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Testing biodegradability with standardized methods.

Laboratory test methods are used by industry laboratories to determine biodegradability, an important parameter for the evaluation of the ecological behaviour of substances. Biodegradability has a key role due to the simple fact that a degradable substance will cause no long term risk in the environment. The great variety of biodegradation processes in the natural environment and in technical plants for treating waste water and solid wastes gave rise to a rather large number of test methods based on different test principles. To guarantee the acceptance of the test results by authorities and customers internationally standardized methods (ISO, OECD) and established quality criteria (GLP, EN 45,000, ISO 9000) are used.

Biodegradation, Environmental↗

Relevance of aquatic biodegradation tests for predicting degradation of polymeric materials during biological solid waste treatment.

The aquatic biodegradability of cellulose and cellulose acetate with degrees of substitution (d.s.) in the range of 1.5 to 3.0, was compared with the mineralization under laboratory controlled composting conditions. In line with previous observations, it was found that cellulose acetates with d.s. < or = 2.5 were readily mineralized to CO2 in the controlled composting test. The degradation rate was clearly affected by the degree of substitution (d.s. 1.5 > d.s. 2.5 > d.s. 3.0). Surprisingly, however, biodegradation of cellulose acetate materials was not observed in the aquatic Strum test. Modifications of the pH and the inoculum source in an attempt to improve the activity of fungi and actinomycetes in the aquatic environment, did not increase CO2-evolution. It is concluded that the relevance of modified Strum tests is limited for predicting complete biodegradation of polymeric materials during biological waste processing. For evaluation of the compostability of polymeric products or packaging materials, more relevant laboratory controlled composting tests should be used.

Actinomycetaceae↗

Biodegradation and toxicity of wastewater from industry producing mineral fibres for thermal insulation.

The water chemistry, toxicity, and biodegradation of wastewater from an industry producing mineral fibres for thermal insulation were studied. Values for COD, BOD5, suspended solids, and phenol exceeded permissible values for the wastewater discharged into a nearby river and acute toxicity was also detected. Consequently, the effluent should be treated in a municipal wastewater treatment plant so its ready biodegradability was investigated. We found that the wastewater was readily biodegradable, therefore we assume it can be treated in the wastewater treatment plant as the ratio of the wastewater flow rate and the minimal total inflow into the sewage treatment plant would be one to at least 30.

Animals↗

Effect of the alkyl chain length on the anaerobic biodegradability and toxicity of quaternary ammonium based surfactants.

The anaerobic biodegradability and toxicity to methanogenic gas production of different alkyl chain length homologs of quaternary ammonium based surfactants were examinated. Two series of these cationic surfactants were selected: alkyl trimethyl ammonium and alkyl benzyl dimethyl ammonium compounds. A simple anaerobic gas production test containing municipal digester solids as a source of anaerobic bacteria was used. Under the applied methanogenic conditions, the cationic surfactants tested showed a very poor primary biodegradation and no evidence of any extent of ultimate biodegradation was observed. The toxicity of quaternary ammonium based surfactants to methanogenic gas production decreased with increasing the alkyl chain length.

Alkylation↗

Toxic effect of surfactants and probable products of their biodegradation on methanogenesis in an anaerobic microbial community.

Surfactants used in household and various industries, are rather toxic; therefore, the accumulation of these compounds in the environment through wastewaters has challenged the problem of their biodegradation. In this research, an attempt was made to assess the toxic effect of various surfactants and the likely products of their biodegradation on the acetoclastic methanogens of an anaerobic microbial community. Among the substances investigated, cationic surfactants were found to be most toxic to methanogens: 154 mg/l alkamon DS and 345 mg/l catamin AB induced a 50% inhibition of methanogenesis. Toxicity studies of some aromatic and cyclic compounds, as the probable products of biodegradation of alkylbenzene sulfonate surfactants, showed that methanogenesis in the microbial community under study are rather tolerant to high concentrations of these compounds.

Anaerobiosis↗

Biodegradability of some antibiotics, elimination of the genotoxicity and affection of wastewater bacteria in a simple test.

