ABSTRACT
Neural transplantation is a promising modality for treatment of neurodegenerative diseases, traumatic brain injury and stroke. Biocompatible scaffolds with optimized properties improve the survival of transplanted neural cells and differentiation of progenitor cells into the desired types of neurons. Silk fibroin is a biocompatible material for tissue engineering. Here, we describe thin-film scaffolds based on photocrosslinked methacrylated silk fibroin (FBMA). These scaffolds exhibit an increased mechanical stiffness and improved water stability. Photocrosslinking of fibroin increased its rigidity from 25 to 480 kPa and the contact angle from 59.7 to 70.8, the properties important for differentiation of neural cells. Differentiation of SH-SY5Y neuroblastoma cells on FBMA increased the length of neurites as well as the levels of neural differentiation markers MAP2 and ßIII-tubulin. Growth of SH-SY5Y cells on the unmodified fibroin and FBMA substrates led to a spontaneous phosphorylation of Src and Akt protein kinases critical for neuronal differentiation; this effect was paralleled by neural cell adhesion molecule elevation. Thus, FBMA is an easily manufactured, cytocompatible material with improved and sustainable properties applicable for neural tissue engineering.
Subject(s)
Cell Differentiation , Fibroins/chemistry , Neurons/physiology , Proto-Oncogene Proteins c-akt/physiology , Proto-Oncogene Proteins pp60(c-src)/physiology , Tissue Scaffolds , Biocompatible Materials , Cell Line, Tumor , Cells, Cultured , HumansABSTRACT
Silk fibroin is a promising biomaterial for tissue engineering due to its valuable mechanical and biological properties. However, being a natural product and a protein, it lacks the processability and uniform quality of an advanced synthetic material. Here we propose a way to overcome this contradiction using novel fibroin photocrosslinkable derivative (FBMA). FBMA was synthesized by methacrylation of native fibroin nucleophilic side groups. It was dissolved in either formic acid (FA) or hexafluoroisopropanol (HFIP), and the obtained solutions were photocrosslinked into hydrogel scaffolds of various structural forms including films, micropatterns, pads and macroporous sponges. UV-exposition of dry FBMA films through a photomask created complex microscaled patterns of the polymer. The nature of the solvent affected the properties of resulting hydrogels. When HFIP was used as the solvent, the resulting hydrogels had a storage modulus â¼4 times higher than that of hydrogels fabricated using FA and â¼20 times higher compared to the reference hydrogel obtained from pristine fibroin. Both FBMA-based hydrogels were biocompatible and supported fibroblast adhesion and growth in vitro. Cells cultivated on FBMA scaffolds produced with HFIP exhibited more spread phenotype at 4 and 24 h of cultivation, consistent with increased stiffness of the hydrogel. Hence, FBMA is an attractive material for fabrication of micropatterned scaffolds of centimeter-scale size with minutely tunable physico-chemical properties via convenient and reproducible technological processes, applicable for rapid prototyping.
Subject(s)
Fibroins/chemistry , Hydrogels/chemistry , Tissue Scaffolds , 3T3 Cells , Actins/chemistry , Animals , Biocompatible Materials/chemistry , Cell Survival , Cross-Linking Reagents/chemistry , Cytoskeleton/chemistry , Formates/chemistry , Methacrylates/chemistry , Mice , Microscopy, Atomic Force , Phenotype , Photochemistry , Polymers/chemistry , Propanols/chemistry , Rheology , Silk/chemistry , Surface Properties , Tissue Engineering/methodsABSTRACT
Novel quaternized polyethyleneimine and cross-linked polyethyleneimine derivatives have been synthesized using both traditional and microwave-assisted techniques to create antimicrobial coatings, with octyl, dodecyl, or hexadecyl bromides as alkylating agent and various bifunctional electrophiles as cross-linkers. Quaternization has been performed using methyl iodide or dimethyl sulfate; it has been shown that methyl iodide has no advantages over dimethyl sulfate. Antimicrobial activity of the polymers against Gram-positive (S. aureus) and Gram-negative (P. aeruginosa) bacteria has been evaluated. Antimicrobial activity declines with increase in the alkylating agent chain length. Equimolar ratio of the alkylating agent and the primary amino groups in polyethyleneimine is optimal. Although cross-linking decreases the antimicrobial activity of quaternized polyethyleneimines, it improves their "non-leaching" properties (i.e. minimizes undesirable water washout of the polymeric coatings).