An clinical and pharmacokinetic study for a drug delivery system (DDS) of gentamycin-loaded chitosan bar were carried out with the purpose to evaluate its efficacy and giving further data for its clinical applications. Eighteen cases of chronic osteomyelitis were treated by surgical necrectomy with implantation of gentamycin-load chitosan bar in the prepared bone cavity. After operation, the concentration of gentamycin in serum and wound drainage fluid were examined at different times and blood urea nitrogen (BUN) and serum creatinine (Cr) as well. The clinical results were evaluated by the conditions of wound healing and clinical and roentgenographic manifestations. The results showed that the serum gentamycin concentration reached its peak level (0.86 microgram/ml) at 24 hours after operation and lasted for 4 days. No increase in the concentrations of BUN and Cr were observed after implantation. The gentamycin concentration in wound drainage fluid was several hundred times higher than the minimum inhibitory concentration (MIC) for staphylococcus aureus. All of the 18 cases were followed up for 24.8 months (in an range of 6-34 months) 16 patients received initial cure and without any recurrence. So, it could be concluded that the gentamycin-loaded chitosan DDS was a simple and effective method for the treatment of chronic osteomylitis without the necessity to carry out a second operation to remove the drug carrier, and it was sound to popularize its clinical application.
Objective To investigate the behavior of rat calvarial osteoblasts cultured on chitosan-gelatin/hydroxyapatite (CSGel/HA) composite scaffolds. Methods The rat calvarial osteoblasts (the 3rd passage) were seeded at a density of 1.01×106 cells/ml onto the CS-Gel/HA composite scaffolds having porosity 85.20%, 90.40% and 95.80%. Cell number was counted after cultured for 3 days,1 week, 2 weeks and 3 weeks. Cell proliferation, bone-like tissue formation, and mineralization were separately detected by HE, von Kossa histological stainingtechniques. Results The CS-Gel/HA composite scaffolds supported the attachmentof seeded rat calvarial osteoblasts. Cells proliferated faster in scaffold withhigher porosity 90.40% and 95.80% than scaffold with lower porosity 85.20%. The osteoblasts/scaffold constructs were feasible for mineral deposition, and bonelike tissue formation in 3 weeks. Conclusion This study suggests the feasibility of using CS-Gel/HA composite scaffolds for bone tissue engineering.
Objective To improve the flexibil ity and hemostatic properties of chitosan (CS)/carboxymethyl chitosan (CMCS) hemostatic membrane by using glycerol and etamsylate to modify CS/CMCS hemostatic membrane. To investigate themechanical properties and hemostatic capabil ity of modified CS/CMCS hemostatic membrane. Methods The 2% CS solution, 2% CMCS solution, 10%, 15%, 20%, 25%, 30% glycerol with or without 0.5% etamsylate were used to prepare CS/CMCS hemostatic membrane with or without etamsylate by solution casting according to ratio of 16 ∶ 4 ∶ 5. The tensile properties were evaluated by tensile test according to GB 13022-1991. Twenty venous incisions and five arterial incisions hemorrhage of 1 cm × 1 cm in rabbit ears were treated by CS/CMCS hemostatic membrane modified by 15% (group A) and 25% (group B) of glycerol, and a combination of them and 0.5% etamsylate (groups C and D). The bleeding time and blood loss were recorded. Results The pH of yellow CS/ CMCS hemostatic membrane with thickness of 30-50 μm was 3-4. The incorporation glycerol into CS/CMCS hemostatic membrane resulted in decreasing in tensile strength (7.6%-60.2%) and modulus (97%-99%). However, elongation at break and water content increased 5.7-11.6 times and 13%-125% markedly. CS/CMCS hemostatic membrane adhered to wound rapidly, absorbed water from blood and became curly. The bleeding time and blood loss of venous incisions were (70 ± 3) seconds and (117.2 ± 10.8) mg, (120 ± 10) seconds and (121.2 ± 8.3) mg, (52 ± 4) seconds and (98.8 ± 5.5) mg, and (63 ± 3) seconds and (90.3 ± 7.1) mg in groups A, B, C, and D, respectively; showing significant differences (P lt; 0.05) between groups A, B and groups C, D. The bleeding time and blood loss of arterial incision were (123 ± 10) seconds and (453.3 ± 30.0) mg in group C. Conclusion CS/CMCS hemostatic membrane modified by glycerol and etamsylate can improve the flexibil ity, and shorten the bleeding time.
