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A kinetic model for surfactant inhibition of pentachlorophenol biodegradation.

A kinetic model is used to describe the effect of the nonionic surfactant Tergitol NP-10 (TNP10) on pentachlorophenol (PCP) biodegradation by Sphingomonas chlorophenolica sp. strain RA2. Different initial biomass to initial substrate ratios ranging from 13 to 418 were tested with 23 TNP10 concentrations ranging from 0 to 1500 mg/L. Tests were also conducted at 10 degrees C and 20 degrees C. No PCP biodegradation inhibition was observed at concentrations below the critical micelle concentration (CMC) of 50 mg/L. TNP10 concentrations above 100 to 200 mg/L were increasingly inhibitory to PCP biodegradation rates. This inhibition was best described by the Monod kinetic equation wherein the effect of TNP10 inhibition is reflected in the half-saturation constant (Ks). The value of the Ks increased from between 1.5 and 13.5 mg/L with no surfactant present to 44 to 131 mg/L at 1000 mg/L TNP10. Using a standard competitive inhibition approach, the inhibition constant for TNP10 was approximately 100 mg/L at both 10 degrees C and 20 degrees C.

Biodegradation, Environmental↗

Yield prediction and stoichiometry of multi-step biodegradation reactions involving oxygenation.

Microorganisms can initiate the degradation of organic compounds by oxygenation reactions that require the investment of energy and electrons. This diversion of energy and electrons away from synthesis reactions leads to decreased overall cell yields. A thermodynamic method was developed that improves the accuracy of cell yield prediction for compounds degraded through pathways involving oxygenation reactions. This method predicts yields and stoichiometry for each step in the biodegradation pathway, thus enabling modeling a multi-step biodegradation process in which oxygenations occur and intermediates may persist. EDTA and benzene biodegradation are presented as examples. The method compares favorably with other yield prediction methods while providing additional information of yields for intermediates produced in the degradation pathway.

Benzene↗

Electrokinetic movement and biodegradation of 2,4-dichlorophenoxyacetic acid in silt soil.

The coupling of electrokinetic movement of an organic contaminant, 2,4-dichlorophenoxyacetic acid (2,4-D), through soil and its biodegradation in situ has been demonstrated. In a first experiment, the direction and rate of movement of 2,4-D were determined using homogeneously contaminated soil (864 mg 2,4-D/kg dry weight soil) compacted into six individual compartments, 6 cm long, 3 cm wide, and 4 cm deep. Each compartment was bordered by a carbon felt anode and a stainless steel cathode. The application of a current density of 3.72 A/m(2) led to migration of 2,4-D towards the anode at a rate of approximately 4 cm/day. In a second experiment, electrokinetic movement and biodegradation were combined in situ. Sterilized silt soil contaminated with ring-labeled 14C-2,4-D (811 mg 2,4-D/kg dry weight soil) was compacted into a single soil compartment, 22 cm long, 7 cm wide, and 4 cm deep, in a 4.5 cm region adjacent to the cathode. The remainder of the compartment was filled with sterilized soil (to a total weight of 1,015 g). Burkholderia spp. RASC c2 (1.88 x 10(11) cells), a tetracycline-resistant bacterium with chromosomally encoded degradative genes for 2,4-D, was inoculated into the soil at a position 14-16 cm from the cathode. The reactor was placed within a sealed perspex box, with a constant air flow connected to sodium hydroxide traps. Under an applied current density of 0.89 A/m(2), the pollutant moved towards the bacteria. As it reached the inoculated region, its concentration decreased in the soil and 14CO2 was recovered in the traps. At the end of the experiment, 87.1% of radiolabel had been removed from the soil, 5.8% of which was recovered as 14CO2. A third, control, experiment showed a significant contrast in the absence of an electric current, where a slow rate of diffusion controlled the movement of both 2,4-D and bacteria in the soil and biodegradation occurred at the interface between the diffusing fronts.

2,4-Dichlorophenoxyacetic Acid↗

Kinetics of biodegradation of p-xylene and naphthalene and oxygen transfer in a novel airlift immobilized bioreactor.

