Different Molecular Weights Of Chitosan Research Articles

Synthesis, characterization, and controlled release of selenium nanoparticles stabilized by chitosan of different molecular weights

Chitosan-stabilized selenium nanoparticles (SeNPs) have been reported, but there is no information on the effect of the chitosan molecular weight on the structure, stability, and selenium release properties of the SeNPs. Herein, we compared the uniform Se0 spherical nanoparticles prepared through the reduction of seleninic acid with ascorbic acid in the presence of chitosan with different molecular weights (Mws). We found that both low and high molecular weight chitosan-stabilized selenium nanoparticles exhibited core–shell microstructures with a size of about 103nm after 30 days growing through the “bottom–up approach” and “top–down approach,” respectively. Moreover, both chitosan SeNPs processed excellent stability towards pH and enzyme treatment. In contrast, selenium was easily released to different extents from these two chitosan SeNPs upon treatment with different free radicals. This makes these materials potentially useful as oral antioxidant supplements.

Development and characterisation of antibacterial suture functionalised with N-halamines

Braided and biodegradable polyglycolide suture was antibacterially functionalised with N-halamines via layer-by-layer assembly technique. Multilayers of chitosan (polycation) and poly-sodium-p-styrenesulfonate (polyanion) were successfully coated via electrostatic assembly, followed by top layer of chitosan on polyglycolide suture. Upon chlorination of coated suture with dilute sodium hypochlorite solution, the amino groups of chitosan were transformed into N-halamine structures. The transformation was assessed by iodometric/thiosulfate titration, Fourier transform infrared spectroscopy and differential scanning calorimeter analysis. The surface morphology of coated suture was observed by scanning electron microscope and atomic force microscope. Chlorine loading, antibacterial efficacy and tensile strength of chlorinated sutures treated with two different molecular weights of chitosan were compared and evaluated. A general trend of linear increase in chlorine loadings of sutures with the increase in number of layers and solution concentration was found. The chlorinated suture with high molecular weight chitosan coating completely inactivated both Escherichia coli O157:H7 and Staphylococcus aureus bacteria within 15 min of contact time. The 3T3 mouse fibroblasts in vitro cell cytocompatibility studies demonstrated that antibacterial sutures have fairly good biocompatibility.

Chitosan-mediated non-viral gene delivery with improved serum stability and reduced cytotoxicity

Polyethyleneimine (PEI) has been widely used in gene delivery systems because of its outstanding ability to promote DNA condensation and endosome escape. However, its use for the introduction of foreign DNA into host cells is compromised by high cytotoxicity and serum reactivity. In this study we examined the possibility of improving the PEI-mediated gene delivery using the serum-stable and biodegradable polymer, chitosan. Among chitosans of different molecular weights, the 45 kD form exhibited significantly higher gene delivery efficiency than the 15 and 100 kD forms. Using the 45 kDa form of chitosan, we formulated composite PEI/DNA/chitosan (PDC) complexes consisting of the core part where DNA chains were tightly condensed by PEI and the outer layer lined with chitosan, and found that an optimal PDC complex exhibits a significantly higher transfection efficiency in a high serum condition and a lower toxicity than monolithic PEI/DNA (PD) complex. Moreover, addition of chitosan to PEI/DNA complex reduced nonspecific interactions with erythrocytes, which may eventually contribute to improved transfection efficiency in high serum concentrations. These results demonstrate that chitosan coating of PD complex might increase the efficiency of PEI-mediated gene delivery in vivo as well as in vitro.

