1,721,252 research outputs found
A biomimetic chitosan composite with improved mechanical properties in wet conditions
No full textChitosan is one of the most widely used structural polymers for biomedical applications because it has many favorable properties. However, one of the most critical drawbacks regarding the use of chitosan as a biomedical material is its poor mechanical properties in wet conditions. Here, we designed a method to improve the mechanical properties of chitosan in wet conditions and minimized the swelling behavior of chitosan film due to water adsorption by mimicking the sclerotization of insect cuticles and squid beaks, that is, catechol-meditated crosslinking. The biomimetic chitosan composite film was prepared by mixing chitosan with l-3,4-dihydroxyphenylalanine (DOPA) as a catecholic crosslinker and sodium periodate as an oxidant. The catechol-meditated crosslinking provided a sevenfold enhancement in the stiffness in wet conditions compared to pure chitosan films and reduced the swelling behavior of the chitosan film. This strategy expands the possible applications for the use of chitosan composites as load-bearing biomaterials. (c) 2013 American Institute of Chemical Engineers Biotechnol. Prog., 29: 505512, 2013X112623sciescopu
Three intrinsically unstructured mussel adhesive proteins, mfp-1, mfp-2, and mfp-3: Analysis by circular dichroism
No full textMussel foot proteins (mfps) mediate fouling by the byssal holdfast and have been extensively investigated as models for versatile polymer-mediated underwater adhesion and coatings. However, insights into the structural properties of mfps have lagged far behind the nanomechanical advances, owing in part to the inability of these proteins to crystallize as well as their limited solubility. Here, solution secondary structures of mfp-1, mfp-2, and mfp-3, localized in the mussel byssal cuticle, adhesive plaque, and plaquesubstratum interface, respectively, were investigated using circular dichroism. All three have significant extended coil solution structure, but two, mfp-1 and mfp-2, appear to have punctuated regions of structure separated by unstructured domains. Apart from its punctuated distribution, the structure in mfp-1 resembles other structural proteins such as collagen and plant cell-wall proteins with prominent polyproline II helical structure. As in collagen, PP II structure of mfp-1 is incrementally disrupted by increasing the temperature and by raising pH. However, no recognizable change in mfp-1's PP II structure was evident with the addition with Ca2+ and Fe3+. In contrast, mfp-2 exhibits Ca2+- and disulfide-stabilized epidermal growth factor-like domains separated by unstructured sequence. Mfp-2 showed calcium-binding ability. Bound calcium in mfp-2 was not removed by chelation at pH 5.5, but it was released upon reduction of disulfide bonds. Mfp-3, in contrast, appears to consist largely of unstructured extended coils.X1123sciescopu
Promotion of osteoblast proliferation on complex coacervation-based hyaluronic acid - recombinant mussel adhesive protein coatings on titanium
No full textMany biological polyelectrolytes are capable of undergoing a fluid-fluid phase separation known as complex coacervation. Coacervates were prepared using hyaluronic acid (HA) and a recombinant fusion protein consisting of mussel adhesive motifs and the RGD peptide (fp-151-RGD). The low interfacial energy of the coacervate was exploited to coat titanium (Ti), a metal widely used in implant materials. The coacervate effectively distributed both HA and fp-151-RGD over the Ti surfaces and enhanced osteoblast proliferation. Approximately half of total fp-151-RGD and HA in the solution transferred to the titanium surface within 2 h. Titanium coated with coacervates having high residual negative surface charge showed the highest cell proliferation of preosteoblast cells (MC-3T3) compared to the treatments tested. Indeed, MC-3T3 cells on complex coacervate coated titanium foils exhibited over 5 times greater cell proliferation than bare, HA coated or fp-151-RGD coated titanium. (C) 2009 Elsevier Ltd. All rights reserved.X116563sciescopu
Tunicate-mimetic nanofibrous hydrogel adhesive with improved wet adhesion
