Postimplantation host tissue response and biodegradation of biologic versus polymer meshes implanted in an intraperitoneal position.

BACKGROUND: This study compared the in vitro and in vivo behaviors at the peritoneal interface of a new polymer material (Bio-A) and of two biologic non-cross-linked materials (Tutomesh [Tuto] and Strattice [St]), all biodegradable. METHODS: Omentum mesothelial cells from rabbits were seeded onto the three prosthetic materials tested. At 1, 4, 8, 16, and 24 h after implantation, mesothelial cover was performed using a scanning electron microscope (SEM). In the in vivo study, 3 × 3 cm mesh fragments were placed on the parietal peritoneum of the same rabbits and fixed at the four corners with individual stitches. The implants were randomized such that six fragments of each material were implanted in nine animals (2 per animal). Adhesion formation was quantified by sequential laparoscopy and image analysis 3, 7, and 14 days after implantation. The animals were killed at 90 days, and the meshes were subjected to microscopy and immunohistochemistry. RESULTS: The in vitro mesothelial cover was significantly greater for St than for Bio-A at each time point. The percentage of cover for St was also higher than for Tuto 16 and 24 h after seeding and higher for Tuto than for Bio-A at all time points. Compared with the biologic meshes, significantly higher adhesion percentages were recorded for Bio-A. At 90 days after implantation, differences in absorption measured as percentage of reduction in mesh thickness were detected among all the meshes. The least absorbed was St. The neoperitoneum thickness was significantly greater for the biologic meshes than for the polymer mesh, although this variable also differed significantly between St and Tuto. Macrophage counts were higher for Bio-A than for the biologic meshes. CONCLUSIONS: Greater mesothelial cover was observed in vitro for St. In vivo, adhesion formation and the macrophage response induced by Bio-A were greater than those elicited by the biologic materials. Bio-A and Tuto showed substantial biodegradation compared with St.

Intraperitoneal behaviour of a new composite mesh (Parietex™ Composite Ventral Patch) designed for umbilical or epigastric hernia repair.

INTRODUCTION: The most common treatment option for ventral and umbilical hernias is the implant of a prosthetic mesh. This study compares the behaviour of a new mesh, Parietex™ Composite Ventral Patch (Ptx), with two commercially available meshes, Ventralex™ ST Hernia Patch and Proceed™ Ventral Patch. MATERIALS AND METHODS: The following meshes were tested in a umbilical-hernia repair model using 54 rabbits: Ventralex™ ST Hernia Patch (Vent) (Bard Davol Inc., USA); Proceed™ Ventral Patch (PVP) (Ethicon, USA) and Ptx (Covidien, Sofradim, France) (n = 18 each). At 3, 7 and 14 days postimplantation, peritoneal behaviour and adhesion formation were assessed by sequential laparoscopy. Adhesions were scored for consistency and quantified by image analysis. The animals were euthanized at 2 (n = 27) and 6 weeks (n = 27) postsurgery. Mesothelial cover of meshes and tissue ingrowth were determined by scanning and light microscopy. RESULTS: Seroma was observed in 1/18 Vent, 7/18 PVP and 4/18 Ptx, mainly between the implant and subcutaneous tissue. Firm omental adhesions between the mesh and parietal peritoneum were noted in 2/9 Vent, 6/9 PVP and 3/9 Ptx at 2 weeks and in 3/9 Vent, 5/9 PVP and 1/9 Ptx at 6 weeks. Three (out of 9) encapsulated PVP implants showed "tissue-integrated" adhesions affecting the intestinal loops. No differences between implants were detected in the surface area occupied by adhesions at 2 weeks, though at 6 weeks, percentages were significantly higher (p < 0.01; Mann-Whitney U test) for PVP compared to Ptx or Vent. At this time point, Ptx and Vent showed good host tissue incorporation and optimal mesothelialization. CONCLUSIONS: The PVP implants showed greater adhesion formation than the other materials. Postimplantation behaviour was comparable for Ptx and Vent including scarce adhesion formation and optimal mesothelialization. Regarding tissue integration, Ptx showed greater long-term collagenization of the neoformed tissue.

