Study of a cross-linked hydrogel of Karaya gum and Starch as a controlled drug delivery system

Abstract

A cross-linked hydrogel was synthesized using a hybrid backbone of karaya gum starch and grafted with polyacrylic acid. It showed a maximum swelling ratio (SR) of 30.5?g/g at pH 10 and was explored as an oral drug delivery carrier using paracetamol and aspirin as model drugs. In vitro release experiments revealed that maximum drug release at pH 7.4 in comparison to pH 1.2 (simulated intestinal vs gastric fluid) and neutral medium. The release profiles of these drugs showed no initial burst. It also showed good hemocompatibilty and non-cytotoxicity for its employment as a site specific drug delivery agent.     

Effect of binder additives on terbutaline hydrogels of alpha-PVA/NaCl/H(2)O system in drug delivery: I. Effect of gelatin and soluble starch

In order to prepare ecologically and biologically safety physical cross-linked hydrogel of a-PVA/NaCl/H(2)O system, we prepared blend hydrogel of natural polymeric binders like gelatin and starch in above system, and effect of these binder additives were evaluated on terbutaline release kinetics. Terbutaline (1%) hydrogels of a-PVA(7%)/NaCl(11%)/H(2)O and a-PVA(7%)/H(2)O system (Cyclic FT process) were prepared along with various concentrations of gelatin and soluble starch by feed-mixture dissolving method. In case of a-PVA/NaCl/H(2)O system only one cycle gelation was done at -20 degrees C for 24 h. On the other hand 3 cycles were done by freezing at -30 degrees C for 16 h followed by thawing at room temperature for 8 h. Drug release was done by paddle method (USP type II) with a rotation of 50 rpm at 37 degrees C in distilled water. Swelling of the gel was done at 37 degrees C for 45 h and melting temperatures of the gel were also studied using the upside-down test tube method. Comparatively lower values of release rate, diffusion coefficient and kinetic constant were found from the blend hydrogel of a-PVA/starch/NaCl/H(2)O system. When % cumulative release was plotted vs. square root of time it showed straight lines, which indicated Higuchi Matrix Dissolution Model. Inclusion of starch binder increased the degree of swelling compared with that of gelatin. 15 h was found as equilibrium swelling time. A Fickian swelling of this blend hydrogel system indicated the swelling controlled Fickian diffusion type of drug release. Melting temperatures of the blend hydrogels were characteristically higher (94-95 degrees C) than that of cyclic FT (72-76 degrees C), resulting a thermostable hydrogel for biological system. SEM morphological studies of gel surface indicated the well-developed interpenetrating macromolecular network structure like fish net in starch blend gel prevails over other hydrogel studied here. Gelatin has got characteristic tunnel (200 microm in diameter) inside the macromolecular network that contributes this system higher release kinetics than that of starch.     

In vitro and in vivo evaluation of gelatin-chondroitin sulphate hydrogels for controlled release of antibacterial proteins

Chemically cross-linked gelatin-chondroitin sulphate (ChS) hydrogels, impregnated in Dacron, were evaluated as drug delivery systems for antibacterial proteins. The gelatin-chondroitin sulphate gels, plain or impregnated in Dacron, were cross-linked with a water-soluble carbodiimide (EDC) and N-hydroxysuccinimide (NHS). The release of lysozyme and recombinant thrombocidin (rTC-1), an antibacterial protein derived from human blood platelets, from the gelatin-ChS gels in Dacron in phosphate-buffered saline at 37 degrees C was determined, and compared to the release from gelatin gels in Dacron and plain gelatin-ChS gels. The incorporation of chondroitin sulphate into gelatin gels, caused a marked increase in lysozyme loading capacity, and a slower release rate. The relative release profiles for rTC-1 and lysozyme were equal for cross-linked gelatin as well as for cross-linked gelatin-ChS gels. Furthermore, rTC-1 showed no loss of antibacterial activity after 1 week of release. The lysozyme concentration profiles in the samples and in the surrounding medium as a function of time were calculated using mathematical solutions for Ficks second law of diffusion for a semi-infinite composite medium, which is a schematic representation of a slab in a surrounding medium. The biocompatibility and degradation of the Dacron matrices impregnated with gelatin-ChS gels was studied after implantation in subcutaneous pockets in rats. Chemically cross-linked gelatin-Ch5 gels showed a mild tissue reaction, and almost complete degradation within 18 weeks of implantation.     