Effect of molecular weight of chitosan on antimicrobial properties and tissue compatibility of chitosan-impregnated bacterial cellulose films

Bacterial cellulose-chitosan (BC-C) films were developed by immersing purified BC pellicles in 1.5 ~ 2.0% (w/v) acetic acid solutions containing chitosan of varying molecular weights. Effects of different molecular weight of chitosan on physical, biological and antimicrobial properties of the composite films were investigated. The cumulative chitosan absorption capacities with Mw of 141,000, 199,000, and 263,000 were 38.43, 24.65, and 23.89 mg/cm3 of dry BC film, respectively. The cumulative release profiles of chitosan from the films strongly depended on molecular weight of chitosan and pH of solution. The order of release of chitosan from the BC-C films was dependent on molecular weight as follows: Mw 141,000 > Mw 199,000 > Mw 263,000. All BC-C films showed the antimicrobial abilities against Staphylococcus aureus and Aspergillus niger but had no inhibitory effect on the growth of Escherichia coli. The BC-C films supported for adhesion, spreading and proliferation of both human skin keratinocytes and fibroblasts. The antibacterial activity against S. aureus of the BC-C with the highest Mw chitosan (263,000) was higher than those of the others. On the other hand, the BC-C films with the lowest Mw chitosan (141,000) promoted the growth of human skin cells more than those of the others.

Preparation of chitosan nanoparticles by spray drying, and their antibacterial activity

Chitosan nanoparticles were prepared from chitosan of different molecular weight by spray drying. The morphology of the particles was characterized by SEM, and size distribution and zeta potential were determined. The effects of chitosan solution concentration, molecular weight of chitosan, and size of the spray dryer nozzles on average size, size distribution and zeta potential of chitosan nanoparticles were investigated. The effects of chitosan nanoparticles and chitosan nanoparticles–amoxicillin complex on Staphylococcus aureus were also tested. The results showed that the average size of chitosan nanoparticles were in the range 95.5–395 nm and zeta potentials were 39.3–45.7 mV, depending on the concentration and molecular weight of the chitosan. The lower the concentration and molecular weight of the chitosan, the smaller the chitosan nanoparticles and the higher the zeta potential. Testing for antibacterial activity against S. aureus indicated that chitosan nanoparticles strongly inhibited growth of the bacteria; the minimum inhibitory concentration, 20 μg/mL, was lower than those of chitosan solution or amoxicillin. The antibacterial capacity of chitosan nanoparticles also depended on the size, zeta potential, and molecular weight of the chitosan. Complexation of chitosan nanoparticles with amoxicillin improved the antibacterial activity of amoxicillin.

Study on Different Molecular Weights of Chitosan as an Immobilization Matrix for a Glucose Biosensor

Two chitosan samples (medium molecular weight (MMCHI) and low molecular weight (LMCHI)) were investigated as an enzyme immobilization matrix for the fabrication of a glucose biosensor. Chitosan membranes prepared from acetic acid were flexible, transparent, smooth and quick-drying. The FTIR spectra showed the existence of intermolecular interactions between chitosan and glucose oxidase (GOD). Higher catalytic activities were observed on for GOD-MMCHI than GOD-LMCHI and for those crosslinked with glutaraldehyde than using the adsorption technique. Enzyme loading greater than 0.6 mg decreased the activity. Under optimum conditions (pH 6.0, 35°C and applied potential of 0.6 V) response times of 85 s and 65 s were observed for medium molecular weight chitosan glucose biosensor (GOD-MMCHI/PT) and low molecular weight chitosan glucose biosensor (GOD-LMCHI/PT), respectively. The apparent Michaelis-Menten constant () was found to be 12.737 mM for GOD-MMCHI/PT and 17.692 mM for GOD-LMCHI/PT. This indicated that GOD-MMCHI/PT had greater affinity for the enzyme. Moreover, GOD-MMCHI/PT showed higher sensitivity (52.3666 nA/mM glucose) when compared with GOD-LMCHI/PT (9.8579 nA/mM glucose) at S/N>3. Better repeatability and reproducibility were achieved with GOD-MMCHI/PT than GOD-LMCHI/PT regarding glucose measurement. GOD-MMCHI/PT was found to give the highest enzymatic activity among the electrodes under investigation. The extent of interference encountered by GOD-MMCHI/PT and GOD-LMCHI/PT was not significantly different. Although the Nafion coated biosensor significantly reduced the signal due to the interferents under study, it also significantly reduced the response to glucose. The performance of the biosensors in the determination of glucose in rat serum was evaluated. Comparatively better accuracy and recovery results were obtained for GOD-MMCHI/PT. Hence, GOD-MMCHI/PT showed a better performance when compared with GOD-LMCHI/PT. In conclusion, chitosan membranes shave the potential to be a suitable matrix for the development of glucose biosensors.