The scope of this study included the biodegradation performance and the rate of oxygen transfer in a pilot-scale immobilized soil bioreactor system (ISBR) of 10-L working volume. The ISBR was inoculated with an acclimatized population of contaminant degrading microorganisms. Immobilization of microorganisms on a non-woven polyester textile developed the active biofilm, thereby obtaining biodegradation rates of 81 mg/L x h and 40 mg/L x h for p-xylene and naphthalene, respectively. Monod kinetic model was found to be suitable to correlate the experimental data obtained during the course of batch and continuous operations. Oxygen uptake and transfer rates were determined during the batch biodegradation process. The dynamic gassing-out method was used to determine the oxygen uptake rate (OUR) and volumetric oxygen mass transfer, K(L) a. The maximum volumetric OUR of 255 mg O(2)/L x h occurred approximately at 720-722 h after inoculation, when the dry weight of biomass concentration was 0.67 g/L.

Biodegradation, Environmental↗

Liquid photocurable biodegradable copolymers: in vivo degradation of photocured poly(epsilon-caprolactone-co-trimethylene carbonate).

Liquid photoreactive poly(epsilon-caprolactone-co-trimethylene carbonate)s endcapped with a coumarin group [coumarinated poly(CL/TMC)s] were prepared using tetra-functional hydroxylated substances such as pentaerythritol or four-branched poly(ethylene glycol), b-PEG. These coumarinated copolymers are tetra-branched and exist as a viscous liquid (MW 5 x10(3) approximately 7 x 10(3)). They were photocured by ultraviolet (UV) light irradiation to obtain a swelling or nonswelling solid under water, depending on the type of initiator used. The resultant films were implanted into the subcutaneous tissues of rats for up to 5 months. The photocured b-PEG-based copolymer was completely degraded and sorbed within a 1 month. On the other hand, surface-eroding degradation of pentaerythritol-based, coumarinated poly(CL/TMC) progressed with implantation time, and minimal recruitment of neutrophils, macrophages, and multinucleated giant cells was observed over the implantation period. Among the pentaerythritol-based copolymers, the fastest surface erosion was observed for the copolymer with the highest epsilon-caprolactone content. Microfabricated films with microarrays in which photoconstructs were stereolithographically prepared, using three different coumarinated copolymers at different regions, showed that upon implantation there was regionally differentiated biodegradation of microarrays, and the degree of region-specific biodegradation depended on the type of photocured copolymer. The observed tendency for biodegradation was in good agreement with that observed during implantation of individual films in vivo. This study also demonstrates that the use of multi-material-arrayed films enables the determination of different responses in vivo using only one sample.

Absorbable Implants↗

In vivo release of vancomycin from biodegradable beads.

The current delivery system of antibiotics for the treatment of osteomyelitis uses polymethylmethacrylate (PMMA) beads as a local drug-release agent. The nonbiodegradable nature of the PMMA, however, necessitates a second operation to remove the beads. This article explores the alternative of using biodegradable polymers as antibiotic beads for a long-term drug release in vivo. To manufacture an antibiotic bead, lactide-glycolide copolymers were mixed with vancomycin. The mixture was compressed and sintered at 55 degrees C to form beads 8 mm in diameter. An in vivo animal model was proposed to characterize the elution rate of antibiotic over a 55-day period. Biodegradable beads released high concentrations of antibiotic (well above the breakpoint sensitivity concentration) in vivo for the period of time needed to treat bone infection; that is, 4-6 weeks. A bacterial inhibition test was also carried out to determine the relative activity of the released antibiotics. The diameter of the sample inhibition zone ranged from 8 to 18 mm, which is equivalent to 9.1 to 100% of relative activity. In addition, the antibiotic concentration of systemic blood was found to be very low. Antibiotic-impregnated biodegradable beads may have a potential role in the prevention and management of surgical infections.

Animals↗

Biodegradation behavior of various calcium phosphate materials in bone tissue.