Transport of biological molecules in surfactant-alginate composite hydrogels

Obstructed transport of biological molecules can result in improper release of pharmaceuticals or biologics from biomedical devices. Recent studies have shown that nonionic surfactants, such as Pluronic? F68 (F68), positively alter biomaterial properties such as mesh size and microcapsule diameter. To further understand the effect of F68 (incorporated at concentrations well above the critical micelle concentration (CMC)) in traditional biomaterials, the transport properties of BSA and riboflavin were investigated in F68-alginate composite hydrogels, formed by both internal and external cross-linking with divalent cations. Results indicate that small molecule transport (represented by riboflavin) was not significantly hindered by F68 in homogeneously (internally) cross-linked hydrogels (up to an 11% decrease in loading capacity and 14% increase in effective diffusion coefficient, D(eff)), while protein transport in homogeneously cross-linked hydrogels (represented by BSA) was significantly affected (up to a 43% decrease in loading capacity and 40% increase in D(eff)). For inhomogeneously cross-linked hydrogels (externally cross-linked by CaCl(2) or BaCl(2)), the D(eff) increased up to 50 and 83% for small molecules and proteins, respectively. Variation in the alginate gelation method was shown to affect transport through measurable changes in swelling ratio (30% decrease) and observable changes in cross-linking structure as well as up to a 3.6- and 11.8-fold difference in D(eff) for riboflavin and BSA, respectively. Aside from the expected significant changes due to the cross-linking method utilized, protein transport properties were altered due to mesh size restrictions (10-25 nm estimated by mechanical properties) and BSA-F68 interaction (DLS). Taken as a whole, these results show that incorporation of a nonionic surfactant at concentrations above the CMC can affect device functionality by impeding the transport of large biological molecules.     

Controlled release of drugs from multi-component biomaterials

In order to control their release, drugs are encapsulated into systems which are expected to provide a certain site with a predetermined amount of drug over a well-defined period of time. Here we report on a multi-component drug delivery biomaterial that consists of a hydrogel matrix in which drug-loaded biodegradable microcarriers are dispersed, and whose potential applications could be found in the design of implantable devices with long-term activity, as required by contraceptive and hormone replacement treatments. The release profile of the drug can actually be tuned by the complex interplay of several release mechanisms, including the permeability and eventually the degradation rate of the microcarriers and the diffusion through the hydrogel. The hydrogel consisted of 2-hydroxyethyl methacrylate cross-linked by ethylene glycol dimethacrylate. The microcarriers were biodegradable poly-epsilon-caprolactone (PCL) microspheres in which active molecules, such as levonorgestrel (LNG), were encapsulated. The hydrogels were characterized by water swelling, thermal properties, LNG diffusion through drug-free and drug-depleted hydrogel membranes and LNG release from devices with drug dispersed in the hydrogel. The PCL microspheres were observed by scanning electron microscopy; their size distribution, LNG loading and release were also investigated. The hydrogel-microsphere assemblies were characterized in terms of the distribution of the microspheres within the hydrogel, water swelling and the release of the encapsulated molecules. The developed device, due to its composite structure, has the ability to combine several release mechanisms, leading to drug release obeying zero-order kinetics for most of the time.     