Role of lysyl oxidases in neointima development in vascular allografts.

BACKGROUND: Extracellular matrix deposition is the main factor inducing stenotic lesions in arterial grafts. Lysyl oxidases (LOX) play a key role in stabilizing collagen and elastin. OBJECTIVE: To examine the repair response to arterial allografts in terms of LOX expression and collagen/elastin deposition using LOX inhibitors. METHODS: Lewis/Fisher-344 rats were used as donors/recipients. Donor segments were grafted to the right iliac artery of recipients and retrieved 14/30 (short-term) or 90/180 days (long-term) after surgery. One group of animals was injected with a potent irreversible LOX inhibitor daily for 30 days. RESULTS: Intimal hyperplasia increased in thickness until 90/180 days postsurgery. Elastin showed great expression in the neointima at 14/30 days and in the media at 90/180 days. LOX/LOXL1 were similarly expressed in the arterial wall during the first month. In the long term, their overexpression was confined to neointimal layers. At 14 days, collagen types I/III were identified in the grafts. The neointima acquired collagen I over time. In the group of animal treated with the LOX inhibitor, intimal hyperplasia was significantly inhibited. CONCLUSION: LOX were overexpressed in late stages of intimal hyperplasia in the allografts. LOX inhibitors prevented the development of the neointimal layer, such that their modulation could reduce the excessive extracellular matrix deposition that leads to stenosis.

Kidney androgen-regulated protein transgenic mice show hypertension and renal alterations mediated by oxidative stress.

BACKGROUND: Kidney androgen-regulated protein (KAP), a proximal tubule androgen-regulated gene, codes for a protein of unknown function. METHODS AND RESULTS: To investigate the consequences of KAP overexpression in kidney, we produced KAP transgenic mice and performed microarray expression analyses in kidneys of control and transgenic males. Downregulation of the androgen-sensitive Cyp4A14 monooxygenase gene in KAP transgenic mice prompted us to analyze blood pressure levels, and we observed that transgenic mice were hypertensive. Inhibition of 20-hydroxyeicosatetraenoic acid synthesis by N-hydroxy-N'-(4-n-butyl-2-methylphenyl) formamidine (HET0016) reduced the increased 20-hydroxyeicosatetraenoic acid levels in urine and normalized arterial pressure in transgenic mice, as did the NADPH oxidase inhibitor apocynin. Increased oxidative stress in transgenic mice was demonstrated by (1) enhanced excretion of urinary markers of oxidative stress, 8-iso-prostaglandin F2alpha, 8-hydroxydeoxyguanosine, and thiobarbituric acid-reacting substances; (2) augmented mitochondrial DNA damage and malondialdehyde levels in kidneys; and (3) diminished catalase and glutathione peroxidase activity in transgenic kidneys. Mice exhibited renal defects that included focal segmental glomerulosclerosis, proteinuria, glycosuria, and fibrosis. CONCLUSIONS: Taken together, these results indicate that KAP expression is critical for cardiovascular-renal homeostasis maintenance and that hypertension is associated with increased oxidative stress. This is the first report showing that overexpression of an androgen-regulated, proximal tubule-specific gene induces hypertension. These observations may shed light on the molecular pathophysiology of gender differences in the prevalence and severity of hypertension and chronic renal disease.

Efficacy of antimicrobial agents delivered to hernia meshes using an adaptable thermo-responsive hyaluronic acid-based coating.

PURPOSE: Mesh-related infection is a critical outcome for patients with hernia defect stabilized with synthetic or biological meshes. Even though bioactive meshes loaded with antibiotics or antiseptics are slowly emerging in the market, the available solutions still lack versatility. Here, we proposed a polymer solution, i.e., hyaluronic acid-poly(N-isopropylacrylamide) (HApN), which forms a hydrogel to be used as coating for meshes only when it reaches body temperature. METHODS: We assessed how the gelation of HApN was influenced by the incorporation of different antibiotic and antiseptic formulations, and how this gel can be used to coat several mesh types. The impact of the coating on the elastic behavior of a macroporous mesh was tested under cyclic elongation condition. Finally, we selected two different coating formulations, one based on antibiotics (gentamicin + rifampicin) and one based on antiseptic (chlorhexidine) and tested in vitro their antimicrobial efficacies. RESULTS: HApN can be used as carrier for different antimicrobial agents, without having a strong influence on its gelation behavior. Porous or dense meshes can be coated with this polymer, even though the stability was not optimal on macroporous meshes such as Optilene when pores are too large. HApN loaded with drugs inhibited in vitro the growth of several Gram-positive and Gram-negative bacteria. CONCLUSION: Compared to the available technologies developed to endow meshes with antibacterial activity, the proposed HApN offers further versatility with potential to prevent mesh-related infection in hernioplasty.

