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Bacterial polyesters: biosynthesis, biodegradable plastics and biotechnology.

The discovery and chemical identification, in the 1920s, of the aliphatic polyester: poly(3-hydroxybutyrate), PHB, as a granular component in bacterial cells proceeded without any of the controversies which marked the recognition of macromolecules by Staudinger. Some thirty years after its discovery, PHB was recognized as the prototypical biodegradable thermoplastic to solve the waste disposal challenge. The development effort led by Imperial Chemical Industries Ltd., encouraged interdisciplinary research from genetic engineering and biotechnology to the study of enzymes involved in biosynthesis and biodegradation. From the simple PHB homopolyester discovered by Maurice Lemoigne in the mid-twenties, a family of over 100 different aliphatic polyesters of the same general structure has been discovered. Depending on bacterial species and substrates, these high molecular weight stereoregular polyesters have emerged as a new family of natural polymers ranking with nucleic acids, polyamides, polyisoprenoids, polyphenols, polyphosphates, and polysaccharides. In this historical review, the chemical, biochemical and microbial highlights are linked to personalities and locations involved with the events covering a discovery timespan of 75 years.

3-Hydroxybutyric Acid↗

Somatic cell cloning in polyester stacks.

Single somatic cells, including fibroblasts, myelomas, and hybridomas, proliferate normally when trapped between a plastic dish and a disc of polyester cloth. Contact between the overlay and the plastic for 8-16 days results in identical colony patterns on the cloth and the plate. When several cloth discs are simultaneously stacked over Chinese hamster ovary cells, three or four-high resolution colony copies can be generated from a single master dish. The colonies on the cloth can be analyzed by radiochemical methods [Esko, J. D. & Raetz, C. R. H. (1978) Proc. Natl. Acad. Sci. USA 75, 1190-1193] or by "replica plating" to a new disc. The use of polyester cloth, singly or in stacks, has several major advantages over previous techniques for somatic cell replica plating, including: (i) broad applicability to diverse cell lines such as fragile membrane mutants of Chinese hamster ovary cells and relatively nonadherent myelomas or hybridomas; (ii) the possibility of generating multiple copies of the same colony population, allowing simultaneous analysis for several enzymes or cellular components; and (iii) superior resolution and transfer efficiency in copying colony patterns from one surface to another. The remarkable capacity of animal cell colonies to proliferate upward through "polyester stacks" may reflect chemotropic movement of individual cells and opens new approaches to somatic cell genetics.

Animals↗

Migration of styrene monomer from thermoset polyester cookware into foods during high temperature applications.

Migration of styrene from thermoset polyester cookware into foods has been studied during normal cooking applications and for the fatty food simulant olive oil under high temperature test conditions. The analysis of foods was by automated headspace gas chromatography/mass spectrometry (GC/MS) in the selected ion mode. Cyclohexane was added to the slurried foods to improve headspace partition and deuterated-styrene was the internal standard. To achieve adequate sensitivity with olive oil, styrene was extracted and concentrated by a steam distillation method and then determined by GC/MS. Styrene in foods cooked in thermoset polyester articles was in the range < 5 to 5 micrograms/kg and 5 to 30 micrograms/kg where the polyester contained 9 and 380 mg/kg residual monomer respectively. Testing for 2 h at 175 degrees C into olive oil resulted in significantly higher migration of styrene than seen for other foods, although there was a marked decrease in migration on repeat-use. Testing of the complete articles as sealed systems filled with olive oil led to higher migration than testing by total immersion of plaques. Of the test methods used with olive oil, testing as filled articles with no lidding gave migration results closest to (but generally higher than) those seen for real foods.

Cooking and Eating Utensils↗

Polyester as a bioimplantable material.

