Protein trace present in chitin/chitosan product has been determined by our proposed method in which protein trace included in chitin/chitosan was extracted by refraction in 10% NaOH solution at 100oC for 5 hours. Chitin/Chitosan precipitate was discarded. protein trace in the solution was restored by neutralizing with HCl 10N to get the yellow powder. This powder was then analyzed for total nitrogen content by Kjeldahl method. The control method was conducted at the same procedure excepted from the presence of 10% NaOH. The results of different chitin/chitosan samples obtained from these two methods showed that amount of protein trace obtained from above proposed method was somewhat higher than that of the control one. Besides, the purified chitosan sample gave the lower protein trace content than that of the technical chitosan sample as well as chitin one. Protein trace bound between macromolecules of chitin/chitosan was separated in the proposed method by boiling water under alkaline condition whereas it was not completely separated in the control method because sodium hydroxide was absent.
Chitin ; Chitosan ; Proteins

This study is to compare the effect of wound healing using three different types of chitin, which include the shapes of sponge, velvet, thick non-woven fabrics, and thin non-woven fabrics. The sponge type had more capacity to absorb the first discharge of a wound than the velvet type and the two non-woven fabrics types. Instead of absorbing the discharge effectively, the velvet type showed a difficulty to take off the dressing stuff from a wound since it was solidly stuck to the wound. The sponge type showed less infiltration of inflammatory cells, producing angiogenesis and fibroblast faster than any other types. Next, the thick non-woven fabrics type was a little more effective than the thin non-woven fabrics type: However, there was no difference between two types. The velvet type sustained the infiltration of inflammatory cells for the longest duration, producing slower angiogenesis and fibroblast. In wound contraction and wound healing, the sponge type was most effective with statistical significance than any other types(p<0.05). The two types of non-woven fabrics did not present any statistical significance in wound contraction, wound epithelization and wound healing(p>0.05). In conclusion, the sponge type showed the best effectiveness to absorb the early discharge, facilitating the progress of inflammatory phase to increase the healing rate. It induced an early healing of wound caused by wound contraction rather than by wound epithelization.
Bandages ; Chitin ; Chitosan* ; Fibroblasts ; Porifera ; Wound Healing* ; Wounds and Injuries*

The effect of chitosan, a carbohydrate biopolymer extracted from chitin, on periodontal regeneration is of particular interest. The purpose of this study was to evaluate the effect of chitosan on primary rat calvarial cells in vitro, with special focus on their proliferative properties by cell activity and the amount of total protein synthesis. The experimental groups were cultured with chitosan in concentration of 0.01, 0.1, 1.0, 2.0 and 5.0 mg/ml for MTT assay. In the experimental groups, cells were cultured with chitosan in concentration of 0.01, 0.1, 1.0 and 2.0 mg/ml. Each group was characterized by examining alkaline phosphatase activity at 3 and 7 days and the ability to produce mineralized nodules of rat calvarial cells at 14 and 21 days. The results were as follows: 1. The cell activity was not reduced in the concentration of 0.01~1.0 mg/ml whereas the cell activity was reduced in the concentration of 5.0 mg/ml than the control at day 1 and 3 (p<0.05). 2. Primary rat calvarial cells treated with chitosan in the concentration 0.01 mg/ml and 0.1 mg/ml showed more protein synthesis than the control at day 3 (p<0.01). But primary rat calvarial cells treated with chitosan showed more protein synthesis than in control but they didn't have statistically difference among groups at day 7. 3. At 3 and 7 days, alkaline phosphatase activity was significantly increased in the concentration of 0.01 mg/ml. 0.1 mg/ml and 1.0 mg/ml (p<0.05). 4. The percentage of mineralized bone nodule was more in the concentration of chitosan 0.1 mg/ml and 1.0 mg/ml than the control. These results suggested that chitosan has a positive effect on the bone formation of primary rat calvarial cells in the concentration of 0.1 mg/ml and 1.0 mg/ml.
Alkaline Phosphatase ; Animals ; Biopolymers ; Chitin ; Chitosan* ; Osteogenesis ; Rats* ; Regeneration