Long term comparative evaluation of two types of absorbable meshes in partial abdominal wall defects: an experimental study in rabbits.

PURPOSE: Synthetic prosthetic materials that are fully absorbable seek to reduce the host foreign body reaction and promote host tissue regeneration. This preclinical trial was designed to analyse, in the long term, the behaviour of two prosthetic meshes, one synthetic and one composed of porcine collagen, in abdominal wall reconstruction. METHODS: Partial defects were created in the abdominal walls of New Zealand rabbits and repaired using a synthetic absorbable mesh (Phasix™) or a non-crosslinked collagen bioprosthesis (Protexa™). After 3, 6, 12 and 18 months, specimens were recovered for light microscopy and collagen expression analysis to examine new host tissue incorporation, macrophage response and biomechanical strength. RESULTS: Both materials showed good host tissue incorporation in line with their spatial structure. At 18 months postimplant, Protexa™ was highly reabsorbed while the biodegradation of Phasix™ was still incomplete. Collagenization of both materials was good. Macrophage counts steadily decreased over time in response to Phasix™, yet persisted in the collagen meshes. At 18 months, zones of loose tissue were observed at the implant site in the absence of herniation in both implant types. The stress-stretch behaviour of Phasix™ implants decreased over time, being more pronounced during the period of 12-18 months. Nevertheless, the abdominal wall repaired with Protexa™ became stiffer over time. CONCLUSION: Eighteen months after the implant both materials showed good compatibility but the biodegradation of Phasix™ and Protexa™ was incomplete. No signs of hernia were observed at 18 months with the stress-stretch relations being similar for both implants, regardless of the more compliant abdominal wall repaired with Protexa™ at short term.

Lysyl oxidase like-1 dysregulation and its contribution to direct inguinal hernia.

OBJECTIVE: The aetiology of inguinal hernia involves changes in collagen turnover and metalloproteinase expression; yet it is not known whether the elastic fibre system could also be affected. This study was designed to compare the expression of tropoelastin (TE), lysyl oxidase-like 1 (LOXL-1) and elastase in the transversalis fascia of patients with and without inguinal hernia. MATERIAL AND METHODS: Transversalis fascia (TF) specimens were obtained from patients undergoing surgery for direct or indirect inguinal hernia (n = 20 each) and from multi-organ donors during organ procurement (controls, n = 16). The specimens were divided according to age (20-40/41-60 years). Tissues were immunohistochemically labelled using anti-tropoelastin, anti-LOXL-1 and anti-elastase antibodies and subjected to Western blot analysis. Relative amounts of LOXL-1 and TE mRNA were determined by real time RT-PCR in cultured cells obtained from the TF of patients and controls. RESULTS: Significantly lower TE and LOXL-1 levels were observed in patients with direct inguinal hernia compared with controls or those with indirect hernia. In contrast, patients with direct inguinal hernia showed significantly higher elastase expression. In fibroblasts isolated from the TF, relative amounts of tropoelastin mRNA were lower for the hernia groups but differences were not significant. LOXL-1 mRNA levels were significantly lower in the direct hernia group compared to controls. CONCLUSIONS: Our findings suggest that impaired elastic fibre function in the transversalis fascia of patients with direct inguinal hernia, reflected by diminished elastin synthesis and its enhanced enzyme degradation, contributes to the development of this type of hernia.

Experimental study on the use of a chlorhexidine-loaded carboxymethylcellulose gel as antibacterial coating for hernia repair meshes.