Insecticidal activity of chitosans of different molecular weights and chitosan-metal complexes against cotton leafworm Spodoptera littoralis and oleander aphid Aphis nerii

As an alternative to synthetic pesticides, chitosan has received much attention as a biopolymer active against some agricultural pests. The, insecticidal activity of chitosans of four molecular weights (2.27 × 10<sup>5</sup>,3.60 × 10<sup>5</sup>,5.97 × 10<sup>5</sup>,and9.47 × 10<sup>5 </sup>g/mol) was investigated against two species of arthropod pests: oleander aphid Aphis nerii and cotton leafworm Spodoptera littoralis. In addition, the most active chitosan of 2.27 × 10<sup>5 </sup>g/mol was chemically modified with metals of Ag(I), Cu(II), Ni(II), and Hg(II) to give corresponding chitosan-metal complexes. Larval mortality, growth inhibition, and antifeedant activities for third instar larvae of S. littoralis were evaluated at 4 g (a.i.) chitosan/kg diet. Chitosan of 2.27 × 10<sup>5 </sup>g/mol and its complexes with Ni and Hg were the most active compounds. The results against A. nerii indicated that chitosans of 3.60 × 10<sup>5</sup> and 5.97 × 10<sup>5 </sup>g/mol showed high activity among the different molecular weights in leaf-dip bioassay after 24 h of treatment with 48 and 49% mortalities, respectively, at 1000 mg/l. All compounds had a systemic effect against A. nerii. Chitosans of 2.27 × 10<sup>5</sup>,3.60 × 10<sup>5</sup>,and5.97 × 10<sup>5 </sup>g/mol showed the highest efficacy at all concentrations tested; however, chitosan-Cu was significantly the most active among the complexes.

The preparation of chitosan nanoparticles by wet media milling

SummaryThree different molecular weights of chitosan were pulverised to nanoparticles by wet media milling. The effects of the milling on properties of chitosan, such as viscosity average molecular weight (Mv), ash content, the degree of deacetylation (DD), particle size, chemical structure, colour, swelling power and apparent viscosity, were investigated. Our results showed that high (578.3 kDa), medium (181.6 kDa) and low (66.3 kDa) molecular weight chitosan with particle size of 342.7, 358.1 and 346.2 μm were milled to nanometre range after 1, 1.5 and 2.5 h of milling, respectively. After 5 h of milling, the particle sizes of the three samples were decreased to 543.8, 366.6 and 308.4 nm, respectively. Chitosan did not show significant increase in ash contents after milling. The milling decreased the Mv of chitosan, but did not change DD and the chemical structure. The suspension of chitosan nanoparticles was of cream‐white colour. The swelling powder and apparent viscosity of chitosan increased after the milling treatment.

Assessment of the effect of chitosan of different molecular weights in controlling Rhizopus rots in tomato fruits

The effect of chitosan was evaluated in three isolates of Rhizopus stolonifer from infected tomatoes that were harvested in different regions of the state of Morelos, México. Changes in the membrane integrity of spores, modifications in pH media and the proteins released were analysed in each isolate against three different molecular weights of chitosan at three fixed concentrations. There was an observed decline in the integrity of spores in the presence of chitosan at all the concentrations evaluated. The protein released was different depending on the isolate, kind of chitosan and used concentration. There was a significant difference in the pH changes on the growth media for all the tested isolates. Chitosan of high molecular weight showed the best results to inhibit the infection caused by R. stolonifer on the tomato fruits. The severity of symptoms of soft rot was not related with the molecular weight of chitosan.

