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PubMed2000

Enzymatic grafting of hexyloxyphenol onto chitosan to alter surface and rheological properties.

An enzymatic method to graft hexyloxyphenol onto the biopolymer chitosan was studied. The method employs tyrosinase to convert the phenol into a reactive o-quinone, which undergoes subsequent nonenzymatic reaction with chitosan. Reactions were conducted under heterogeneous conditions using chitosan films and also under homogeneous conditions using aqueous methanolic mixtures capable of dissolving both hexyloxyphenol and chitosan. Tyrosinase was shown to catalyze the oxidation of hexyloxyphenol in such aqueous methanolic solutions. Chemical evidence for covalent grafting onto chitosan was provided by three independent spectroscopic approaches. Specifically, enzymatic modification resulted in (1) the appearance of broad absorbance in the 350-nm region of the UV/vis spectra for chitosan films; (2) changes in the NH bending and stretching regions of chitosan's IR spectra; and (3) a base-soluble material with (1)H-NMR signals characteristic of both chitosan and the alkyl groups of hexyloxyphenol. Hexyloxyphenol modification resulted in dramatic changes in chitosan's functional properties. On the basis of contact angle measurements, heterogeneous modification of a chitosan film yielded a hydrophobic surface. Homogeneously modified chitosan offered rheological properties characteristic of associating water-soluble polymers.

Chitin↗

PubMed2000

Chitosan supports the expression of extracellular matrix proteins in human osteoblasts and chondrocytes.

The search for biocompatible materials that can support the growth and phenotypic expression of osteoblasts and chondrocytes is a major challenge in the application of tissue engineering techniques for the repair of bone and cartilage defects. Chitosan, a copolymer of glucosamine and N-acetylglucosamine, may provide an answer to this search. Chitosan is the deacetylated product of chitin, a ubiquitous biopolymer found in the exoskeleton of insects and marine invertebrates. Little is known about the utility of chitosan in propagating human osteoblasts and chondrocytes. In this study, we test the hypothesis that chitosan promotes the survival and function of osteoblasts and chondrocytes. Chitosan (4%, w/v in 2% HAc) was coated onto plastic coverslips that had been fitted into 24-well plates. Human osteoblasts and articular chondrocytes were seeded on either uncoated or chitosan-coated coverslips at 1 x 10(5)/cells per well. Cultures were incubated at 37 degrees C, 5% CO(2) for a period of 7 days. Cell viability was assessed at that time using a fluorescent molecular probe. The phenotypic expression of osteoblasts and chondrocytes was analyzed by reverse transcriptase-polymerase chain reaction and immunocytochemistry. Osteoblasts and chondrocytes appeared spherical and refractile on chitosan-coated coverslips. In contrast, greater than 90% of cells on plastic coverslips were elongated and spindle shaped after 7 days of culture. Similar to cells propagated on uncoated control wells, greater than 90% of human osteoblasts and chondrocytes propagated on chitosan remained viable. Human osteoblasts propagated on chitosan films continued to express collagen type I whereas chondrocytes expressed collagen type II and aggrecan, as shown by reverse transcriptase-polymerase chain reaction analysis and immunostaining. The present in vitro work demonstrates the biocompatibility of chitosan as a substrate for the growth and continued function of human osteoblasts and chondrocytes. Chitosan may have potential use as a tissue engineering tool for the repair of osseous and chondral defects.

Aggrecans↗

PubMed1992

Gel electrophoretic analysis of chitosan hydrolysis products.

