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.
Biodegradable polymers are compatible, permeable and nontoxic, thus they can provide a useful tool for drug delivery or tissue engineering. These polymers can form hydrogels, which are suitable vehicles for different types of materials e.g. drugs, bioactive molecules or cells. In the case of dentistry, photopolymerization is an obvious method to obtain in situ useable devices which can provide a more efficient way of tailoring drug release. A hydrogel system was developed based on poly-gamma-glutamic acid that was modified with methacryloyl groups to achieve this purpose. The resulting new reactive structure was proved by NMR spectroscopy. The swelling ratio of this type of hydrogel has been found remarkable, over 300 % after 24 h, and it can release 5 ng/mm(2) metronidazole. The prepared hydrogels were nontoxic as viability, cytotoxicity tests and cell morphology investigations proved it. These results render this model system an excellent candidate for use as an in situ curing local drug delivery device. The new photoactive system can be utilized in the treatment of periodontal diseases or raising the effectiveness of drugs used only in the minimal effective dose.
The main objective of this work was to combine the positive characteristics of transparent photopolymers and light-sensitive chalcogenide glasses, with aim to improve the amplitude-phase modulation characteristics of in situ optically recorded photonic elements on the surface, and in the bulk of thick composite layer on a given substrate. The positive results were obtained due to the developed technology routes of nanocomposite (NC) fabrication by intermixing selected, optically tunable, VIS-NIR transparent and high refractive index As-S (Se) nanoparticles (NPs) produced by chemical dissolution, and acrylate monomers with initiators. Subsequent photopolymerization of such nanocomposite occurs during optical recording photonic elements and is supplemented by mass-transport processes, which enhance relief parameters. Structure, optical parameters of the new light-sensitive media and conditions of one step recording of optical elements in it were investigated.
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