Kulkarni Gaurav, Guha Ray Preetam, Das Shreyasi, Biswas Souvik, Dhara Santanu, Das Soumen
School of Medical Science & Technology, IIT Kharagpur, West Bengal 721302, India.
School of Nano Science & Technology, IIT Kharagpur, West Bengal 721302, India.
Spectrochim Acta A Mol Biomol Spectrosc. 2021 Aug 5;257:119760. doi: 10.1016/j.saa.2021.119760. Epub 2021 Apr 1.
Biochemical evaluation of cell-matrix interaction using conventional labelling techniques often possesses limitations due to dye entrapment. In contrast, Raman spectroscopy guided approach offers label-free determination of cell-matrix biochemistry. Herein, gelatin (Gel) matrices modified with 1-Ethyl-3-(3-dimethylaminopropyl) carbodiimide/ N-Hydroxysuccinimide (EDC/NHS) and glutaraldehyde (GTA) was used as standards for comparative evaluation. Raman spectroscopy was deployed as a label-free approach to investigate interaction of cells with Gel hydrogels. Raman-based approach assisted in evaluation of cell-matrix interactions by identifying key biomolecular signatures retrospecting the fact that L929 fibroblast cells portrayed excellent growth and proliferation kinetics in crosslinked Gel as compared to its bare counterpart. EDC crosslinked hydrogels exhibited superior cell proliferation than its GTA counterparts. Cell proliferation on differentially crosslinked gel was also confirmed using standard MTT Assay and Rhodamine-DAPI staining thus corroborating the fact that Raman spectroscopy can be deployed as a superior label-free alternative towards real-time determination of cell proliferation and growth.
使用传统标记技术对细胞 - 基质相互作用进行生化评估时，由于染料截留，常常存在局限性。相比之下，拉曼光谱引导的方法提供了无标记的细胞 - 基质生物化学测定。在此，用1-乙基-3-(3-二甲基氨基丙基)碳二亚胺/ N-羟基琥珀酰亚胺（EDC/NHS）和戊二醛（GTA）修饰的明胶（Gel）基质用作比较评估的标准。拉曼光谱作为一种无标记方法，用于研究细胞与Gel水凝胶的相互作用。基于拉曼的方法通过识别关键生物分子特征，辅助评估细胞 - 基质相互作用，回顾这一事实，即与未交联的Gel相比，L929成纤维细胞在交联的Gel中表现出优异的生长和增殖动力学。EDC交联的水凝胶比GTA交联的水凝胶表现出更好的细胞增殖。使用标准MTT法和罗丹明 - DAPI染色也证实了细胞在不同交联凝胶上的增殖，从而证实了拉曼光谱可作为一种优越的无标记替代方法用于实时测定细胞增殖和生长。
