Optimization of biodegradable sponges as controlled release drug matrices. I. Effect of moisture level on chitosan sponge mechanical properties

Cross-linked chitosan sponges as controlled release drug carrier systems were developed. Tramadol hydrochloride, a centrally acting analgesic, was used as a model drug. The sponges were prepared by freeze-drying 1.25% and 2.5% (w/w) high and low M.wt. chitosan solutions, respectively, using glutaraldehyde as a cross-linking agent. The hardness of the prepared sponges was a function of glutaraldehyde concentration and volume where the optimum concentration that offered accepted sponge consistency was 5%. Below or above 5%, very soft or very hard and brittle sponges were obtained, respectively. The determined drug content in the prepared sponges was uniform and did not deviate markedly from the calculated amount. Scanning electron microscopy (SEM) was used to characterize the internal structures of the sponges. The SEM photos revealed that cross-linked high M.wt. chitosan sponges have larger size surface pores that form connections (channels) with the interior of the sponge than cross-linked low M.wt. ones. Moreover, crystals of the incorporated Tramadol hydrochloride were detected on the lamellae and within pores in both chitosan sponges. Differences in pore size and dissolution medium uptake capacity were crucial factors for the more delayed drug release from cross-linked low M.wt. chitosan sponges over high M.wt. ones at pH 7.4. Kinetic analysis of the release data using linear regression followed the Higuchi diffusion model over 12 hours. Setting storage conditions at room temperature under 80-92% relative humidity resulted in soft, elastic, and compressible sponges.     

Optimization of Biodegradable Sponges as Controlled Release Drug Matrices. I. Effect of Moisture Level on Chitosan Sponge Mechanical Properties

Cross‐linked chitosan sponges as controlled release drug carrier systems were developed. Tramadol hydrochloride, a centrally acting analgesic, was used as a model drug. The sponges were prepared by freeze‐drying 1.25% and 2.5% (w/w) high and low M.wt. chitosan solutions, respectively, using glutaraldehyde as a cross‐linking agent. The hardness of the prepared sponges was a function of glutaraldehyde concentration and volume where the optimum concentration that offered accepted sponge consistency was 5%. Below or above 5%, very soft or very hard and brittle sponges were obtained, respectively. The determined drug content in the prepared sponges was uniform and did not deviate markedly from the calculated amount. Scanning electron microscopy (SEM) was used to characterize the internal structures of the sponges. The SEM photos revealed that cross‐linked high M.wt. chitosan sponges have larger size surface pores that form connections (channels) with the interior of the sponge than cross‐linked low M.wt. ones. Moreover, crystals of the incorporated Tramadol hydrochloride were detected on the lamellae and within pores in both chitosan sponges. Differences in pore size and dissolution medium uptake capacity were crucial factors for the more delayed drug release from cross‐linked low M.wt. chitosan sponges over high M.wt. ones at pH 7.4. Kinetic analysis of the release data using linear regression followed the Higuchi diffusion model over 12 hours. Setting storage conditions at room temperature under 80–92% relative humidity resulted in soft, elastic, and compressible sponges.     

壳聚塘-明胶共混膜的制备及机械性能测试

以共混方法制备了壳聚糖-明胶共混薄膜,并对共混薄膜的机械性能进行了测试。研究表明:壳聚糖-明胶共混膜的混合比例可以改变共混膜的力学强度,共混膜随明胶含量增加其弹性模量和拉伸断裂应力逐渐减小,壳聚糖溶液与明胶共混的体积比为4∶6时,拉伸强度达到最小值,而断裂伸长率在壳聚糖与明胶体积比为6∶4的时候达到了最大值。     

Studies on gelatin-based sponges. Part III: A comparative study of cross-linked gelatin/alginate, gelatin/hyaluronate and chitosan/hyaluronate sponges and their application as a wound dressing in full-thickness skin defect of rat

Novel cross-linked sponges composed of gelatin/alginate and gelatin/hyaluronate and chitosan/hyaluronate (GH, GA and CH, respectively) were prepared and compared. Six different sponges with or without silver sulfadiazine (AgSD) were applied on the full-thickness dorsal skin defect of Wistar rat. The histology and epidermal wound healing rates of the skin defects were investigated by light microscopy and computerized morphometry 5 and 12 days post-operatively. In our full-thickness wound model (diameter 1 cm), the AgSD-impregnated sponges showed good wound healing performances on the whole. However, there appeared meaningful differences of wound healing between the gelatin-based sponges (GH, GA) and the CH. GH with AgSD was found to show the best wound healing properties as a wound dressing resulting from histological findings and computerized morphometric analysis of epidermal healing.     

