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Ultrasound detection of visceral adhesion after intraperitoneal ventral hernia treatment: a comparative study of protected versus unprotected meshes.

Intraperitoneal (IP) ventral hernia repair has been proposed with the advantages of reducing dissection, operative time, and postoperative pain. The IP position of the mesh is suspected of increasing the risk of visceral adhesion and inducing complications. To overcome these drawbacks, a mesh protected on one side by a hydrophilic resorbable film (Parietex Composite) has been validated. Using a previously described ultrasound procedure, the purpose of this study was to compare the rate of visceral adhesion after intraperitoneal placement of a polyester mesh versus this protected mesh. Fifty-one patients who received a Parietex Composite mesh were prospectively compared to a retrospective series of 22 consecutive asymptomatic patients who received a Mersilene mesh. To objectively assess visceral adhesion toward the abdominal wall, an ultrasound (US) specific examination was firstly validated and secondly used to evaluate the adhesion incidence in both groups. Both groups were equivalent in terms of inclusion criteria and body mass index (BMI). Pre-operative US versus perioperative macroscopical findings determined the following parameters: sensitivity 83%, accuracy 78%, negative predictive value 81%. Using this procedure, 77% of the patients exhibited visceral adhesion to the mesh in the Mersilene group, against 18% in the Parietex Composite group (P<0.001, chi-square). US examination represents a suitable tool to evaluate postoperative adhesions to the abdominal wall. Using this procedure, a significant reduction of visceral adhesion in the Parietex Composite group was shown.

Coated Materials, Biocompatible↗

Comparison of two composite meshes using two fixation devices in a porcine laparoscopic ventral hernia repair model.

INTRODUCTION: Laparoscopic ventral hernia repair uses tacks to secure mesh. The mesh is designed to maximize tissue ingrowth while minimizing adhesions. We hypothesized: (1) a collagen-coated polyester mesh (PCO) will form fewer adhesions than an ePTFE-polypropylene composite (BC) and (2) absorbable tacks are equivalent to metal tacks. METHODS: In a porcine model of adhesion formation, three pieces of 10x15-cm mesh were placed on the anterior abdominal wall. PCO was secured with absorbable (PLA) or metal tacks (PT), BC with PT. At 28 days, adhesion formation, abdominal-wall adherence, and tissue ingrowth were analyzed. RESULTS: PCO induced fewer adhesions (14.5% vs 53.4%, P = 0.007). On an adhesion scale (0 5), BC scored 3.6 vs 1.75 for PCO (P < 0.03). There was no difference in adhesion strength, tack adhesions, or abdominal-wall peel force. Histology showed equal ingrowth. CONCLUSIONS: PCO induces fewer adhesions than BC. There is no difference in the ingrowth of the two mesh types. The PLA achieves equivalent mesh incorporation to the PT.

Animals↗

Late intestinal fistula formation after incisional hernia using intraperitoneal mesh.

Enteric fistulas are a rare but serious complication following the repair of an incisional hernia using a prosthesis. We report the case of a 52-year-old man who developed an enterocolocutaneus fistula after incisional hernia repair with intra-abdominal polyester mesh. This case shows that one may want to avoid placing the parietal prostheses in direct contact with intestinal loops.

Cicatrix↗

Biosynthesis of poly-beta-hydroxyalkanoates by Sphingopyxis chilensis S37 and Wautersia sp. PZK cultured in cellulose pulp mill effluents containing 2,4,6-trichlorophenol.

Poly-beta-hydroxyalkanoates (PHA) polymer is synthesized by different bacterial species. There has been considerable interest in the development and production of biodegradable polymers; however, the high cost of PHA production has restricted its applications. Kraft cellulose industry effluents containing 2,4,6-trichlorophenol (10 or 20 microg ml(-1)) were used by the bacteria Sphingopyxis chilensis S37 and Wautersia sp. PZK to synthesize PHA. In this condition, S. chilensis S37 was able to grow and degrade 2,4,6-trichlorophenol (ca. 60%) and 80% of these cells accumulated PHA. Wautersia PZK completely degraded 2,4,6-TCP and more than 90% of the cells accumulated PHA in 72 h. The PHA detection was performed by flow cytometry and polyester composition was characterized by gas chromatography-mass spectroscopy (GC-MS), indicating that these polymers are made by 3-hydroxybutyric acid and 3-hydroxyhexadecanoic acid for S37 and PZK strains, respectively. Results demonstrated that strains' growth and PHA production and composition are not modified in cellulose effluents with or without 2,4,6-TCP (10-20 microg ml(-1)). Therefore, our results indicate that S. chilensis S37 and Wautersia sp. PZK are able to degrade a toxic compound such as a 2,4,6-TCP and simultaneously produce a valuable biopolymer using low-value substrates.