Most antibiotics and their metabolites are excreted by humans after administration and therefore reach the municipal sewage with the excretions. Only little is known about their biodegradability in aquatic environments. It was recognised that genotoxic substances may represent a health hazard to humans but also may affect organisms in the environment. Therefore, the biodegradability of some clinically important antibiotic drugs (ciprofloxacin, ofloxacin, metronidazole) and hereby the elimination of their genotoxicity was investigated as the first step of an environmental risk assessment using the Closed Bottle test (CBT) (OECD 301 D) and the SOS chromotest. Additionally, to assess toxicity of the antibiotics tested against aquatic bacteria (i) a growth inhibition test (GIT) with Pseudomonas putida was conducted, (ii) a toxicity control was used in the CBT and (iii) the colony forming units (CFUs) were monitored in the test vessels. Worst case concentrations of the antibiotics in hospital effluents were estimated and compared with minimum inhibitory concentrations for susceptible pathogenic bacteria and with the genotoxic potency in the SOS chromotest. Both the concentrations calculated for hospital effluents and the adverse effects in bacteria were in the same order of magnitude. None of the test compounds were biodegraded. The genotoxicity was not eliminated.

Anti-Bacterial Agents↗

Biodegradability of ethoxylated fatty amines: detoxification through a central fission of these surfactants.

Non-ionic surfactants are initially degraded by a central fission of the molecule or by the oxidation of the far end of the polyoxyethylene (EO) chain. Alcohol ethoxylates are metabolized via a central fission, whereas alkylphenol ethoxylates are degraded by a stepwise shortening of the polyoxyethylene moiety. The biodegradation curves of ethoxylated fatty amines suggest a 'rapid' mineralization via an oxidation of the alkyl chain. The intermediates formed, viz., secondary ethoxylated amines, are 'slowly' biodegraded. This 'total' mineralization of the ethoxylated fatty amines was demonstrated in 'prolonged' Closed-Bottle tests. Decisive evidence for a central fission of ethoxylated fatty amines was obtained in a pure culture study. An isolated Pseudomonas sp. cleaved the C(alkyl)-N bond of octadecyl-bis(2-hydroxyethyl) amine and utilized the alkyl chain as sole carbon and energy source. Biodegradation products of both alcohol ethoxylates and ethoxylated fatty amines, formed through a central fission of the molecule, are non-toxic.

Amines↗

Comparison of the fuel oil biodegradation potential of hydrocarbon-assimilating microorganisms isolated from a temperate agricultural soil.

Strains of hydrocarbon-degrading microorganisms (bacteria and fungi) were isolated from an agricultural soil in France. In a field, a portion was treated with oily cuttings resulting from the drilling of an onshore well. The cuttings which were spread at the rate of 600 g HC m-2 contained 10% of fuel oil hydrocarbons (HC). Another part of the field was left untreated. Three months after HC spreading, HC adapted bacteria and fungi were isolated at different soil depths in the two plots and identified. The biodegradation potential of the isolated strains was monitored by measuring the degradation rate of total HC, saturated hydrocarbons, aromatic hydrocarbons and resins of the fuel. Bacteria of the genera Pseudomonas, Brevundimonas Sphingomonas, Acinetobacter, Rhodococcus, Arthrobacter, Corynebacterium and fungi belonging to Aspergillus, Penicillium, Beauveria, Acremonium, Cladosporium, Fusarium, and Trichoderma were identified. The most active strains in the assimilation of saturates and aromatics were Arthrobacter sp., Sphingomonas spiritivorum, Acinetobacter baumanii, Beauveria alba and Penicillum simplicissimum. The biodegradation potential of the hydrocarbon utilizing microorganisms isolated from polluted or unpolluted soils were similar. In laboratory pure cultures, saturated HC were more degraded than aromatic HC, whereas resins were resistant to microbial attack. On an average, individual bacterial strains were more active than fungi in HC biodegradation.

Bacteria↗

Biodegradable implants in fracture fixation: early results of treatment of fractures of the ankle.

