Raman and fluorescence micro-spectroscopy applied for the monitoring of sunitinib-loaded porous silicon nanocontainers in cardiac cells

Abstract

Nanomaterials are a central pillar in modern medicine. They are thought to optimize drug delivery, enhance therapeutic efficacy, and reduce side-effects. To foster this technology, analytical methods are needed to validate not only the localization and distribution of these nanomaterials, but also their compatibility with cells, drugs, and drug release. In the present work, we assessed nanoparticles based on porous silicon (pSiNPs) loaded with the clinically used tyrosine kinase inhibitor sunitinib for their effectiveness of drug delivery, release, and toxicity in colon cancer cells (HCT 116 cells) and cardiac myoblast cells (H9c2) using Raman micro-spectroscopy, high-resolution fluorescence microscopy, along with biological methods for toxicological effects. We produced pSiNPs with a size of about 100 nm by grinding mesoporous silicon layers. pSiNPs allowed an effective loading of sunitinib due to their high porosity. Photoluminescence properties of the nanoparticles within the visible spectrum allowed the visualization of their uptake in cardiac cells. Raman micro-spectroscopy allowed not only the detection of the uptake and distribution of pSiNPs within the cells via a characteristic silicon Raman band at about 518–520 cm−1, but also the localization of the drug based on its characteristic molecular fingerprints. Cytotoxicity studies by Western blot analyses of apoptotic marker proteins such as caspase-3, and the detection of apoptosis by subG1-positive cell fractions in HCT 116 and MTT analyses in H9c2 cells, suggest a sustained release of sunitinib from pSiNPs and delayed cytotoxicity of sunitinib in HCT 116 cells. The analyses in cardiac cells revealed that pSiNPs are well tolerated and that they may even protect from toxic effects in these cells to some extent. Analyses of the integrity of mitochondrial networks as an early indicator for apoptotic cellular effects seem to validate these observations. Our study suggests pSiNPs-based nanocontainers for efficient and safe drug delivery and Raman micro-spectroscopy as a reliable method for their detection and monitoring. Thus, the herein presented nanocontainers and analytical methods have the potential to allow an efficient advancement of nanoparticles for targeted and sustained intracellular drug release that is of need, e.g., in chronic diseases and for the prevention of cardiac toxicity.

Raman Imaging—A Valuable Tool for Tracking Fatty Acid Metabolism—Normal and Cancer Human Colon Single-Cell Study

“…The usefulness of RI for single-cell analysis at the level of individual organelles has been proven in our previous papers [66][67][68][69]. Moreover, fluorescence staining confirmed the effectiveness of RI for individual structure visualization and correctness of Raman data interpretation [70][71][72][73][74][75]. Scheme S1 in the Supplementary Materials shows the simplified illustration of the RI experiment idea and the comparison of RI and fluorescence data for one cell chosen from our database.…”

Identification of mitophagy-related hub genes during the progression of spinal cord injury by integrated multinomial bioinformatics analysis

“…Second, we found that there are 11 and 8 TFs regulating the hub genes SKP1 and BAP1, respectively, using the NetworkAnalyst database. Finally, based on the DGIdb database, we found that histone deacetylase inhibitors, such as vorinostat [ 62 ], panobinostat [ 63 , 64 ], mTOR pathway inhibitor everolimus [ 65 ], apitolisib [ 66 , 67 ], tyrosine kinase inhibitor sunitinib [ 68 ] and poly (ADP-ribose) polymerase inhibitor olaparib [ 69 ] may target the BAP1 gene and promote neural recovery after SCI.…”

Efficacy of tyrosine kinase inhibitors examined by a combination of Raman micro-spectroscopy and a deep wavelet scattering-based multivariate analysis framework

“…For instance, the distribution of tyrosine kinase inhibitors such as sorafenib and sunitinib was monitored in cells using Raman bands located at 1028 cm −1 and 1320/1570 cm −1 , respectively. 31,75 In SRS imaging study, Fu et al 40 used also the Raman bands of nilotinib, chloroquine, and GNF and GNF at 1300 cm, 1370, and 1600 cm −1 to display their distribution in the cells. Besides, the distribution of doxorubicin and squalene-doxorubicin in cancer cells was detected through several Raman bands in the fingerprint region.…”