3-Hydroxybutyric Acid↗

Perineal puborectalis sling operation for fecal incontinence: preliminary report.

PURPOSE: This study was designed to evaluate the safety efficacy, and impact on quality of life of the perineal puborectalis sling operation for fecal incontinence. METHODS: Since August 2001, we performed the perineal puborectalis sling operation on eight patients with idiopathic fecal incontinence. A specially designed polyester mesh sling was introduced along the puborectalis muscle, from a posterior perianal incision, running to a small suprapubic incision. The ends were tied together with moderate tension. Patients were evaluated with the Fecal Incontinence Severity Index, the Cleveland Clinic Score of Incontinence, and the Fecal Incontinence Quality of Life Scale. Manometry and defecography were performed before and six months after the operation. RESULTS: Eight patients (7 females; mean age, 63 (range, 44-77) years) were evaluated. A wound infection developed in one patient, which subsided with antibiotics. A rectal ulcer developed in one patient, necessitating sling removal. In the remaining seven patients, the Fecal Incontinence Severity Index improved from 27 to 9, and the Cleveland Clinic Score of Incontinence improved from 13 to 5 (P < 0.05). All parameters in the Fecal Incontinence Quality of Life Scale improved: lifestyle from 2.1 to 3.6; coping/behavior from 1.5 to 3.4; depression/self perception from 2.3 to 3.7; and embarrassment from 2 to 3.6 (P < 0.05). No significant difference was found between preoperative and postoperative maximum resting pressure and maximum squeeze pressure. However, the median anorectal angle on defecography after the operation was significantly reduced (P < 0.05). CONCLUSIONS: We believe that the perineal puborectalis sling operation is technically feasible, with low morbidity, and can be an effective procedure for idiopathic fecal incontinence.

Anal Canal↗

Osteoblast behaviour on in situ photopolymerizable three-dimensional scaffolds based on D, L-lactide, epsilon-caprolactone and trimethylene carbonate.

Polymer networks formed by photocrosslinking of multifunctional oligomers have great potential as injectable and in situ forming materials for bone tissue engineering. Porous scaffolds varying in polyester type and crosslinking density were prepared from methacrylate-endcapped oligomers based on D,L-lactide, epsilon -caprolactone and trimethylene carbonate: LA/CL-hexanediol, LA/CL-dipentaerythritol and LA/TMC-HXD. The biocompatibility and bone formation were related with the degradation time and mechanical properties. The viability of fibroblasts was evaluated after incubation with extraction medium by MTT-assay. All scaffolds showed a good biocompatibility. Rat bone marrow cells were cultured on the scaffolds for 21 days and were able to attach and differentiate on the scaffolds. The cells expressed high alkaline phosphatase activity, have formed a mineralized extracellular matrix and secreted osteocalcin. TEM of the polymer interface revealed osteoblasts which secreted an extracellular matrix containing matrix vesicles loaded with apatite crystals.LA/TMC-HXD, LA/CL-HXD and LA/CL-DPENT had a 50% mass loss at 3,5 months respectively 6 and 7, 5 months. The mechanical properties improve by increasing the branching of the precursor methacrylates (by replacing HXD by DPENT) but do not depend on their chemical composition. Hence, scaffolds with high elastic properties and variable degradation time can be obtained, which are promising for bone tissue engineering.

Alkaline Phosphatase↗

A thermostable beta-ketothiolase of polyhydroxyalkanoates (PHAs) in Thermus thermophilus: purification and biochemical properties.

