ABSTRACT
Surface enhanced Raman-based sensors are widely used for chemical and biological species analysis; but to date the high cost, long production time, hazardous, and toxic content as well as small sensing area and opacity are limiting their capabilities for widespread applications in the medical and environmental fields. We present a novel cost-effective method for fast laser-based fabrication of affordable large-area and transparent periodic arrays of ligand-free metallic nanoparticles, offering a maximum possibility for the adsorption/immobilization of molecules and labeling. Further, we demonstrate a remarkable detection limit in the picomolar range by means of Raman scattering, thus evidencing a superior signal-to-noise ratio compared to other sensor substrates. The high sensitivity performance along with a fast and cheap fabrication procedure of reusable large-area transparent plasmonic devices opens the route for direct, in situ multimodal optical analysis with broad applications in the biomedical/analytical fields.
Subject(s)
Biosensing Techniques/methods , Adsorption , Biosensing Techniques/instrumentation , Glass/chemistry , Gold/chemistry , Hair/chemistry , Humans , Hydrophobic and Hydrophilic Interactions , Lasers , Metal Nanoparticles/chemistry , Spectrum Analysis, RamanABSTRACT
The development of bioactive materials is in the premise of tissue engineering. For several years, surface functionalization of scaffolds has been one of the most promising approaches to stimulate cellular activity and finally improve implant success. Herein, we describe the development of a bioactive composite scaffold composed of a biodegradable photopolymer scaffold and titanate nanotubes (TNTs). The biodegradable photopolymer scaffolds were fabricated by applying mask-projection excimer laser photocuring at 308 nm. TNTs were synthesized and then spin-coated on the porous scaffolds. Upon culturing fibroblast cells on scaffolds, we found that nanotubes coating affects cell viability and proliferation demonstrating that TNT coatings enhance cell growth on the scaffolds by further improving their surface topography.
Subject(s)
Coated Materials, Biocompatible/chemistry , Nanotubes/chemistry , Polymers/chemistry , Tissue Scaffolds/chemistry , Titanium/chemistry , 3T3 Cells , Animals , Cell Proliferation/drug effects , Cell Survival/drug effects , Cells, Cultured , Mice , Microscopy, Electron, Transmission , Porosity , Tissue EngineeringABSTRACT
Rigid, biodegradable photopolymer scaffolds were coated with titanate nanotubes (TNTs) by using a spin-coating method. TNTs were synthesized by a hydrothermal process at 150 °C under 4.7 bar ambient pressure. The biodegradable photopolymer scaffolds were produced by mask-assisted excimer laser photocuring at 308 nm. For scaffold coating, a stable ethanolic TNT sol was prepared by a simple colloid chemical route without the use of any binding compounds or additives. Scanning electron microscopy along with elemental analysis revealed that the scaffolds were homogenously coated by TNTs. The developed TNT coating can further improve the surface geometry of fabricated scaffolds, and therefore it can further increase the cell adhesion.
Subject(s)
Coated Materials, Biocompatible/chemistry , Light , Nanotubes/chemistry , Polymers/chemistry , Tissue Scaffolds/chemistry , Titanium/chemistry , Biodegradation, Environmental , Fumarates/chemistry , Nanotubes/ultrastructure , Polymerization/radiation effects , Polypropylenes/chemistry , Porosity , Powders , X-Ray DiffractionABSTRACT
Fluorescence-activated cell sorting (FACS) applying flow cytometry to separate cells on a molecular basis is a widespread method. We demonstrate that both fluorescent and unlabeled live cells in a Petri dish observed with a microscope can be automatically recognized by computer vision and picked up by a computer-controlled micropipette. This method can be routinely applied as a FACS down to the single cell level with a very high selectivity. Sorting resolution, i.e., the minimum distance between two cells from which one could be selectively removed was 50-70 micrometers. Survival rate with a low number of 3T3 mouse fibroblasts and NE-4C neuroectodermal mouse stem cells was 66 ± 12% and 88 ± 16%, respectively. Purity of sorted cultures and rate of survival using NE-4C/NE-GFP-4C co-cultures were 95 ± 2% and 62 ± 7%, respectively. Hydrodynamic simulations confirmed the experimental sorting efficiency and a cell damage risk similar to that of normal FACS.
Subject(s)
Cell Separation/instrumentation , Cell Separation/methods , Image Processing, Computer-Assisted/methods , 3T3 Cells , Animals , Animals, Newborn , Astrocytes/cytology , Cell Line , Cell Survival , Cells, Cultured , Coculture Techniques , Flow Cytometry/instrumentation , Flow Cytometry/methods , Green Fluorescent Proteins/genetics , Green Fluorescent Proteins/metabolism , HEK293 Cells , Humans , Keratinocytes/cytology , Mice , Mice, Transgenic , Microglia/cytology , Microscopy, Fluorescence , Microscopy, Phase-Contrast , Microscopy, Video , Reproducibility of ResultsABSTRACT
We demonstrate high-resolution photocross-linking of biodegradable poly(propylene fumarate) (PPF) and diethyl fumarate (DEF) using UV excimer laser photocuring at 308 nm. The curing depth can be tuned in a micrometre range by adjusting the total energy dose (total fluence). Young's moduli of the scaffolds are found to be a few gigapascal, high enough to support bone formation. The results presented here demonstrate that the proposed technique is an excellent tool for the fabrication of stiff and biocompatible structures on a micrometre scale with defined patterns of high resolution in all three spatial dimensions. Using UV laser photocuring at 308 nm will significantly improve the speed of rapid prototyping of biocompatible and biodegradable polymer scaffolds and enables its production in a few seconds, providing high lateral and horizontal resolution. This short timescale is indeed a tremendous asset that will enable a more efficient translation of technology to clinical applications. Preliminary cell tests proved that PPF : DEF scaffolds produced by excimer laser photocuring are biocompatible and, therefore, are promising candidates to be applied in tissue engineering and regenerative medicine.