PURPOSE: Biomaterials with an antimicrobial coating could avoid mesh-associated infection following hernia repair. This study assesses the use of a chlorhexidine-loaded carboxymethylcellulose gel in a model of Staphylococcus aureus mesh infection. METHODS: A 1% carboxymethylcellulose gel containing 0.05% chlorhexidine was prepared and tested in vitro and in vivo. The in vitro tests were antibacterial activity (S. aureus; agar diffusion test) and gel cytotoxicity compared to aqueous 0.05% chlorhexidine (fibroblasts; alamarBlue). For the in vivo study, partial abdominal wall defects (5 × 2 cm) were created in New Zealand white rabbits (n = 15) and inoculated with 0.25 mL of S. aureus (106 CFU/mL). Defects were repaired with a lightweight polypropylene mesh (Optilene) without coating (n = 3) or coated with a carboxymethylcellulose gel (n = 6) or chlorhexidine-loaded carboxymethylcellulose gel (n = 6). Fourteen days after surgery, bacterial adhesion to the implant (sonication, immunohistochemistry), host tissue incorporation (light microscopy) and macrophage reaction (immunohistochemistry) were examined. RESULTS: Carboxymethylcellulose significantly reduced the toxicity of chlorhexidine (p < 0.001) without limiting its antibacterial activity. While control and gel-coated implants were intensely contaminated, the chlorhexidine-gel-coated meshes showed a bacteria-free surface, and only one specimen showed infection signs. The macrophage reaction in this last group was reduced compared to the control (p < 0.05) and gel groups. CONCLUSIONS: When incorporated in the carboxymethylcellulose gel, chlorhexidine showed reduced toxicity yet maintained its bactericidal effect at the surgery site. Our findings suggest that this antibacterial gel-coated polypropylene meshes for hernia repair prevent bacterial adhesion to the mesh surface and have no detrimental effects on wound repair.

[Importance of PSA velocity and PSA density in the prediction of prostate cancer in TURP or open prostatectomy specimen of patients with previous negative prostate biopsy]. / Importancia de la velocidad y la densidad de PSA en la predicción de cáncer de próstata en la pieza de RTU o adenomectomía de pacientes con biopsia prostática previa negativa.

PURPOSE: To determine variables related to the finding of prostate cancer (PC) in patients who underwent surgery following at least one negative prostate biopsy (PB). MATERIALS AND METHODS: A retrospective study of 170 patients who underwent transurethral resection of the prostate (TURP) or open prostatectomy between 1999 and 2007, following one or more negative PB sets. A multivariate logistic regression analysis was carried out in order to determine variables related to the finding of PC. The predictive capacity of PSA, PSA-density and PSA-velocity was assessed by means of ROC curves and the area under the curve (AUC). Sensitivity, specificity and predictive values were determined for several PSA-density and PSA-velocity cut-off points. RESULTS: Open prostatectomy was carried out on 104 patients (61.18%) and TURP on 66 (38.82%). PC was detected in the surgical specimen of 16 patients (9.41%). Variables associated with the finding of PC in the surgical specimen were PSA-density (OR: 1.47; 95% CI: 1.22-6.64; p: 0.007) and PSA-velocity (OR: 2.87; 95% CI: 1.60-5.12: p < 0.001). The AUCs were 0.746, 0.793 and 0.832, for PSA, PSA-density and PSA-velocity, respectively. The most sensitive PSA-density and PSA-velocity cut-off points in detecting PC were 0.15 and 1 ng/ml/year, respectively. Patients without PC showed a median PSA reduction of 9.35 ng/ml (-2.40 - 35.40), following surgery. CONCLUSIONS: PSA-density and PSA-velocity in particular, allow for the prediction of the presence of PC in the TURP or open prostatectomy specimen of patients with previously negative PBs. Diagnostic TURP could prove useful in patients with clinical suspicion of PC, susceptible to curative treatment, with PSA-velocity > 1l ng/ml/year and one or more negative saturation biopsies.

Peroxisome proliferator-activated receptor gamma-dependent repression of the inducible nitric oxide synthase gene.