In order to study the biodegradation behavior of calcium phosphate materials, cylinders of standard size were implanted in the tibiae of rabbits. Material parameters were stoichiometry (hydroxyapatite with a Ca/P ratio of 1.67 versus tricalcium phosphate with a Ca/P ratio of 1.50), crystallographic structure (apatite versus beta-whitlockite), microporosity, and macroporosity. The extent of biodegradation was evaluated by radiography, light and fluorescence microscopy, microradiography, and porosity measurements. All calcium phosphate materials were biocompatible in bone tissue. Hydroxyapatite ceramics had a higher osteogenic potential than beta-whitlockite materials. Depending on their porosities, sintered tricalciumphosphate (beta-whitlockite) materials were more or less biodegradable, in contrast to sintered hydroxyapatite materials, which showed no detectable resorption over a period of 9 months of implantation.

Animals↗

Biodegradation of and tissue reaction to 50:50 poly(DL-lactide-co-glycolide) microcapsules.

The biodegradation of the copolymer 50:50 poly(DL-lactide-co-glycolide)-lypressin microcapsules was studied by light and electron microscopic methods and 14C release. Intramuscular injection sites of microcapsules in rats were studied by dissecting and conventional light microscopy as well as scanning (SEM) and transmission electron microscopy. A minimal localized acute myositis was seen initially at the injection sites. By Day 4, a few small foreign body giant cells were present participating in the minimal foreign body response. Later the inflammatory cells decreased and the individual microcapsules were walled off by immature fibrous connective tissue and large syncytial foreign body giant cells. By Day 35, definitive changes in some microcapsules, consisting of a granular and slightly eroded appearance of the internal matrix, were seen by SEM. By Day 42, the outer rims of the microcapsules were extensively eroded. At Day 56, the inflammatory and connective tissue reactions were almost completely resolved and biodegradation continued so that only remnant pieces of the microcapsules were present at Day 63. The morphologic picture correlated well with loss of 14C radioactivity, which could no longer be detected at the injection sites on Day 56. Phagocytosis did not seem to be an important factor in the biodegradation.

Animals↗

Water-curable and biodegradable prepolymers.

In an attempt to develop biodegradable polymers which can be shaped in situ and adhere to living tissues, we synthesized esterurethane prepolymers which can be cured upon contact with water in living tissues. First, D,L-lactide polymerization or D,L-lactide-epsilon-caprolactone (50:50) copolymerization was carried out using ethylene glycol or poly(ethylene glycol) as initiator to obtain hydroxyl-terminated biodegradable polyesters. They were then reacted with an excess of diisocyanate such as hexamethylene diisocyanate, toluylene diisocyanate, and diphenylmethane diisocyanate to introduce a reactive isocyanate group to both of the end groups of the polyesters. The isocyanate-terminated prepolymers could be cured in the presence of water and the cured polymers were degraded by hydrolysis both in vitro and in vivo. It was found that the presence of appropriate amounts of hydrophilic units in the main chain was essential for giving a high curing rate and a high degradation rate for the biodegradable urethane prepolymers. The tissue responses to the cured polymers were not severe.

Animals↗

Introducing a selectively biodegradable filament wound arterial prosthesis: a short-term implantation study.

This article introduces a new compliant and selectively biodegradable filament wound vascular graft and reports the findings of a short-term implantation study. A basic feature of filament winding is its ability to tailor and better control the mechanical properties of the prosthesis, so that a closer match with the anisotropic properties of native arteries is achieved. The elastomeric vascular grafts comprise poly(ether urethane urea) fibers (Lycra) embedded in a two-component matrix consisting of poly(ether urethane) (Pellethane) and a highly flexible poly(ethylene glycol)/poly(lactic acid) biodegradable segmented copolymer (PELA). Typical tensile modulus values fall in the few megapascals (MPa) range, this being comparable to that of natural arteries. The wound graft exhibits excellent handling and suturability characteristics as well as enhanced burst strength. Furthermore, due to its biodegradable constituent, the prosthesis combines minimal intraoperative blood loss and high healing porosity. The graft displays initially negligible in vitro water permeation, which increases gradually with time. In this short-term study, the prostheses were implanted in the canine carotid, and their biological performance was compared to that of expanded Gore-Tex. The luminal surface of the wound grafts was coated with a thin layer of pseudointima, strongly adhered to the prosthesis surface. Contrasting with the very stiff Gore-Tex grafts, the filament wound prostheses retained their high compliance, being highly pulsatile upon explanation. Histological studies fully corroborated these findings, underscoring the healing properties of these new filament wound vascular prostheses.