Studied on a novel human keratinocyte membrane delivery system in vitro

The culture of keratinocytes on flexible membranes has been proposed as a means to simplify, accelerate and improve the efficiency with which proliferating cells are delivered to full thickness or non-healing skin defects. The purpose of this article was to study the ability of chitosan-gelatin manbranes to facilitate the growth of human keratinocytes. The membranes with different chitosan contents were studied. The surface properties of chitosan-gelatin membranes were investigate by SEM, and water contact angle test. The mechanical property of the membranes was tested. Data implied that gelatin could make the membranes more flexible and hydrophilic than chitosan membranes, which may regulate the seeded cells behavior. Loading human keratinocytes on chitosan-gelatin membranes, cells attachment, spread, and growth were investigated by light microscopy, SEM, and MTT test. The results suggested that the adhesion and proliferation of keratinocytes seeded on chitosan-gelatin membranes were same as on tissue culture plate, in which gelatin could modify the interaction between keratinocytes and chitosan membranes. Therefore, chitosan-gelatin membrane is a good candidate for keratinocytes delivery system.     

Evaluation of selected polysaccharide excipients in buccoadhesive tablets for sustained release of nicotine

Some naturally occurring biocompatible materials were evaluated as mucoadhesive controlled release excipients for buccal drug delivery. A range of tablets were prepared containing 0-50% w/w xanthan gum, karaya gum, guar gum, and glycol chitosan and were tested for swelling, drug release, and mucoadhesion. Guar gum was a poor mucoadhesive and lacked sufficient physical integrity for buccal delivery. Karaya gum demonstrated superior adhesion to guar gum and was able to provide zero-order drug release, but concentrations greater than 50% w/w may be required to provide suitable sustained release. Xanthan gum showed strong adhesion to the mucosal membrane and the 50% w/w formulation produced zero-order drug release over 4 hours, about the normal time interval between daily meals. Glycol chitosan produced the strongest adhesion, but concentrations greater than 50% w/w are required to produce a nonerodible matrix that can control drug release for over 4 hours. Swelling properties of the tablets were found to be a valuable indicator of the ability of the material to produce sustained release. Swelling studies also gave an indication of the adhesion values of the gum material where adhesion was solely dependent upon penetration of the polymer chains into the mucus layer.     

Preparation and bioactivity evaluation of hydroxyapatite-titania/chitosan-gelatin polymeric biocomposites

Biocomposites consisting of hydroxyapatite (HA) and natural polymers such as collagen, chitosan, chitin,and gelatin have been extensively investigated. However, studies on the combination of HA and titania with chitosan and gelatin have not been conducted yet. Novel biodegradable hydroxyapatite-titania/chitosan-gelatin polymeric composites were fabricated. In this work, our results are concerning with the preparation and characterization of HA powder and HA filler containing titania powder (10 and 30%) with a chitosan and gelatin copolymer matrix. The present research focuses on characterizing the structure of this novel class of biocomposites. Thermogravimetric analysis (TGA), X-ray diffraction (XRD), and Fourier Transformed Infrared Spectroscopy (FT-IR), Scanning electron microscopy (SEM-EDAX) were employed to assess the produced composites. The mechanical properties in terms of compressive strength and hardness test were also investigated. The in vitro study in simulated body fluid (SBF) was performed to assess the bioactivity of composites. The results proved that apatite resembling natural bone are formed faster and greater in the case the composite of HA containing 10% titania into chitosan-gelatin polymeric matrix when they are soaked in a simulated body fluid (SBF) than the composite containing 30% titania. The biocomposites containing HA with 10% titania are expected to be attractive for bioapplications as bone substitutes and scaffolds for tissue engineering in future.     

Preparation and in vitro characteristics of lactoferrin-loaded chitosan microparticles

In order to achieve the delivery and controlled release of lactoferrin (LF), a biologically multifunctional protein, chitosan microparticles loaded with LF were prepared. Several types of chitosan microparticles containing LF were prepared by the w/o emulsification-solvent evaporation method, and the particle characteristics and release properties in JP 2nd fluid, pH 6.8, were examined. All kinds of microparticles were obtained at a yield of more than 75% (w/w). LF-loaded microparticles prepared by nonsonication and nonaddition of sulfate, named Ch-LF(N), showed high drug content, small particle size and spherical particle shape. Also, for release properties, Ch-LF(N) exhibited gradual drug release over 7 hr with less remaining in the microparticles. Considering the mucoadhesive properties of chitosan microparticles, Ch-LF(N) are suggested to be useful for gradual supply to topical diseased sites or for effective delivery to intestinal areas with abundant LF receptors.     