Differential blood lipid-lowering effects of alkylsulfonated chitosan of different molecular weights in Syrian hamsters in vivo

This study investigated the effects of alkylsulfonated chitosan of different molecular weights on intestinal lipid absorption, blood lipid profiles and circulating adhesion molecules. Syrian hamsters were fed an AIN-93G-based high‑fat diet (HFD) and were orally administered 5 or 10 mg/kg BW of oligomer (6 kDa) chitosan (OC), low‑molecular-weight (70 kDa) chitosan (LMC) or high‑molecular-weight (200 kDa) chitosan (HMC) four times per week for 12 weeks. Animals receiving 2.5 mg/kg BW lovastatin (LOVA) served as a positive control. The blood lipid profiles of these control animals revealed that all chitosans and LOVA significantly decreased total triglyceride, total cholesterol, low‑density lipoprotein (LDL)-cholesterol and very‑low‑density lipoprotein (VLDL)-cholesterol levels in a dose‑dependent manner compared to the HFD-fed controls (P<0.05). The blood lipid lowering effectiveness of the three chitosans followed the order of LMC>OC>HMC. Hamsters receiving 5 and 10 mg/kg LMC (P<0.05) exhibited an increase in fecal fat content. Immunoblot assay revealed that acyl‑coenzyme A:cholesterol acyltransferase-2 (ACAT-2) expression was suppressed in all chitosan-fed animals compared to the HFD-fed controls (P<0.05). These results suggest that chitosan effectively decreases blood lipid content, and its effectiveness depends on the molecular size of chitosan. The hypolipidemic effect of chitosan is partly attributed to its suppression of intestinal lipid absorption and hepatic ACAT-2 expression.

Phosphorylatable short peptide conjugated low molecular weight chitosan for efficient siRNA delivery and target gene silencing

Small interfering RNA (siRNA) has been widely investigated as a potential therapeutic approach for diseases with genetic defects. However, its application was greatly hampered by the rapid degradation and poor cellular uptake. Recently, chitosan (CS) and its derivant have been considered as a promising siRNA transporter with the advantages of low toxicity, good biodegradability and biocompatibility. Chitosan of different molecular weight (Mw) and degrees of deacetylation (DD) showed significantly varied target gene silencing efficacy, and it is still not well clarified how these characteristics influence CS mediated siRNA transfection. To compare the aspects of cell permeability and intracellular unpacking of CS/siRNA complex on the effect of CS/siRNA transfection. A radiolabeled siRNA, targeting firefly luciferase gene, was loaded by chitosan of different molecular weight (varying from 2000 to 800,000 Da) and subjected to the transfection against MDA-MB-231/Luc human breast cancer cell line which stably expressed knocked in firefly Luciferase reporter gene. Following transfection intracellular radioactivity was measured to represent cell entrance ability of the CS/siRNA, while, luciferase activity in the cell lysate was also determined to reflect target gene silencing effect. The results revealed that although low molecular weight chitosan (LMWC) condensed siRNA has the highest cell permeability of almost two folds of medium molecular weight chitosan and lipofactamine, its target gene silencing effect is really low of almost eight times less than lipofactamine. This conspicuous contradiction gave us the hypothesis that LMWC generated more condensed CS/siRNA complex to facilitate cell entrance but the tight electrostatic interaction probably limited intracellular siRNA unpacking as well and unfavorably hindered target gene silencing as the final consequence. To approve this hypothesis a phosphorylatable short peptide conjugated LMWC was adopt to promote intracellular siRNA unpacking. Which was demonstrated of perfect target gene knock down ability to the extent of being even superior to lipofactamine 2000. In a conclusion, low molecular weight chitosan has the great potential to be an ideal siRNA vehicle if the issue of siRNA unpacking could be properly resolved.