Animals↗

Novel bioactive and biodegradable glass ceramics with high mechanical strength in the CaO--SiO2--B2O3 system.

Novel bioactive and biodegradable glass ceramics with high mechanical strength in the (50-x/2)CaO. SiO(2)--xB(2)O(3) (4.2 < or = x < or = 17.2) system were investigated. The systems consisted of three phases: monoclinic wollastonite, calcium metaborate, and amorphous borosilicate matrix. The glass ceramics containing 4.2 mol% and 8.4 mol% B(2)O(3) showed high bulk density and a dense microstructure. Mechanical strengths of the glass ceramics were higher than those of other bioactive ceramics: high compressive strength (2813 MPa), bending strength of 212 MPa, and fracture toughness of 3.12 MPa. m(1/2). The glass-ceramic formed apatite layer on their surface in the simulated body fluid and showed significant biodegradation. The degree of apatite formation in the glass ceramics depended on the calcium metaborate content and borosilicate glassy matrix. Additional calcium metaborate and borosilicate glassy matrix increased the apatite formation rate on the surface. It might be likely that calcium metaborate causes supersaturation of Ca ions, for its high solubility in SBF and the water-reactive borosilicate glassy matrix formed Sibond;OH groups on the surface to provide nucleation sites for apatite formation. Also, through in vitro test for the biocompatibility of the CaO--SiO(2)--B(2)O(3) glass ceramics, no cytotoxicity of the glass ceramics were found. The results on bioactivity and noncytotoxicity indicated that glass ceramics in the (50-x/2)CaO. SiO(2)--xB(2)O(3) (4.2 < or = x < or = 17.2) system could be useful as a biodegradable bone replacement material.

Biocompatible Materials↗

Microbial biodegradation of cellophase.

The microbial biodegradation of cellophane (U.C.B.--Division Sidac) was studied. Preliminary experiments with pure cultures of seven cellulolytic microorganisms (Aspergillus sp., Penicillium sp., Chaetomium crispatum, Ch. globosum, Sclerotium rolfsii and two actinomycetes) revealed that the substrate as such was very recalcitrant, probably due to the occurrence of insoluble coating agents. Therefore, mixed cultures of the above mentioned cellulolytic microorganisms were used as inoculum. The cellophane showed a slow microbial degradation which starts only after 37 days of incubation. This long lag-phase is due to the unaltered presence of the coating agents. However, when the coating agents are extracted with tetrahydrofuran, the biodegradation starts after 10 days, resulting in a biodegradation rate of 85% after 52 days of incubation and a protein content of 30%. The endproduct (30% protein, 60% soluble sugars, 10% residual substrate) will probably be useful as compost.

Actinomycetales↗

Novel biodegradable polymers as gene carriers.

This study investigated two new biodegradable polymers as gene controlled-released coatings for gene transfer. Poly(ethylene glycol)-co-poly(D,L-lactic acid) (PELA) and poly(ethylene glycol)-co-poly(lactic acid)-co-poly(glycolic acid) random copolymer (PELGA) were synthesized and used as microspheres matrices with encapsulated plasmid pCH110. The plasmid loading efficiency, cytotoxicity, transfection efficiency and in vitro degradation and release profiles of microsphere complexes were evaluated in details. The biodegradable polymers showed high DNA loading efficiency and low cytotoxicity as gene controlled-released coatings, and the poly(ethylene glycol) (PEG) contents of polymer matrices influenced the diameter, loading efficiency and transfection efficiency of plasmid DNA within the microspheres. The average diameters of PELA and PELGA microspheres were between 0.5 and 1.5 microm, and the plasmid loading efficiency was 62 and 73% for PELA and PELGA microspheres with 10% PEG content, respectively. In vitro testing showed a gradual release profile of DNA from polymeric matrices. The polymers/DNA microspheres had high transfection efficiency and early gene expression and maintenance of gene expression level for up to 96 h, although transfection efficiency were slightly lower than that of liposome in the initial 24 h. The biodegradable polymeric materials possess potential superiority as gene carriers.