Enzymatic hydrolysis of commercial crustacean chitosan by barley chitosanases was analyzed by subjecting chitosan to electrophoresis in a 10% w/v polyacrylamide slab gel in the presence of 7 M urea and 5.5% v/v acetic acid. Chitosan migrated as a polycation. Chitosan was stained with Coomassie Brilliant Blue R-250 or visualized by ultraviolet transillumination after staining with Calcofluor White M2R. Some chitosan molecules were retarded by gel electrophoresis while small chitosan molecules migrated at the bottom of a 10% w/v polyacrylamide gel. Such analysis revealed that 96 h were necessary to convert all chitosan to oligosaccharides under our assay conditions. Chitosan oligosaccharides generated by enzymatic or chemical hydrolysis were further analyzed by electrophoresis in a 33% w/v polyacrylamide gel containing urea and acetic acid. Coomassie Brilliant Blue R-250 was found to be better than Calcofluor White M2R for staining chitosan oligosaccharides. Chitosan oligomers of four residues (tetramers) or more were easily resolved in such a polyacrylamide gel system. To our knowledge, this is the first report of a gel electrophoretic separation of chitosan and its oligosaccharides.

Chitin↗

PubMed2005

Insecticidal and fungicidal activity of new synthesized chitosan derivatives.

Chitosan, the N-deacetylated derivative of chitin, is a potential biopolysaccharide owing to its specific structure and properties. In this paper, we report on the synthesis of 24 new chitosan derivatives, N-alkyl chitosans (NAC) and N-benzyl chitosans (NBC), that are soluble in dilute aqueous acetic acid. The different derivatives were synthesized by reductive amination and analyzed by 1H NMR spectroscopy. A high degree of substitution (DS) was obtained with N-(butyl)chitosan (DS 0.36) at a 1:1 mole ratio for NAC derivatives and N-(2,4-dichlorobenzyl)chitosan (DS 0.52) for NBC derivatives. Their insecticidal and fungicidal activities were tested against larvae of the cotton leafworm Spodoptera littoralis (Boisduval) (Lepidoptera: Noctuidae), the grey mould Botrytis cinerea Pers (Leotiales: Sclerotiniaceae) and the rice leaf blast Pyricularia grisea Cavara (Teleomorph: Magnaporthe grisea (Hebert) Barr). The oral feeding bioassay indicated that all the derivatives had significant insecticidal activity at 5 g kg(-1) in artificial diet. The most active was N-(2-chloro-6-fluorobenzyl)chitosan, which caused 100% mortality at 0.625 g kg(-1), with an estimated LC50 of 0.32 g kg(-1). Treated larvae ceased feeding after 2-3 days; the mechanism of action remains unknown. In a radial hyphal growth bioassay with both plant pathogens, all derivatives showed a higher fungicidal action than chitosan. N-Dodecylchitosan, N-(p-isopropylbenzyl)chitosan and N-(2,6-dichlorobenzyl)chitosan were the most active against B cinerea, with EC50 values of 0.57, 0.57 and 0.52 g litre(-1), respectively. Against P grisea, N-(m-nitrobenzyl)chitosan was the most active, with 77% inhibition at 5 g litre(-1). The effect of different substitutions is discussed in relation to insecticidal and fungicidal activity.

Animals↗

PubMed2006

Efficacy and mechanism of action of chitosan nanocapsules for oral peptide delivery.

PURPOSE: We have previously shown that high molecular weight (MW > 100 kDa) chitosan nanocapsules are efficient vehicles for improving the oral absorption of salmon calcitonin (sCT). In the present work, our objectives were, first, to investigate the influence of some formulation parameters on the efficacy of chitosan nanocapsules as carriers for the oral administration of sCT and, second, to elucidate the mechanism of interaction of chitosan nanocapsules with intestinal model cell lines. METHODS: sCT-loaded chitosan nanocapsules were prepared by the solvent displacement technique. They were characterized for their size, zeta potential, and sCT loading. The ability of chitosan nanocapsules to enhance the oral absorption of sCT was investigated in rats by monitoring the serum calcium levels. Finally, the mechanism of interaction of chitosan nanocapsules with the Caco-2 cell model or in the coculture of Caco-2 with HT29-M6 cells was investigated by confocal fluorescence microscopy. RESULTS: Chitosan nanocapsules presented a particle size in nanometer range, a positive surface charge, and an efficient encapsulation of sCT. Following oral administration to rats, all formulations of nanocapsules exhibited the ability to reduce calcemia levels; however, the intensity of the response varied depending on the formulation conditions. With regard to the mechanism of interaction of chitosan nanocapsules with cell culture, the xz images evidenced that chitosan nanocapsules interact and remain associated to the apical side of both model cell cultures. In addition, chitosan nanocapsules showed a preferable association to the mucus-secreting cells (HT29-M6). CONCLUSIONS: Chitosan nanocapsules are able to enhance and prolong the intestinal absorption of sCT and this effect could be mainly ascribed to their mucoadhesive character and intimate interaction with the intestinal barrier.