Novel hyaluronic acid–chitosan nanoparticles as non-viral gene delivery vectors targeting osteoarthritis

Gene therapy is a promising new treatment strategy for common joint-disorders such as osteoarthritis. The development of safe, effective, targeted non-viral gene carriers is important for the clinical success of gene therapy. The present work describes the use of hybrid hyaluronic acid (HA)/chitosan (CS) nanoparticles as novel non-viral gene delivery vectors capable of transferring exogenous genes into primary chondrocytes for the treatment of joint diseases. HA/CS plasmid–DNA nanoparticles were synthesized through the complex coacervation of the cationic polymers with pEGFP. Particle size and zeta potential were related to the weight ratio of CS to HA, where increases in nanoparticle size and decreases in surface charge were observed as HA content increased. The particle size and the zeta potential varied according to pH. Transfection of primary chondrocytes was performed under different conditions to examine variations in the pH of the transfection medium, different N/P ratios, different plasmid concentrations, and different molecular weights of chitosan. Transfection efficiency was maximized for a medium pH of approximately 6.8, an N/P ratio of 5, plasmid concentration of 4 μg/ml, and a chitosan molecular weight of 50 kDa. The transfection efficiency of HA/CS-plasmid nanoparticles was significantly higher than that of CS-plasmid nanoparticles under the same conditions. The average viability of cells transfected with HA/CS-plasmid nanoparticles was over 90%. These results suggest that HA/CS-plasmid nanoparticles could be an effective non-viral vector suitable for gene delivery to chondrocytes.

Physicochemical and structural characteristics of chitosan nanopowders prepared by ultrafine milling

Two different molecular weights of chitosan were pulverized to nanopowders by ultrafine milling. The nanopowders were characterized by viscometry small angle X-ray scattering (SAXS), transmission electron microscopy (TEM), X-ray diffraction (XRD), thermogravimetric analysis (TGA), FT-IR spectroscopy and UV-vis spectroscopy. Our results showed that ultrafine milling effectively reduced the particle size of chitosan to a nanoscale. The viscosity average molecular weight (Mv) of chitosan was decreased by the milling treatment. The crystalline structure of chitosan was destroyed by the milling since the nanopowder exhibited an amorphous XRD pattern. In addition, thermal stability of the low molecular weight chitosan was decreased after the milling treatment. FT-IR and UV-vis spectra showed that the milling process did not cause significant changes in the chemical structure of chitosan.

Chitosan as a flocculant for pre-hydrolysis liquor of kraft-based dissolving pulp production process

In this work, the flocculation concept was employed to recover the dissolved lignocellulosic materials of industrially produced pre-hydrolysis liquor (PHL) using two different molecular weights of chitosan. The analysis revealed the maximum turbidity of 1000NTU or 1900NTU via adding 2.2mg/g low molecular weight (LMW) or 1.7mg/g high molecular weight (HMW) chitosan to the PHL, respectively. Also, the maximum furfural removal of 55% or 50% was obtained by adding 0.7mg/g or 0.5mg/g LMW or HMW chitosan to the PHL, respectively. The maximum recovery of oligomeric sugars was 20% or 25% by adding 1.5 or 0.5mg/g LMW or HMW chitosan, while that of lignin was 40% or 35% by applying 2.2mg/g or 1.7mg/g LMW or HMW chitosan to the PHL, respectively. The removal of monomeric sugars and acetic acid was rather limited. The particle size and nitrogen analyses were also hired to characterize the properties of the formed complexes.