No full textThe main impediment to medical application of biomaterial-based adhesives is their poor wet adhesion strength due to hydration-induced softening and dissolution. To solve this problem, we mimicked the wound healing process found in tunicates, which use a nanofiber structure and pyrogallol group to heal any damage on its tunic under sea water. We fabricated a tunicate-mimetic hydrogel adhesive based on a chitin nanofiber/gallic acid (a pyrogallol acid) composite. The pyrogallol group-mediated cross-linking and the nanofibrous structures improved the dissolution resistance and cohesion strength of the hydrogel compared to the amorphous polymeric hydrogels in wet condition. The tunicate-mimetic adhesives showed higher adhesion strength between fully hydrated skin tissues than did fibrin glue and mussel-mimetic adhesives. The tunicate mimetic hydrogels were produced at low cost from recyclable and abundant raw materials. This tunicate-mimetic adhesive system is an example of how natural materials can be engineered for biomedical applications. (C) 2015 Acta Materialia Inc. Published by Elsevier Ltd. All rights reserved.113627sciescopu
Role of Dopamine Chemistry in the Formation of Mechanically Strong Mandibles of Grasshoppers
No full text1154sciescopu
Molecular interactions of mussel protective coating protein, mcfp-1, from Mytilus californianus
No full textProtective coating of the byssus of mussels (Mytilus sp.) has been suggested as a new paradigm of medical coating due to its high extensibility and hardness co-existence without their mutual detriment. The only known biomacromolecule in the extensible and tough coating on the byssus is mussel foot protein-1 (mfp-1), which is made up with positively charged residues (similar to 20 mol%) and lack of negatively charged residues. Here, adhesion and molecular interaction mechanisms of Mytilus californianus foot protein-1 (mcfp-1) from California blue mussel were investigated using a surface forces apparatus (SFA) in buffer solutions of different ionic concentrations (0.2-0.7 M) and pHs (3.0-5.5). Strong and reversible cohesion between opposed positively charged mcfp-1 films was measured in 0.1 M sodium acetate buffer with 0.1 M KNO3. Cohesion of mcfp-1 was gradually reduced with increasing the ionic strength, but was not changed with pH variations. Oxidation of 3,4-dihydroxyphenylalanine (DOPA) residues of mcfp-1, a key residue for adhesive and coating proteins of mussel, didn't change the cohesion strength of mcfp-1 films, but the addition of chemicals with aromatic groups (i.e., aspirin and 4-methylcatechol) increased the cohesion. These results suggest that the cohesion of mcfp-1 films is mainly mediated by cation-it interactions between the positively charged residues and benzene rings of DOPA and other aromatic amino acids (similar to 20 mol% of total amino acids of mcfp-1), and pi-pi interactions between the phenyl groups in mcfp-1. The adhesion mechanism obtained for the mcfp-1 proteins provides important insight into the design and development of functional biomaterials and coatings mimicking the extensible and robust mussel cuticle coating. (C) 2011 Elsevier Ltd. All rights reserved.X1146Nsciescopu
Glycosylated Hydroxytryptophan in a Mussel Adhesive Protein from Perna viridis
No full textThe 3,4-dihydroxyphenyl-L-alanine (Dopa)-containing proteins of mussel byssus play a critical role in wet adhesion and have inspired versatile new synthetic strategies for adhesives and coatings. Apparently, however, not all mussel adhesive proteins are beholden to Dopa chemistry. The cDNA-deduced sequence of Pvfp-1, a highly aromatic and redox active byssal coating protein in the green mussel Perna viridis, suggests that Dopa may be replaced by a post-translational modification of tryptophan. The N-terminal tryptophan-rich domain of Pvfp-1 contains 42 decapeptide repeats with the consensus sequences ATPKPW(1)TAW(2)K and APPPAW(1)TAW(2)K. A small collagen domain (18 Gly-X-Y repeats) is also present. Tandem mass spectrometry of isolated tryptic decapeptides has detected both C-2-hexosylated tryptophan(W-1) and C-2-hexosylated hydroxytryptophan (W-2), the latter of which is redox active. The UV absorbance spectrum of W-2 is consistent with 7-hydroxytryptophan, which represents an intriguing new theme for bioinspired opportunistic wet adhesion.X112424sciescopu
Expression of functional human transferrin in stably transfected Drosophila S2 cells