Preparation and In Vitro Characteristics of Lactoferrin-loaded Chitosan Microparticles

ABSTRACT

In order to achieve the delivery and controlled release of lactoferrin (LF), a biologically multifunctional protein, chitosan microparticles loaded with LF were prepared. Several types of chitosan microparticles containing LF were prepared by the w/o emulsification-solvent evaporation method, and the particle characteristics and release properties in JP 2nd fluid, pH 6.8, were examined. All kinds of microparticles were obtained at a yield of more than 75% (w/w). LF-loaded microparticles prepared by nonsonication and nonaddition of sulfate, named Ch-LF(N), showed high drug content, small particle size and spherical particle shape. Also, for release properties, Ch-LF(N) exhibited gradual drug release over 7 hr with less remaining in the microparticles. Considering the mucoadhesive properties of chitosan microparticles, Ch-LF(N) are suggested to be useful for gradual supply to topical diseased sites or for effective delivery to intestinal areas with abundant LF receptors.     

Development of chitosan/gelatin/keratin composite containing hydrocortisone sodium succinate as a buccal mucoadhesive patch to treat desquamative gingivitis

The aim of this research was to develop chitosan/gelatin/keratin composite containing hydrocortisone sodium succinate as a buccal mucoadhesive patch to treat desquamative gingivitis, which was fabricated through an environmental friendly process. Mucoadhesive films increase the advantage of higher efficiency and drug localization in the affected region. In this research, mucoadhesive films, for the release of hydrocortisone sodium succinate, were prepared using different ratios of chitosan, gelatin and keratin. In the first step, chitosan and gelatin proportions were optimized after evaluating the mechanical properties, swelling capacity, water uptake, stability, and biodegradation of the films. Then, keratin was added at different percentages to the optimum composite of chitosan and gelatin together with the drug. The results of surface pH showed that none of the samples were harmful to the buccal cavity. FTIR analysis confirmed the influence of keratin on the structure of the composite. The presence of a higher amount of keratin in the composite films resulted in high mechanical, mucoadhesive properties and stability, low water uptake and biodegradation in phosphate buffer saline (pH?=?7.4) containing 10⁴?U/ml lysozyme. The release profile of the films ascertained that keratin is a rate controller in the release of the hydrocortisone sodium succinate. Finally, chitosan/gelatin/keratin composite containing hydrocortisone sodium succinate can be employed in dental applications.     

Gelatin sponges (Gelfoam ® ) as a scaffold for osteoblasts

Gelatine sponge because of its flexibility, biocompatibility, and biodegradability, has the potential to be used as a scaffold to support osteoblasts and to promote bone regeneration in defective areas. This study aimed to determine osteoblast proliferation, differentiation, and integration in modified and un-modified gelatine sponges. Three scaffolds were studied: gelatine sponge (Gelfoam), gelatin sponge/mineral (hydroxyapatite) composite, and gelatin sponge/polymer (poly-lactide-co-glycolide) composite. 2-D plastic coverslip was used as control. The gelatin sponges were modified using PLGA coating and mineral deposition to increase biodegradation resistance and osteoblast proliferation respectively. The scaffolds were characterized using Scanning Electron Microscopy (SEM) and X-ray diffraction. Cell number (DNA content), cell-replication rate (thymidine assay), and cell differentiation (alkaline phosphatase activity) were measured 24 h, 3 days, and 1, 2, 3 weeks after the osteoblast-like cells were cultured onto the scaffolds. Cell penetration into the sponges was determined using haematoxylin-eosin staining. Both modified and unmodified gelatine sponges demonstrated ability to support cell growth and cells were able to penetrate into the sponge pores. In a comparison of different scaffolds, cell number and cell replication were highest in sponge/hydroxyapatite composite and lowest in sponge/PLGA composite.     