Polyhydroxyalkanoates (PHAs) are polyesters of hydroxyalkanoates (HAs) synthesised by numerous bacteria as intracellular carbon and energy storage compounds which accumulate as granules in the cytoplasm of the cells. The biosynthesis of PHAs, in the thermophilic bacterium T. thermophilus grown in a mineral medium supplemented with sodium gluconate as sole carbon source has been recently reported. Here, we report the purification at apparent homogeneity of a beta-ketoacyl-CoA thiolase from T. thermophilus, the first enzyme of the most common biosynthetic pathway for PHAs. B-Ketoacyl-CoA thiolase appeared as a single band of 45.5-kDa molecular mass on SDS/PAGE. The enzyme was purified 390-fold with 7% recovery. The native enzyme is a multimeric protein of a molecular mass of approximately of 182 kDa consisting of four identical subunits of 45.5 kDa, as identified by an in situ renaturation experiment on SDS-PAGE. The enzyme exhibited an optimal pH of approximately 8.0 and highest activity at 65 degrees C for both direction of the reaction. The thiolysis reaction showed a substrate inhibition at high concentrations; when one of the substrates (acetoacetyl CoA or CoA) is varied, while the concentrations of the second substrates (CoA or acetoacetyl CoA respectively) remain constant. The initial velocity kinetics showed a pattern of a family of parallel lines, which is in accordance with a ping-pong mechanism. beta-Ketothiolase had a relative low Km of 0.25 mM for acetyl-CoA and 11 microM and 25 microM for CoA and acetoacetyl-CoA, respectively. The enzyme was inhibited by treatment with 1 mM N-ethylmaleimide either in the presence or in the absence of 0.5 mM of acetyl-CoA suggesting that possibly a cysteine is located at/or near the active site of beta-ketothiolase.

Acetyl-CoA C-Acyltransferase↗

Insulin containing nanocomplexes formed by self-assembly from biodegradable amine-modified poly(vinyl alcohol)-graft-poly(L-lactide): bioavailability and nasal tolerability in rats.

PURPOSE: The bioavailability and local tolerability of insulin containing nanocomplexes from amine-modified poly(vinyl alcohol)-graft-poly(L-lactide) were studied in rats. Histology of the nasal epithelium was studied to document integrity of the mucosa. METHODS: Nanocomplexes (NC) were prepared by spontaneous self-assembly of insulin and the water-soluble amphiphilic polymer. Changes in blood glucose and insulin blood concentration were monitored in anesthetized rats using a glucose meter and enzyme-linked immunosorbent assay, respectively. Histological sections of the nasal cavity were examined after H&E staining by light microscopy. RESULTS: NC reduced blood glucose level in fasted healthy rats by 20% after 50-80 min and in streptozotocin induced diabetic rats by 30% within 75-95 min compared to basal levels. In both animal models significant concentrations of human insulin were detected, with relative bioavailabilities F(rel) of 2.8 up to 8.3%. The more hydrophobic, lactic acid grafted polyester were more effective at a threefold higher polymer concentration, increasing the relative bioavailability F(rel) of a 5 IU/kg dose from 2.8 to 5.7%. Histological examination of the nasal mucosa after 4 h showed no signs of toxicity at the site of nasal administration. CONCLUSIONS: These results demonstrate that the NC significantly enhanced insulin absorption, suggesting that amphiphilic biodegradable comb-polymers offer a promising approach for nasal peptide delivery.

Administration, Intranasal↗

Fast and sensitive silver staining of DNA in polyacrylamide gels.

The photochemically derived silver stain of nucleic acids in polyacrylamide gels originally described by Merril et al. (1981, Science 211, 1437-1438) was modified to reduce unspecific background staining and increase sensitivity (down to 1 pg/mm2 band cross-section). Detection limits for double-stranded DNA fragments from HaeIII endonuclease digests of phage phi X174 were maintained despite eliminating oxidation pretreatment of fixed gels and reducing silver nitrate concentration. Preexposure to formaldehyde during silver impregnation enhanced sensitivity and the inclusion of the silver-complexing agent sodium thiosulphate in the image developer decreased background staining. Higher formaldehyde concentration during image development resulted in darker bands with good contrast. The procedure almost halves the number of steps, solutions and experimental time required and can be used for the staining of DNA fragments in polyacrylamide gels bound to a polyester backing film by controlling temperature during image development. We have applied this improved staining procedure for the routine analysis of complex DNA profiles generated by DNA amplification fingerprinting (DAF).

