place of resected bile duct segments in rats. The rats were killed at 2, 6, and 12 weeks postimplantation. The implantation site was histologically evaluated for bile duct regeneration. At postoperative 2 weeks, migrating cells were observed in the ABD pores. The implanted ABD was mostly degraded and replaced by collagen fibers at 6 weeks. Ki67-positive bile duct epithelial cells appeared within the implanted ABD. These were most abundant within the central part of the ABD after 6 weeks. The percentages of Ki67-positive cells were 31.7 ± 9.1% in the experimental group and 0.8 ± 0.6% in the sham operation group at 6 weeks (p< 0.05), indicating that mature biliary epithelial cells at the stump proliferated to regenerate the biliary epithelium. Biliary epithelial cells had almost completely covered the bile duct lumen at 12 weeks (epithelialization ratios: 10.4 ± 6.9% at 2 weeks, 93.1 ± 5.1% at 6 weeks, 99.2 ± 1.6% at 12 weeks). The regenerated epithelium was positive for the bile duct epithelium marker cytokeratin 19. Bile duct regeneration was accompanied by angiogenesis, as evidenced by the appearance of CD31-positive vascular structures. Capillaries were induced 2 weeks after implantation. The number of capillaries reached a maximum at 6 weeks and decreased to the same level as that of normal bile ducts at 12 weeks. These results showed that an ABD of GHNF contributed to successful bile duct regeneration in rats by facilitating the cell migration required for extracellular matrix synthesis, angiogenesis, and epithelialization.Impact StatementDevelopment of an artificial bile duct (ABD) enables physiological biliary reconstruction and may solve clinical problems associated with choledochojejunostomy. In this study, we created ABDs with gelatin hydrogel nonwoven fabric and implanted them in place of resected bile duct in rats. We evaluated the process of bile duct regeneration as well as decomposition of the ABD and demonstrated successful regeneration of resected bile duct, highlighting the possibility of this novel biliary reconstruction method to replace choledochojejunostomy.