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PubMed2001

Oriented Schwann cell growth on micropatterned biodegradable polymer substrates.

This paper investigates the influence of substrate-mediated chemical and physical guidance on the growth and alignment of Schwann cells in vitro. Novel techniques were developed to fabricate microgrooves with adsorbed proteins on biodegradable polymer substrates made of poly(D,L-lactic acid). Compression molding and solvent-casting were used to transfer micropatterns from quartz and silicon substrates onto biodegradable polymer films. Laminin was selectively adsorbed onto the grooves and rat sciatic Schwann cells were seeded on the substrates. Laminin was found to improve adhesion of Schwann cells on the substrates. The microgrooves were found to cause the Schwann cells to align along the direction of the grooves. The groove width influenced Schwann cell alignment the most, while groove depth did not seem to play a significant role. The degradation of the grooves in the solvent cast films was much slower than those in the compression-molded films, making them the preferred substrates for Schwann cell culture.

Biocompatible Materials↗

PubMed2002

Characterization of biodegradable drug delivery vehicles with the adhesive properties of leukocytes.

The site-specific expression of selectins (E- and P-selectin) on endothelial cells of blood vessels during inflammation provides an opportunity for the targeted delivery of anti-inflammatory drugs to inflammatory sites. Previous work in our laboratory has shown that artificial capsules with the adhesive properties of leukocytes can be made by attaching leukocyte adhesive ligands to polystyrene microspheres. In this work, we have adapted this technology to create a targeted delivery system using biodegradable, poly lactic-co-glycolic-acid (PLGA) microspheres. Biotinylated-Sialyl Lewis(x) (sLe(x)), a carbohydrate that serves as a ligand to selectins, was attached to the surface of avidin-linked PLGA microspheres. These carbohydrate-coated microspheres mimic the adhesive behavior of leukocytes on selectins in flow chambers, displaying slow rolling under flow. The rolling velocities displayed by sLe(x)-coated microspheres were similar to those displayed by leukocytes rolling on P- or E-selectin coated surfaces, and these rolling velocities, which relate to the residence time of the capsules, can be tuned by changing the density of carbohydrate residues on microsphere surfaces. We have also demonstrated that these microspheres will release model drugs on a time scale of several days. Therefore, we have made a targeted drug delivery vehicle that mimics the adhesive properties of leukocytes and is biodegradable.

Animals↗

PubMed2002

Pegylated recombinant human epidermal growth factor (rhEGF) for sustained release from biodegradable PLGA microspheres.

Recombinant human epidermal growth factor (rhEGF) was conjugated with polyethylene glycol (PEG) to improve its physical stability during microencapsulation in biodegradable poly(lactic-co-glycolic acid) microspheres. rhEGF was conjugated with N-hydroxysuccimide (NHS)-derivatized methoxy-PEG (mPEG) of MW 2000 and 5000 under various reaction conditions to optimize the extent of pegylation. Pegylated rhEGF showed much enhanced physical stability against homogenization. Pegylated rhEGF was encapsulated in PLGA microspheres by a double emulsion solvent evaporation method to achieve a sustained release. Pegylated rhEGF exhibited a tri-phasic release profile with a reduced initial burst, compared with unpegylated rhEGF. This study demonstrated that protein pegylation enhanced physical stability of protein and could be a good approach to achieve a sustained protein release profile from biodegradable microspheres.

Amino Acid Sequence↗

PubMed2002

Production and evaluation of biodegradable composites based on PHB-PHV copolymer.