Biodegradable polylactide-glycolide copolymer implants for fracture fixation were developed and tested in rabbits. In a prospective clinical study 44 patients with a displaced fracture of the ankle were randomly allocated to two groups; one was treated with conventional metallic implants and the other with the biodegradable implants. There were no differences between the two groups in the early results, but the biodegradable fixation method is advantageous because the removal procedure associated with metallic implants is avoided.

Adult↗

Placebo-controlled trial of safety and efficacy of intraoperative controlled delivery by biodegradable polymers of chemotherapy for recurrent gliomas. The Polymer-brain Tumor Treatment Group.

Chemotherapy for brain tumours has been limited because of difficulty in achieving adequate exposure to the tumour without systemic toxicity. We have developed a method for local sustained release of chemotherapeutic agents by their incorporation into biodegradable polymers. Implantation of the drug-impregnated polymer at the tumour site allows prolonged local exposure with minimal systemic exposure. We conducted a randomised, placebo-controlled, prospective study to evaluate the effectiveness of biodegradable polymers impregnated with carmustine to treat recurrent malignant gliomas. In 27 medical centres, 222 patients with recurrent malignant brain tumours requiring re-operation were randomly assigned to receive surgically implanted biodegradable polymer discs with or without 3.85% carmustine. Randomisation balanced the treatment groups for all of the prognostic factors examined. Median survival of the 110 patients who received carmustine polymers was 31 weeks compared with 23 weeks for the 112 patients who received only placebo polymers (hazard ratio = 0.67, p = 0.006, after accounting for the effects of prognostic factors). Among patients with glioblastoma, 6-month survival in those treated with carmustine-polymer discs was 50% greater than in those treated with placebo (mortality = 32 of 72 [44%] vs 47 of 73 [64%], p = 0.02). There were no clinically important adverse reactions related to the carmustine polymer, either in the brain or systemically. Interstitial chemotherapy delivered with polymers directly to brain tumours at the time of surgery seems to be a safe and effective treatment for recurrent malignant gliomas.

Biodegradation, Environmental↗

A new approach based on injection moulding to produce biodegradable starch-based polymeric scaffolds: morphology, mechanical and degradation behaviour.

One of the present challenges in polymer scaffold processing is the fabrication of three-dimensional (3D) architectures with an adequate mechanical performance to be used in the tissue engineering of hard tissues. This paper describes a preliminary study on the development of a new method to produce biodegradable scaffolds from a range of corn-starch-based polymers. In some cases, hydroxlapatite was also used as a reinforcement of the biodegradable polymers. The developed methodology consists of a standard conventional injection moulding process, on which a solid blowing agent based on carboxylic acids is used to generate the foaming of the bulk of the moulded part. The proposed route allows for the production of scaffolds with a compact skin and a porous core, with promising mechanical properties. By using the developed method it is possible to manufacture biodegradable polymer scaffolds in an easy (melt-based processing) and reproducible manner. The scaffolds can be moulded into complex shapes, and the blowing additives do not affect the non-cytotoxic behaviour of the starch-based materials. The materials produced using this method were evaluated with respect to the morphology of the porous structure, and the respective mechanical properties and degradation behaviour. It was demonstrated that it is possible to obtain, by a standard melt based processing route, 3D scaffolds with complex shapes that exhibit an appropriate morphology, without decreasing significantly the mechanical properties of the materials. It is believed that the optimisation of the proposed processing methodology may lead to the production of scaffolds that might be used on the regeneration of load-bearing tissues.

Biocompatible Materials↗

A new nerve guide conduit material composed of a biodegradable poly(phosphoester).