Periodontal therapy has dealt primarily with attempts at arresting progression of disease, however, more recent techniques have focused on regenerating the periodontal ligament having the capacity to regenerate the periodontium. The effect of chitosan(poly-N-acetyl glucosaminoglycan), a carbohydrate biopolymer extracted from chitin, on periodontal ligament regeneration is of particular interest. The purpose of this study was to evaluate the effect of chitosan on the human periodontal ligament fibroblasts(hPDLFs) in vitro, with special focus on their proliferative properties by MTT assay, the synthesis of type I collagen by reverse transcription-polymerase chain reaction(RT-PCR) and the activity of alkaline phosphatase(ALP). Fibroblast populations were obtained from individuals with a healthy periodontium and cultured with alpha-MEM as the control group. The experimental groups were cultured with chitosan in concentration of 0.01, 0.1, 1, 2mg/ml. The results are as follows; 1. Chitosan-induced proliferative responses of hPDLFs reached a plateau at the concentration of 0.1mg/ml (p<0.05). 2. When hPDLFs were stimulated with 0.1mg/ml chitosan, mRNA expression of type I collagen was up-regulated. 3. When hPDLFs were stimulated with 0.1mg/ml chitosan, ALP activity was significantly upregulated(p<0.05). In summary, chitosan(0.1mg/ml) enhanced the type I collagen synthesis in the early stage, and afterwards, facilitated differentiation into osteogenic cells. The results of this in vitro experiment suggest that chitosan potentiates the differentiation of osteoprogenitor cells and may facilitate the formation of bone.
Biopolymers ; Chitin ; Chitosan* ; Collagen Type I ; Fibroblasts* ; Humans* ; Periodontal Ligament* ; Periodontium ; Regeneration ; RNA, Messenger

Polyurethane foam dressings are becoming widely used in partial and full thickness skin wounds since they are safe and convenient for both patients and physicians. However, the influence of polyurethane foam dressings on epithelialization has not been fully determined yet. This study was designed to compare 2 internationally available polyurethane foams and 3 locally made polyurethane foams as to their effects on the epithelialization in vivo. Twenty white rats were used. A 5 mm punch was employed to excise skin and subcutaneous tissue in a round fashion at six sites in the back area of 10 rats. After creating 6 open wounds, above mentioned 5 polyurethane foams and gauze dressings were applied. On the 5th day the lengths of the nonepithelialized area were measured under light microscope. In addition, partial thickness wounds were created using carbon dioxide laser at six sites on the back of the another 10 rats. Six dressings were applied on the wounds as same as above. On the 4th day the degree of the epithelialization was measured and compared. In full thickness wound study the best wound closing was also seen in chitin mixed Medifoam-N group. In partial thickness wound study the best epithelialization was seen in chitin mixed Medifoam-N and chitosan mixed Medifoam-N groups. These results demonstrated that polyurethane foam dressings influenced the epithelialization, and chitin mixed Medifoam-N showed the best result.
Animals ; Bandages* ; Chitin ; Chitosan ; Humans ; Lasers, Gas ; Polyurethanes* ; Rats* ; Skin ; Subcutaneous Tissue ; Wounds and Injuries

PURPOSE: Chitin and chitosan are nontoxic natural chelators that chelate radiostrontium effectively. The purpose of this study was to compare radiostrontium chelation of chitin and chitosan with that of well known chemical chelators, namely EDTA and DTPA. MATERIALS AND METHODS: The chelaton rates of chitin, chitosan, EDTA and DTPA were compared using a column chromatography method (Sephadex G-25M, Sweden). Three kinds of chitins and four kinds of chitosans were used. All of them were water soloble. RESULTS: Phosphated chitosan showed the highest chelation yield of more than 97% at pH 7. All of chitins, chitosans, EDTA and DTPA showed chelation yield of more than 90% independent of varing pH level. CONCLUSION: Chitin and chitosan have similar chelation rate as compared with EDTA and DTPA.
Chelating Agents ; Chitin* ; Chitosan* ; Chromatography ; Edetic Acid* ; Hydrogen-Ion Concentration ; Pentetic Acid*

OBJECTIVETo optimize the preparation of genistein chitosan microspheres with central composite design (CCD).