In recent years, emphasis in biomaterials engineering has moved from materials that remain stable in the biological environment to materials that can degrade in the human body. Biodegradable materials are designed to degrade gradually and be replaced eventually by newly formed tissue in the body. In this investigation, two particulate bioactive ceramics, i.e., hydroxyapatite (HA) and tricalcium phosphate (TCP), were incorporated into polyhydroxybutyrate-polyhydroxyvalerate (PHB-PHV), which is a biodegradable copolymer. to produce new biomaterials for potential medical applications. All raw materials were commercially available and they were characterised prior to composite production. HA/PHB-PHV and TCP/PHB-PHV composites containiing up to 30 vol% of the bioceramics were produced through an established procedure. Compounded and compression moulded materials were evaluated using various techniques including thermogravimatric analysis, scanning electron microscopy, differential scanning calorimetry and dynamic mechanical analysis. The results showed that intended compositions of composites had been achieved and bioceramic particles were well distributed in the polymer. The degradation temperature of PHB-PHV was significantly reduced by the incorporation of bioceramics, while the melting temperature was slightly affected by the addition of bioceramics. The crystallinity of PHB-PHV was also varied with the presence of HA or TCP particles. The storage modulus and loss modulus of the composites increased with the increase in HA or TCP content. Composites containing the highest percentage of bioceramics exhibited the highest stiffness. Preliminary in vitro study indicated enhanced ability of the composites to induce the formation of bone-like apatite on their surfaces.

Apatites↗

PubMed2002

Localized delivery of paclitaxel in solid tumors from biodegradable chitin microparticle formulations.

Paclitaxel (Taxol)-containing chitin and chitin-Pluronic F-108 microparticles were formulated as biodegradable systems for localized administration in solid tumors. The microparticles were characterized by Fourier transform infrared (FT-IR) spectroscopy, scanning electron microscopy (SEM), and swelling studies in phosphate-buffered saline (PBS, pH 7.4). Lysozyme-induced degradation and in vitro release of paclitaxel was examined in PBS at 37 degrees C. The percent change in tumor volume was used to assess efficacy of the Formulations after local administration in murine Lewis lung carcinoma model. FT-IR confirmed higher degree of acetylation in chitin microparticles from the starting chitosan sample and the SEM showed that the chitin-Pluronic F-108 microparticles were significantly more porous than chitin microparticles. Due to higher porosity, chitin-Pluronic microparticles were able to imbibe higher swelling medium and degraded much faster in the presence of lysozyme than chitin microparticles. After 48 h. 51% of incorporated paclitaxel was released from chitin-Pluronic microparticles as compared to 28% from chitin microparticles. In vivo studies in Lewis lung carcinoma-bearing mice showed that the tumor volumes after 6 days using paclitaxel-loaded chitin and chitin-Pluronic F-108 microparticles was 458 and 307 mm3, respectively. In contrast, the tumor volume was 997 mm3 for the untreated control. The results of this study show that chitin and chitin-Pluronic F-108 microparticles are biodegradable drug delivery systems that can be useful for localized delivery of paclitaxel in solid tumors.

Acetylglucosamine↗

PubMed2003

Extracellular matrix production by human osteoblasts cultured on biodegradable polymers applicable for tissue engineering.

The nature of the extracellular matrix (ECM) is crucial in regulating cell functions via cell-matrix interactions, cytoskeletal organization, and integrin-mediated signaling. In bone, the ECM is composed of proteins such as collagen (CO), fibronectin (FN), laminin (LM), vitronectin (VN), osteopontin (OP) and osteonectin (ON). For bone tissue engineering, the ECM should also be considered in terms of its function in mediating cell adhesion to biomaterials. This study examined ECM production, cytoskeletal organization, and adhesion of primary human osteoblastic cells on biodegradable matrices applicable for tissue engineering, namely polylactic-co-glycolic acid 50:50 (PLAGA) and polylactic acid (PLA). We hypothesized that the osteocompatible, biodegradable polymer surfaces promote the production of bone-specific ECM proteins in a manner dependent on polymer composition. We first examined whether the PLAGA and PLA matrices could support human osteoblastic cell growth by measuring cell adhesion at 3, 6 and 12h post-plating. Adhesion on PLAGA was consistently higher than on PLA throughout the duration of the experiment, and comparable to tissue culture polystyrene (TCPS). ECM components, including CO, FN, LM, ON, OP and VN, produced on the surface of the polymers were quantified by ELISA and localized by immunofluorescence staining. All of these proteins were present at significantly higher levels on PLAGA compared to PLA or TCPS surfaces. On PLAGA, OP and ON were the most abundant ECM components, followed by CO, FN, VN and LN. Immunofluorescence revealed an extracellular distribution for CO and FN, whereas OP and ON were found both intracellularly as well as extracellularly on the polymer. In addition, the actin cytoskeletal network was more extensive in osteoblasts cultured on PLAGA than on PLA or TCPS. In summary, we found that osteoblasts plated on PLAGA adhered better to the substrate, produced higher levels of ECM molecules, and showed greater cytoskeletal organization than on PLA and TCPS. We propose that this difference in ECM composition is functionally related to the enhanced cell adhesion observed on PLAGA. There is initial evidence that specific composition of the PLAGA polymer favors the ECM. Future studies will seek to optimize ECM production on these matrices for bone tissue engineering applications.

