Study On The Interaction Between Osteosarcoma-cell And Cisplatin Based On Raman Micro-spectral Imaging

Confocal Raman Microspectroscopy Imaging(CRMI)is a non-destructive,label-free,realtime characterization vibrational spectroscopy technique that can gain insight into tissue and cellular structure and obtain high-resolution data.It has been widely used in biomedical fields such as toxicology,microbiology,disease prediction and diagnosis.In this paper,based on the CRMI technology combined with multivariate analysis method,the interaction between osteosarcoma cells cultured in vitro and the anticancer drug-cisplatin was studied.The specific work was as follows:Firstly,based on the Raman scattering principle,Raman spectroscopy technology and its wide application in biomedicine were introduced.The Confocal Raman imaging system and multivariate data processing methods used in this paper were described and analyzed,and the experimental methods and data analysis methods in the follow-up research were determined.Secondly,the CRMI technique was used to study the interaction of 24-h cultured osteosarcoma cells with cisplatin drug doses(0 μM,20 μM,and 40 μM,respectively).Nucleic acid and protein peak intensities decreased with increasing drug dose,indicating that CRMI technology was able to respond to changes in biochemical content due to cisplatin dose.Principal component analysis(PCA)and linear discriminant analysis(LDA)were used to identify differences in spectral information caused by drug doses,and to establish paraclinical models of drug responses.The above experimental results preliminarily show that Raman imaging technology combined with multivariate analysis method may provide a preliminary experimental basis for dose optimization of patients with osteosarcoma.Finally,the effects of cisplatin dose(0 μM,20 μM and 40 μM)and drug action time(24and 48 hours)on cell biochemical composition were deeply analyzed,and the effect of cisplatin treatment was evaluated.The results of spectral analysis showed that the main changes in biochemical components caused by cisplatin came from nucleic acids and proteins.Through PCA dimensionality reduction,the most significant spectral variables were obtained and used as input variables of Support Vector Machines(SVM)to build a PCA-SVM model to highlight weak spectral information changes.Meanwhile,the obtained spectral results were analyzed chemically based on Multivariate Curve ResolutionAlternating Least Squares(MCR-ALS).The structural changes of subcellular regions induced by cisplatin were visualized by Raman imaging,and information about the content changes of proteins and nucleic acids in subcellular regions was extracted and obtained.The research work in this paper preliminary confirms the reliability of CRMI technology combined with multivariate data analysis method to study the reaction between osteosarcoma and cisplatin and reflects the potential of CRMI technology to study the response mechanism of anticancer drugs.Provide experimental basis and data support for designing new therapeutic strategies based on cisplatin.