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- W1255668061 abstract "INTRODUCTION: To date, no real-time tool exists for assessing the in-situ fate of biomaterials for surgical implants or scaffolds in tissue engineering. The aim of this work is to establish the framework for a real-time monitoring system to follow the degradation of scaffolds in-vitro and invivo. The strategy is based on the covalently binding of a fluorescent probe into the backbone of the polymers used to produce scaffolds. As the polymer degrades, fluorescent probe molecules will be released and the fluorescence intensity of the scaffolds will decrease. Thus, at a given point the mass of the scaffold can be derived from intensity measurements. METHODS: Production of labelled scaffolds: Chitosan scaffolds with 80µmx1.5cmx1.5cm were produced by casting a 1% chitosan solution in 2% acetic acid. After drying and neutralization the membranes were immersed in a 0.1 mg/ml solution of Tetramethylrhodamine Isothiocyanate (TRITC) in ethanol/water/bicarbonate buffer for 24 hours. The free dye was removed using an extraction system. Scaffolds’ degradation: Chitosan scaffolds were incubated for 10 days in a 0.2 mg/ml lipase solution (in Phosphate Buffer Saline (PBS)), pH 7.4 at 37 oC. At days 3, 7 and 10 the scaffolds were rinsed with distilled water, dried to a constant weight and the weight loss was determined. Quantification of fluorescence intensity (FI): The FI of the scaffolds was assessed by Confocal Microscopy with a laser line at 543 nm. Measurements were done before and after a given incubation period. The samples FI was normalized with the FI of standards prepared from InSpeck Red and InSpeck Green (6μm) (Molecular Probes). The same settings were used for all measurements. Cell viability, morphology and extra-cellular matrix deposition: MG63 osteoblast-like cells were seeded on the scaffolds and cultured for 7 days. Cell viability and morphology were evaluated by confocal microscopy after Calcein AM staining. At the same time, collagen deposition was assed by light microscopy after Picro-sirius red staining." @default.
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- W1255668061 date "2008-01-01" @default.
- W1255668061 modified "2023-09-26" @default.
- W1255668061 title "Fluorescence Unravelling the Fate of Scaffolds: Real-time Monitoring of Scaffold Degradation" @default.
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