In response to the recent controversy over silicone gel implants, alternative substances are being sought to offer new options in bioimplantable materials. Polyester is a Food and Drug Administration-approved prosthetic material utilized in vascular grafts for decades without significant problems in body tolerance. Working with textile scientists, polyester (both in its woven and fibrous forms) was engineered to influence differential fibrous ingrowth. Implants of medical-grade polyester were constructed in a 3-cm2 diskette configuration. Four combinations of material with varying layers of weave and fiber were created to influence tissue ingrowth differentially. Four implants were placed in separate dorsal pockets beneath the panniculus carnosus of each of 12 adult female New Zealand White rabbits. Care was taken to sort the implant types equally among the four quadrants such that a similar distribution of implants existed between animals. Implants with intact capsules were harvested at 1, 3, and 6 months, and were analyzed for histology, capsular thickness, and tissue ingrowth. A phantom mammogram was taken through the implant material that did not obscure interpretation. The materials were further tested to determine fiber fill-to-wet implant weight ratios to determine postimplantation weight predictably. Tensile strength was also determined.

Animals↗

The adherence of Staphylococcus aureus, Staphylococcus epidermidis and Escherichia coli on cotton, polyester and their blends.

The adherent behaviour of the Gram-positive Staphylococcus aureus and Staphylococcus epidermidis and the Gram-negative Escherichia coli on cotton, polyester and their blends through contact in aqueous suspensions was studied. Staphylococcus epidermidis was found to adhere to fabrics much more so than Staph. aureus. The adherence of both Staph. epidermidis and Staph. aureus to fabrics increased as the content of polyester fibres in the fabrics increased. The attachment of E. coli to all fabrics was very low and was not affected by the fibre contents. Total numbers of adherent bacteria on cotton and polyester fabrics were related directly to the concentrations of the bacterial suspensions. The extents of adherence, expressed by the percentage of adherent bacteria from the suspension, however, were independent of the concentration. The length of contact with bacteria was also found to affect the adherence of bacteria on fabrics studied.

Absorption↗

Analysis of the aliphatic monomer composition of polyesters associated with Arabidopsis epidermis: occurrence of octadeca-cis-6, cis-9-diene-1,18-dioate as the major component.

Although the surface waxes from Arabidopsis thaliana leaves and stems have been thoroughly characterized, the monomer composition of the polyesters of the cuticular membrane has not been analyzed. Delipidated Arabidopsis leaves or stems, when depolymerized under conditions to cleave polyesters, produced typical omega-hydroxy fatty acid cutin monomers such as 16-hydroxy-palmitate, 10,16-dihydroxy-palmitate and 18-hydroxy-9,10-epoxy-stearate. However, the major monomer was octadeca-cis-6, cis-9-diene-1,18-dioate, with lesser amounts of octadec-cis-9-ene-1,18-dioate and hexadeca-1,16-dioate. These dicarboxylates were found predominantly in epidermal peels from Arabidopsis stems and are therefore likely to be associated with the cuticular membrane. They were also found in analyses of canola leaves but were absent in tomato and apple fruit cutins. In the fad2-1 mutant line of Arabidopsis, which has reduced levels of linoleate and linolenate and elevated oleate in cytosolic phospholipids, the amount of octadeca-cis-6, cis-9-diene-1,18-dioate was 50% reduced, with a concomitant increase in octadec-cis-9-ene-1,18-dioate. In a fatb-ko line of Arabidopsis, where the availability of cytosolic palmitate is impaired, there was an 80% loss of C16 monomers and a compensating increase in C18 monomers. The presence of substantial amounts of dicarboxylates in cuticular membranes is unexpected. High amounts of aliphatic dicarboxylates are usually considered as an indicator of suberin, and are reported only as very minor components of cutin. The high level of polyunsaturation is also unusual in cuticles; saturated fatty acid monomers usually predominate, with lesser amounts of monounsaturates. These novel findings for Arabidopsis demonstrate that a broad range of monomer compositions are possible for polyesters of the epidermis.

Arabidopsis↗

Is the guideline the cause of structural failure in current polyester vascular prostheses? An experimental study.