Alginate-based microencapsulation of retinal pigment epithelial cell line for cell therapy

The goals of this study were to evaluate human retinal pigment epithelial cell line (ARPE-19) for cell encapsulation and to optimize the alginate-based microencapsulation. We used immortalized ARPE-19 cells and the transfected sub-line that expresses secreted alkaline phosphatase (SEAP) reporter enzyme. Alginate was cross-linked with different divalent cations (Ca(2+), Ba(2+), Sr(2+) and combination of Ca(2+) and Ba(2+)), coated first with poly-l-lysine (PLL), and then with alginate. Microcapsules with different pore sizes and stability were generated. The pore size of the microcapsules was assessed by the release of encapsulated fluorescein isothiocyanate (FITC)-dextrans. The viability of the cells in the microcapsules was studied in vitro by assessing the secretion rates of SEAP and oxygen consumption by the cells. The best microcapsule morphology, durability and cellular viability were obtained with alginate microcapsules that were cross-linked with Ca(2+) and Ba(2+) ions and then coated with PLL and alginate. Based on FITC-dextran release these microcapsules have porous wall that enables the rapid contents release. The ARPE-19 cells maintained viability in the Ca(2+) and Ba(2+) cross-linked microcapsules for at least 110 days. The alginate microcapsules cross-linked with Ca(2+) and Ba(2+) have sufficiently large pore size for prolonged cell viability and for the release of secreted SEAP model protein (Mw 50 kDa; radius of gyration of 3 nm). ARPE-19 cells show long-term viability and protein secretion within alginate microcapsules cross-linked with Ca(2+) and Ba(2+). This combination may be useful in cell therapy.     

Secondary cytotoxicity of cross-linked dermal sheep collagens during repeated exposure to human fibroblasts.

We investigated commercially available dermal sheep collagen either cross-linked with hexamethylenediisocyanate, or cross-linked with glutaraldehyde. In previous in vitro studies we could discriminate primary, i.e. extractable, and secondary cytotoxicity, due to cell-biomaterial interactions, i.e. enzymatic actions. To develop dermal sheep collagen for clinical applications, we focused in this study on the release, e.g. elimination, of secondary cytotoxicity over time. We used the universal 7 d methylcellulose cell culture with human skin fibroblasts as a test system. Hexamethylenediisocyanate-cross-linked dermal sheep collagen and glutaraldehyde-cross-linked dermal sheep collagen were tested, with intervals of 6 d, over a culture period of 42 d. With hexamethylenediisocyanate-cross-linked dermal sheep collagen, cytotoxicity, i.e. cell growth inhibition and deviant cell morphology, was eliminated after 18 d of exposure. When testing glutaraldehyde-cross-linked dermal sheep collagen, the bulk of cytotoxic products was released after 6 d, but a continuous low secondary cytotoxicity was measured up to 42 d. As a control, non-cross-linked dermal-sheep collagen was tested over a period of 36 d, but no secondary cytotoxic effects were observed. The differences in release of secondary cytotoxicity between hexamethylenediisocyanate-cross-linked dermal sheep collagen, glutaraldehyde-cross-linked dermal sheep collagen and non-cross-linked dermal sheep collagen are explained from differences in cross-linking agents and cross-links obtained. We hypothesize that secondary cytotoxicity results from enzymatic release of pendant molecules from hexamethylene-diisocyanate-cross-linked dermal sheep collagen, e.g. formed after reaction of hydrolysis products of hexamethylenediisocyanate with dermal sheep collagen.(ABSTRACT TRUNCATED AT 250 WORDS)     

Volume change of double cross-linked poly(aspartic acid) hydrogels induced by cleavage of one of the crosslinks

In the present paper we report for the first time the development of redox-responsive biocompatible polymer gels. Double cross-linked poly(aspartic acid) hydrogels were prepared using two different cross-linking agents simultaneously. One of the cross-linkers was diaminobutane (DAB), the other cystamine (CYS). The relative amounts of DAB and CYS molecules were varied over a wide range while the total amount of cross-linker molecules (DAB + CYS) was kept constant. DAB provides stable cross-links, whereas CYS contains disulfide bonds, which can be broken by reduction. The cleavage of disulfide cross-links results in enhanced swelling and a significant decrease in the elastic modulus of the gels. These novel types of stimuli-responsive gels are promising candidates for new swelling controlled release matrices.     