缓冲包装用明胶-淀粉泡沫的性能研究

目的以明胶和预胶化淀粉为原料制备具有缓冲效果的生物质可降解泡沫材料,为缓冲包装用生物质泡沫提供一种新的选择。方法通过对不同明胶-淀粉质量比、固含量、十二烷基硫酸钠(SDS)用量进行实验研究,并进行结构表征及静态压缩性能测定对泡沫材料进行综合评价。结果得到了明胶-淀粉缓冲泡沫材料的最优条件,固含量(用质量分数表示)为20%,表面活性剂质量分数为0.75%,明胶-淀粉质量比为70∶30。在此最优条件下的明胶-淀粉缓冲泡沫材料发泡倍率为5.14倍,表观密度为0.064 g/cm³,弹性模量为36.64 kPa,50%抗压强度为2.49 kPa。结论以明胶和预胶化淀粉为原料制备的复合泡沫材料具有表观密度低、缓冲性能较好的特点。单因素实验结果表明,预胶化淀粉对泡沫的力学性能有增强作用。     

Interpenetrating polymer networks (IPN) based on gelatin/poly(ethylene glycol) dimethacrylate/clay nanocomposites: Structure–properties relationship

The effects of cross-linking sequence (simultaneous or sequential) and incorporation of exfoliated sodium-montmorillonite (Na⁺-MMT) nanoclay on the structure and properties of interpenetrating polymer networks (IPNs) based on gelatin/poly(ethylene glycol)dimethacrylate were studied by means of different complementary techniques. Gelatin and PEGdmA phases were cross-linked via chemical and in-situ UV curing, respectively. 2,2-dimethoxy-2-phenylacetophenone (DMPA) (1.5% w/w) was used as photo-initiator to cross-link PEGdmA. The results showed that the incorporation of small amount of Na⁺-MMT nanoplatelets accelerates the kinetics of chemical cross-linking of gelatin by glutaraldehyde (1.0% w/w). This led to a new hypothesis concerning the tuning structural evolution of the IPNs by the Na⁺-MMT content. In the case of simultaneous IPNs, in which both phases cross-linked at the same time, the accelerated cross-linking of gelatin in the presence of exfoliated sodium-montmorillonite led to increased structural homogeneity, improved mechanical and thermal properties. Incorporation of nanoclay did not show any significant effect on the structure and properties of the IPNs synthesized via sequential method in which gelatin and PEGdma phases were cross-linked separately. For the semi-IPNs, however, Na⁺-MMT induced macroscopic phase separation and resulted in lower mechanical properties. These results might shed light on the mechanisms underlying structure–property relationship in biohybrid IPNs based on gelatin as promising candidates for tissue engineering and drug delivery applications.     

Microstructure and characteristic properties of gelatin/chitosan scaffold prepared by a combined freeze-drying/leaching method

A combined freeze-drying and particulate leaching method for scaffold synthesis showed an improvement in the horizontal microstructure of the gelatin/chitosan scaffolds. Type and concentration of the cross-linking agent, freezing temperature, concentration of the polymeric solution and gelatin/chitosan weight ratio were the variables affecting the scaffold properties. Assessment of the tensile properties of the scaffolds revealed that for a scaffold with 50% chitosan, glutaraldehyde, as a cross-linking agent, created much tighter polymeric network compared to N,N-(3-dimethylaminopropyl)-N′-ethyl carbodiimide (EDC). However, in the case of gelatin scaffolds, EDC was identified as the stronger cross-linker. Compressive behavior of the scaffolds satisfied formulations obtained from the theoretical modeling of the low-density, elastomeric foams. The investigation of the scaffold degradation indicated that the increase in the mechanical strength of the scaffolds would not always reduce their degradation rate.     

Promotion of fibrovascular tissue ingrowth into porous sponges by basic fibroblast growth factor

Fibrovascular tissue ingrowth into poly(vinyl alcohol) (PVA) sponges of different pore sizes was investigated by incorporating basic fibroblast growth factor (bFGF) into the sponges. The average pore size of PVA sponges used in this study was 30, 60, 110, 250, 350, and 700 m and gelatin microspheres were employed as release carrier of bFGF. The sponges were subcutaneously implanted into the back of mice after incorporating free bFGF or gelatin microspheres containing bFGF into the sponges. Fibrovascular tissue infiltrated with time into the sponge pores and the extent of fibrous tissue ingrowth showed a maximum at a pore size around 250 m 1 and 6 weeks after implantation. Significant promotion of the growth of fibrous tissue by bFGF was observed only at 3 weeks post-implantation (p < 0.05). New capillaries were formed in the tissue at any time, as long as bFGF was given to the sponges. Both empty gelatin microspheres and phosphate buffered solution neither promoted tissue ingrowth nor induced capillary formation in the sponges. It was concluded that bFGF was essential to induce the fibrovascular tissue ingrowth into the pores of PVA sponges. ©2000 Kluwer Academic Publishers     

