ABSTRACT
Laser direct write techniques represent a prospective alternative for engineering a new generation of hybrid biomaterials via the creation of patterns consisting of biological proteins onto practically any type of substrate. In this paper we report on the characterization of fibronectin features obtained onto titanium substrates by UV nanosecond laser transfer. Fourier-transform infrared spectroscopy measurements evidenced no modification in the secondary structure of the post-transferred protein. The molecular weight of the transferred protein was identical to the initial fibronectin, no fragment bands being found in the transferred protein's Western blot migration profile. The presence of the cell-binding domain sequence and the mannose groups within the transferred molecules was revealed by anti-fibronectin monoclonal antibody immunolabelling and FITC-Concanavalin-A staining, respectively. The in vitro tests performed with MC3T3-E1 osteoblast-like cells and Swiss-3T3 fibroblasts showed that the cells' morphology and spreading were strongly influenced by the presence of the fibronectin spots.
Subject(s)
Fibronectins/chemistry , Lasers, Excimer , Microtechnology , Tissue Engineering/instrumentation , Tissue Scaffolds/chemistry , Animals , Cell Adhesion/drug effects , Cells, Cultured , Coated Materials, Biocompatible/chemical synthesis , Coated Materials, Biocompatible/chemistry , Coated Materials, Biocompatible/pharmacology , Fibroblasts/cytology , Fibroblasts/drug effects , Fibroblasts/physiology , Fibronectins/pharmacokinetics , Fibronectins/pharmacology , Humans , Mice , Microtechnology/instrumentation , Microtechnology/methods , Models, Biological , Osteoblasts/cytology , Osteoblasts/drug effects , Osteoblasts/physiology , Surface Properties/radiation effects , Swiss 3T3 CellsABSTRACT
Active protein and bioceramic calcium hydroxyapatite (HA) bilayers were grown by combining conventional pulsed laser deposition (PLD) and matrix-assisted pulsed laser evaporation (MAPLE) techniques. A pulsed UV KrF* excimer laser was used for the irradiations. The HA layers were grown by PLD. Proteins with antimicrobial action were attached to the bioceramic layers using MAPLE. The composite MAPLE targets were obtained by dissolving the proteins powder in distilled water. The crystalline status and chemical composition of the obtained structures were studied by X-ray diffractometry and Fourier transform infrared spectroscopy. The layers were grown for the design of advanced future metal implants coatings, ensuring both enhanced bone formation and localized antimicrobial therapy. Our results demonstrated that protein coatings improve bone cell proliferation in vitro. Immunofluorescence experiments show that actin filaments stretch throughout bone cells and sustain their optimal spreading.
Subject(s)
Biocompatible Materials/chemistry , Durapatite/chemistry , Proteins/chemistry , Alloys , Cell Line , Cell Proliferation , Coated Materials, Biocompatible/chemistry , Humans , Lasers , Materials Testing , Muramidase/chemistry , Osseointegration , Osteoblasts/cytology , Papain/chemistry , Prostheses and Implants , Surface Properties , TitaniumABSTRACT
Creatinine thin films were synthesised by matrix assisted pulsed laser deposition (PLD) techniques for enzyme-based biosensor applications. An UV KrF* (lambda=248 nm, tau approximately 10 ns) excimer laser source was used for the irradiation of the targets at incident fluence values in the 0.3-0.5 J/cm2 range. For the matrix assisted PLD the targets consisted on a frozen composite obtained by dissolving the biomaterials in distilled water. The surface morphology, chemical composition and structure of the obtained biomaterial thin films were investigated by scanning electron microscopy, Fourier transform infrared spectroscopy, and electron dispersive X-ray spectroscopy as a function of the target preparation procedure and incident laser fluence.