Animals↗

A novel approach to biodegradable block copolymers of epsilon-caprolactone and delta-valerolactone catalyzed by new aluminum metal complexes.

The chemical preparation of structurally well-defined biodegradable amphiphilic block copolymers is now of great interest for biomedical applications and the fundamental mimetic study of biomacromolecule self-assembly. For this purpose, in this study, (R,R)-N,N'-bis(3-tert-butylsalicylidene)-1,2-cyclohexanediamine 2 as a ligand was first synthesized from 1,2-cyclohexanediamine (DACH) and was allowed to further react with AlMe3, leading to a precursor compound 3. Then, the novel five-coordinated aluminum metal complexes 4-6 and 7-8 were prepared with good yields of 80-90%, bearing various molar mass monofunctional methoxy-poly(ethylene glycol) MPEG and difunctional poly(ethylene glycol) PEG as the alkoxy moieties, respectively. By means of nuclear magnetic resonance spectrometry (NMR), mass spectrometry (MALDI-FTMS) and Fourier Transform infrared spectrometry (FT-IR), new metal aluminum complexes 4-8 were characterized as having distinct chemical structures. Utilizing the synthesized metal complexes 4-8 as novel coordination polymerization catalytic templates, biodegradable amphiphilic MPEG-b-PCL, MPEG-b-PVL, PCL-b-PEG-b-PCL and PVL-b-PEG-b-PVL were synthesized with good control of the molecular weight distribution via the ring opening polymerization of epsilon-caprolactone and delta-valerolactone monomers at 100 degrees C in toluene. In addition, the chemical and crystalline structures and the thermal properties of these block biodegradable copolymers were analyzed by means of NMR, gel permeation chromatography (GPC), wide-angle X-ray diffraction (WAXD), differential scanning calorimetry (DSC) and thermal gravimetric analysis (TGA). It was found that the melting points and crystallinities of the block copolymers synthesized strongly depended on the molecular structures of the polyether and polyester building blocks. Only one glass transition stage was detected, indicating good chain/segmental miscibility between the hydrophilic MPEG/PEG and hydrophobic PCL/PVL blocks in the non-crystalline regions. Moreover, TGA analysis exhibited typical two-step decomposition profiles with the weight-loss percentages in good agreement with block compositions from NMR calculations.

Aluminum↗

Effects of an avidin-biotin binding system on chondrocyte adhesion and growth on biodegradable polymers.

Cell adhesion to a scaffold is a prerequisite for tissue engineering. Many studies have been focused on enhancing cell adhesion to synthetic materials that are used for scaffold fabrication. In this study, we applied an avidin-biotin binding system to enhance chondrocyte adhesion to biodegradable polymers. Biotin molecules were conjugated to the cell membrane of chondrocytes, and mediated cell adhesion to avidin-coated surfaces. We demonstrated that immobilization of biotin molecules to chondrocyte surfaces enhanced cell adhesion to avidin-coated biodegradable polymers such as poly(L-lactic acid), poly(D,L-lactic acid), and polycaprolactone, compared to the adhesion of normal chondrocytes to the same type of biodegradable polymer. The biotinylated chondrocytes still maintained their proliferation ability. This study showed the promise of applying the avidin-biotin system in cartilage tissue engineering. [diagram in text].

Avidin↗

Nerve regeneration through a two-ply biodegradable nerve guide in the rat and the influence of ACTH4-9 nerve growth factor.