To evaluate the impact of the guideline on the structural failure of current polyester vascular prostheses, we studied seven unsealed polyester vascular prostheses. An accelerated time test was used for assessing degradation of polymeric medical materials at pH 7.4 in phosphate buffer at a temperature of 95 degrees C. Probe burst strength was determined on the guideline and on standard parts of the prostheses during the accelerated time test. Weight-average molecular weights of the guideline and of the standard parts in all knitted grafts during the accelerated time test were measured. The guideline was significantly weaker than the standard parts in most prostheses. The guideline had significantly lower values of weight-average molecular weights than the standard parts in most knitted grafts. This study showed that guideline degeneration was one factor responsible for the structural failure of some current polyester vascular prostheses.

Blood Vessel Prosthesis↗

Formation of polyesters consisting of medium-chain-length 3-hydroxyalkanoic acids from gluconate by Pseudomonas aeruginosa and other fluorescent pseudomonads.

Pseudomonas aeruginosa PAO and 15 other strains of this species synthesized a polyester with 3-hydroxydecanoate as the main constituent (55 to 76 mol%) if the cells were cultivated in the presence of gluconate and if the nitrogen source was exhausted; 3-hydroxyhexanoate, 3-hydroxyoctanoate, and 3-hydroxydodecanoate were minor constituents of the polymer. The polymer was deposited in granules within the cell and amounted to 70% of the cell dry matter in some strains. Among 55 different strains of 41 Pseudomonas species tested, P. aureofaciens (21.6% of cellular dry matter), P. citronellolis (78.0%), P. chlororaphis (8.5%), P. marginalis (11.4%), P. mendocina (50.7%), P. putida (33.5%), and Pseudomonas sp. strain DSM 1650 (54.6%) accumulated this type of polymer at significant levels (greater than 5%) during cultivation on gluconate. In two strains of P. facilis and P. fluorescens, as well as in one strain of P. syringae, this polymer was detected as a minor constituent (much less than 5%). All other strains accumulated either poly(3-hydroxybutyrate) or a polymer consisting mainly of 3-hydroxyoctanoate with octanoate but no polyester with gluconate as the carbon source. Only a few species (e.g., P. stutzeri) were unable to accumulate poly(hydroxyalkanoic acids) (PHA) at all. These results indicated that the formation of PHA depends on a pathway which is distinct from all other known PHA-biosynthetic pathways. The polyesters accumulated by gluconate- or octanoate-grown cells of recombinant strains of P. aeruginosa and P. putida, which harbored the Alcaligenes eutrophus poly(3-hydroxybutyrate)biosynthetic genes, contained 3-hydroxybutyrate as an additional constituent.

Gluconates↗

Activity and distribution of superoxide dismutase in the layers of polyester grafts.

The aim of the present study was to evaluate superoxide dismutase (Cu, Zn-SOD) activity in the principal layers of polyester grafts implanted into the abdominal aorta of dogs. The grafts were examined 7 days and 1, 4 and 12 months after the operation. It was found that SOD activity occurred in all polyester graft layers during the whole experiment. The neointima activity ws 1.5-fold lower during the 1st month; with the passage of time, it became 2-fold lower than in the normal aorta (p < 0.01). SOD activity in the medial and outer graft layers behaved similarly: it was lowest after 7 days, increased after 1 month and significantly decreased after 4 and 12 months (p < 0.01). Our study shows that the activity of SOD is low in polyester grafts and it can thus be inferred that protection against the superoxide anion is inadequate.

Animals↗

In vitro and in vivo hemocompatibility evaluation of graphite coated polyester vascular grafts.