Bacteriophages↗

In vivo guidance of regenerating nerve by laminin-coated filaments.

Laminin is reportedly an extremely potent neurite-promoting agent in vitro. We investigated the effect of laminin on regeneration of axons in vivo, using a cord of polyester filaments coated with laminin as a guide, replacing a 10-mm segment of the rat sciatic nerve. After 4 weeks the cross section of the nerve guide at its midportion was examined under an electron microscope. Many regenerated axons were seen in the laminin-coated group, whereas no axon was observed in the controls. We conclude that laminin guides the regeneration of axons in vivo.

Animals↗

Immobilization of antibodies on a new solid phase for use in ELISA.

Antibodies to myoglobin were immobilized by covalent linkage to polyester film for use in a solid-phase ELISA. The covalent linkage of antibody to this new solid phase was accomplished by partial acid hydrolysis of the film followed by periodate oxidation. About 60 ng of protein could be immobilized per cm2 of film and the binding was stable. Antimyoglobin IgM immobilized on the films was used in the ELISA technique to detect myoglobin within the range 0.25-10 ng. The assay procedure was found to be very accurate and the coefficient of variation of each concentration ranged from 0.63 to 2.1%. Furthermore the immobilized film could be re-used after dissociating the antigen antibody complexes.

Animals↗

Biosynthesis of poly(3-hydroxyalkanoates) from threonine.

A series of copolyesters of 3-hydroxybutyrate (3HB) and 3-hydroxyvalerate (3HV) was biosynthesized by Alcaligenes eutrophus from an amino acid, threonine. The 3HV content of these polyesters ranged from less than 0.1% to 30%.

3-Hydroxybutyric Acid↗

Production of heteropolymeric polyhydroxyalkanoate in Escherichia coli from a single carbon source.

Poly[beta-hydroxybutyrate-co-beta-hydroxyvalerate] co-polymer, PHBV, is a polyhydroxyalkanoate (PHA) that has greater utility as a biodegradable thermoplastic polyester than poly-beta-hydroxybutyrate, PHB. In order to produce PHBV, a system of pathways is required to produce both hydroxybutyrate (HB) and hydroxyvalerate (HV) monomers from the sources of carbon. A working model for conversion of glucose to PHBV via acetyl- and propionyl-coenzyme A was constructed by expressing the PHA biosynthesis genes from Alcaligenes eutrophus in Escherichia coli strain K-12 under novel growth conditions. When 1 mM valine was added to 1% glucose medium, growth ceased and up to 2.5% of the incorporated monomers were HV; up to 4% were HV when 1 mM threonine was added as well. Threonine dehydratase (TD) converts threonine to alpha-ketobutyrate; TD is required for HV to be incorporated into PHA unless its transaminated reaction product, alpha-aminobutyrate, is added to the medium. Intracellular alpha-ketobutyrate accumulates when valine is added to the medium because valine, which cannot be metabolized to HV by E. coli strain K-12, stimulates TD and inhibits acetolactate synthase. In turn, alpha-ketobutyrate is converted to propionyl-CoA by the E. coli pyruvate dehydrogenase complex. This constitutes a defined system of pathways for synthesis of a heteropolymeric PHA from a single carbon source, which in the future could be transferred to other organisms including plants.

Acyl Coenzyme A↗

Tissue reaction to fabrics coated with turbostratic carbon: subcutaneous versus vascular implants.

