Graduate School of Engineering, Kobe University, 1-1 Rokkodai-cho, Nada-ku, Kobe 657-8501, Japan.
Graduate School of Engineering, Kobe University, 1-1 Rokkodai-cho, Nada-ku, Kobe 657-8501, Japan.
Acta Biomater. 2016 Jul 1;38:163-7. doi: 10.1016/j.actbio.2016.04.022. Epub 2016 Apr 20.
Enzymatic degradation of poly(ε-caprolactone) (PCL) thin films was analyzed by reflectometric interference spectroscopy (RIfS)-based sensing system, and validated by attenuated total reflection infrared spectroscopy (ATR-IR) imaging. The degradation of the PCL thin film spin-coated on the silicon substrate on which 65-nm silicon nitride layer was deposited as an interference layer was easily monitored by shifting the peak bottom of reflectance spectra (Δλ) that is known to be proportional to the thickness of thin films. The Δλ values decreased with increasing the concentration of lipase from Pseudomonas cepacia, and the obtained sensorgrams were applied for kinetic analysis using a curve fitting software. ATR-IR spectra and imaging analysis on the surface of the PCL film revealed that carbonyl groups on the surface decreased with time, resulting from proceeding with the enzymatic hydrolysis, and importantly, extinction of the carbonyl group was declined with proportional to the decrease in the film thickness measured by the RIfS system. Consequently, the present RIfS-based label-free monitoring system can provide a simple and reliable way for evaluating biodegradability on synthetic materials.
A RIfS-based sensing system in combination with ATR-IR measurements can be an analytical method for evaluation of biodegradability of polymeric thin films. This study demonstrates the utility of the RIfS-based sensing approach for analyzing the lipase-catalyzed degradation of PCL. Despite the RIfS is known as an inexpensive label-free detection method for biological interaction, the RIfS applications as monitoring methods for enzymatic degradation of biodegradable polymers had not been systematically explored. This study additionally demonstrated the capability of combined analysis of the biodegradation with ATR-IR spectra/imaging and RIfS measurements, which could be broadly applied towards evaluating biodegradability of various biodegradable polymers in environmental protection research.
通过基于反射干涉光谱(RIfS)的传感系统分析了聚(ε-己内酯)(PCL)薄膜的酶促降解,并通过衰减全反射红外光谱(ATR-IR)成像进行了验证。PCL 薄膜在沉积有 65nm 氮化硅层的硅衬底上旋涂,作为干涉层,其薄膜的降解很容易通过反射光谱的峰底(Δλ)的移动来监测,已知该峰底与薄膜的厚度成正比。随着假单胞菌脂肪酶浓度的增加,Δλ 值减小,并且使用曲线拟合软件对获得的传感器图进行了动力学分析。ATR-IR 光谱和表面的成像分析PCL 薄膜表明,表面上的羰基基团随着时间的推移而减少,这是由于酶促水解的进行,重要的是,羰基基团的消光与通过 RIfS 系统测量的薄膜厚度的减少成正比。因此,本研究中基于 RIfS 的无标记监测系统为评估合成材料的生物降解性提供了一种简单可靠的方法。
基于 RIfS 的传感系统与 ATR-IR 测量相结合,可以作为评估聚合物薄膜生物降解性的分析方法。本研究证明了基于 RIfS 的传感方法在分析脂肪酶催化的 PCL 降解中的应用。尽管 RIfS 是一种用于生物相互作用的廉价无标记检测方法,但 RIfS 在可生物降解聚合物的酶促降解监测方法中的应用尚未得到系统的探索。本研究还展示了与 ATR-IR 光谱/成像和 RIfS 测量相结合的生物降解分析的能力,这可以广泛应用于评估环境保护研究中各种可生物降解聚合物的生物降解性。