Cross-linked hemoglobin as a potential membrane for an artificial red blood cell

A cross-linked hemoglobin membrane has been created with discerning permeability between dissolved hemoglobin and small molecules. Such a membrane could be used to enclose a sphere of hemoglobin solution thereby allowing the entire "cell" to transport oxygen. The hemoglobin membrane was cross-linked on a polycarbonate support; the mechanical support was necessary for diffusion experiments in this study and would not be used during any sphere preparation. A 30% methemoglobin solution in phosphate buffer was used to fill the pores of the 10 microm polycarbonate support, then cross-linked with a homobifunctional cross-linking agent. The cross-linked hemoglobin within the support was evaluated for hemoglobin and benzoic acid permeability in a side-by-side diffusion cell. Disuccinimidyl glutarate, disuccinimidyl suberate and disuccinimidyl tartrate were used as cross-linking agents. Disuccinimidyl glutarate, 4.65 mM, created a hemoglobin-impermeable membrane after cross-linking for 20 minutes, reducing the concentration to 0.46 mM required a cross-linking time to 60 minutes. Benzoic acid, representing a typical small molecule, was capable of diffusing through the disuccinimidyl glutarate cross-linked hemoglobin membrane at 87.2% of its diffusion through buffer.     

Immobilization of hormones for drug targeting

Biological active compounds such as insulin, heparin, progesterone and labeled-LH were entrapped in glutaraldehyde cross-linked bovine serum albumin (BSA) and human serum albumin (HAS) microspheres. Studies were carried out for their binding capacity and biodegradability using new proteolytic enzymes. Effects of proteolytic enzymes such as trypsin, chymotrypsin, papain and pronase-E on microspheres were studied in order to understand the biodegradability of the cross-linked proteins. It has been observed that labeled-LG was entrapped 60% in BSA and HAS microspheres. Labelled-LH-BSA, Labelled-LH-HAS and insulin microspheres were injected into mice and rabbits. It was observed that these cross-linked microspheres were biodegradable and the process appeared to be slow one, useful for sustained release of hormones. It was also observed that these albumin microspheres exhibit fluorescence at 495 nm.     

Controlled release of plasmid DNA from photo-cross-linked pluronic hydrogels

Chemically cross-linked hydrogels composed of Pluronic, water-soluble tri-block copolymers of poly(ethylene oxide)-b-poly(propylene oxide)-b-poly(ethylene oxide), were synthesized by a photo-polymerization method to achieve controlled DNA release. Pluronic F127 was di-acrylated to form a macromer and cross-linked to form a hydrogel structure in the presence and absence of vinyl group-modified hyaluronic acid (HA). UV irradiation time and the presence of the vinyl group-modified HA affected the mechanical property of Pluronic hydrogels to a great extent. Swelling ratio, degradation, and rheological behaviors of Pluronic hydrogels were investigated. When plasmid DNA was loaded in the hydrogels for sustained delivery, various release profiles were attained by varying UV irradiation time and modified HA amounts. Entrapped DNA was gradually damaged with increasing the UV exposure time as evidenced by decreasing the transfection efficiency. The DNA fractions released from the HA/Pluronic hydrogels, however, exhibited considerable transfection efficiencies commensurate with the UV exposure time, suggesting that they were not chemically degraded during the release period and substantially maintained functional gene expression activities despite the UV irradiation.     

Novel carboxymethylcellulose-based microporous hydrogels suitable for drug delivery

Several materials capable of acting as structures for controlled release were analysed for the fabrication of matrices. Among those used, hydrophilic polysaccharides appeared to be the most suitable materials. Carboxymethylcellulose (a semi-synthetic polysaccharide) was chemically cross-linked with a 60% and 90% cross-linking degree in order to obtain hydrogels and utilised as matrix for the realisation of controlled drug release systems. The morphology of the gels was changed in order to obtain a microporous structure with different porosity (14, 30 and 40 μm). The obtained porous matrices were characterised in terms of pore density, dimension and swelling behaviour. The influence of both the pore dimension and technique of loading on the release kinetics was analysed. By increasing the pore dimension the release of ibuprofen-lysin was slower. Inducing the microporous structure after the loading of the hydrogel with the drug resulted in a slower release.     