Study of Flocs Morphology of Chitosan Coagulation Aid Treatment and Fractal Dimensions

Flocculation water treatment is one of the important aspects of water treatment .The aim of this work is to use PFS for treating Yellow River surface water and different molecular weights of chitosan as coagulant aid. Electron microscope was used to observe flocs morphology and study CTS coagulation aid morphological change, floc image with microscope camera system analysis, calculation of floc fractal dimension. The results show that the different molecular weights of CTS in dealing with surface raw water to play a good coagulation aid effect. Microscope and image analysis system of flocs morphology show that fractal dimension of morphology of PFS / CTS were significantly higher ,Among them，high molecular weight of CTS as coagulant aid，and fractal dimension of morphology of PFS/ CTS were 1.1026,1.5593 , low molecular weight of CTS as coagulant aid，and fractal dimension of morphology of PFS/ CTS were 1.0904,1.5440 respectively. Floc fractal dimension changes to better reflect the flocculation level and treatment effect for the establishment of floc fractal dimension number of changes in the coagulation control is provided for reference.

Chitosan physico–chemical properties modulate defense responses and resistance in tobacco plants against the oomycete Phytophthora nicotianae

Enzymatic defense responses and protection against Phytophthora nicotianae were studied in tobacco plants treated with chitosan of different molecular weights (MW) and degrees of acetylation (DA). The concentration and mode of chitosan application affected enzymatic induction in tobacco leaves. β-1,3-Glucanase (EC 3.2.1.6) activity required 10 times the polymer concentration relative to the oligochitosan mixture to induce the highest activity above control when treated by foliar spray, indicating the influence of molecular weight in this response. PAL (EC 4.3.1.5) and POD (EC 1.11.1.6) activities increased above control as a result of the influence of polymer degree of acetylation when treatments were applied by foliar spray. A higher DA favored PAL activity, whereas a lower DA induced higher POD activity. Using an in vitro bioassay, it was found that the three chitosan compounds caused a reduction of the infection index of P. nicotianae in tobacco plants that was dependent on elicitor concentrations. There was a significant relationship between the reduction of the infection index and an increase in PAL activity when chitosan was applied by foliar spray and by substrate drench to the plant rhizosphere. These results demonstrate the influence of chitosan physico–chemical properties in plant-induced resistance and the relevance of particular responses in plant protection against pathogens.

The Effect of Polymer Molecular Weight on Citrate Crosslinked Chitosan Films for Site-Specific Delivery of a Non-Polar Drug

Purpose: To develop citrate crosslinked chitosan films using chitosan of different molecular weights (MW) in order to achieve site-specific delivery of a model non-polar drug, indomethacin.Methods: Films prepared with different molecular weights of chitosan and incorporating indomethacin as a non-polar model drug were obtained by a casting/solvent evaporation method. The chitosan films were crosslinked by dipping in varying concentrations of sodium citrate solution and for different crosslinking times. The films were assessed by, amongst others, scanning electron microscopy (SEM), dissolution studies and differential scanning calorimetry (DSC) for surface morphology, drug release and ingredient compatibility, respectively.Results: Crosslinking time and concentration of crosslinking agent significantly (p < 0.05) influenced the in vitro release of indomethacin as well as swelling of the films. Also, the higher the molecular weight (MW) of chitosan the lower the drug release rate (p < 0.05). Furthermore, film swelling index rose as chitosan MW decreased (p < 0.05). The practical absence of the sharp endothermic peak characteristic of indomethacin in the films suggests that crosslinking may have transformed the drug from the crystalline to the amorphous state.Conclusion: The citrate-crosslinked chitosan films can be modulated to vary swelling and drug release at pH 3.5 and 6.2; this feature makes them useful tools for designing site-specific delivery systems.Keywords: Chitosan film, Sodium citrate, Site-specific delivery, Crosslinking, IndomethacinTropical Journal of Pharmaceutical Research December 2010; 9 (6): 525-531

Stabilisation of SWNTs by alkyl-sulfate chitosan derivatives of different molecular weight: towards the preparation of hybrids with anticoagulant properties

