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通过具有耗散功能的石英晶体微天平监测不同共混比例的聚己内酯-壳聚糖薄膜上的细胞粘附情况。

Monitoring Cell Adhesion on Polycaprolactone-Chitosan Films with Varying Blend Ratios by Quartz Crystal Microbalance with Dissipation.

作者信息

Özdabak Sert Ayşe Buse, Bittrich Eva, Uhlmann Petra, Kok Fatma Nese, Kılıç Abdulhalim

机构信息

Department of Molecular Biology and Genetics, Istanbul Technical University, 34469 Istanbul, Turkey.

Leibniz-Institut für Polymerforschung Dresden e.V., 01069 Dresden, Germany.

出版信息

ACS Omega. 2023 May 5;8(19):17017-17027. doi: 10.1021/acsomega.3c01055. eCollection 2023 May 16.

Abstract

A detailed understanding of the cell adhesion on polymeric surfaces is required to improve the performance of biomaterials. Quartz crystal microbalance with dissipation (QCM-D) as a surface-sensitive technique has the advantage of label-free and real-time monitoring of the cell-polymer interface, providing distinct signal patterns for cell-polymer interactions. In this study, QCM-D was used to monitor human fetal osteoblastic (hFOB) cell adhesion onto polycaprolactone (PCL) and chitosan (CH) homopolymer films as well as their blend films (75:25 and 25:75). Complementary cell culture assays were performed to verify the findings of QCM-D. The thin polymer films were successfully prepared by spin-coating, and relevant properties, , surface morphology, ζ-potential, wettability, film swelling, and fibrinogen adsorption, were characterized. The adsorbed amount of fibrinogen decreased with an increasing percentage of chitosan in the films, which predominantly showed an inverse correlation with surface hydrophilicity. Similarly, the initial cell sedimentation after 1 h resulted in lesser cell deposition as the chitosan ratio increased in the film. Furthermore, the QCM-D signal patterns, which were measured on the homopolymer and blend films during the first 18 h of cell adhesion, also showed an influence of the different interfacial properties. Cells fully spread on pure PCL films and had elongated morphologies as monitored by fluorescence microscopy and scanning electron microscopy (SEM). Corresponding QCM-D signals showed the highest frequency drop and the highest dissipation. Blend films supported cell adhesion but with lower dissipation values than for the PCL film. This could be the result of a higher rigidity of the cell-blend interface because the cells do not pass to the next stages of spreading after secretion of their extracellular matrix (ECM) proteins. Variations in the QCM-D data, which were obtained at the blend films, could be attributed to differences in the morphology of the films. Pure chitosan films showed limited cell adhesion accompanied by low frequency drop and low dissipation.

摘要

为提高生物材料的性能,需要深入了解细胞在聚合物表面的黏附情况。石英晶体微天平耗散技术(QCM-D)作为一种表面敏感技术,具有无标记和实时监测细胞-聚合物界面的优势,能为细胞-聚合物相互作用提供独特的信号模式。在本研究中,QCM-D用于监测人胎儿成骨细胞(hFOB)在聚己内酯(PCL)和壳聚糖(CH)均聚物膜及其共混膜(75:25和25:75)上的黏附情况。进行了补充细胞培养试验以验证QCM-D的结果。通过旋涂成功制备了聚合物薄膜,并对其相关性能,如表面形态、ζ电位、润湿性、薄膜溶胀和纤维蛋白原吸附进行了表征。随着薄膜中壳聚糖百分比的增加,纤维蛋白原的吸附量减少,这主要与表面亲水性呈负相关。同样,随着薄膜中壳聚糖比例的增加,1小时后初始细胞沉降导致的细胞沉积减少。此外,在细胞黏附的前18小时内在均聚物膜和共混膜上测量的QCM-D信号模式也显示出不同界面性质的影响。通过荧光显微镜和扫描电子显微镜(SEM)监测发现,细胞在纯PCL薄膜上完全铺展并具有伸长的形态。相应的QCM-D信号显示出最高的频率下降和最高的耗散。共混膜支持细胞黏附,但耗散值低于PCL薄膜。这可能是由于细胞-共混物界面具有更高刚性,因为细胞在分泌细胞外基质(ECM)蛋白后不会进入铺展的下一阶段。在共混膜上获得的QCM-D数据变化可归因于薄膜形态的差异。纯壳聚糖薄膜显示出有限的细胞黏附,伴随着低频下降和低耗散。

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