Chinese Journal of Tissue Engineering Research ›› 2018, Vol. 22 ›› Issue (34): 5477-5483.doi: 10.3969/j.issn.2095-4344.0683
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Gu Shaodong1, Zhou Yun2, Xiong Qianwei2, Zhang Ya3, Yang Jinlong4
Received:
2018-07-10
Online:
2018-12-08
Published:
2018-12-08
Contact:
Zhou Yun, Doctoral supervisor, Department of Pediatric Urology, Children Hospital of Soochow University, Suzhou 215025, Jiangsu Province, China
About author:
Gu Shaodong, Master, Associate chief physician, Department of Pediatric Surgery, Lianyungang No. 1 Hospital, Lianyungang 222002, Jiangsu Province, China
Supported by:
the Jiangsu Provincial Clinical Medical Science and Technology Special Project - New Clinical Diagnostic Technology Research Project, No. BL2012051
CLC Number:
Gu Shaodong, Zhou Yun, Xiong Qianwei, Zhang Ya, Yang Jinlong. Bladder acellular matrix combined with vascular endothelial growth factor repairs rabbit urethral defects[J]. Chinese Journal of Tissue Engineering Research, 2018, 22(34): 5477-5483.
2.5 术后不同时间点各组尿道标本苏木精-伊红染色观察 模型组、对照组和实验组中均有尿路上皮层、平滑肌组织和血管的再生,新生尿道组织层次清,黏膜层、浆膜层、肌层排列规律,上皮细胞形态正常,无明显畸形。实验组较模型组、对照组上皮细胞排列更规律,局部血供更丰富,再生肌肉形态更成熟。 术后4周,模型组黏膜上皮厚度较周边薄;黏膜下有较多胶原纤维细胞生长,纤维排列紊乱,其间少量淋巴细胞浸润;对照组和实验组黏膜厚度基本正常,黏膜下胶原基质内有较多淋巴细胞浸润;实验组上皮修复良好,基质表面可见完整的黏膜覆盖,黏膜下层可见未完全降解的胶原基质,基质内有新的胶原纤维细胞生长,可见新生血管长入,对照组基质内未见明显新生血管。术后12周,模型组胶原纤维细胞进一步增多,纤维排列紊乱,有较多淋巴细胞浸润;实验组胶原基质基本降解,新生胶原纤维排列规律,其间血管较多,未见血管瘤等;对照组见团块状胶原沉积,视野内血管较少(图5)。"
2.6 术后不同时间点各组尿道标本Masson染色胶原退化情况观察 术后4周,模型组黏膜下见粗大的胶原纤维,纤维间可见少量血管新生;实验组修补的移植物部分胶原含量较高,其内有较多红细胞分布;对照组胶原纤维粗大,纤维间红细胞数量明显较实验组少。术后12周,模型组胶原纤维较前更粗大,纤维间血管量少,瘢痕倾向明显;对照组黏膜下胶原沉积明显,可见团块状胶原分布,其内有少量红细胞;实验组胶原沉积较少,视野内见密集血管分布(图6)。 用IPP 6.0软件统计进行统计学处理,结果如下:术后4周,模型组、对照组、实验组胶原含量均高于正常组和假手术组(P < 0.05);术后12周,实验组胶原组织沉积明显少于对照组、模型组(P < 0.05),接近正常组和假手术组(P > 0.05),见表2。"
2.7 术后不同时间点各组尿道标本CD31染色新生血管情况观察 术后4周,模型组内纤维间少量血管分布,对照组胶原块周边可见少量血管分布,实验视野下见较多血管分布。术后12周,模型组纤维较4周时间更粗大,排列紧凑,纤维间见少量血管,分布不均匀;对照组胶原团块缩小,血管分布较均匀,数量较少;实验组血管数量较多,分布均匀(图7)。 由镜下观察发现,无论术后4周还是12周,不同组间新生血管的形态学均接近成熟,血管壁完整,血管通透性良好,无明显红细胞外溢,水肿等其他炎性表现。 经CD31染色剂染色后,用200倍视野下,每切片取3个以上视野对镜下的血管数目进行统计,观察统计各组尿道组织中血管数量,统计学分析可看出,同周期实验组明显优于对照组、模型组(P < 0.05),见表3。"
2.8 术后不同时间点各组尿道标本α-SMA免疫组织化学染色观察 α-SMA免疫组织化学染色后,肌肉呈淡黄色。术后4周,各组均存在未分化成熟的肌肉组织,表现为肌肉含量少,肌肉形态不规则,肌成纤维细胞假阳性率高等。术后12周,实验组诱导的再生肌肉分化成熟度明显高于对照组和模型组,表现为肌肉成规则状排列,形态结构完整,成条束状,肌肉组织层次清楚,考虑在VEGF诱导作用下,肌纤维细胞假阳性率降低,向肌肉细胞方向分化加强,肌肉含量较之前增多(图8)。 用IPP 6.0软件统计200倍放大视野内再生平滑肌所占面积百分比,可观察到,实验组再生肌肉含量明显高于对照组和模型组(P < 0.05),见表4。 "
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