Objective To study hemostasis of a new chitosan hemostatic powder. Methods Twenty-four adult SD rats were made the models of l iver injury, male or female, and weighing 210-240 g. They were divided into three groups randomly (n=8) depending on different hemostatic powders. The incision of the l iver was treated with 300 mg Yunnan baiyao (group A1), chitosan hemostatic powder of pH6.5 (group B1) and pH7.5 (group C1), respectively. The bleeding time and bleeding amount were recorded. In vitro, with the modified Ree-White method, 2 mL artery blood from New Zealand whiterabbit was added into the 0.2 mL solution of Yunnan baiyao, chitosan hemostatic powder of pH6.5 and pH7.5 (concentration of 0.2 mg/mL), respectively. The blood coagulation time was recorded. The chitosan blood clots of group B2 and group C2 were observed with scanning electron microscopy (SEM). Results The bleeding time of group A1, group B1 and group C1 was (292 ± 31), (261 ± 23), and (224 ± 28) s, respectively, the bleeding amount was (1.63 ± 0.21), (1.47 ± 0.18), and (1.18 ± 0.17) g, respectively, showing statistically significant differences between groups B1, C1, and group A1 (P lt; 0.05), between group C1 and group B1 (P lt; 0.05). The blood clotting time of group A2, group B2, and group C2 was (653 ± 41), (255 ± 20), and (202 ± 11) s, respectively, showing statistically significant differences between groups B2, C2, and group A2 (P lt; 0.05), between group C2 and group B2 (P lt; 0.05). The SEM showed that the blood cells of group B2 and group C2 gathered around the chitosan. Conclusion Chitosan hemostatic powder of pH7.5 has good hemostasis.
OBJECTIVE: To investigate the effects of chitosan membrane on tendon adhesion and healing and obtain experimental data for clinical use in preventing postoperative tendon adhesion. METHODS: The long flexor tendon of 55 adult legborn hens were cut and sutured encapsulated by chitosan membrane. Movement and anti-tension capability of tendon were assessed at 2, 4, 6, 8 and 10 weeks after operation by SWD-10 type tendon stretcher. Tendon healing and adhesion were observed with light microscope. RESULTS: Tendon healing could be effected by chitosan membrane within 4 weeks, and tendon anti-tension strength was increased after 4 weeks. CONCLUSION: Chitiosan membrane possesses the following characteristics: no side effects, good permeability, resolvable, absorbable and selective inhibition the growth of fibroblast. It is a desirable biological material to prevent tendon adhesion.
ObjectiveTo study the ectopic osteogenesis and biocompatibility of bone morphogenetic protein 2 (BMP-2)-derived peptide P24 loaded chitosan-4-thio-butylamidine (CS-TBA) hydrogel.MethodsFirst, the CS-TBA/hydroxyapatite (HA) solution was prepared by using chitosan, 2-iminothiolane hydrochloride, and HA. Then, the different amount of P24 peptides were added to the CS-TBA/HA to prepare the CS-TBA/5%P24/HA and CS-TBA/10%P24/HA solutions. Finally, β-glycerophosphate disodium (β-GP) was added to the CS-TBA/HA, CS-TBA/5%P24/HA, and CS-TBA/10%P24/HA to prepare the CS-TBA/HA/β-GP, CS-TBA/5%P24/HA/β-GP, and CS-TBA/10%P24/HA/β-GP hydrogels, respectively. Eighteen Sprague Dawley female rats were randomly divided into 3 groups (n=6), which were injected into the back muscle pouches with equal volume CS-TBA/HA/β-GP hydrogel (group A), CS-TBA/5%P24/HA/β-GP hydrogel (group B), and CS-TBA/10%P24/HA/β-GP hydrogel (group C). The animals were sacrificed at 4 and 8 weeks and conducted micro-CT. The ability of biodegradation and osteogenesis of hydrogl was detected by trabecular thickness (Tb.Th), trabecular number (Tb.N), bone mineral density (BMD), and histological staining (HE and Masson).ResultsAll the rats survived to the time point of the harvest. Micro-CT results showed that the new bones gradually increased in each group after operation. At the same time, the new bone formation was more obvious in groups B and C than in group A, and with the increase of P24 concentration, new bone formation in group C was much more than that in group B. The Tb.Th, Tb.N, and BMD increased gradually in 3 groups, and the differences between 4 and 8 weeks were significant (P<0.05) except the Tb.Th in group A. At different time points, the Tb.Th, Tb.N, and BMD were significantly higher in groups B and C than in group A (P<0.05), and in group C was higher than in group B (P<0.05), showing significant differences between groups. Histological staining showed that the materials of groups B and C were biodegradable, and the osteogenic effect was increased with the increase of P24 concentration.ConclusionP24 peptide can improve the ectopic osteogenesis of CS-TBA hydrogel, and the 10% concentration is more effective.
