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表面改性的聚己内酯电纺支架,提高了光学透明度和生物活性,用于受损眼表的重建。

Surface-modified electrospun poly(epsilon-caprolactone) scaffold with improved optical transparency and bioactivity for damaged ocular surface reconstruction.

机构信息

Department of Ophthalmology, Dr. Rajendra Prasad Centre for Ophthalmic Sciences, All India Institute of Medical Sciences, New Delhi, India.

出版信息

Invest Ophthalmol Vis Sci. 2014 Feb 12;55(2):899-907. doi: 10.1167/iovs.13-12727.

Abstract

PURPOSE

The purpose of this study was to modify and functionalize the surface of synthetic poly-ε-caprolactone (PCL) nanofibrous scaffolds to improve their biocompatibility in order to provide better "cell-substrate" interaction.

METHODS

Poly-ε-caprolactone solution was electrospun and its surface functionality was modified by helium-oxygen (He/O2) plasma discharge. Scaffolds were characterized for their morphology, wetting ability, mechanical strength, and optical properties by using scanning electron microscopy (SEM), water contact angle measurement, tensile strength, and ultraviolet-visible (UV-Vis) spectrophotometer, respectively. The biocompatibility of nanofibers was explored by culturing human corneal epithelial (HCE-T) cell line. Subsequently, human limbal epithelial cells (LECs) were cultured to evaluate the bioactivity. Cell proliferation was checked by MTT (3-(4, 5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay. Immunofluorescent staining and reverse transcription-polymerase chain reaction were done to check the gene expression; SEM was used to study the morphology.

RESULTS

Plasma-treated and untreated scaffolds showed almost similar morphology and tensile strength. Water contact angle measurement and optical transparency data showed that the plasma-treated PCL (pPCL) exhibited significantly improved wettability and transparency as compared to the untreated PCL scaffolds. Biocompatibility results indicated that both scaffolds are biocompatible in terms of cell survival and proliferation. However, pPCL showed better cell adhesion and proliferation. Results supported that LEC cultured on pPCL scaffolds had enhanced cell adhesion and proliferation, in comparison to untreated PCL. Gene expression study showed cultures were able to retain their normal phenotype on both scaffolds.

CONCLUSIONS

The hydrophilicity of the surface achieved by plasma treatment effectively enhanced the transparency and promoted the biocompatibility of scaffolds. These nanofibers may act as biological cues for endorsing ocular surface engineering.

摘要

目的

本研究旨在对合成聚己内酯(PCL)纳米纤维支架的表面进行修饰和功能化,以提高其生物相容性,从而提供更好的“细胞-基底”相互作用。

方法

通过氦氧(He/O2)等离子体放电对聚己内酯溶液进行电纺,并对其表面功能进行修饰。通过扫描电子显微镜(SEM)、水接触角测量、拉伸强度和紫外可见分光光度计(UV-Vis)分别对支架的形貌、润湿性、机械强度和光学性能进行了表征。通过培养人角膜上皮(HCE-T)细胞系来探索纳米纤维的生物相容性。随后,培养人角膜缘上皮细胞(LEC)以评估其生物活性。通过 MTT(3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四氮唑溴盐)检测法检查细胞增殖。通过免疫荧光染色和逆转录-聚合酶链反应(RT-PCR)检查基因表达;通过扫描电子显微镜(SEM)观察细胞形态。

结果

未经处理和经等离子体处理的支架的形态和拉伸强度几乎相同。水接触角测量和光学透明度数据表明,与未经处理的 PCL 支架相比,等离子体处理的 PCL(pPCL)表现出显著改善的润湿性和透明度。生物相容性结果表明,两种支架在细胞存活和增殖方面均具有生物相容性。然而,pPCL 显示出更好的细胞黏附和增殖。结果表明,与未经处理的 PCL 支架相比,在 pPCL 支架上培养的 LEC 具有增强的细胞黏附和增殖能力。基因表达研究表明,在两种支架上培养的细胞均能够保持其正常表型。

结论

等离子体处理所达到的表面亲水性有效提高了透明度,并促进了支架的生物相容性。这些纳米纤维可能作为促进眼表工程的生物线索。

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