There is a resurgence of interest in the development of degradable and biocompatible polymers for fabrication of nerve guide conduits (NGCs) in recent years. Poly(phosphoester) (PPE) polymers are among the attractive candidates in this context, in view of their high biocompatibility, adjustable biodegradability, flexibility in coupling fragile biomolecules under physiological conditions and a wide variety of physicochemical properties. The feasibility of using a biodegradable PPE, P(BHET-EOP/TC), as a novel NGC material was investigated. Two types of conduits were fabricated by using two batches of P(BHET-EOP/TC) with different weight-average molecular weights (Mw) and polydispersity indexes (PI). The polymers as well as conduits were non-toxic to all six types of cells tested, including primary neurones and neuronally differentiated PC12 cells. After in situ implantation in the sciatic nerve of the rat, two types of conduits triggered a similar tissue response, inducing the formation of a thin tissue capsule composed of approximately eight layers of fibroblasts surrounding the conduits at 3 months. Biological performances of the conduits were examined in the rat sciatic nerve model with a 10 mm gap. Although tube fragmentation, even tube breakage, was observed within less than 5 days post-implantation, successful regeneration through the gap occurred in both types of conduits, with four out of 10 in the Type I conduits (Mw 14,900 and PI 2.57) and 11 out of 12 in the Type II conduits (Mw 18,900 and PI 1.72). The degradation of conduits was further evidenced by increased roughness on the tube surface in vivo under scanning electron microscope and a mass decrease in a time-dependent manner in vitro. The Mw of the polymers dropped 33 and 24% in the Type I and II conduits, respectively, in vitro within 3 months. Among their advantages over other biodegradable NGCs, the PPE conduits showed negligible swelling and no crystallisation after implantation. Thus, these PPE conduits can be effective aids for nerve regeneration with potential to be further developed into more sophisticated NGCs that have better control of the conduit micro-environment for improved nerve regeneration.

Animals↗

Isolation of methylene dianiline and aqueous-soluble biodegradation products from polycarbonate-polyurethanes.

Polycarbonate-polyurethanes (PCNUs) have provided the medical device industry with practical alternatives to oxidation-sensitive polyether-urethanes (PEUs). To date, many studies have focused on PCNUs synthesized with 4,4'-methylene diphenyl-diisocyanate (MDI). The relative hydrolytic stability of this class of polyurethanes is actually quite surprising given the inherent hydrolytic potential of the aliphatic carbonate group. Yet, there has been little information reporting on the rationale for the material's demonstrated hydrolytic stability. Recent work has shown that PCNU materials have a strong sensitivity towards hydrolysis when changes are made to their hard segment content and/or chemistry. However, knowledge is specifically lacking in regards of the identification of cleavage sites and the specific nature of the biodegradation products. Using high-performance liquid chromatography, radiolabel tracers and mass spectrometry, the current study provides insight into the distribution of biodegradation products from the enzyme-catalyzed hydrolysis of five different PCNUs. The hydrolytic sensitivity of the materials is shown to be related to the distribution of products, which itself is a direct consequence of unique micro-structures formed within the different materials. While an MDI-based polymer was shown to be the most hydrolytically stable material, it was the only PCNU that produced its diamine analog, in this case 4,4'-methylene dianiline (MDA), as a degradation product. Given the concern over aromatic diamine toxicity, this finding is important and highlights the fact that relative biostability is a distinct issue from that of degradation product toxicity, and that both must be considered separately when assessing the impact of biodegradation on biomaterial in vivo compatibility.

Aniline Compounds↗

Biodegradation and distribution of water-soluble chitosan in mice.

Randomly 50% deacetylated chitin, called Chi, was examined on the biodegradability, body distribution and urinary excretion after the intraperitoneal (ip) administration to mice. These characteristics were investigated using fluorescein isothiocyanate (FITC)-labeled Chi (FTC-Chi). The in vitro biodegradability was investigated by incubation with lysozyme and murine plasma and urine. The degradation of Chi or FTC-Chi was accelerated by lysozyme, plasma and urine. The molecular weight was checked by gel-chromatography. The degradation product showed a fairly small molecular weight and contained no FTC-Chi of a large one. The body distribution and urinary excretion of FTC-Chi were investigated at 1, 14 and 24 h after the ip injection to mice. FTC-Chi moved fast to the kidney and urine, and was scarcely distributed to the liver, spleen, abdominal dropsy and plasma. Most of FTC-Chi was excreted into urine after 14 h, and the molecular weight of the excreted FTC-Chi was as small as that of the product obtained by the long in vitro incubation. Therefore, Chi is considered to be highly biodegradable and easily excreted in urine, and further it is suggested to have no problem on accumulation in the body; however, at the same time, Chi is found not to operate as a polymer support showing long retention in the body.