Objective To prepare chitosan microcarriers and to use it to cultivate rat primary hepatocytes. Methods The crosslinked chitosan microcarrier was prepared by the reaction of glutaraldehyde with chitosan. Various factors that influence the preparation were studied and the reaction conditions were optimized. Rat primary hepatocytes cultured on chitosan microcarrier were observed by using phase contrast microscope and scanning electron microscope. Results Chitosan microcarriers with good properties could be prepared by adjusting the concentration of chitosan solution and the quantity of glutaraldehyde. Rat hepatocytes cultured on chitosan microcarriers retained the spherical shape as they have in vivo. And albumin secretion can last over one week. The highest albumin secretion rate reached 26.7μg/24h/ml. Conclusion Chitosan microcarriers is a promising scaffold for hepatocyte attachment, which can be used in bioartificial liver support system.
OBJECTIVE: To investigate the feasibility of the chitosan-collagen membrane (CCM) as a dermal substitute. METHODS: Fresh bovine tendo calcaneus collagen was dispersed in 0.5 mol/L acetic acid, co-precipitated with chitosan and lyophilized. Dry membranes were cross-linked in 0.05% glutaraldehyde for 24 hours. In vitro its degrading rate was measured by use of collagenase degrading test. The chitosan-collagen membrane was implanted to subcutaneous dorsal sites of SD rats. After implantation, histocompatibility, vascularity and degradation were observed in vivo. RESULTS: The chitosan-collagen membrane was yellowish, translucent, and porous. Pore size ranged 50-250 microns, and collagen fiber bundles were reticular arrangement in the membrane. It had slower degradation than pure collagen membrane by collagenase in vitro. Subcutaneous implantation test showed the minimal inflammation, good histocompatibility and earlier vascularization. The membrane degradation was slower in vivo. Eight weeks after implantation, organized collagen structure was retained. CONCLUSION: The chitosan-collagen membrane has better physical and biological properties, ideal histocompatibility, earlier vascularization and slower degradation. Therefore, It is an optimum substitute for dermal scaffold.
In order to study the effect of chitin and chitosan on the growth of Schwann cell (SC) of rats in vitro, the SC was isolated from sciatic nerve and brachial plexus of new-born rats. After the enzymatic and mechanical dissociation, the cell suspension was vaccinated on chitin membrane and chitosan fluid-coated glass coverslips. Then, the growth of SC was examined at 1, 3, 7 days after culture under light microscope and scanning electron microscope. The results showed that 94 percent of the cell grown from was SC and only 6% was fibroblast (FB), while that of the control SC 71% and FB 29% in population. The number of SC in chitosan suspension was more than that in chitin. Therefore, the conclusion was that the chitin and chitosan was histocompatible to SC, and chitosan suspension was superior to chitin, and both could inhibit the growth of fibroblast.
Objective To explore a way to make a new kind of chitosan-basedmicrosphere (MS), which can be used as a novel biodegradable haemostatic powder, and to confirm its haemostatic efficiency. MethodsChitosan(CTS), a haemostatic polysaccharide, was selected as a main material for the haemostatic powder; alginate (ALG), another haemostatic polysaccharide that has been found to be effective in promoting haemostasis in surgical procedures, was selected to be thecostar. The emulsification and the cross-link were chosen as a preparation process based on the interaction between the polysaccharides. The diameter of the prepared MS was determined by SPOS, and the surface of MS was observed under SEM. The swelling characteristics of MS in the simulative wound efflusion were investigated. In a splenic bleeding model in 6 rabbits, MS and Yunnanbaiyao were randomly used as a haemostatic agent, and the corresponding bleeding time was recorded. Results The MS prepared in the above-mentioned process was well proportioned and was similarly shaped. It became a kind of white powder after dehydration, and had a coralloid surface under SEM. The diameter of the MS was 4.05±2.55 μm, which was determined by SPOS. The swelling ratio of the MS was 280.139% within 5 min. The bleeding time was significantly decreased in the MStreated group (2.83±0.17 min) when compared with that in the control group (5.33±0.49 min)(P＜0.01). Conclusion The CTS/ALG-MS, which is made from haemostatic biomaterials (CTS, ALG) by emulsification and the cross-link processes, can be provided with favorable haemostatic efficiency. It can be used as a novel haemostaticpowder.However, its biodegrading rate and mode still remain to be further studied.
