不同质量比SF/PLLA复合纤维支架上共培养的上皮细胞和成纤维细胞增殖和表型的变化

doi:10.13481/j.1671-587x.20150420

吉林大学学报(医学版) ›› 2015, Vol. 41 ›› Issue (04): 774-778.doi: 10.13481/j.1671-587x.20150420

不同质量比SF/PLLA复合纤维支架上共培养的上皮细胞和成纤维细胞增殖和表型的变化

张笑^1,2, 王苹², 于姝媛², 王洪艳³, 汤勇⁴, 周余来⁵, 祝威²

1. 吉林大学中日联谊医院神经内科, 吉林长春 130033;
2. 吉林大学第一医院耳鼻咽喉-头颈外科, 吉林长春 130021;
3. 吉林大学化学学院化工与应用化学系, 吉林长春 130012;
4. 吉林省人民医院耳鼻咽喉科, 吉林长春 130021;
5. 吉林大学药学院生物工程实验中心, 吉林长春 130021

收稿日期:2014-11-18 发布日期:2015-08-01
通讯作者: 祝威,教授,博士研究生导师(Tel:0431-81875506,E-mail:zhuwei30000@163.com);周余来,教授,硕士研究生导师(Tel:0431-85619299,E-mail:zhouyl@jlu.edu.cn) E-mail:zhuwei30000@163.comzhouyl@jlu.edu.cn
作者简介:张笑(1987-),女,吉林省长春市人,医学硕士,主要从事生物医学工程方面的研究。
基金资助:
吉林省科技厅自然科学基金资助课题(201115052,201015223)

Changes of proliferation and phenotypes of epithelial cells and fibroblasts co-cultured on SF/PLLA composite fibrous scaffolds

ZHANG Xiao^1,2, WANG Ping², YU Shuyuan², WANG Hongyan³, TANG Yong⁴, ZHOU Yulai⁵, ZHU Wei²

1. Departmwnt of Neurology, China-Japan Union Hospital, Jilin University, Changchun 130033, China;
2. Department of Otorhinolaryngology and Head-Neck Surgery, First Hospital, Jilin University, Changchun 130021, China;
3. Department of Chemical Engineering and Applied Chemistry, School of Chemistry, Jilin University, Changchun 130012, China;
4. Department of Otorhinolaryngology, People's Hospital of Jilin Province, Changchun 130021, China;
5. Bioengineering Experimental Center, School of Pharmacy, Jilin University, Changchun 130021, China

Received:2014-11-18 Published:2015-08-01

摘要/Abstract

摘要：

目的: 观察不同质量配比的丝素蛋白(SF)和聚乳酸(PLLA)构建的SF/PLLA复合纤维支架对上皮细胞和成纤维细胞增殖和表型的影响,阐明SF/PLLA生物工程膜的细胞相容性。方法: 使用SF和PLLA作为支架材料,以SF与PLLA不同质量比(S50/P50、S40/P60和S30/P70)进行混合纺丝,制备出不同比例的SF/PLLA复合纤维支架,采用SF/PLLA复合纤维支架与皮肤组织中分离的原代上皮细胞和成纤维细胞进行共培养。采用CCK8方法检测细胞在不同质量比的支架材料上的增殖活性,免疫荧光染色观察共培养细胞的特异标记蛋白表达。结果: 共培养第5和6天时,在S50/P50组复合纤维支架上的成纤维细胞增殖活性高于S40/P60组和S30/P70组(P<0.01),在S40/P60组复合纤维支架上的上皮细胞增殖活性明显高于S50/P50和S30/P70组(P<0.05或P<0.01)。接种于SF/PLLA复合纤维支架的上皮细胞表达CK19蛋白,接种的成纤维细胞表达Vimentin 蛋白。2种细胞与SF/PLLA复合纤维支架共培养,S40/P60和S50/P50组复合纤维支架上的细胞生长状态优于S30/P70组。结论: 支架中SF的浓度增加可以明显提高细胞增殖活性,促进细胞存活,SF/PLLA复合纤维支架对细胞的生物表型无明显影响。

关键词: 上皮细胞, 成纤维细胞, 丝素蛋白, 聚乳酸, 复合支架

Abstract:

Objective To observe the influence of SF/PLLA composite fibrous scaffolds with different quality ratios of SF and PLLA in the proliferation and phenotypes of epithelial cells and fibroblasts, and to clarify the cytocompatibility of SF/PLLA bioengineering membrane. Methods The SF and PLLA were served as materials to construct composite scaffolds.The different proportions of SF/PLLA nanofibers membrane were compounded with different ratios of SF and PLLA(S50/P50,S40/P60 and S30/P70) and mixed spinning.The primary epithelial cells and fibroblasts from rat skin tissue were cultured on the SF/PLLA composite scaffolds.CCK8 test was used to evaluate the cell proliferation,and immunofluorescence staining was used to determine the expressions of the specific marker proteins of cultured cells. Results The proliferation activities of fibroblasts in S50/P50 group were significantly higher than those in S40/P60 and S30/P70 groups,and the proliferation activities of epithelial cells on the composite scaffolds in S40/P60 group were higher than those in S50/P50 and S30/P70 groups (P<0.01) on the 5th and 6th days.The epithelial cells cultured on the SF/PLLA composite scaffolds expressed CK 19 protein and the fibroblasts expressed Vimentin protein.When two kinds of cells were co-cultured on SF/PLLA composite fibrous scaffolds,the cell growth status in S40/P60 and S50/P50 groups were superior to that in S30/P70 group. Conclusion The increasing of SF concentration in composite scaffolds can obviously increase the cell proliferation activity and promote the cell survival;SF/PLLA scaffolds has no significant influence in the cell phenotype.

Key words: epithelial cells, fibroblasts, silk fibroin, polylactic acid, composite scaffold

中图分类号:

张笑, 王苹, 于姝媛, 王洪艳, 汤勇, 周余来, 祝威. 不同质量比SF/PLLA复合纤维支架上共培养的上皮细胞和成纤维细胞增殖和表型的变化[J]. 吉林大学学报(医学版), 2015, 41(04): 774-778.

ZHANG Xiao, WANG Ping, YU Shuyuan, WANG Hongyan, TANG Yong, ZHOU Yulai, ZHU Wei. Changes of proliferation and phenotypes of epithelial cells and fibroblasts co-cultured on SF/PLLA composite fibrous scaffolds[J]. Journal of Jilin University Medicine Edition, 2015, 41(04): 774-778.

参考文献

[1] Tang X,Xue C,Wang Y,et al. Bridging peripheral nerve defects with a tissue engineered nerve graft composed of an in vitro cultured nerve equivalent and a silk fibroin-based scaffold[J].Biomaterials, 2012,33(15):3860-3867.

[2] Shalumon KT,Chennazhi KP,Tamura H,et al.Fabrication of three-dimensional nano,micro and micro/nano scaffolds of porous poly(lactic acid) by electrospinning and comparison of cell infiltration by Z-stacking/three-dimensional projection technique[J].IET Nanobiotechnol,2012,6(1):16-25.

[3] 邢帅,夏亚一,袁凌伟,等,聚乳酸/丝素蛋白复合组织工程支架的制备及生物学性能的评价[J].吉林大学学报:医学版,2008,34(3):405-410.

[4] 孙皎,宁丽,顾国珍,等,医用丝素蛋白皮肤再生膜的细胞相容性评价[J].生物医学工程学杂志,2000,17(4):393-395.

[5] 张晓燕,李正强,孙莹,等丝素蛋白/左旋聚乳酸复合组织工程纳米材料的生物相容性及安全性评价[J].吉林大学学报:医学版,2014,40(3):578-582.

[6] Stoppato M,Stevens HY,Carletti E,et al.Effects of silk fibrion fiber incorporation on mechanical propenties,endothelial cell colonization and vascularization of PDLLA scaffolds[J].Biomaterials,2013,34(19):4573-4581.

[7] Li SL,Liu Y,Hui L.Construction of engineering adipose-like tissue in vivo utilizing human insulin gene-modified umbilical cord mesenchymal stromal cells with silk fibroin 3D scaffolds[J].J Tissue Eng Regen Med,2013,doi:10.1002/term.1695.

[8] Cui M,Liu L,Guo N,et al.Preparation,cell compatibility and degradability of collagen-modified Poly(lactic acid)[J].Molecules,2015,20(1):595-607.

[9] Tawakkal IS,Cran MJ,Miltz J,et al.A review of poly(lactic acid)-based materials for antimicrobial packaging[J].J Food Sci,2014,79(8):R1477-1490.

[10] Zeng C,Yang Q,Zhu M,et al.Silk fibroin porous scaffolds for nucleus pulposus tissue engineering[J].Mater Sci Eng C Mater Biol Appl, 2014,37:232-240.dio:10.1016/j.msec.2014.01.012.

[11] 张丽斯,强琳辉,颜静,等.静电纺丝法制备丝素蛋白/聚乳酸纤维膜及其性能表征[C].郑州:中国化学会第十二届全国应用化学年会论文集,2011.

[12] Hakki SS,Korkusuz P,Purali N,et al.Attachment,proliferation and collagen type I mRNA expression of human gingival fibroblasts on different biodegradable membranes[J].Connect Tissue Res,2013,54(4/5):260-266.

[13] DalPrà I,Petrini P,Chiarini A,et al.Silk fibroin-coated three-dimensional polyurethane scaffolds for tissue engineering:interactions with normalhuman fibroblasts[J].Tissue Eng,2003,9(6):1113-1121.