Administration, Oral↗

PubMed2004

The interaction of Schwann cells with chitosan membranes and fibers in vitro.

The bridging of nerve gaps is still one of the major problems in peripheral nerve regeneration. A promising alternative for the repair of peripheral nerve injuries is the bioartificial nerve graft, comprised of a biomaterial pre-seeded with Schwann cells (SCs), which is an effective substrate for enhancing nerve regeneration. Interaction between cultured SCs and biomaterials is of importance. For the purposes of this study, culture systems of normal SCs were used. The biocompatibility of chitosan, including chitosan membranes and chitosan fibers, was evaluated in vitro. The growth of SCs was observed by light and scanning electron microscopy at regular intervals. SCs were identified by immunocytochemical staining and the viability of SCs was measured by MTT assay. The experimental results indicated that SCs could grow onto chitosan materials with two different shapes: spherical and long olivary. They contacted with the extensions. The long olivary cells inclined to encircle chitosan fibers up. It was also found that the cells on the chitosan fibers migrated faster than those on the chitosan membranes. There was a good biological compatibility between chitosan and SCs. Compared with the chitosan membranes, SCs migrated more easily onto the stereoframe of chitosan fibers. These studies contribute information necessary to enhancing our understanding of biocompatibility of chitosan.

Animals↗

PubMed2005

Synthesis and in vitro evaluation of a novel thiolated chitosan.

In order to achieve the same properties as chitosan-4-thio-butyl-amidine and to overcome at the same time its insufficient stability, the aim of this study was to evaluate the imidoester reaction of isopropyl-S-acetylthioacetimidate for the chemical modification of chitosan and to study the properties of the resulting chitosan-thioethylamidine (TEA) derivative. The thioalkylamidine substitute was introduced without the formation of N-substituted non-thiol products. The resulting conjugates exhibited 1.05+/-0.17% or 139.68+/-17.13 micromol immobilized free thiol groups per gram polymer and a total amount of reduced and oxidized thiol groups of 1.81+/-0.65% or 179.46+/-67.95 micromol/g polymer. By the immobilization of thiol groups mucoadhesion was strongly improved due to the formation of disulfide bonds with mucus glycoproteins. Chitosan-TEA was investigated regarding to its mucoadhesive properties via tensile studies and the rotating cylinder method. In tensile studies the total work of adhesion of chitosan-TEA was increased 3.3-fold in comparison to unmodified chitosan. Results from the rotating cylinder method showed an improvement ratio of 8.9 for chitosan-TEA compared with unmodified chitosan. In spite of the immobilization of thiol groups onto chitosan its swelling behavior in aqueous solutions was not significantly altered. Cumulative release studies out of matrix tablets comprising the chitosan-TEA and the model compound fluorescence labeled dextrane (FD(4)) demonstrated a controlled release over 3h with a trend toward a pseudo-zero-order kinetic. Because of these features the new chitosan thioamidine conjugate might represent a promising new polymeric excipient for various drug delivery systems.

Absorbable Implants↗

PubMed2006

Novel design of osmotic chitosan capsules characterized by asymmetric membrane structure for in situ formation of delivery orifice.