Attempts have been made in this study to prepare a homogeneous and stable coating of graphite on polyester vascular grafts (GPVG) using an electrophoresis method to evaluate thromboresistant and blood compatibility of GPVG in comparison to non-coated PVG and InterGard (collagen sealed PVG) as control. Lactate dehydrogenase (LDH) activity measurement was carried out on all PVG types to evaluate platelet adhesion. To examine tissue reaction GPVG and non-coated sheets of knitted polyester fabric were implanted simultaneously in the dorsal flank of rats subcutaneously. The GPVG, non-coated and control were implanted in descending aorta as end-to-end or end-to-side implantation substitution in 25 sheep for 4-60 weeks. Results showed that the graphite coating on polyester vascular grafts reduced the number of adherent platelets and prevent platelet activation and spreading on the surface in comparison with non-coated and control. Pathological investigation showed inflammatory reactions were totally resolved after 12 weeks and there was no difference in the tissue reaction between graphite coated, non-coated and control patches. All grafts remained patent and there was no significant difference in patency rate between these three types of PVG. We found that GPVG has no need for pre-clotting and it showed lower platelet aggregation, thinner capsule formation and lower calcification after 15 months. However, suturing of GPVG was more difficult in comparison with the other types.

Animals↗

Custom-made stent-grafts for aortic aneurysm repair using gianturco Z stents and woven polyester: healing in an animal model.

The purpose of this study was to evaluate the healing at 6 months of aortic stent-grafts custom-made by using Z stents and woven polyester in an animal model. Stent-grafts were built by a published method using autoexpandable stainless steel stents continuously compiled with polyester sutures and covered with a woven polyester membrane. Fourteen stent-grafts of 3 different designs were deployed under fluoroscopic control into the thoracic and the abdominal aorta of 7 adult sheep. At 6 months, all the implants of the sheep that survived the implantation were angiographed and harvested for macroscopy and microscopy. All stent-grafts were implanted successfully and remained patent from then to the explantation procedure. All stent-grafts implanted among the 6 of 7 (86%) animals that survived after the implantation remained patent, stayed free from local complications, and did not migrate during more than 6 months. In all these cases during this period, the implant functioned as a satisfactory aortic substitute while the aortic segment containing the graft kept a normal structure. There was no adverse effects of the presence of the implant upon the aorta or the animal. Stent-grafts home-made according to the described methods gave results at 6 months in this animal model compatible with a safe clinical application among humans.

Animals↗

Graft infectivity of rifampin and silver-bonded polyester grafts to MRSA contamination.

The purpose of this study was to evaluate the ability of vascular polyester grafts with antibacterial properties to resist colonization following surface contamination by methicillin-resistant Staphylococcus aureus (MRSA) in an experimental canine model or aortic graft infection. Twenty-four pathogen-free dogs underwent replacement of the infrarenal aorta with either a rifampin-soaked (30 mg/mL) or silver-impregnated (Ag-acetate) woven polyester graft. Following implantation, the external graft surface was inoculated with 2 mL of 10(7) colony-forming units/mL (CFU) of MRSA. Preoperative antibiotic prophylaxis consisted of a single intravenous dose of 500 mg of sodium cefazolin. Four grafts of each type were explanted at 3, 7, and 14 days after implantation. Quantitative cultures (CFU/specimen) of perigraft fluid (1 mL), graft material (1 cm segment), and adjacent aorta (1 cm segment) were performed. Differences between grafts are expressed as % mean log reduction in recoverable CFU compared to the inoculation solution concentration of 10(7) CFUs. At 3 days, explanted rifampin-soaked grafts exhibited no MRSA growth (4 of 4 grafts) and a > or =97% mean log reduction of MRSA CFUs from the adjacent aorta and perigraft fluid (PGF). At 3 days, all silver-bonded grafts exhibited signs of infection and a mean log CFU reduction of MRSA ranging from 68% (absolute range 10(1)-10(3) recoverable CFU) for the graft, 79% (absolute range 10(1)-10(3) recoverable CFU) for the aorta, and 86% (absolute range 10(1)-10(4) recoverable CFU) for PGF. The 7-day rifampin group had an average log reduction in MRSA CFU of 72% (graft), 58% (PGF), 75% (aorta). Quantitative cultures of 14-day rifampin grafted demonstrated continued bacterial growth suppression with mean MRSA CFU log reductions of 82%, graft; 72%, PGF; 89%, aorta. Silver-bonded grafts demonstrated <50% mean CFU reduction in MRSA growth at 7 days (absolute range 10(5)-10(7) recoverable CFU) and 14 days (absolute range 10(3)-10(7) recoverable CFU). No animal died from sepsis or anastomotic hemorrhage. Neither rifampin- nor silver-bonded grafts demonstrated prolonged resistance to surface MRSA contamination. Rifampin-soaked polyester grafts exhibited a marked but transient resistance MRSA colonization likely the result of high antibiotic concentration in the perigraft tissue. While both types of grafts failed to eradicate the MRSA infection future research with silver-bonded grafts that have an additional antibiotic attached may have a place in the treatment of MRSA infection.