A technique allowing the deposition of an adherent thin film of turbostratic, high-density carbon on heat-sensitive polymers was recently developed. The biological response to this biomaterial on yarns and fabrics of the type used in cardiovascular surgery has been studied. Polyester yarns, knitted Dacron sheets and knitted uncrimped Dacron vascular grafts were coated with a thin film (less than 1 micron) of turbostratic carbon using a physical vapour deposition process. Coated and control discs of knitted material, as well as coated and uncoated yarns, were implanted in pairs in the subcutaneous tissue of mice, using for each type of implant two cohorts of 12 animals, with observation periods of 4 and 8 wks respectively. Vascular grafts (8 cm long, 8 mm i.d.) coated with carbon on the luminal side only, were implanted for 12 wks in the infrarenal aortic position in six dogs, and compared to uncoated Dacron grafts of the same dimensions inserted in the same location and for the same duration in the infrarenal aortic position in six control animals. With subcutaneous implants, there was no significant difference in the tissue reaction to either coated or uncoated patches. In contrast, the vascular grafts, all of which were patent upon retrieval, showed a much lower extent of thrombosis on the blood-exposure surface in the case of carbon-coated Dacron, as compared to the luminal surface of control prostheses. The internal capsule (tissue formed between the polymer fabric and the blood interface) was thinner in carbon-coated grafts than in control grafts.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Degradation of poly(ester) microspheres.

Biodegradable polymeric microspheres have been prepared by spray drying, precipitation, rotary evaporation and press grinding methods. Erosion of microspheres of poly(lactide), poly(3-hydroxybutyrate), copolymers of lactide and glycolide, and copolymers of 3-hydroxybutyrate and 3-hydroxyvalerate at 85 degrees C and 37 degrees C have been studied using ion chromatography, nuclear magnetic resonance, residual mass measurements, viscometry and gel permeation chromatography. Such studies demonstrated that these polyester matrices degraded via (1) random chain scission and (2) release of soluble monomeric and oligomeric products. Protein release from microspheres prepared by these methods indicated that most of the protein is released before the polymer matrix loses weight.

Biodegradation, Environmental↗

Degradation of and tissue reaction to biodegradable poly(L-lactide) for use as internal fixation of fractures: a study in rats.

Samples of high-molecular-weight poly(L-lactide) (PLLA) (Mv = 9.0 x 10(5), a biomaterial developed for plates and screws used in internal fixation of jaw fractures, were implanted subcutaneously in the backs of rats to study tissue reaction to PLLA and to follow the degradation process. The PLLA seemed to follow the degradation pattern typical of biodegradable polyesters. After pure hydrolysis up to about 104 wk, phagocytic activity of macrophages was found at about 143 wk. Full resorption of PLLA was not demonstrated in this study. Except for the early and final parts of the implant period, no acute or chronic inflammatory reaction was observed. No implant was rejected. It is estimated that more than 3 yr will be required for total resorption of PLLA. For bone-healing this long period is of no practical importance. There is no need for removal of PLLA after fracture healing as is the case with metal fixation devices. Thus, PLLA has potential application in internal fixation of fractures and osteotomies in the maxillofacial region and other fractures that are not too heavily loaded in the human body.

Animals↗

Gel casting of resorbable polymers. 1. Processing and applications.

A gel casting technique based on resorbable, synthetic, alpha-polyesters (or lactide-glycolide polymers) is described for producing medical implants such as bone graft substitutes and timed-release carriers for medication. This solution-based method enables production of thick-section solid and microporous materials, and blending of polymers and particulate fillers. Implant degradation rate, for example, may be adjusted by variation of polymer type, molecular weight range, crystallinity and morphology. Gel casting conditions are reported for solid and microporous materials and processing characteristics are interpreted in terms of established crystallization and dissolution behaviour of polymers.

Biodegradation, Environmental↗

Biodegradation of PLA/GA polymers: increasing complexity.

The degradation of aliphatic polyesters derived from lactic and glycolic acids (PLA/GA) depends on many factors. It has been found recently that the interior of large size devices degrades faster than the outer zone. A qualitative model has been proposed to account for this heterogeneous degradation. It is based on diffusion-reaction phenomena combined with the well-known autocatalytic effect of carboxylic chain ends. This contribution recalls the present understanding of the hydrolytic degradation of PLA/GA polymers and emphasizes its complexity on the basis of the influence of secondary factors such as the presence of a basic load, namely, gentamycin, in poly(lactic acid) matrices, and the presence of long stereoregular sequences in poly(DL-lactic acid) macromolecules.

Biocompatible Materials↗