In this study, chitosan capsules with asymmetric membrane to induce osmotic effects and in situ formation of the delivery orifice were optimally prepared and characterized. Chitosan capsules were formed on stainless steel mold pins by dipping the pins into a chitosan solution followed by forming asymmetric structure by dipping into a quenching solution containing tripolyphosphate (TPP) to cause an ionic cross-linking reaction between the outer layer of chitosan and TPP. Factors influencing the properties of the capsule membrane, such as the molecular weight of chitosan, the dipping solution and dipping time, and the quenching solution and time, were optimized to successfully produce osmotic chitosan capsules with asymmetric membrane using chitosans that possessed different viscosities. In situ formation of a delivery orifice on the asymmetric membrane of the chitosan capsule was proven by the observation of a jet stream of chlorophyll being released from the capsule. Drugs with different solubility were selected, and a linear correlation between drug solubility and the initial drug release rate calculated from the slope of the drug release profile was used to verify that the delivery orifices that were in situ formed on the asymmetric membrane of the chitosan capsules induced by osmotic effect was responsible for the drug release. Water permeability across the optimally produced asymmetric membrane of the capsule from chitosan of 500 cps (300-700 cps) quenched with TPP for 30 min (C500/TPP30) was determined to be 1.40 x 10(-6)cm(2)h(-1)atm(-1) at 37.0+/-0.5 degrees C. The encapsulation of poorly water-soluble drugs, felodipine (FE) and nifedipine (NF), in such an asymmetric chitosan capsule was capable of creating a sufficient osmotic effect to activate the release of the drug with the addition of SLS and HPMC. The multiple regression equations of maximal release percent at 24h for FE and NF confirmed that both sodium lauryl sulfate (SLS) and hydroxypropyl methylcellulose (HPMC) positively influenced this response factor, and the effect of SLS was greater than that of HPMC.

Capsules↗

PubMed2000

A simple preparation of half N-acetylated chitosan highly soluble in water and aqueous organic solvents.

A simple and improved method of preparing highly soluble chitosan (half N-acetylated chitosan) was developed using a series of chitosan samples of low molecular weights, and the solubility of the half N-acetylated chitosan in water and organic solvents was investigated in detail. To reduce the molecular weight, chitosan was treated with NaBO3 under the condition that chitosan was homogeneously dissolved in aqueous acetic acid. Weight-average molecular weights of the obtained chitosan samples were determined using a size-exclusion chromatography system equipped with a low-angle laser light-scattering photometer. Each chitosan sample was then N-acetylated with acetic anhydride under the condition that chitosan was homogeneously dissolved in aqueous acetic acid again. The water solubility of the half N-acetylated chitosan thus prepared increased with decreasing molecular weight. From 1H NMR spectroscopy, it was suggested that the sequence of N-acetylglucosamine and glucosamine residues was random. The solubility of the half N-acetylated chitosan of low molecular weight was rather high even in aqueous dimethylacetamide and dimethylsulfoxide.

Acetylation↗

PubMed2001

Chitosan microparticles for oral vaccination: preparation, characterization and preliminary in vivo uptake studies in murine Peyer's patches.