The peroxisome proliferator-activated receptor gamma (PPARgamma) is a member of the nuclear receptor superfamily that activates target gene transcription in a ligand-dependent manner. In addition, liganded PPARgamma can inhibit transcription of genes induced by gamma interferon (IFN-gamma) and/or lipopolysaccharides (LPSs), including the inducible nitric oxide synthase (iNOS) gene. Inhibition of the iNOS promoter is achieved partially through antagonizing the activities of NF-kappaB, AP-1, and STAT1, which are known to mediate effects of LPS and IFN-gamma. Previous studies have suggested that transrepression of these factors by nuclear receptors involves competition for limiting amounts of the general coactivators CREB-binding protein (CBP) and p300. CBP and p300 are thought to be recruited to nuclear receptors through bridging factors that include SRC-1, although CBP also interacts directly with PPARgamma through its amino terminus. These observations have raised questions concerning the involvement of SRC-1-like factors in CBP recruitment and transrepression. We here provide evidence that PPARgamma's ability to repress iNOS transcription requires the ligand-dependent charge clamp that mediates interactions with CBP and SRC-1. Single amino acid mutations in PPARgamma that abolished ligand-dependent interactions with SRC-1 and CBP not only resulted in complete loss of transactivation activity but also abolished transrepression. Conversely, a CBP deletion mutant containing the SRC-1 interaction domain but lacking the N-terminal PPARgamma interaction domain was inactive as a PPARgamma coactivator and failed to rescue transrepression. Together, these findings are consistent with a model in which transrepression by PPARgamma is achieved by targeting CBP through direct interaction with its N-terminal domain and via SRC-1-like bridge factors.

IkappaB kinase (IKK)-associated protein 1, a common component of the heterogeneous IKK complex.

Activation of the transcription factor NF-kappaB is controlled by the sequential phosphorylation, ubiquitination, and degradation of its inhibitory subunit, IkappaB. We recently purified a large multiprotein complex, the IkappaB kinase (IKK) signalsome, which contains two regulated IkappaB kinases, IKK1 and IKK2, that can each phosphorylate IkappaBalpha and IkappaBbeta. The IKK signalsome contains several additional proteins presumably required for the regulation of the NFkappaB signal transduction cascade in vivo. In this report, we demonstrate reconstitution of IkappaB kinase activity in vitro by using purified recombinant IKK1 and IKK2. Recombinant IKK1 or IKK2 forms homo- or heterodimers, suggesting the possibility that similar IKK complexes exist in vivo. Indeed, in HeLa cells we identified two distinct IKK complexes, one containing IKK1-IKK2 heterodimers and the other containing IKK2 homodimers, which display differing levels of activation following tumor necrosis factor alpha stimulation. To better elucidate the nature of the IKK signalsome, we set out to identify IKK-associated proteins. To this end, we purified and cloned a novel component common to both complexes, named IKK-associated protein 1 (IKKAP1). In vitro, IKKAP1 associated specifically with IKK2 but not IKK1. Functional analyses revealed that binding to IKK2 requires sequences contained within the N-terminal domain of IKKAP1. Mutant versions of IKKAP1, which either lack the N-terminal IKK2-binding domain or contain only the IKK2-binding domain, disrupt the NF-kappaB signal transduction pathway. IKKAP1 therefore appears to mediate an essential step of the NF-kappaB signal transduction cascade. Heterogeneity of IKK complexes in vivo may provide a mechanism for differential regulation of NF-kappaB activation.

Biomechanical and histologic evaluation of two application forms of surgical glue for mesh fixation to the abdominal wall.