We have previously demonstrated that chitosan derivative N-octyl-O-sulfate chitosan (NOSC), which presents important pharmacological properties, can suspend single walled carbon nanotubes (SWNTs) up to 20 times more effectively than other chitosan derivatives in an aqueous environment. In an attempt to further investigate the impact of different molecular weights of chitosan to the solubilization and anticoagulant properties of these hybrids an array of NOSC derivatives varying their molecular weight (low, medium and high respectively) was synthesised and characterised by means of FT-IR spectroscopy, NMR spectroscopy and thermal gravimetric analysis (TGA). Microwave and nitric acid purified SWNTs, characterised by FT-IR spectroscopy, transmission electron microscopy (TEM) and Raman spectroscopy, were colloidally stabilised by these polymers and their anticoagulant activity was assessed. The results revealed that the low molecular weight NOSC coated SWNTs exhibit the highest activity when 0.5 mg mL(-1) NOSC solutions are used, activity which is similar to that of the free polymer. Preliminary studies by exposure of these hybrids to Brine Shrimp (Artemia) cysts revealed no effect on the viability of sub-adult Artemia. Our findings suggest the possibility of tailoring these nanomaterials to bear the required properties for application as biocompatible building blocks for nanodevices including biosensors and biomaterials.

Preparation, charactarization and anti-inflammatory activity of celecoxib chitosan gel formulations

This study was designed to evaluate the suitability of chitosan polymer as a vehicle for topical delivery system. Celecoxib, which is a nonsteroidal anti-inflammatory drug, was incorporated into the gel vehicles in a concentration of 0.5 % w/v. Gels were prepared using three different concentrations and different molecular weights of chitosan. Viscosity, drug release from gels, permeation of drug through rat skin and anti-inflammatory activity of the drug were studied. In vitro release characteristics of the drug from different gels were carried out using dialysis membrane in phosphate buffer using a pH of 6.8. The results showed that, the gel form containing 1.0 % w/v medium molecular weight chitosan has superior drug release than other forms, whilst the gel form containing 2.0 % w/v high molecular weight chitosan shows the lowest amount of drug release. The release data were treated with various kinetic principles to assess the relevant parameters. The results revealed an inverse correlation between the percent drug release and the polymer concentration used. The results also showed that the release of drug from the prepared gels obeyed the Higuchi’s diffusion model. The permeation of drug through rat skin was carried out. The flux of drug is independent on the viscosity of the formulae. The anti-inflammatory activity of the drug in different gel formulations was studied using carrageenan-induced rat paw edema method. The results obtained show that there is excellent anti-inflammatory activity of the gel forms on rat paw edema.

The osteoinductive effect of chitosan-collagen composites around pure titanium implant surfaces in rats

The enhancing effects of chitosan on activation of platelets and differentiation of osteoprogenitor cells have been demonstrated in vitro. The purpose of this study was to evaluate the in vivo osteoinductive effect of chitosan-collagen composites around pure titanium implant surfaces. Chitosan-collagen composites containing chitosan of different molecular weights (450 and 750 kDa) were wrapped onto titanium implants and embedded into the subcutaneous area on the back of 15 Sprague-Dawley rats. The control consisted of implants wrapped with plain collagen type I membranes. Implants and surrounding tissues were retrieved 6 wks after surgery and identified by Alizarin red and Alcian blue whole mount staining. The newly formed structures in the test groups were further analyzed by Toluidine blue and Masson-Goldner trichrome staining, and immunohistochemical staining with osteopontin and alkaline phosphotase. The bone formation parameters of the new bone in the two test groups were measured and compared. New bone formed ectopically in both chitosan-collagen groups, whereas no bone induction occurred in the negative control group. These newly formed bone-like structures were further confirmed by immunohistochemical staining. Comparison of bone parameters of the newly induced bone revealed no statistically significant differences between the 450 and 750 kDa chitosan-collagen groups. Our results demonstrated that chitosan-collagen composites might induce in vivo new bone formation around pure titanium implant surfaces. Different molecular weights of chitosan did not show significantly different effects on the osteoinductive potential of the test materials.