Although oral vaccination has numerous advantages over parenteral injection, degradation of the vaccine in the gut and low uptake in the lymphoid tissue of the gastrointestinal tract still complicate the development of oral vaccines. In this study chitosan microparticles were prepared and characterized with respect to size, zeta potential, morphology and ovalbumin-loading and -release. Furthermore, the in vivo uptake of chitosan microparticles by murine Peyer's patches was studied using confocal laser scanning microscopy (CLSM). Chitosan microparticles were made according to a precipitation/coacervation method, which was found to be reproducible for different batches of chitosan. The chitosan microparticles were 4.3+/-0.7 microm in size and positively charged (20+/-1 mV). Since only microparticles smaller than 10 microm can be taken up by M-cells of Peyer's patches, these microparticles are suitable to serve as vaccination systems. CLSM visualization studies showed that the model antigen ovalbumin was entrapped within the chitosan microparticles and not only associated to their outer surface. These results were verified using field emission scanning electron microscopy, which demonstrated the porous structure of the chitosan microparticles, thus facilitating the entrapment of ovalbumin in the microparticles. Loading studies of the chitosan microparticles with the model compound ovalbumin resulted in loading capacities of about 40%. Subsequent release studies showed only a very low release of ovalbumin within 4 h and most of the ovalbumin (about 90%) remained entrapped in the microparticles. Because the prepared chitosan microparticles are biodegradable, this entrapped ovalbumin will be released after intracellular digestion in the Peyer's patches. Initial in vivo studies demonstrated that fluorescently labeled chitosan microparticles can be taken up by the epithelium of the murine Peyer's patches. Since uptake by Peyer's patches is an essential step in oral vaccination, these results show that the presently developed porous chitosan microparticles are a very promising vaccine delivery system.

Administration, Oral↗

PubMed2002

A new lipid emulsion formulation with high antimicrobial efficacy using chitosan.

The antimicrobial activity of chitosan in lipid emulsions as well as in aqueous solutions was investigated. Two types of long-chained chitosan were used differing in the molecular weights, degree of the deacetylation and their viscosity: type I, mol. weight 8.7 x 10(4) g/mol, 92% degree of deacetylation and a viscosity of 14 mPa s, type II, mol. weight of 5.32 x 10(5) g/mol, 73% degree of deacetylation and a viscosity of 461 mPa s. In order to assess the pH optimum of the antimicrobial activity of the biopolymer, suspensions of the microorganisms Pseudomonas aeruginosa, Staphylococcus aureus, Candida albicans and Aspergillus niger were incubated at different pH-values in lactic acid solution (1% w/v) containing different concentrations of chitosan up to 1.5% (w/v). Emulsion formulations containing either 0.25%, 0.5% or no chitosan, respectively, were inoculated with the same microorganisms and were incubated at 25 degrees C. The aqueous solutions as well as the emulsions were examined for microbial counts on agar plates after different periods of incubation. After 24 h of incubation in aqueous solutions only the cfu numbers of the bacteria were reduced. Both types of chitosan revealed a pH optimum of their antibacterial activity at pH 5.0-5.1 for P. aeruginosa, and at pH 5.3 for S. aureus. In addition, chitosan with a mol. weight of 8.7 x 10(4) g/mol, high degree of deacetylation and low viscosity showed a higher antimicrobial activity than the other chitosan type of this study. It was found that lipid emulsions containing 0.5% chitosan (type I) conformed to the requirements of the preservation efficacy test for topical formulations according to the European Pharmacopoeia while the emulsion without chitosan and a lactic acid solution with and without the biopolymer did not conform. In hemolysis studies on human erythrocytes, the hemolytic activity of the lipid emulsions with chitosan was assessed. These emulsions showed a negligible hemolytic behavior. The results indicate a use of chitosan as antimicrobial preservative in emulsion formulations for mucosal as well for parenteral applications.

Anti-Bacterial Agents↗

PubMedDate not supplied

Synthesis and characterization of a novel chitosan-gelatin bioconjugate with fluorescence emission.

