改性PET材料编织物引导小鼠BMSC迁移模型的建立

The Cell Migration Model for Modified Knitted PET-guided Mouse BMSC

目的:壳聚糖接枝改性的聚对苯二甲酸乙二醇酯(PET)纤维编织材料是一种具有良好组织相容性的新型生物材料,小鼠骨髓间充质干细胞(BMSC)为主要的人工韧带种子细胞。本研究通过建立一种可应用于生物材料上的细胞迁移模型,旨在观察BMSC在改性PET材料编织物上的迁移能力,探讨该材料作为人工韧带应用的可行性。方法:利用316L医用不锈钢自主设计制作细胞迁移模具,置于六孔板,在模具加样孔内接种小鼠BMSC,培养24 h后撤出模具,分别继续培养48 h、72 h和96 h;在证实模具可有效用于量化细胞迁移过程后,将不同线密度的PET材料编织物平铺于六孔板底,利用本模型观察BMSC在3000D组和5000D组经改性的PET材料编织物上的迁移能力,吉姆萨染色法计算不同时间点BMSC的迁移面积。结果:细胞接种24 h后撤出模具,可见六孔板底中央出现与模具底面大小相当的无细胞空白区,周围接种BMSC,随培养时间延长,BMSC逐渐由周围向中央区域迁移;在经改性的PET材料编织物上成功建立细胞迁移模型后,随培养时间增加,两组细胞迁移面积均逐渐增大,其中3000D组细胞迁移面积显著大于5000D组(P<0.05)。结论:利用316L医用不锈钢模具建立的细胞迁移模型能够应用于平铺编织物引导细胞迁移的基础研究,本模型可作为传统模型无法观察人工材料对细胞迁移可能影响的良好补充;3000D经改性的PET材料编织物的线密度较5000D更有利于小鼠BMSC在材料上的迁移和生长。

Objective： Chitosan grafted Polyethylene terephthalate （PET） woven fabrics are new biomaterials with good histocom- patibility. Mouse bone marrow mesenchymal stem cells （BMSC） are the major source of seed cells. This study aimed to establish a cell migration model applied to biological material, and to investigate the migration ability of BMSC guided by knitted PET on the model. Methods： 316L medical stainless steel was designed to produce the cell migration model. The model was placed into 6-well cell culture plates, mouse BMSC was seeded through the sample hole of model. After cultured for 24 h, the model was withdrawaled and continued to culture for 48 h, 72 h and 96 h; when the model was confmned that it can be effectively used to quantify the cell migration process, the model then was utilized to observe the migration ability of BMSC on group 3000D and 5000D modified knitted PET. The PET was stained by giemsa and the migration area was calculated. Results： The model was removed after the cells cultured for 24 h, the central area of 6-well plate was blank which is equal to the bottom of the model, BMSC were cultured around the blank area. The migration ar- eas were gradually increased with time between groups, area of 3000D group was significantly larger than 5000D （P〈0.05）. Conclusion： 316L medical stainless steel model could be applied to basic research for fiat contexture -guided cell migration, our model might be a beneficial supplement for traditional model which can not observe the artificial material-guided cell migration; the 3000D-line density of the modified knitted PET has an effect on migration ability of mouse BMSC.