Novel carboxymethylcellulose-based microporous hydrogels suitable for drug delivery

Several materials capable of acting as structures for controlled release were analysed for the fabrication of matrices. Among those used, hydrophilic polysaccharides appeared to be the most suitable materials. Carboxymethylcellulose (a semi-synthetic polysaccharide) was chemically cross-linked with a 60% and 90% cross-linking degree in order to obtain hydrogels and utilised as matrix for the realisation of controlled drug release systems. The morphology of the gels was changed in order to obtain a microporous structure with different porosity (14, 30 and 40 microm). The obtained porous matrices were characterised in terms of pore density, dimension and swelling behaviour. The influence of both the pore dimension and technique of loading on the release kinetics was analysed. By increasing the pore dimension the release of ibuprofen-lysin was slower. Inducing the microporous structure after the loading of the hydrogel with the drug resulted in a slower release.     

Preparation and characterization of cross-linked microspheres C(Dex-g-PSSS) and their drug-carrying and colon-specific drug delivery properties

The graft polymer Dex-g-PSSS was obtained through poly(sodium 4-styrene sulfonate) (PSSS) grafted on dextran(Dex) by using the cerium salt–hydroxyl group redox initiation system. The cross-linked microspheres C(Dex-g-PSSS) were synthesized by suspension polymerization with epichlorohydrin as the cross-linking agent. The chemical structure and physicochemical characteristics of C(Dex-g-PSSS) microspheres were represented by infrared spectroscopy (FTIR), optical microscope, and zeta potential analysis. The aim of the study is to constitute a colon-specific drug delivery system via molecular design, using C(Dex-g-PSSS) microspheres as the drug-carrying material and taking 5-fluorouracil (5-FU) as the model drug. The drug-carrying ability and mechanism of the cross-linked microspheres C(Dex-g-PSSS) for 5-FU were investigated. Finally, in vitro release tests for the drug-carrying microspheres were conducted. The experimental results show that in the medium with pH 2, the cross-linked microspheres C(Dex-g-PSSS) exhibit a strong adsorption ability for 5-FU because of strong electrostatic interactions and have an adsorption capacity of 154 ± 7.5 mg/g, displaying high drug-carrying efficiency. The in vitro release behavior of the drug-carrying microspheres is highly dependent on pH and dextranase. In the medium with pH 2, the drug-carrying microspheres do not release the drug and in the medium with pH 1, they release a little, whereas in the medium with pH 7.4, a sudden delivery phenomenon of the drug will occur, and in the presence of dextranase, a more sudden delivery phenomenon of the drug will occur, displaying an excellent colon-specific drug delivery behavior.     

Thiolated Glycol Chitosan Bearing α-Cyclodextrin for Sustained Delivery of PEGylated Human Growth Hormone

Abstract

The extensive use of human growth hormone (hGH), emerging as protein therapeutics, has been limited by its instability in biological fluids and short biological half-life. In this study, thiolated glycol chitosan bearing α-cyclodextrin (TGC-CD), in situ cross-linked by poly(ethylene glycol)-diacrylate (PEG-DA), was synthesized to develop a sustained release system for PEGylated hGH (PEG-hGH). TGC-CD could form a stable inclusion complex with PEG-hGH by the physical interaction between the inner cavity of CD and PEG. Such a complex was readily cross-linked in the presence of PEG-DA via a Michael-type addition reaction. From the in vitro release experiments of PEG-hGH, it was confirmed that PEG-hGH was completely released from the complex for 12 h in PBS (pH 7.4), whereas the release rate of PEG-hGH was significantly reduced by the chemical cross-linking of the complex. PEG-hGH, released from the cross-linked complexes, maintained its structural integrity, which was demonstrated using circular dichroism spectroscopy. Overall, TGC-CD might be useful for sustained delivery of PEG-hGH.     

