Herein, a microfluidic device (MD) containing immobilized trypsin for rapid and efficient proteolysis was described. Trypsin was immobilized via non-specific protein adsorption onto the hydrophobic poly(dimethylsiloxane) (PDMS) channel wall of the MD. Peptide mapping of bovine serum albumin (BSA) samples was carried out to estimate the stability of trypsin adsorbed on PDMS surface. Peptide maps of BSA samples were obtained by capillary zone electrophoresis (CZE), the RSD% for migration times were under 1%. Several proteins (hemoglobin, myoglobin, lysozyme, and BSA) in a wide molecular size range (15-70 kDa) were digested efficiently with ∼50 s contact time. The number of separated peaks correlated well with the expected number of peptides formed in the complete tryptic digestion of the proteins. Peptide mass fingerprinting of BSA and human serum was carried out. Trypsin retained its activity for 2 h; within this period, the MD can be used for multiple digestions. The main properties of this device are simple channel pattern, simple immobilization procedure, regenerability, and disposability; all these features make this MD one of the simplest yet applicable enzymatic microreactors. Graphical abstract Development of microfluidic device including a serpentine channel as an enzyme reactor for protein digestion.
In this study, the possibility of preparation and application of highly porous silica aerogel-based bioactive materials are presented. The aerogel was combined with hydroxyapatite and β-tricalcium phosphate as bioactive and osteoinductive agents. The porosity of aerogels was in the mesoporous region with a maximum pore diameter of 7.4 and 12.7 nm for the composite materials. The newly developed bioactive materials were characterized by SEM. The in vitro biological effect of these modified surfaces was also tested on SAOS-2 osteogenic sarcoma cells by confocal laser scanning microscopy.
Nanoparticles and hydrogels have gained notable attention as promising potential for fabrication of scaffolds and delivering materials. Visible light-curable systems can allow for the possibility ofin situfabrication and have the advantage of optimal applicability. In this study nanogel was created from methacrylated poly-gamma-glutamic acid nanoparticles by visible (dental blue) light photopolymerization. The average size of the particles was 80 nm by DLS, and the NMR spectra showed that the methacrylation rate was 10%. Polymerization time was 3 minutes, and a stable nanogel with a swelling rate of 110% was formed. The mechanical parameters of the prepared structure (compression stress 0.73 MPa, and Young’s modulus 0.93 MPa) can be as strong as necessary in a real situation, for example, in the mouth. A retaining effect of the nanogel was found for ampicillin, and the biocompatibility of this system was tested by Alamar Blue proliferation assay, while the cell morphology was examined by fluorescence and laser scanning confocal microscopy. In conclusion, the nanogel can be used for drug delivery, or it can be suitable for a control factor in different systems.
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