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Nano-Silicon Dioxide particles (100 μg/ml) were incubated in 5 ml culture medium supplemented with 1.25% serum for 4 h at 37°C. The suspension was then centrifuged at 5700 g for 10 min and the pellet re-suspended in 1 ml PBS, transferred to an Eppendorf microcentrifuge tube (ca 1.5 ml) and centrifuged at 10,000×g for 5 min. The supernatant was removed and the pellet re-suspended again in 1 ml PBS and centrifuged as before. This washing process was repeated a further 3-times. Finally, the pellet was re-suspended in 0.1 ml 9 M urea/2% CHAPS by vortex mixing for 1 min before centrifugation as before to remove any insoluble material. The supernatant was collected, the protein content was determined using the BCA method and 5 μg of protein was loaded onto an SDS-PAGE gel in preparation for proteomics as described above. The data derived from LC-MS/MS analysis of each gel slice was analysed using SEQUEST by searching a database comprising Bos taurus and Homo sapiens proteins from Refseq release 56. In addition, entire procedure was repeated in the presence of A549 cells by applying the same nano-SiO2–containing suspension to a T25 flask of confluent A549 cells. After 4 h the medium was removed and processed as described above.

Raw abundance intensities for each protein were assessed for possible differences in levels between samples from control and nano-Silicon Dioxide treated cells. Software was developed to aid the identification of trends in the Progenesis output. This software combined all the results for one gel divided into a series of slices (analysed by LC-MS/MS separately) into a single table of identified proteins. The program combined and used data from adjacent slices to aid the process. Protein isoforms were automatically combined, and proteins with shared peptides grouped together. Finally, the program sorted the protein groups by size and showed each group with a summary of the detected abundance of each peptide.

Ethanol as a dispersion medium of nano Silicon Dioxide surface modified with a coupling agent KH-570, its modification effect were characterized by transmission electron microscopy and photoelectron spectroscopy, the researchers found that nano-Silicon Dioxide nanometer in ethanol dispersion in nano level, and coupling agents and nano silica surface have chemical reactions.


 

 

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