首页|壳聚糖水凝胶/聚丙烯网片复合材料修复大鼠腹壁缺损的实验研究

壳聚糖水凝胶/聚丙烯网片复合材料修复大鼠腹壁缺损的实验研究

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目的 探究复合壳聚糖(chitosan,CS)水凝胶对于传统聚丙烯(polypropylene,PP)网片修复腹壁缺损的改善效果与机制.方法 通过物理交联方式制备CS水凝胶,并将其与PP网片复合,制备CS水凝胶/PP网片复合材料,采用大体观察、正置金相显微镜观察、扫描电镜观察及水接触角检测方法对复合材料内部结构、亲水性等进行表征.通过成纤维细胞(NIH-3T3)浸润实验和人脐静脉内皮细胞(human umbilical vein endothelial cells,HUVECs)管形成实验检测复合材料(实验组)抗细胞黏附和支持细胞长入的性能,以单纯细胞作为对照组.最后取18只8~10周龄SD大鼠建立双侧腹壁缺损模型(1.5 cm×1.0 cm),两侧分别使用复合材料(实验组)和PP网片(对照组)修补缺损,通过大体观察、组织学染色(HE染色和Masson染色)和免疫组织化学染色(CD31、CD68)探究其促进伤口愈合、抗粘连、血管生成和抗炎等性能.结果 复合材料呈浅黄色透明固态,厚度2~3 mm,PP网片牢固包裹在水凝胶中;扫描电镜观察示,水凝胶具有100~300 pm的连通孔隙,形成多孔结构;表面接触角测量结果示,CS水凝胶表现出良好亲水性,而PP网片则高度疏水.在体外细胞培养实验中,DAPI染色示,实验组阳性细胞数量在培养1 d时较少,而对照组细胞覆盖了整个孔板;培养3 d,实验组材料能够减少细胞黏附同时支持细胞长入.HUVECs管形成实验示,实验组细胞数量增加且有形成管的趋势,而对照组细胞分布稀疏,未见迁移.大鼠腹壁缺损修复实验结果示,术后1周,实验组已有组织和血管长入,术后4周完整性恢复良好,肌肉和血管再生明显;而对照组则有粘连且缺损未愈合.HE染色示,实验组细胞浸润较弱,且细胞密度在术后2、4周显著高于对照组(P<0.05).Masson染色示,与对照组相比,实验组胶原纤维排列整齐,术后2周胶原含量显著增高(P<0.05),而术后4周两组胶原含量相近(P>0.05).免疫组织化学染色示,实验组CD31阳性细胞均匀分布于肌肉层间,而对照组缺损明显,术后2周CD31阳性细胞比明显高于对照组(P<0.05);术后2、4周,实验组CD68阳性细胞比显著低于对照组(P<0.05).结论 CS水凝胶对于抗粘连和伤口愈合有积极作用,可以在愈合过程中抗炎和促血管生成,为解决与腹部粘连和重建问题提供了有希望的策略.
Experimental study on repairing rat abdominal wall defect with chitosan hydrogel/polypropylene mesh composite
Objective To investigate the improvement effects and mechanisms of composite chitosan(CS)hydrogel on traditional polypropylene(PP)mesh for repairing abdominal wall defects.Methods CS hydrogel was prepared via physical cross-linking and then combined with PP mesh to create a CS hydrogel/PP mesh composite.The internal structure and hydrophilicity of the composite were characterized using macroscopic observation,upright metallographic microscope,scanning electron microscopy,and water contact angle measurements.The performance of the composite(experimental group)in resisting cell adhesion and supporting cell infiltration was assessed through fibroblast(NIH-3T3)infiltration experiments and human umbilical vein endothelial cells(HUVECs)tube formation assays,and simple cells were used as control group.Finally,a bilateral abdominal wall defect model(1.5 cm× 1.0 cm)was established in 18 Sprague Dawley rats aged 8-10 weeks,with the composite used on one side(experimental group)and PP mesh on the other side(control group).The effects on promoting wound healing,preventing adhesion,angiogenesis,and anti-inflammation were investigated through macroscopic observation,histological staining(HE and Masson staining),and immunohistochemical staining(CD31,CD68).Results The composite appeared as a pale yellow,transparent solid with a thickness of 2-3 mm,with the PP mesh securely encapsulated within the hydrogel.Scanning electron microscopy revealed that the hydrogel contained interconnected pores measuring 100-300 pm,forming a porous structure.Contact angle measurements indicated that CS hydrogel exhibited good hydrophilicity,while PP mesh was highly hydrophobic.In vitro cell culture experiments showed that DAPI staining indicated fewer positive cells in the experimental group after 1 day of culture,while the cells in control group covered the entire well plate.After 3 days of culture,the cells in experimental group were spherical and displayed uneven fluorescence,suggesting that the material could reduce cell adhesion while supporting cell infiltration.HUVECs tube formation experiments demonstrated an increase in cell numbers in experimental group with a trend towards tube formation,while cells in control group were sparsely distributed and showed no migration.In the rat abdominal wall defect repair experiment,results showed that after 1 week post-surgery,the experimental group had tissue and blood vessels infiltrating,and by 4 weeks,the integrity was well restored with significant regeneration of muscle and blood vessels,while the control group exhibited adhesions and incomplete healing.HE staining results indicated weaker cell infiltration in the experimental group,with cell density significantly higher than that of the control group at 2 and 4 weeks post-surgery(P<0.05).Masson staining revealed that collagen fibers in the experimental group were arranged neatly,with significantly increased collagen content at 2 weeks post-surgery(P<0.05),while collagen content was similar in both groups at 4 weeks(P>0.05).Immunohistochemical staining showed that CD31-positive cells were evenly distributed between muscle layers in the experimental group,whereas the control group exhibited notable defects.At 2 weeks after operation,the CD31-positive cell ratio was significantly higher than that in the control group(P<0.05);at 2 and 4 weeks after operation,the CD68-positive cell ratio in the experimental group was significantly lower than that in the control group(P<0.05).Conclusion CS hydrogel has a positive effect on preventing adhesions and promoting wound healing,exhibiting anti-inflammatory and pro-angiogenic properties during the healing process.This provides a promising strategy to address challenges related to abdominal adhesions and reconstruction.

Abdominal wall repairchitosanpolypropylenehydrogelsurgical meshrat

张明浩、贺唯、于素香、邸运涛、李晓明

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保定市第四中心医院普外二科(河北保定 072350)

北京航空航天大学生物与医学工程学院生物力学与力生物学教育部重点实验室北京市生物医学工程高精尖创新中心(北京 100083)

保定市第四中心医院病理科(河北保定 072350)

保定市第四中心医院神经外一科(河北保定 072350)

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腹壁修复 壳聚糖 聚丙烯 水凝胶 手术网片 大鼠

国家自然科学基金资助项目河北省自然科学基金资助项目

32171345C2022104003

2024

中国修复重建外科杂志
中国康复医学会,四川大学华西医院

中国修复重建外科杂志

CSTPCD北大核心
影响因子:1.239
ISSN:1002-1892
年,卷(期):2024.38(10)