Biodegradation, Environmental↗

PubMed1999

Delivering neuroactive molecules from biodegradable microspheres for application in central nervous system disorders.

Nerve growth factor (NGF) may enhance axonal regeneration following injury to the central nervous system (CNS), such as after spinal cord injury. The release profile of NGF, co-encapsulated with ovalbumin, was tailored from biodegradable polymeric microspheres using both polymer degradation and protein loading. Biodegradable polymeric microspheres were prepared from PLGA 50/50, PLGA 85/15, PCL and a blend of PCL/PLGA 50/50 (1:1, w/w), where the latter was used to further tailor the degradation rate. The amount of protein loaded in the microspheres was varied, with PCL encapsulating the greatest amount of protein and PLGA 50/50 encapsulating the least. A two-phase release profile was observed for all polymers where the first phase resulted from release of surface proteins and the second phase resulted predominantly from polymer degradation. Polymer degradation influenced the release profile most notably from PLGA 50/50 and PLGA 85/15 microspheres. The amount and bioactivity of released NGF was followed over a 91 d period using a NGF-ELISA and PC12 cells, respectively. NGF was found to be bioactive for 91 d, which is longer than previously reported.

Animals↗

PubMed1999

Effect of filler content on the profile of released biodegradation products in micro-filled bis-GMA/TEGDMA dental composite resins.

This study assesses the effect of the filler content, in a micro-filled composite (0.04 microm), on the liberation of biodegradation products derived from two model composite systems. The materials were based on bis-phenyl glycidyl dimethacrylate (bis-GMA) and triethylenene glycol dimethacrylate (TEGDMA) monomers. The composites were produced using silica filler concentrations of 20 and 40%) by weight. Samples were incubated with either cholesterol esterase (CE) or phosphate buffer solutions (PBS) for 8, 16 and 32 days. Products were isolated by high-performance liquid chromatography (HPLC) and identified by mass spectrometry. The identified products included TEGDMA, 2,2-bis[4(2,3-hydroxypropoxy)-phenyl]propane (bis-HPPP) and triethylene glycol methacrylate (TEGMA). Bis-HPPP was only produced in the presence of enzyme. The amount of isolated TEGMA, in both composite systems, was shown to be significantly higher for materials incubated with enzyme than their buffer counterparts (P < 0.05). Between 0 and 8 days incubation with enzyme, significantly higher amounts of Bis-HPPP and TEGMA were generated with the lower filler model material (composite-20) than the higher filled composite (composite-40), while the opposite effect was observed between 8 and 16 days. The data indicate that biodegradation product release profiles are dependent on the filler/resin ratios, and suggests that this parameter should be considered when assessing product release for biocompatibility issues pertaining to dental composite systems.

Biodegradation, Environmental↗

PubMed1999

Vascularization effect of basic fibroblast growth factor released from gelatin hydrogels with different biodegradabilities.