Animals↗

Tandem mass spectrometry characteristics of polyester anions and cations formed by electrospray ionization.

Electrospray ionization of polyesters composed of isophthalic acid and neopentyl glycol produces carboxylate anions in negative mode and mainly sodium ion adducts in positive mode. A tandem mass spectrometry (MS/MS) study of these ions in a quadrupole ion trap shows that the collisionally activated dissociation pathways of the anions are simpler than those of the corresponding cations. Charge-remote fragmentations predominate in both cases, but the spectra obtained in negative mode are devoid of the complicating cation exchange observed in positive mode. MS/MS of the Na(+) adducts gives rise to a greater number of fragments but not necessarily more structural information. In either positive or negative mode, polyester oligomers with different end groups fragment by similar mechanisms. The observed fragments are consistent with rearrangements initiated by the end groups. Single-stage ESI mass spectra also are more complex in positive mode because of extensive H/Na substitutions; this is also true for matrix-assisted laser desorption ionization (MALDI) mass spectra. Hence, formation and analysis of anions might be the method of choice for determining block length, end group structure and copolymer sequence, provided the polyester contains at least one carboxylic acid end group that is ionizable to anions.

Anions↗

Anterior cruciate ligament replacement. Biocompatibility and biomechanics of polyester and carbon fibre in rabbits.

The anterior cruciate ligament was replaced in rabbits, using implants of carbon or polyester filaments with known mechanical properties. The biocompatibility of the implants was assessed in detail using light microscopy, and scanning and transmission electron microscopy. Mechanical tests were made of stability, in comparison with normal joints and controls after excision of the ligament. Some carbon fibre implants broke down in vivo, allowing instability; the fragments caused chronic inflammation. Intact carbon implants did not induce the formation of neoligaments; they were covered by tissue, but there was no ingrowth. Polyester did not degrade mechanically and supported early collagenous ingrowth within the implant, even in the mid-joint space. It was concluded that there was no justification for the use of carbon fibres as anterior cruciate replacements; polyester appeared to be suitable.

Animals↗

Polyester meshes and adhesive materials in the brain: comparative research in rats to optimize surgical strategy.

OBJECT: The goal of this study was to determine the biocompatibility of polyester mesh electrode carriers for auditory brainstem implants with and without adhesives in a rat model. METHODS: Physical properties of the meshes were evaluated within the fourth ventricle region, both without (Group A) and with adhesives (muscle, Group B; oxidized regenerated cellulose [ORC], Group C; and fibrin glue, Group D). The stability of the mesh position, the healing process, and host defense reaction after 2 to 60 days were examined in series of tissue sections in which meshes were preserved in situ. The cellular reaction was further evaluated using electron microscopy. Although otherwise pliable, polyester meshes were too rigid when used with adhesives, especially fibrin glue or muscle. Also, the sharp edges of the meshes presented a risk of brainstem and cerebellar lesions. Regardless of the material, meshes induced persistent inflammatory tissue reactions characterized by numerous macrophages and foreign-body giant cells. After 14 days, the cellular response had resulted in sufficient fibroblast and collagen fiber encapsulation of the meshes and remained essentially unchanged thereafter. No influence of adhesives on the healing process was observed, and, unexpectedly, these substances did not reduce the risk of dislocation prior to adequate cellular encasement. In some rats in Groups A and C, purulent inflammation, in part with Gram-positive bacteria, occurred after 2 to 14 days. The ORC exhibited persistent swelling, introducing the risk of occlusive hydrocephalus and/or brainstem compression. CONCLUSIONS: Polyester meshes and various adhesives exhibited acceptable biocompatibility in terms of local tissue reaction. Adhesives reduced pliability of the meshes, however, and were ineffective in reducing the risk of dislocation. Handling characteristics could be improved by better mesh designs, and risk of infection could be reduced by both improved designs and surface treatment of the meshes with antibacterial agents.