Formulation and comparative characterization of chitosan, gelatin, and chitosan-gelatin-coated liposomes of CPT-11-HCl

Liposomes containing phosphatidylcholine and cholesterol (uncoated) and coated by chitosan, gelatin, and combination of chitosan and gelatin were prepared by the modified ethanol injection method. The aim of this work was to formulate and characterize liposomes of camptothecin (CPT)-11-HCl (Irinotecan HCl) containing chitosan, gelatin, and both polymers as coating materials; and also to increase its circulation longevity when compared with the free drug while maintaining the agent in its active lactone form. Size, shape, zeta potential, encapsulation efficiency, stability study, in vitro, and in vivo release study were used for characterization of liposomes. The size of liposomes was in the order of uncoated < chitosan coated < gelatin coated < combination of chitosan and gelatin coated. The zeta potential of liposomes was in the order of combination of chitosan and gelatin coated > chitosan coated > gelatin coated > uncoated. The formulations showed the long-term stability. The encapsulation efficiency of liposomes was in order of combination of chitosan and gelatin coated > gelatin coated > chitosan coated > uncoated. The in vitro and in vivo release of drug was observed in the order of combination chitosan and gelatin coated > gelatin coated > chitosan coated > uncoated.     

多级开孔壳聚糖海绵的细胞行为分析

使用冰滴为致孔剂制备表面大孔、内部孔洞相连的新型壳聚糖(HPCS)支架,将其与聚乳酸复合制备出三维蜂窝状孔洞结构的复合支架(THCP).对HPCS和THCP进行了表面形貌、力学性能、细胞相容性等方面的表征,并与常规冻干法所制备的壳聚糖(CS)海绵进行了对比.结果表明,在HPCS海绵表面均匀分布着大而开放的孔洞,大孔内部...     

Comparison of various mixtures of β-chitin and chitosan as a scaffold for three-dimensional culture of rabbit chondrocytes

With the use of a recently created chitosan neutral hydrogel, we have been able to create various mixtures of chitin and chitosan without changing their characteristics even at room temperature. The aim of this study was the initial comparison of various mixtures of β-chitin and chitosan as a scaffold for rabbit chondrocyte culture. We created five types of sponges: pure β-chitin, pure chitosan, 3:1, 1:1, and 1:3 β-chitin-chitosan. The absorption efficiencies of chondrocytes in all five types of sponges were found to be around 98%. The mean concentrations of chondroitin sulfate were statistically different neither at week 2 nor at week 4 postculture between the types of sponges. The content of hydroxyproline in the β-chitin sponge was significantly greater than in other sponges at week 4 postculture. From the histochemical and immunohistochemical findings, the cartilage-like layer in the chondrocytes-sponge composites of all five types of sponges was similar to hyaline cartilage. However, only immunohistochemical staining of type II collagen in the pure β-chitin sponge was closer to normal rabbit cartilage than other types of sponges. The pure β-chitin sponge was superior to other sponges concerning the content of extracellular matrices of collagen.     

The effect of cross-linking on the microstructure,mechanical properties and biocompatibility of electrospun polycaprolactone–gelatin/PLGA–gelatin/PLGA–chitosan hybrid composite

Abstract

In this study, multilayered scaffolds composed of polycaprolactone (PCL)–gelatin/poly(lactic-co-glycolic acid) (PLGA)–gelatin/PLGA–chitosan artificial blood vessels were fabricated using a double-ejection electrospinning system. The mixed fibers from individual materials were observed by scanning electron microscopy. The effects of the cross-linking process on the microstructure, mechanical properties and biocompatibility of the fibers were examined. The tensile stress and liquid strength of the cross-linked artificial blood vessels were 2.3 MPa and 340 mmHg, respectively, and were significantly higher than for the non-cross-linked vessel (2.0 MPa and 120 mmHg). The biocompatibility of the cross-linked artificial blood vessel scaffold was examined using the MTT assay and by evaluating cell attachment and cell proliferation. The cross-linked PCL–gelatin/PLGA–gelatin/PLGA–chitosan artificial blood vessel scaffold displayed excellent flexibility, was able to withstand high pressures and promoted cell growth; thus, this novel material holds great promise for eventual use in artificial blood vessels.