A comparative study of releasing and nonreleasing human basophils: nonreleasing basophils lack an early component of the signal transduction pathway that follows IgE cross-linking.

Basophils from approximately one fifth of the population were found to be unresponsive (nonreleasers), in terms of both histamine and leukotriene release, to an IgE cross-linking stimulus, such as anti-IgE antibody. Although unresponsive to any IgE-mediated stimulation, these basophils responded to non-IgE-mediated stimuli, such as the phorbol ester, 12-o-tetradecanoyl phorbol-13 acetate, the calcium ionophore, A23187, and to formyl-methionyl-leucyl-phenylalanine peptide. These stimuli produced equal dose-response curves in both releaser (basophils able to respond with greater than 5% histamine release to anti-IgE antibody) and nonreleaser basophils. Nonreleaser basophils possessed statistically similar densities of cell-surface IgE antibody (287,000 versus 400,000 IgE molecules per basophil for releaser and nonreleaser basophils, respectively), and with 12-o-tetradecanoyl phorbol-13 acetate as a probe of anti-IgE-induced cross-linking, the IgE on nonreleaser basophils was found to be cross-linked by the polyclonal anti-IgE antibody used for these studies. Interleukin-3 (IL-3) has previously been demonstrated to enhance markedly both histamine and leukotriene release in human basophils. However, IL-3 was unable to convert nonreleasing basophils into releasing basophils, as measured by anti-IgE antibody. IL-3 equivalently enhanced formyl methionine peptide-induced release in both releaser and nonreleaser basophils, suggesting that the lack of an effect on anti-IgE-induced release was not due to a lack of IL-3 receptors. Although there are several possible interpretations of these data, these results and results of our previous studies of protein kinase C activation and cytosolic Ca++ elevations in human basophils suggest that nonreleasing basophils have a defect in early signal transduction, possibly involving the influx of Ca++.     

Drug release from starch-acetate microparticles and films with and without incorporated alpha-amylase

Acetylation of starch considerably decreases its swelling and enzymatic degradation. Thus, starch-acetate (SA) based delivery systems may be suitable for controlled drug delivery. The aim of the present study was to evaluate drug release from the SA microparticles (SA mps) and SA films. The average degree of acetyl substitution (DS) per glucose residue in the starch was either 1.9 (SA DS 1.9) or 2.6 (SA DS 2.6). Timolol (mw 332), calcein (mw 623) and bovine serum albumin (BSA, mw 68,000) were used as model drugs. A continuous timolol release from the both SA mps was observed in phosphate buffer solution (PBS) pH 7.4 (50-days incubation). The release of timolol was faster from the SA DS 1.9 mps than from the SA DS 2.6 mps. Calcein release from both SA mps was continuous in PBS pH 7.4 (5-days incubation). But, calcein release profile from the SA DS 2.6 film in PBS pH 7.4 showed discontinuities. However, the release of calcein from both SA films was continuous in human serum in vitro during the 7-day incubation, i.e. enzymes enhanced calcein release. Thus, alpha-amylase was incorporated into the SA films in order to enhance drug release from the films. However, the effects of incorporation of alpha-amylase on the model macromolecule (BSA) release from the SA films were modest. In conclusion, this study demonstrates the achievement of slow release of different molecular weight model drugs from the SA mps and films as compared to fast release from the native starch preparations. DS of SA, physicochemical properties of a drug and the presence of enzymes can all affect drug release profiles from SA based preparations.     