Biodegradable gelatin hydrogels were prepared through the glutaraldehyde crosslinking of acidic gelatin with an isoelectric point (IEP) of 5.0 and the basic gelatin with an IEP of 9.0. The hydrogel water content was changed by the concentration of both gelatin and glutaraldehyde, used for hydrogel preparation. An aqueous solution of basic fibroblast growth factor (bFGF) was sorbed into the gelatin hydrogel freeze-dried to obtain a bFGF-incorporating gelatin hydrogel. Irrespective of the hydrogel water content, approximately 30% of the incorporated bFGF was released from the bFGF-incorporating acidic gelatin hydrogel, within the first day into phosphate-buffered saline solution at 37 degrees C, followed by no substantial release. Probably, the basic bFGF complexed with the acidic gelatin through poly-ion complexation would not be released under the in vitro non-degradation condition of gelatin. On the contrary, almost 100% of the incorporated bFGF was initially released from all types of basic gelatin hydrogels. This is due to the simple diffusion of bFGF because of no complexation between bFGF and the basic gelatin. When implanted subcutaneously into the mouse back, bFGF-incorporating acidic and basic gelatin hydrogels with higher water contents were degraded with time faster than those with lower water contents. Significant neovascularization was induced around the implanted site of the bFGF-incorporating acidic gelatin hydrogel. The induction period prolonged with the decrease in hydrogel water content. On the other hand, such a prolonged vascularization effect was not achieved by the bFGF-incorporating basic gelatin hydrogel and the hydrogel initially exhibited less enhanced effect, irrespective of the water content. These findings indicate that the controlled release of biologically active bFGF is caused by biodegradation of the acidic gelatin hydrogel, resulting in induction of vascularization effect dependent on the water content. It is possible that only the transient vascularization by the basic gelatin hydrogel is due to the initial large burst in bFGF release, probably because of the down regulation of bFGF receptor.

Animals↗

PubMed2000

Biodegradable photo-crosslinked poly(ether-ester) networks for lubricious coatings.

Biodegradable poly(ether-ester) networks were synthesized by UV photopolymerization and their lubrication performances were evaluated. Polyethers such as poly(ethylene glycol) (PEG), poly(propylene glycol) (PPG), and poly(tetramethylene glycol) (PTMG) were copolymerized with oligomers of D,L-lactic acid and terminated with acrylate groups to form photopolymerizable macromers. 1H NMR and IR spectra confirmed the chemical structures of copolymers and diacrylated macromers. Crosslinked polymer networks were formed upon UV-initiated free-radical polymerization. Gel contents, water contents, and contact angles were measured to characterize the crosslinked networks. In vitro degradation times of the crosslinked networks at 37 degrees C in 1 N NaOH varied from 20 min to 7 days depending on the crosslinking density (molecular weight of macromer) and the hydrophilic susceptibility (types of polyethers). The crosslinked polymers were coated on stainless-steel needles to investigate the lubrication properties by measuring penetration and drag forces through rubber stoppers. The maximum improvement in penetration force over control was 41% in the needle coated with PPG-based polymer networks (molecular weight of PPG = 4000). These materials can potentially be used as biodegradable lubricants for coating various medical products to replace the existing non-degradable silicone-based materials currently used.

Biodegradation, Environmental↗

PubMed2003

An assessment of the toxicity of pyridinium chlorides and their biodegradation intermediates.

Toxicity investigations were conducted for four pyridinium chlorides belonging to cationic surface-active substances (CSAS), which differed from each other in the numbers of methyl groups (CH(3)) in pyridinium ring. The crustacean Daphnia magna, the fish Lebistes reticulatus and the alga Scenedesmus quadricauda were chosen as biotests. Toxicity of examined preparations appeared to be very high but did not depend on their chemical structure. S. quadricauda was the most sensitive organism. Toxicity of intermediate products obtained in biological oxidation process was also examined. Biodegradation was conducted according to the "river water test". It was found that only partial degradation took place while pyridinium chlorides constituted main energy and carbon source. Presence of biodegradation intermediate products was shown on the basis of 1H NMR analysis. Intermediates were not toxic to any biotests.

Animals↗

PubMed2002

Tolerance to biodegraded crude oil in marine invertebrate embryos and larvae is associated with expression of a multixenobiotic resistance transporter.