The use of an adhesive for mesh fixation in hernia repair reduces chronic pain and minimizes tissue damage in the patient. This study was designed to assess the adhesive properties of a medium-chain (n-butyl) cyanoacrylate glue applied as drops or as a spray in a biomechanical and histologic study. Both forms of glue application were compared to the use of simple-loose or continuous-running polypropylene sutures for mesh fixation. Eighteen adult New Zealand White rabbits were used. For mechanical tests in an ex vivo and in vivo study, patches of polypropylene mesh were fixed to an excised fragment of healthy abdominal tissue or used to repair a partial abdominal wall defect in the rabbit respectively. Depending on the fixation method used, four groups of 12 implants each or 10 implants each respectively for the ex vivo and in vivo studies were established: Glue-Drops, Glue-Spray, Suture-Simple and Suture-Continuous. Biomechanical resistance in the ex vivo implants was tested five minutes after mesh fixation. In vivo implants for biomechanical and histologic assessment were collected at 14 days postimplant. In the ex vivo study, the continuous suture implants showed the highest failure sample tension, while the implants fixed with glue showed lower failure sample tension values. However, the simple and continuous suture implants returned the highest stretch values. In the in vivo implants, failure sample tension values were similar among groups while the implants fixed with a continuous running suture had the higher stretch values, and the glue-fixed implants the lower stretch values. All meshes showed good tissue integration within the host tissue regardless of the fixation method used. Our histologic study revealed the generation of a denser, more mature repair tissue when the cyanoacrylate glue was applied as a spray rather than as drops.

Influence of the structure of new generation prostheses on shrinkage after implant in the abdominal wall.

When a biomaterial is used to repair an abdominal wall defect, wound contraction can cause the prosthesis to shrink, and the tension generated can provoke recurrence of the defect. This study was designed to determine whether the structure of a prosthesis can directly influence prosthetic shrinkage. Abdominal wall defects (7 x 5 cm) in rabbits were repaired using the laminar prosthesis DualMesh (DM), the composites Sepramesh (Se) and Vypro II (Vy), and the reticular prosthesis Surgipro (PP). The animals were sacrificed 14 and 90 days after surgery, at which time implant specimens were morphologically and immunohistochemically examined to establish the presence of myofibroblasts and macrophages. The size of each prosthesis was measured at the end of the study through image analysis. Morphometric measurements revealed greatest prosthesis shrinkage for Se, PP, and Vy (16.05% +/- 5.08%; 13.75% +/- 4.22%; 16.16% +/- 6.34%), while the DM prostheses only showed a 7.57% +/- 0.62% decrease in size (p < 0.05). In the DM implants, the macrophage response and myofibroblast labeling were reduced. Both biomaterial structure and the macrophage reaction induced at the implant site modulate prosthetic shrinkage, laminar prostheses of the ePTFE type undergoing less shrinkage than reticular meshes. Reduced DM shrinkage was linked to decreased myofibroblast numbers 2 weeks postimplant.

[The utility of artificial neural networks in the prediction of prostate cancer on transrectal biopsy]. / Utilidad de las redes neuronales artificiales en la predicción de cáncer de próstata en la biopsia transrectal.

OBJECTIVE: To determine whether the development of an artificial neural network (ANN) made up of clinical variables allows for the prediction of prostate biopsy (PB) outcome. MATERIALS AND METHODS: Patients (n=953) underwent PB at the Arquitecto Marcide Hospital in Ferrol (Spain), between january 2000 and june 2005. The variables studied were age, PSA, digital rectal examination (DRE) and prostate volume, data for all of which were available in 843 cases. In order to determine factors related to prostate cancer (PC) diagnosis, a logistic regression analysis and a feed-forward neural network were developed, including three hidden layer nodes and an output node, representing the probability of PC. Both models were constructed from a random sample of n=643 patients (derivation set). The predictive capacity was assessed with the remaining 200 patients (validation set), by means of ROC curves and the area under the curve (AUC). RESULTS: PC was detected in 500 (59.3%) cases. Adjusting for age, PSA, digital rectal examination and prostate volume, in a multivariate logistic regression model it was observed that all the variables were independent predictors of PC. The AUC were 0.693 for PSA, 0.707 for prostate volume, 0.815 for logistic regression and 0.819 for ANN. The predictive capacity of the ANN was significantly higher than that of the PSA (p=0.002) and prostate volume (p<0,001) and similar to that of logistic regression (p=0.760). CONCLUSIONS: The ANN shows a PC prediction capacity that is significantly higher than unimodal diagnosis methods, and similar to that of logistic regression.

Tissue integration and inflammatory reaction in full-thickness abdominal wall repair using an innovative composite mesh.