Polysaccharide-protein conjugations have generated increasing interests for biomedical applications in recent years. A naturally occurring cross-linking reagent, genipin, which has been used in herbal medicine, was employed to cross-link chitosan and gelatin for the preparation of a novel chitosan-gelatin conjugate. The primary amine groups on chitosan and gelatin were covalently linked with genipin, leading to the formation of a chitosan-gelatin conjugate with nitrogen-containing heterocycle units, the pyrindine-like derivatives. The FT-IR and UV-vis studies revealed that chitosan could react with genipin via a nucleophilic ring-opening reaction to construct more sufficient and extensive cross-link networks, as compared with its gelatin counterpart. The UV-vis absorption properties of the chitosan-gelatin conjugates were strongly related to the chitosan-to-gelatin weight ratio in the compositions. It is worth noting that the conjugation process endows the special emission properties of the chitosan-gelatin conjugates, which depends on the cross-linking reaction and the formation of hydrogen bonding involved chitosan-gelatin complex. Fluorescence quenching or enhancement was observed from the chitosan-gelatin conjugates upon coordinated with a wide variety of heavy metal ions (Ag+, Cu2+, Fe2+, and Co2+). This study also examined the possibility of covalent coupling the capture chelator (chitosan) with bioactive protein (e.g., albumin, alpha-globulin, and fibrinogen) to create fluorescence emission. These findings may provide a novel way to deliver therapeutic radionuclides for immuno-targeting purposes in the future.

Adjuvants, Immunologic↗

PubMed2001

Experimental evaluation of photocrosslinkable chitosan as a biologic adhesive with surgical applications.

BACKGROUND: In various surgical cases, effective tissue adhesives are required for both hemostasis (eg, intraoperative bleeding) and air sealing (eg, thoracic surgery). We have designed a chitosan molecule (Az-CH-LA) that can be photocrosslinked by ultraviolet (UV) light irradiation, thereby forming a hydrogel. The purpose of this work was to evaluate the effectiveness and safety of the photocrosslinkable chitosan hydrogel as an adhesive with surgical applications. METHODS: The sealing ability of the chitosan hydrogel, determined as a bursting pressure, was assessed with removed thoracic aorta, trachea, and lung of farm pigs and in a rabbit model. The carotid artery and lung of rabbits were punctured with a needle, and the chitosan hydrogel was applied to, respectively, stop the bleeding and the air leakage. In vivo chitosan degradability and biologic responses were histologically assessed in animal models. RESULTS: The bursting pressure of chitosan hydrogel (30 mg/mL) and fibrin glue, respectively, was 225 +/- 25 mm Hg (mean +/- SD) and 80 +/- 20 mm Hg in the thoracic aorta; 77 +/- 29 mm Hg and 48 +/- 21 mm Hg in the trachea; and in the lung, 51 +/- 11 mm Hg (chitosan hydrogel), 62 +/- 4 mm Hg (fibrin glue, rubbing method), and 12 +/- 2 mm Hg (fibrin glue, layer method). The sealing ability of the chitosan hydrogel was stronger than that of fibrin glue. All rabbits with a carotid artery (n = 8) or lung (n = 8) that was punctured with a needle and then sealed with chitosan hydrogel survived the 1-month observation period without any bleeding or air leakage from the puncture sites. Histologic examinations demonstrated that 30 days after application, a fraction of the chitosan hydrogel was phagocytosed by macrophages, had partially degraded, and had induced the formation of fibrous tissues around the hydrogel. CONCLUSIONS: A newly developed photocrosslinkable chitosan has demonstrated strong sealing ability and a great potential for use as an adhesive in surgical operations.

Animals↗

PubMed1994

Effect of chitin and chitosan on nutrient digestibility and plasma lipid concentrations in broiler chickens.