No full textHuman transferrin (hTf) is a serum glycoprotein involved in Fe3+ transport. Here, a plasmid encoding the hTf gene fused with a hexahistidine (His(6)) epitope tag under Drosophila metallothionein promoter (pMT) was stably transfected into Drosophila melanogaster S2 cells as a nonlytic plasmid-based system. Following 3 days of copper sulfate induction, transfected S2 cells were found to secrete hTf into serum-free culture medium at a competitively high expression level of 40.8 mug/mL, producing 6.8 mug/mL/ day in a 150-mL spinner flask culture. Purification of secreted recombinant hTf using immobilized metal affinity chromatography (IMAC) yielded 95.5% pure recombinant hTf with a recovery of 32%. According to MALDI-TOF mass spectrometry analysis, purified S2 cell-derived His(6)-tagged recombinant hTf had a molecular weight (76.4 kDa) smaller than that of native apo-hTf (78.0 kDa). 2-Dimensional gel electrophoresis patterns showed recombinant hTf had a simpler and less acidic profile compared to that of native hTf. These data suggest recombinant hTf was incompletely (noncomplex) glycosylated and lacked sialic acids on N-glycans. However, this difference in N-glycan structure compared to native hTf had no effect on the iron-binding activity of recombinant hTf. The present data show that a plasmid-based stable transfection S2 cell system can be successfully employed as an alternative for producing secreted functional recombinant hTf.X111815sciescopu
Chitosan and hydroxyapatite composite cross-linked by dopamine has improved anisotropic hydroxyapatite growth and wet mechanical properties
No full textThree of the major impediments to using hydroxyapatite (HAp)-collagen composites for hard tissue repair are the difficulties in anisotropic growth of HAp, in functional collagen production, and in their cross-linking. To solve these problems, we fabricated HAp-based composites for hard tissue repair by using chitosan as a collagen matrix substitute, and dopamine as a replacement for aldehyde-based cross-linkers. In the presence of chitosan and dopamine, the HAp particles grew anisotropically in a needle shape with an aspect ratio of similar to 4.4. The needle-shaped HAp particles were dispersed well in the chitosan matrix, and dopamine-mediated cross-linking enhanced the stiffness and reduced swelling in the presence of water. The composite is too weak for use in hard tissue repair, but could be used for curing dentin sensitivity by blocking and remineralization on dentinal tubules, and in drug-delivery applications.X1133sciescopu
Strong Reversible Fe3+-mediated Bridging between Dopa-Containing Protein Films in Water
No full textMetal-containing polymer networks are widespread in biology, particularly for load-bearing exoskeletal biomaterials. Mytilus byssal cuticle is an especially interesting case containing moderate levels of Fe3+ and cuticle protein-mussel foot protein-1 (mfp-1), which has a peculiar combination of high hardness and high extensibility. Mfp-1, containing 13 mol % of dopa (3, 4-dihydroxyphenylalanine) side-chains, is highly positively charged polyelectrolyte (pI similar to 10) and didn't show any cohesive tendencies in previous surface forces apparatus (SFA) studies. Here, we show that Fe3+ ions can mediate unusually strong interactions between the positively charged proteins. Using an SFA, Fe3+ was observed to impart robust bridging (W-ad approximate to 4.3 mJ/m(2)) between two noninteracting mfp-1 films in aqueous buffer approaching the ionic strength of seawater. The Fe3+ bridging between the mfp-1-coated surfaces is fully reversible in water, increasing with contact time and iron concentration up to 10 mu M; at 100 mu M, Fe3+ bridging adhesion is abolished. Bridging is apparently due to the formation of multivalent dopa-iron complexes. Similar Fe-mediated bridging (W-ad approximate to 5.7 mJ/m(2)) by a smaller recombinant dopa-containing analogue indicates that bridging is largely independent of molecular weight and posttranslational modifications other than dopa. The results suggest that dopa-metal interactions may provide an energetic new paradigm for engineering strong, self-healing interactions between polymers under water.open11203201sciescopu
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