PURPOSE: When composite meshes are used in abdominal wall repair, seroma formation may persist and delay the desired integration leading to recurrence. This study compares tissue integration and inflammatory response in abdominal wall repair with composites with different absorbable synthetic barriers. METHODS: Full-thickness defects created in the abdominal wall of rabbits were repaired using polypropylene prosthesis or the following composites: Physiomesh™ (Phy); Ventralight™ (Vent) and "new composite mesh" (Ncm) not yet used clinically in humans. The collected seroma was evaluated for IFN-γ/IL-4 by ELISA. Tissue integration, anti- (IL-13/TGFß-1/IL-10/IL-4) and pro-inflammatory (TNF-α/IL-6/IFN-γ/VEGF) cytokine mRNA expression and TGFß/VEGF immunolabeling were evaluated at 14 and 90 days post-implant. RESULTS: Seroma was observed in 10 of 12 Phy/Vent and 4 of 12 Ncm. Wound fluid IFN-γ showed a time-dependent significant increase in Vent and tendency to decrease in Ncm, while all composites exhibited IL-4 upward trend. Prostheses were fully infiltrated by an organized connective tissue at end time although the area had shown prior seroma. A stable mesothelium was developed, except in adhesion areas. Vent/Phy displayed a significant increase in TNF-α/IFN-γ-mRNA over time. Significant decrease in VEGF mRNA was observed in Phy/Ncm, while a significant increase of TGFß-1 mRNA was evident in all composites over time. Ncm exhibited the highest TGFß protein expression area at short term and the greatest percentage of VEGF positive vessels at end time. CONCLUSION: Ncm could be an appropriate candidate to improve clinical outcome showing the lower development of seroma and optimal tissue integration with minimal pro-inflammatory cytokine response over time and consistent pro-wound healing cytokine expression.

Biaxial Mechanical Evaluation of Absorbable and Nonabsorbable Synthetic Surgical Meshes Used for Hernia Repair: Physiological Loads Modify Anisotropy Response.

The aim of this study was to obtain information about the mechanical properties of six meshes commonly used for hernia repair (Surgipro(®), Optilene(®), Infinit(®), DynaMesh(®), Ultrapro™ and TIGR(®)) by planar biaxial tests. Stress-stretch behavior and equibiaxial stiffness were evaluated, and the anisotropy was determined by testing. In particular, equibiaxial test (equal simultaneous loading in both directions) and biaxial test (half of the load in one direction following the Laplace law) were selected as a representation of physiologically relevant loads. The majority of the meshes displayed values in the range of 8 and 18 (N/mm) in each direction for equibiaxial stiffness (tangent modulus under equibiaxial load state in both directions), while a few achieved 28 and 50 (N/mm) (Infinit (®) and TIGR (®)). Only the Surgipro (®) mesh exhibited planar isotropy, with similar mechanical properties regardless of the direction of loading, and an anisotropy ratio of 1.18. Optilene (®), DynaMesh (®), Ultrapro (®) and TIGR (®) exhibited moderate anisotropy with ratios of 1.82, 1.84, 2.17 and 1.47, respectively. The Infinit (®) scaffold exhibited very high anisotropy with a ratio of 3.37. These trends in material anisotropic response changed during the physiological state in the human abdominal wall, i.e. T:0.5T test, which the meshes were loaded in one direction with half the load used in the other direction. The Surgipro (®) mesh increased its anisotropic response (Anis[Formula: see text] = 0.478) and the materials that demonstrated moderate and high anisotropic responses during multiaxial testing presented a quasi-isotropic response, especially the Infinit(®) mesh that decreased its anisotropic response from 3.369 to 1.292.

Biomechanical and morphological study of a new elastic mesh (Ciberlastic) to repair abdominal wall defects.