Animals↗

Evaluation of the healing process after implantation of synthetic material called Tegmentum consisting of polyester mesh covered with polyurethane in rat peritoneum.

Polyester meshes are one of the synthetic prosthetic materials widely applied for the reconstruction of abdominal layers in child and neonatal surgery, cardiosurgery or vascular surgery. Because of the lack or insufficiency of the own, natural material multiple researches are performed in order to find the best material able to replace natural tissue. The most suitable prosthesis ought to be: sterile, non-toxic, soft, flexible, elastic, not very stretchy, durable, easy to operate (cutting, suturing); to create desired shape required while reconstracting, totally resistant to contagion, without any complication after being implanted. Since early seventies many scientists following above requirements have carried out researches to create the most similar prosthesis morphologically and functionally to human tissue but still there is no such an ideal material on the market. Polyester with its derivatives is one of the most commonly applied synthetic substances in surgery. The type of biomaterial highly depends on its purpose e.g.: for vascular prosthesis materials that are resorptive in organism and support tightness (gelatine, albumin, collagen); while for temporal replacement abdominal layers with prosthesis the most suitable is impervious to systemic fluids bacteria etc material, that prevents penetrating the surface of prosthesis by surrounding tissues, adhesions with intestinum or other organs. Tegmentum is a material that posses these features. This is a polyester mesh covered with polyurethane on one side produced by Tricomed S.A. (Łódź). Tegmentum has already been applied to neonates with congenital eventration in Child and Neonatal Ward in the "Szpital Pomnik Matki Polki" hospital in Łódź.

Animals↗

Cytosolic free Ca2+ concentration in canine aortic endothelial cells lining the polyester arterial prosthesis.

A rise in baseline cytosolic free Ca2+ in canine vascular endothelial-like cells (VEC) lining the luminal surface of the polyester arterial prosthesis is described. In one, three and six month implantation experiments we employed six adult mongrel dogs, polyester arterial prostheses Arteknit Ra K, fluorescent Ca2+ indicator Fura-2 and digital imaging microscopy to study cytosolic free Ca2+ in cultured VEC. The electron microscopy scanning of the luminal surface in different regions of the graft were also performed. A rise in cytosolic free Ca2+ in the VEC lining the luminal surface of the prosthesis is probably the result of the immunologic reaction and mechanical stress which stimulate the proliferation activity of the endothelial cells. It seems that the baseline cytosolic free Ca2+ reflects the course of the endothelization process on the polyester arterial prosthesis.

Animals↗

In vitro studies of the influence polyester materials with a different degree of surface wettability have on blood haematological parameters and coagulation and fibrinolysis system parameters.

The paper presents the studies of the influence polyester knitings with a different degree of surface wettability have on the coagulation system, fibrinolytic system and blood haematological parameters in in vitro studies. The studies of Human whole blood were performed for 5, 15, 30, 60, 90, 180 and 240 min. In view of the obtained results in has been reported that a hydrophobic surface knitting does not cause important quantitative changes in any of the selected parametes of the coagulation system and fibrinolysis system. A hydrophilic surface polyester knit causes important lengthening of aPTT value and reduced factor XII, IX and VIII activity. A hydrophilic surface polyester knitting causes a bigger decrease in leukocytes and blood platelets and a bigger increase in beta thromboglobulin in relation to the hydrophobic surface knitting. Neither hydrophobic surface knit nor hydrophilic surface knitting cause changes in the erythrocyte system.

Blood Coagulation↗