The toxicity of water-soluble fractions of biodegraded crude oil (BWSF) to embryos and larvae of two marine invertebrates, the white sea urchin (Lytechinus anamesus) and the fat innkeeper (Urechis caupo), was studied. Santa Barbara Channel crude oil was artificially weathered and subjected to biodegradation using a mixed microbe culture obtained from natural oil seep sites. The degradation culture inoculated with seep sediment microbes accumulated 43.7 microg/l water-soluble hydrocarbons. In contrast water-soluble fractions from the non-degraded cultures (NWSF) only accumulated 3.05 microg/l. BWSF proved deleterious to Lytechinus embryo development at low concentrations (EC50 = 0.33 mg/l) but was essentially non-toxic to Urechis embryos/larvae up to 3.0 mg/l. An established mechanism for handling of a wide array of xenobiotics in Urechis embryos is the multixenobiotoic resistance transporter multixenobiotic response (MXR, also known as multidrug resistance, MDR). This mechanism is primarily mediated by ATP-dependent, efflux pumps that extrude a wide array of xenobiotic compounds. In this study, we show that Lytechinus larvae do not appear to express MXR efflux protein nor MXR mediated dye efflux capacity. In contrast, BWSF acts as a competitive inhibitor of MXR transport-mediated dye efflux in Urechis larvae. These results suggest that MXR may be an important mechanism for extrusion of the by-products of crude oil degradation by microbes, and that the level of its expression may determine the susceptibility of organisms to degraded oil hydrocarbons.

Adaptation, Physiological↗

PubMed2001

Versatility of biodegradable biopolymers: degradability and an in vivo application.

Biodegradable materials have various important applications in the biomedical field. There are basically two groups of polyesters which have significant importance in this field. These are polylactides and polyhydroxybutyrates. Both groups degrade via hydrolysis with the rates of degradation depending on medium properties such as pH, temperature, solvent and presence of biocatalysts, as well as on chemical compositions. In order for these biomaterials to be suitable for use in load bearing applications without deformation or warping their strengths and their capability to maintain their form must be improved. To insure dimensional stability during degradation and to match modulus and strength to that of bone, introduction of a reinforcing structure for those applications to plate fixation through the creation of an interpenetrating network might be a feasible approach. In this study, poly(lactide-co-glycolide) (PLGA), was the major structural element to be strengthened by a three-dimensional network or "scaffold" of another biodegradable polymer, poly(propylene fumarate) (PPF). PPF would be crosslinked with a biocompatible vinyl monomer, vinylpyrrolidone (VP). Three different approaches were tested to create dimensionally stable bone plates. First, via in situ crosslinking of PPF in the presence of PLGA. Secondly, by blending of precrosslinked PPF with PLGA. Finally, by simultaneous crosslinking and molding of the PLGA, PPF and VP. These were compared against extruded or compression molded PLGA controls. Results showed that compression molding at room temperature followed by crosslinking under pressure at elevated temperature and subsequently by gamma-irradiation appeared to yield the most favorable product as judged by swelling, hardness and flexural strength data. The composition of the implant material, PLGA(3):PPF(1):VP(0.7), appeared to be suitable and formed the compositional and procedural basis for in vivo biocompatibility studies.

Absorbable Implants↗

PubMed2001

Mathematical model for evaluation of mass transfer limitations in phenol biodegradation by immobilized Pseudomonas putida.

A mathematical model is proposed to analyze the mass transfer limitations in phenol biodegradation using Pseudomonas putida immobilized in calcium alginate. The model takes into account internal and external mass transfer limitations, substrate inhibition kinetics and the dependence of the effective diffusivity of phenol in alginate gel on cell concentration. The model is validated with the experimental data from batch fermentation. The effect of various operating conditions such as initial phenol concentration, initial cell loading, alginate gel loading on the biodegradation of phenol is experimentally demonstrated. Phenol degradation time is found to decrease initially and reach stationary value with increase in cell loading as well as gel loading. The model predicts these trends reasonably well and shows the presence of external mass transfer limitations. A new concept of effectiveness factor is introduced to analyze the overall performance of batch fermentation.

Biodegradation, Environmental↗

PubMed2003

Estimation of viable biomass in aerobic biodegradation processes of organic fraction of municipal solid waste (MSW).

2-(p-Iodophenyl)-3-(p-nitrophenyl)-5-phenyltetrazolium chloride (INT) dehydrogenase test and RNA assay were introduced to evaluate biomass in the processes of aerobic biodegradation of the organic fraction of municipal solid waste (MSW) in bioreactors. It was found that RNA quantification by KOH/UV method delivered reliable and repeatable results. Relative standard deviation (RSD) for INT test was significantly higher than for RNA assay and achieved values of 3-15%. Moreover, it occurred that the optimum temperature for the growth of autochthonic biomass, which takes part in the biodegradation process, was in the range from 25 to 37 degrees C.