Broiler chickens were fed on a control diet based on maize and maize starch or diets containing chitin, or 94, 82 or 76% deacetylated chitin (chitosans) with different viscosities (360, 590 and 620 m Pa.s respectively) at an inclusion level of 30 g/kg. Animals had free access to feed and water for the whole experimental period. On days 10 and 18 of the experiment chickens given the control and chitin-containing diets weighed more, had consumed more feed and had lower feed conversion ratios (g feed/g weight gain) than chitosan-fed birds. Feeding of chitosan-containing diets generally reduced total plasma cholesterol and high-density-lipoprotein (HDL)-cholesterol concentrations and gave an increased HDL:total cholesterol ratio in comparison with chickens given the control and chitin-containing diets. However, no significant reductions in plasma triacylglycerol concentrations resulting from feeding of the chitosan-containing diets were observed. The reduction in total cholesterol concentration and increased HDL:total cholesterol ratio were probably caused by enhanced reverse cholesterol transport in response to intestinal losses of dietary fats. The suggestion that dietary fat absorption was impeded by the chitosans was strengthened by the observation that ileal fat digestibility was reduced by 26% in comparison with control and chitin-fed animals. In a plasma triacylglycerol response study on day 21, feeding of 94 and 76%-chitosan-containing diets generally reduced postprandial triacylglycerol concentrations compared with chickens given the chitin-containing diet. Duodenal digestibilities of nutrients amongst chickens given the chitin-containing diet were generally lower than those of control and chitosan-fed birds indicating decreased intestinal transit time. The reduced caecal short-chain fatty acid concentrations of chickens given chitosan diets compared with the control diet illustrates the antimicrobial nature of chitosan. The fact that the three chitosan-containing diets affected the registered variables similarly indicated that the level of inclusion of chitosans in the diet exceeded the level at which the effect of the different viscosities could be significant.

Animals↗

PubMed2003

Laminin-1 peptide-conjugated chitosan membranes as a novel approach for cell engineering.

Laminin, a major component of the basement membrane, has diverse biological activities. Recently, we identified various biologically active sequences on laminin-1 by using a large set of synthetic peptides. Chitosan, a polysaccharide, is biodegradable and has been used as a biomaterial. Here, we conjugated several biologically active laminin peptides onto chitosan membranes and measured the cell attachment activity of peptide-conjugated chitosan membranes with various cell types. The active laminin peptide-conjugated chitosan membranes promoted cell attachment with cell type specificity. A99 (AGTFALRGDNPQG)-chitosan membrane promoted cell attachment with well-organized actin stress fibers. This adhesion was inhibited by EDTA but not by heparin. AG73 (RKRLQVQLSIRT)-chitosan membrane promoted cell attachment with filopodia formation, and this adhesion was inhibited by heparin but not by EDTA. These data suggest that the A99-chitosan membrane interacted with an integrin cellular receptor and that the AG73-chitosan membrane promoted proteoglycan-mediated cell attachment, as previously reported. Furthermore, both AG73-chitosan and A99-chitosan membranes effectively promoted neurite outgrowth with PC12 rat pheochromocytoma cells. We conclude that conjugation on a chitosan membrane is applicable for testing quantitatively the biological activity of synthetic peptides and that these constructs have a potential ability to serve as bioadhesive materials for tissue regeneration and engineering.

Amino Acid Sequence↗

PubMed1998

Effect of low-molecular-weight chitosans on the adhesive properties of oral streptococci.

It was previously shown that a low-molecular-weight chitosan and its derivatives N-carboxymethyl chitosan and imidazolyl chitosan inhibit Streptococcus mutans adsorption to hydroxyapatite. The ability of the same molecules to interfere with adhesive properties of other oral streptococci (Streptococcus sanguis, Streptococcus gordonii, Streptococcus constellatus, Streptococcus anginosus, Streptococcus intermedius, Streptococcus oralis, Streptococcus salivarius, Streptococcus vestibularis) was tested. When saliva-coated or -uncoated hydroxyapatite beads were treated with N-carboxymethyl chitosan, a reduction varying from 60% to 98% depending on strains was observed. Low-molecular-weight chitosans and imidazolyl chitosan did not have any effect. Growth in N-carboxymethyl chitosan-supplemented medium (final concentrations ranging from 20 to 500 micrograms.ml-1) caused a dose related reduction in the adsorption of all strains to hydroxyapatite and in their affinity towards xylene. No effect was observed with low-molecular-weight chitosans and imidazolyl chitosan. In contrast to what observed with S. mutans, the three polysaccharides did not affect detachment from hydroxyapatite beads and adherence to cheek epithelial cells of the other streptococci. These results suggest that low-molecular-weight chitosans and/or imidazolyl chitosan, selectively affecting S. mutans adsorption to hydroxyapatite, may be very interesting as potential anti-dental caries agents.