Biodegradable polyurethane-based (PU) nerve guides, instilled with or without ACTH4-9 analog (a melanocortin) were used for bridging an 8 mm gap in the rat sciatic nerve and were evaluated for function and histological appearance after 16 weeks of implantation. Autologous nerve grafts functioned as controls. The guides successfully enabled the sciatic nerve to regenerate across the 8 mm gap, thus effectively reestablishing the contact between the proximal and distal nerve ends. The mean conduction velocity, motor latency, and muscle action potentials of all the nerve guides did not differ significantly from the autografts. The histological quality of the regeneration in the nerve guides was significantly better than in the autografts; in the nerve guides, a well-defined nerve cable of normal architecture had regenerated without extensive endoneural scarring as seen in the autografts. ACTH4-9 instilled in the nerve guides showed a slight, but significant, increase in the number of myelinated axons. It is concluded that biodegradable PU nerve guides result in similar functional recovery when compared with autografts, but their histological quality is significantly better. ACTH4-9 showed only slight, but significant, improved nerve growth promoting activity. Therefore biodegradable PU nerve guides with ACTH4-9 would appear to be promising alternatives to autografts for bridging nerve defects.

Adrenocorticotropic Hormone↗

Stream periphytic biodegradation of the anionic surfactant C12-alkyl sulfate at environmentally relevant concentrations.

The effects of continuous exposure to C12-alkyl sulfate on a periphytic microbial community were determined in an 8-week stream mesocosm study. C12-alkyl sulfate concentrations ranged from environmentally relevant (< 10-20 micrograms/liter) to unrealistically high concentrations (> 1500 micrograms/liter). Endpoints evaluated included turnover rates, bacterial cell density, heterotrophic mixed amino acid uptake, and fatty acid profile evaluations. Predosed periphyton demonstrated a mean turnover rate for C12-alkyl sulfate of 0.08/hr. During the 8-week dosing period, a significant increase in mean turnover rates was observed in streams dosed with > or = 61 micrograms C12-alkyl sulfate/liter, despite a 10 degrees C drop in stream temperature. A significant correlation between turnover rate and C12-alkyl sulfate concentration was also observed. While bacterial cell density increased during the study, it was determined that the biodegradation acclimation to C12-alkyl sulfate was not biomass-specific. Likewise, bacterial activity generally increased over the study, but it did not correlate with either biodegradation or bacterial cell density. Lastly, phospholipid fatty acid profiles indicate that a shift in the microbial community occurred in the high-dose stream as opposed to the control stream. This study demonstrates that C12-alkyl sulfate is rapidly degraded and induces a biodegradative acclimation response at environmentally relevant concentrations.

Amino Acids↗

Probability of biodegradation, a novel concept for improving chemical classification and risk assessment.

In this article biodegradability is considered as a combination of an inherent substance property, defined as the maximum specific growth rate, mumax and a condition of the environment defined as a specific fraction (fs) of the total viable biomass. By proper analysis of test results it is possible to quantify both parameters by one single standard test. Calculations with literature data indicate that for the majority of the degradable substances, mumax may vary between 0.5 and 10 per day. The specific fractions, however, may vary 5 or 6 orders of magnitude and can be as low as 10(-8). This concept gives a valuable tool in environmental risk assessment. As the results will be less influenced by test conditions, data will be more reproducible and can be more predictive for a specific environment. The results allow predicting the time needed to achieve adaptation in a treatment plant and, in particular, the behavior under conditions with discontinuous discharge. By using threshold criteria for mumax, fs, and percentage mineralization, a new classification scheme with eight different classes for biodegradability is proposed. Compared to the currently used system with two classes, the prediction of biodegradation will be more sophisticated. The proposed system differentiates for six types of inherently degradable substances and identifies those substances with an abnormal growth curve, due to inhibition or toxicity, poor water solubility, or incomplete mineralization. For these classes the proposed equations are not directly applicable and more research will be required to predict their behavior.

Animals↗