Bacteria, Aerobic↗

PubMed1998

Biodegradable products by lipase biocatalysis.

The interest in the applications of biocatalysis in organic syntheses has rapidly increased. In this context, lipases have recently become one of the most studied groups of enzymes. We have demonstrated that lipases can be used as biocatalyst in the production of useful biodegradable compounds. A number of examples are given. 1-Butyl oleate was produced by direct esterification of butanol and oleic acid to decrease the viscosity of biodiesel in winter use. Enzymic alcoholysis of vegetable oils without additional organic solvent has been little investigated. We have shown that a mixture of 2-ethyl-1-hexyl esters can be obtained in a good yield by enzymic transesterification from rapeseed oil fatty acids for use as a solvent. Trimethylolpropane esters were also similarly synthesized as lubricants. Finally, the discovery that lipases can also catalyze ester syntheses and transesterification reactions in organic solvent systems has opened up the possibility of enzyme catalyzed production of biodegradable polyesters. In direct polyesterification of 1,4-butanediol and sebacic acid, polyesters with a mass average molar mass of the order of 56,000 g mol-1 or higher, and a maximum molar mass of about 130,000 g mol-1 were also obtained by using lipase as biocatalyst. Finally, we have demonstrated that also aromatic polyesters can be synthesized by lipase biocatalysis, a higher than 50,000 g mol-1 mass average molar mass of poly(1,6-hexanediyl isophthalate) as an example.

Biodegradation, Environmental↗

PubMed1999

Biodegradation of benzene, toluene, ethylbenzene, and o-xylene by a coculture of Pseudomonas putida and Pseudomonas fluorescens immobilized in a fibrous-bed bioreactor.

A fibrous-bed bioreactor containing the coculture of Pseudomonas putida and P. fluorescens immobilized in a fibrous matrix was developed to degrade benzene (B), toluene (T), ethylbenzene (E), and o-xylene (X) in synthetic waste streams. The kinetics of BTEX biodegradation by immobilized cells adapted in the fibrous-bed bioreactor and free cells grown in serum bottles were studied. In general, the BTEX biodegradation rate increased with increasing substrate concentration and then decreased after reaching a maximum, showing substrate-inhibition kinetics. However, for immobilized cells, the degradation rate was much higher than that of free cells. Compared to free cells, immobilized cells in the bioreactor tolerated higher concentrations (> 1000 mg l-1) of benzene and toluene, and gave at least 16-fold higher degradation rates for benzene, ethylbenzene, and o-xylene, and a 9-fold higher degradation rate for toluene. Complete and simultaneous degradation of BTEX mixture was achieved in the bioreactor under hypoxic conditions. Cells in the bioreactor were relatively insensitive to benzene toxicity; this insensitivity was attributed to adaptation of the cells in the bioreactor. Compared to the original seeding culture, the adapted cells from the fibrous-bed bioreactor had higher specific growth rate, benzene degradation rate, and cell yield when the benzene concentration was higher than 100 mg l-1. Cells in the fibrous bed had a long, slim morphology, which is different from the normal short-rod shape found for suspended cells in solution.

Benzene↗

PubMed2003

Biodegradable microspheres for protein delivery.

In a very short time, since their emergence, the field of controlled delivery of proteins has grown immensely. Because of their relatively large size, they have low transdermal bioavailabilities. Oral bioavailability is generally poor since they are poorly absorbed and easily degraded by proteolytic enzymes in the gastrointestinal tract. Ocular and nasal delivery is also unfavorable due to degradation by enzymes present in eye tissues and nasal mucosa. Thus parenteral delivery is currently most demanding and suitable for delivery of such molecules. In systemic delivery of proteins, biodegradable microspheres as parenteral depot formulation occupy an important place because of several aspects like protection of sensitive proteins from degradation, prolonged or modified release, pulsatile release patterns. The main objective in developing controlled release protein injectables is avoidance of regular invasive doses which in turn provide patient compliance, comfort as well as control over blood levels. This review presents the outstanding contributions in field of biodegradable microspheres as protein delivery systems, their methods of preparation, drug release, stability, interaction with immune system and regulatory considerations.

Biodegradation, Environmental↗