METHODThe chitosan microspheres were prepared by the O/W/O multiple emulsion method. Second-order polynomial and linear equations were fitted to the data, and the resulting equations were used to produce response surface graphs and the best experiment conditions.

RESULTThe theoretical drug content was 13%-15%, the concentration of organic phase was 30%-40% and the concentration of oil phase was 68%-72%.

CONCLUSIONThe best experiment conditions can be obtained by central composite design and response surface methodology. The observed values agree well with model predicted values.

Antineoplastic Agents ; administration & dosage ; Chitin ; analogs & derivatives ; Chitosan ; Delayed-Action Preparations ; Genistein ; administration & dosage ; Mathematics ; Microspheres

Chitosanases represent a class of glycoside hydrolases with high catalytic activity on chitosan but nearly no activity on chitin. Chitosanases can convert high molecular weight chitosan into functional chitooligosaccharides with low molecular weight. In recent years, remarkable progress has been made in the research on chitosanases. This review summarizes and discusses its biochemical properties, crystal structures, catalytic mechanisms, and protein engineering, highlighting the preparation of pure chitooligosaccharides by enzymatic hydrolysis. This review may advance the understandings on the mechanism of chitosanases and promote its industrial applications.
Chitosan/chemistry* ; Chitin ; Glycoside Hydrolases/genetics* ; Protein Engineering ; Oligosaccharides/chemistry* ; Hydrolysis

The biofilms formed by pathogenic microorganisms seriously threaten human health and significantly enhance drug resistance, which urgently call for developing drugs specifically targeting on biofilms. Chitooligosaccharides extracted from shrimp and crab shells are natural alkaline oligosaccharides with excellent antibacterial effects. Nevertheless, their inhibition efficacy on biofilms still needs to be improved. Spirulina (SP) is a microalga with negatively charged surface, and its spiral structure facilitates colonization in the depth of the biofilm. Therefore, the complex of Spirulina and chitooligosaccharides may play a synergistic role in killing pathogens in the depth of biofilm. This research first screened chitooligosaccharides with significant bactericidal effects. Subsequently, Spirulina@Chitooligosaccharides (SP@COS complex was prepared by combining chitooligosaccharides with Spirulina through electrostatic adsorption. The binding of the complex was characterized by zeta potential, z-average size, and fluorescence labeling. Ultraviolet-visible spectroscopy (UV-Vis) showed the encapsulation efficiency and the drug loading efficiency reached up to 90% and 16%, respectively. The prepared SP@COS2 exhibited a profound synergistic inhibition effect on bacterial and fungal biofilms, which was mainly achieved by destroying the cell structure of the biofilm. These results demonstrate the potential of Spirulina-chitooligosaccharides complex as a biofilm inhibitor and provide a new idea for addressing the harm of pathogenic microorganisms.
Humans ; Spirulina ; Anti-Bacterial Agents/chemistry* ; Chitosan/pharmacology* ; Biofilms ; Chitin/pharmacology*

Chitin is a natural product while chitosan is synthetic product from chitin. A study on the activity of chitin and chitosan in dogs has shown that the use of intradermal chitosan at dose of 200 mg/kg killed all experimental dogs because of pulmonary hemorrhage while this dose of chitin did not kill any dogs. Until now, it was limited to use chitin and chitosan in human as a food complementary, anti obesity and scar healing.
chotosan ; chitin ; toxicity