[1]	李里, 宫亮, 吴玉斌. 沉默WNT4基因对稳定转染PAX2基因大鼠肾小管上皮细胞迁移和侵袭能力的影响[J]. 吉林大学学报(医学版), 2018, 44(06): 1212-1217.
[2]	楚爱景, 程旭锋, 赵慧朵, 王丰莲. 柴芍乳癖颗粒对乳腺增生模型大鼠血清激素水平和乳腺组织中VEGF和bFGF蛋白表达水平的影响及其意义[J]. 吉林大学学报(医学版), 2018, 44(05): 1005-1009.
[3]	张啸环, 段明华, 闫玉礼, 潘新, 李海清, 周长龙, 赵天倚. 人羊膜成纤维细胞的分离和鉴定[J]. 吉林大学学报(医学版), 2018, 44(04): 845-848.
[4]	阎慧, 娄石磊, 苗永迪, 岂蕊, 郭子硕, 邱悦, 孙聪. 大鼠结肠原代成纤维细胞的分离、培养和鉴定[J]. 吉林大学学报(医学版), 2018, 44(04): 849-852.
[5]	王孝芸, 蓝楠, 唐红梅, 袁谢芳, 王星, 李国平. Lyn对屋尘螨诱导的人支气管上皮细胞中MUC5AC表达的影响及其机制[J]. 吉林大学学报(医学版), 2018, 44(02): 211-215.
[6]	宋效庆, 刘红, 陈天杰, 刘称称, 路政宽, 秦爽, 黄山. 淫羊藿苷/载明胶纳米复合物-PLGA缓释系统的制备及工艺优化[J]. 吉林大学学报(医学版), 2018, 44(02): 438-443.
[7]	王丽萍, 王建辉, 李树民, 杨秀红. N-乙酰半胱氨酸通路对AGT-REN双转基因高血压小鼠心肌成纤维细胞中电导钙激活钾离子通道蛋白表达的影响及其意义[J]. 吉林大学学报(医学版), 2017, 43(05): 881-886.
[8]	赛依帕, 宋元春, 刘璟, 赵海燕. 淫羊藿苷联合胚胎干细胞对造影剂肾病大鼠模型肾上皮细胞凋亡的影响[J]. 吉林大学学报(医学版), 2017, 43(05): 953-957.
[9]	陈楠楠, 马宁, 车鸿泽, 焦鹏, 陈飞, 张莉. 复方总黄酮聚乳酸-羟基乙酸共聚物微囊的制备及体外释药特性[J]. 吉林大学学报(医学版), 2017, 43(03): 593-599.
[10]	车鸿泽, 车彦海, 卢晴, 陈楠楠, 陈飞, 金权, 马宁. PLGA涂层处理的nHA-Mg多孔复合材料对兔颌骨缺损的修复作用[J]. 吉林大学学报(医学版), 2017, 43(02): 276-280.
[11]	刘微, 姚杨, 马萧萧, 邓裕宣, 梅迪, 刘磊, 王会岩. 融合表达载体pET22b-SUMO-FGFR4的构建及其在大肠杆菌中表达条件的优化[J]. 吉林大学学报(医学版), 2016, 42(04): 642-647.
[12]	李凤娥, 向国艳, 孔繁利, 郝峰, 杨紫嫣, 郑岩, 方芳. pEGFP-N1脂质体转染方法的优化及其应用[J]. 吉林大学学报(医学版), 2016, 42(03): 612-616.
[13]	卢晴, 车鸿泽, 陈楠楠, 陈飞, 焦鹏, 马宁. 野菊花总黄酮-PLGA缓释微球的制备及其工艺优化[J]. 吉林大学学报(医学版), 2016, 42(03): 617-621.
[14]	齐慧慧, 陈冰, 陈晓娜, 王瑶琪, 佟青玥, 孙丽光, 杨永广. 几种成年小鼠胸腺上皮细胞分离和富集方法效果的比较[J]. 吉林大学学报(医学版), 2016, 42(01): 24-30.
[15]	陈为, 张葵, 张宸豪, 李强, 李妍. OX-LDL介导内皮细胞损伤及其对TLR-4和EpCAM表达的影响[J]. 吉林大学学报(医学版), 2015, 41(06): 1144-1149.

不同质量比SF/PLLA复合纤维支架上共培养的上皮细胞和成纤维细胞增殖和表型的变化

Changes of proliferation and phenotypes of epithelial cells and fibroblasts co-cultured on SF/PLLA composite fibrous scaffolds

RichHTML

PDF (PC)

赞

可视化

摘要/Abstract

引用本文

使用本文

参考文献

相关文章 15

Metrics

本文评价

推荐阅读 0