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参考体外生物传感器,探讨肝细胞与化学修饰多孔硅的兼容性。

The compatibility of hepatocytes with chemically modified porous silicon with reference to in vitro biosensors.

机构信息

Department of Chemistry and Biochemistry, University of California, San Diego, La Jolla, CA 92093-0358, USA.

出版信息

Biomaterials. 2009 Jan;30(1):26-34. doi: 10.1016/j.biomaterials.2008.09.005. Epub 2008 Oct 8.

Abstract

Porous Si is a nanostructured material that is of interest for molecular and cell-based biosensing, drug delivery, and tissue engineering applications. Surface chemistry is an important factor determining the stability of porous Si in aqueous media, its affinity for various biomolecular species, and its compatibility with tissues. In this study, the attachment and viability of a primary cell type to porous Si samples containing various surface chemistries is reported, and the ability of the porous Si films to retain their optical reflectivity properties relevant to molecular biosensing is assessed. Four chemical species grafted to the porous Si surface are studied: silicon oxide (via ozone oxidation), dodecyl (via hydrosilylation with dodecene), undecanoic acid (via hydrosilylation with undecylenic acid), and oligo(ethylene) glycol (via hydrosilylation with undecylenic acid followed by an oligo(ethylene) glycol coupling reaction). Fourier Transform Infrared (FTIR) spectroscopy and contact angle measurements are used to characterize the surface. Adhesion and short-term viability of primary rat hepatocytes on these surfaces, with and without pre-adsorption of collagen type I, are assessed using vital dyes (calcein-AM and ethidium homodimer I). Cell viability on undecanoic acid-terminated porous Si, oxide-terminated porous Si, and oxide-terminated flat (non-porous) Si are monitored by quantification of albumin production over the course of 8 days. The stability of porous Si thin films after 8 days in cell culture is probed by measuring the optical interferometric reflectance spectra. Results show that hepatocytes adhere better to surfaces coated with collagen, and that chemical modification does not exert a deleterious effect on primary rat hepatocytes. The hydrosilylation chemistry greatly improves the stability of porous Si in contact with cultured primary cells while allowing cell coverage levels comparable to standard culture preparations on tissue culture polystyrene.

摘要

多孔硅是一种纳米结构材料,在分子和基于细胞的生物传感、药物输送和组织工程应用中很有意义。表面化学是决定多孔硅在水介质中稳定性、对各种生物分子种类的亲和力以及与组织相容性的重要因素。在这项研究中,报告了含有各种表面化学物质的多孔硅样品中主要细胞类型的附着和活力,并且评估了多孔硅薄膜保留其与分子生物传感相关的光学反射率特性的能力。研究了四种化学物质接枝到多孔硅表面:氧化硅(通过臭氧氧化)、十二烷基(通过与十二烯的硅氢加成)、十一烷酸(通过与十一烯酸的硅氢加成)和聚(乙二醇)(通过与十一烯酸的硅氢加成,然后进行聚(乙二醇)偶联反应)。傅里叶变换红外(FTIR)光谱和接触角测量用于表征表面。使用活细胞染料(钙黄绿素-AM 和 ethidium homodimer I)评估这些表面上的原代大鼠肝细胞的粘附和短期活力,包括有无预先吸附胶原蛋白 I。在 8 天的过程中,通过白蛋白产量的定量监测,监测终止于十一烷酸的多孔硅、终止于氧化硅的多孔硅和终止于氧化硅的平面(无孔)硅上的细胞活力。通过测量光学干涉反射率谱来探测多孔硅薄膜在细胞培养 8 天后的稳定性。结果表明,涂有胶原蛋白的表面上肝细胞的附着更好,化学修饰对原代大鼠肝细胞没有有害影响。硅氢加成化学大大提高了多孔硅在与培养的原代细胞接触时的稳定性,同时允许细胞覆盖率与组织培养聚苯乙烯上的标准培养制剂相当。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3a18/2784167/f2d17d4699a3/nihms-78322-f0001.jpg

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