Controlled release of DNA from poly(vinylpyrrolidone) capsules using cleavable linkers

The design of polymer carriers with tunable degradation and cargo release is fundamental for applications in drug and gene delivery. In this study, we report low-fouling poly(N-vinyl pyrrolidone) (PVPON) capsules assembled via hydrogen bonding and stabilized using covalent cross-linking. We first investigated the effects of pH and ionic strength to optimize the assembly conditions. A model therapeutic cargo (plasmid DNA) was then loaded in the capsules and used for encapsulation and release studies. Two bisazide cross-linkers that contain a disulfide bond, termed PEG? (poly(ethylene glycol)) and PEG(16), were employed to stabilize the multilayer films, and used to tune the degradation and cargo release behavior of the capsules in simulated cytoplasmic conditions. The results suggest that PEG?-stabilized capsules were more efficiently cross-linked, and hence displayed higher plasmid encapsulation. Consequently, the capsules cross-linked with PEG? also showed a two-fold reduction in degradation rate. This ability to achieve controlled carrier degradation and cargo release makes these capsules of potential interest for drug and gene delivery.     

Heparin-modified dendrimer cross-linked collagen matrices for the delivery of basic fibroblast growth factor (FGF-2)

Tissue integration between a tissue-engineered corneal equivalent and the host eye is of critical importance in ensuring long-term implant success. A novel dendrimer cross-linked collagen scaffold has previously shown good corneal epithelial cell compatibility in vitro particularly when the highly functional dendrimer cross-linkers were functionalized to introduce additional biological groups. Herein we investigated heparinization of these materials and their potential to facilitate the delivery of basic fibroblast growth factor (FGF-2) in an active form, ultimately for use as a corneal tissue scaffold. Collagen gels cross-linked with 1-ethyl-3-(3-dimethyl aminopropyl) carbodiimide hydrochloride (EDC) and N-hydroxysuccinimide (NHS) chemistry, and varying amounts of polypropyleneimine octaamine generation 2 (G2) dendimer and heparin were synthesized. Swelling studies and differential scanning calorimetry characterization indicated higher gel stability with the introduction of dendrimer cross-linking, which was not compromised by heparin integration. Dendrimer cross-linked gels with or without heparin gave multiple denaturation peaks, as did the heparinized EDC gels. This is thought to be the result of the heterogeneous cross-linking possible between collagen, the dendrimer and heparin. Release of FGF-2 from collagen gels showed typical first-order kinetics, with an initial burst followed by a prolonged gradual release. Heparinized dendrimer cross-linked gels released approx. 40% of the growth factor over a 2-week period, with significance maintenance of growth factor activity. Incorporation of heparin resulted in somewhat prolonged release from these systems.     

Cross-linked chitosan microspheres as carriers for prolonged delivery of macromolecular drugs

Bovine serum albumin (BSA) and diphtheria toxoid (DT) were loaded by passive absorption from aqueous solutions into preformed glutaraldehyde cross-linked chitosan microspheres. In vitro release of BSA under sink conditions at 37°C showed that even though there was a large burst effect, there was a more or less steady increase with time thereafter for several days. Coating the BSA-loaded particles with paraffin oil or with a polymer, such as polylactic acid, modulated drug release. After the initial burst from PLA coated particles, the release rate increased with time for nearly 2 months. Preliminary immunogenicity studies on Wistar rats using DT loaded chitosan spheres showed that the antibody titres were fairly constant over a 5-month period, although very low compared to DT given on alum as control. Histological studies of placebo microspheres intramuscularly injected into rats demonstrated their tissue compatibility. Biodegradation was not complete in 6 months demonstrating the potential of cross-linked chitosan spheres as a long-acting drug delivery vehicle. The study demonstrated the possibility of incorporating biological macromolecules which are very sensitive to organic solvents, pH, temperature, ultrasound, etc. by a passive absorption technique to degradable biopolymer matrices thereby preserving their biological integrity. It is also shown that drugs passively absorbed into such matrices by taking advantage of their swelling behaviour need not necessarily be released completely in the initial 'burst' and a sustained release may be possible for macromolecules thus incorporated.