Adult↗

PubMed2006

Releasing growth factors from activated human platelets after chitosan stimulation: a possible bio-material for platelet-rich plasma preparation.

INTRODUCTION: Thrombin is commonly used for activating the platelets and releasing the growth factors on the application of platelet-rich plasma (PRP). We have previously reported that chitosan can enhance rabbit platelet aggregation. In this study, the effects of chitosan on the subsequent growth factors release after human platelets activation were examined to evaluate the possibility of chitosan being used as a substitute for thrombin during PRP preparation. MATERIAL AND METHODS: Human platelet activation was determined by aggregation, adhesion and alpha-granule membrane glycoprotein expression. Platelet aggregation was measured by the turbidimetric method, the adhesion was directly examined on chitosan-coated glass plates under light microscope and scanning electron microscope (SEM), and the alpha-granule membrane glycoprotein was detected by fluorescent isothiocyanate (FITC)-conjugated anti-CD61 antibody through flow cytometry. The subsequent epidermal growth factor (EGF), platelet-derived growth factor (PDGF)-AB and transforming growth factor (TGF)-beta1 release from platelets were assayed by ELISA after mixing with chitosan. RESULTS: The enhancing effects on the platelet adhesion and the aggregation from chitosan were observed. Under both microscopes, the adhesive platelets on the chitosan-coated plates were not only greater in number but also earlier in activation than those on the control plates. With flow cytometry, increased glycoprotein IIIa expression in platelets was detected after chitosan treatment. Greater concentrations of growth factors were measured from PRP after chitosan treatment than after the solvent treatment. CONCLUSION: Because of the observations of growth factors releasing from activated human platelets after chitosan stimulation, we suggest that chitosan may be an appropriate substitute for thrombin in PRP preparation.

Biocompatible Materials↗

PubMed1997

Inhibition of Streptococcus mutans adsorption to hydroxyapatite by low-molecular-weight chitosans.

The role of Streptococcus mutans in the initiation of dental caries has been recognized and attributed, at least in part, to its ability to colonize the tooth surface. Therefore, factors which prevent S. mutans attachment to hydroxyapatite (HA) are of considerable interest for the prophylaxis of this infectious disease. Chitosan, a chitin derivative by N-deacetylation, is an interesting candidate in this respect, since it stimulates the ordered regeneration of oral soft tissues, prevents the deleterious action of organic acid, and exhibits bactericidal action against several pathogens. In the present work, the efficacy of a low-molecular-weight chitosan (LMWC) and its derivatives N-carboxymethyl chitosan (NCMC) and imidazolyl chitosan (IMIC) in preventing S. mutans attachment to HA beads was assessed. The effects of chitosan on both sucrose-dependent and -independent adherence were evaluated. In both cases, when saliva-coated or uncoated HA beads were treated with any of the chitosans, a reduction in S. mutans adsorption ranging from 47 to 66% was observed. When HA beads were coated with saliva after the treatment with chitosan, neither carbohydrate caused a statistically significant reduction in S. mutans adsorption, suggesting that saliva deposition restores HA binding properties. Bacteria grown in the presence of chitosan subminimal inhibitory concentrations (sub-MICs) ranging from 12 to 500 micrograms mL-1 adsorbed poorly to HA and exhibited a lower affinity toward xylene than untreated controls. In the presence of chitosan sub-MICs up to 60 micrograms mL-1, an increase in the percentage of detached bacteria from two- to nine-fold was observed. The desorptive effect of chitosan was weaker when S. mutans had adhered to saliva-coated HA in the presence of sucrose. These results demonstrate that the presence of minor amounts of modified chitosans prevents S. mutans adsorption to HA and suggest that colonization of the tooth surface might be impaired by the use of toothpastes, mouthrinses, or chewing gums containing any of the tested polysaccharides.

Adsorption↗