Acetylation↗

Integrated assessment for anaerobic biodegradability of organic compounds using the analytical hierarchy process.

Anaerobic biodegradability of organic compounds can be assessed from the changes in organic compounds, the end products, and the activity of microorganisms. In this study, all three of these variables were considered to integrate assessment of the anaerobic biodegradability of organic compounds. The analytical hierarchy process (AHP) was introduced to integrate the assessment. Real values between 0 and 10 were determined in the pairwise comparison matrix according to the values of assessment indices of the three variables. A new adjustment method, which can obtain minimum adjustment of the values in matrix, was proposed to reach the acceptable consistency of the matrix. Finally, the anaerobic biodegradability of 14 organic compounds was assessed using AHP.

Anaerobiosis↗

Biodegradation of bioaccessible textile azo dyes by Phanerochaete chrysosporium.

Azo dyes are important chemical pollutants of industrial origin. Textile azo dyes with bioaccessible groups for lignin degrading fungi, such as 2-methoxyphenol (guaiacol) and 2,6-dimethoxyphenol (syringol), were synthesised using different aminobenzoic and aminosulphonic acids as diazo components. The inocula of the best biodegradation assays were obtained from a pre-growth medium (PAM), containing one of the synthesised dyes. The results of the dye biodegradation assays were evaluated every 7 days, by the decrease of the absorbance at the maximum wavelength of the dye, by the decrease of the sucrose concentration in the culture medium and by the increase of the biomass during the 28 days of assay. It was observed that the extent of dye biodegradation depended on the sucrose concentration, on the degraded dye structure and, on the dye present in the PAM medium.

Azo Compounds↗

New biodegradable polymers for injectable drug delivery systems.

Many biodegradable polymers were used for drug delivery and some are successful for human application. There remains fabrication problems, such as difficult processability and limited organic solvent and irreproducible drug release kinetics. New star-shaped block copolymers, of which the typical molecular architecture is presented, results from their distinct solution properties, thermal properties and morphology. Their unique physical properties are due to the three-dimensional, hyperbranched molecular architecture and influence microsphere fabrication, drug release and degradation profiles. We recently synthesized thermosensitive biodegradable hydrogel consisting of polyethylene oxide and poly(L-lactic acid). Aqueous solution of these copolymers with proper combination of molecular weights exhibit temperature-dependent reversible sol-gel transition. Desired molecular arrangements provide unique behavior that sol (at low temperature) form gel (at body temperature). The use of these two biodegradable polymers have great advantages for sustained injectable drug delivery systems. The formulation is simple, which is totally free of organic solvent. In sol or aqueous solution state of this polymer solubilized hydrophobic drugs prior to form gel matrix.

Animals↗

Branched biodegradable polyesters for parenteral drug delivery systems.

Continuous, 'infusion-like' drug release profiles from biodegradable parenteral delivery systems are difficult to achieve for proteins and other hydrophilic macromolecular drugs with commonly used linear polyesters from lactic acid (PLA) and its random copolymers with glycolic acid (PLG). Drug release rates can be modified either by increasing the hydrophilicity of polyesters or by manipulating the polymer architecture to adjust polymer degradation rates and thus drug release. Therefore, we investigated different branching concepts for biodegradable polyesters of PLA and PLG. For one four- and eight-arm poly(ethylene oxide)s (PEO) were grafted with shorter polyester chains leading to star-branched structures. Secondly we obtained comb-like polyesters using both charged and uncharged dextrans or poly(vinyl alcohol)s (PVA) as hydrophilic backbones. The star-shaped and brush-like grafted polymers were intensively characterized by methods, such as NMR, IR, SEC-SLS, DSC and viscosity measurements. Tailor-made properties make these novel biodegradable polyesters promising candidates for parenteral protein delivery systems. While the star-branched polyesters have shown some interesting properties with respect to their degradation behavior, retaining the PEO blocks longer than ABA triblock copolymers, their release properties need further optimization. Brush-like branched polyesters on the other hand seem to possess both degradation and release properties meriting further investigations for parenteral protein delivery systems.

Biodegradation, Environmental↗