The aim of this study was to conduct a preclinical evaluation of the behaviour of a new type of abdominal LW prosthesis (Ciberlastic), which was designed with a non-absorbable elastic polyurethane monofilament (Assuplus, Assut Europe, Italy) to allow greater adaptability to mechanical area requirements and higher bio-mimicking with the newly formed surrounding tissues. Our hypothesis was that an increase in the elasticity of the mesh filament could improve the benefits of LW prostheses. To verify our hypothesis, we compared the short- and long-term behaviour of Ciberlastic and Optilene(®) elastic commercial meshes by repairing the partially herniated abdomen in New Zealand White rabbits. The implanted meshes were mechanically and histologically assessed at 14 and 180 days post-implant. We mechanically characterized the partially herniated repaired muscle tissue and also determined mesh shrinkage at different post-implant times. This was followed by a histological study in which the tissue incorporation process was analysed over time. The new prosthesis designed by our group achieved good behaviour that was similar to that of Optilene(®), one of the most popular LW prostheses on the market, with the added advantage of its elastic property. The mechanical properties are significantly lower than those of the polypropylene Optilene(®) mesh, and the new elastic mesh meets the basic mechanical requirements for positioning in the abdominal wall, which was also demonstrated by the absence of recurrences after implantation in the experimental model. We found that the growth of a connective tissue rich in collagen over the hernial defect and the proper deposit of the collagen fibres in the regenerated tissue substantially modified the original properties of the mesh, thereby increasing its biomechanical strength and making the whole tissue/mesh stiffer.

In vitro assessment of an antibacterial quaternary ammonium-based polymer loaded with chlorhexidine for the coating of polypropylene prosthetic meshes.

PURPOSE: This study assesses the use of an absorbable polymer loaded with chlorhexidine (CHX) as an antibacterial coating for polypropylene (PP) meshes employed in hernia repair. METHODS: The polymer N,N-dimethyl-N-benzyl-N-(2-methacryloyloxyethyl) ammonium bromide was loaded with CHX (1 % w/w). Fragments (1 cm2) of Optilene® Mesh Elastic were coated either with the unloaded (POL) or CHX-loaded polymer (POL-CHX). Uncoated fragments (PP) served as controls. The release kinetics of the POL-CHX coating was monitored by HPLC. Sterile fragments were placed on agar plates previously contaminated with 106 CFU of Staphylococcus aureus (Sa) ATCC25923, Staphylococcus epidermidis (Se) ATCC12228, or Escherichia coli (Ec) ATCC25922 and incubated at 37 °C for 1/2/7 days. At each time point, inhibition halos were measured and bacterial adhesion to the meshes quantified by sonication and scanning electron microscopy. Coating cytotoxic effects were examined on cultured fibroblasts. RESULTS: The polymer coating gradually released CHX over 3 days. Inhibition halos were produced only around the POL-CHX-coated meshes and these were significantly smaller for Ec than Sa or Se (p < 0.01). While POL-CHX prevented bacterial adhesion to the mesh, the reduced bacterial yields over time were observed for the POL-coated versus control PP meshes (p < 0.001). By day 7, only Ec remained attached to the surface of control meshes. The POL coating was not cytotoxic, yet POL-CHX reduced the viability of cultured fibroblasts. CONCLUSIONS: When loaded with the antiseptic CHX, this quaternary ammonium-based polymer coating released its contents in a controlled manner indicating its potential prophylactic use to reduce the risk of infection following PP mesh implantation.

Muscle-derived stem cells in tissue engineering: defining cell properties suitable for construct design.

The terms construct or tissue equivalent refer to neotissue produced by tissue engineering techniques. The elements forming the construct are scaffolds on which cells are "recreated" to form an engineered-tissue sensitive to certain cell signals. The ability of the cells to expand and differentiate on the scaffold is determined by properties such as fixation, adhesion, proliferation and migration. Among the cell types that seem to be most promising for designing constructs are tissue-residing, or adult, stem cells, which show two main features: a capacity to differentiate into many cell lineages and the power of self-renewal. These features make them good candidates for cell replacement therapies. Here, we report the identification, isolation and culture of muscle stem cells aimed at establishing the ideal culture in terms of defining when the cultured cell population would show optimal characteristics for transfer to the scaffold to obtain a particular construct. Stem cells harvested from the dorsal muscle of white New Zealand rabbits were cultured in vitro and characterized 5 to 14 days after the start of culture. Fibroblasts obtained from the same experimental animal served as controls. The stem cells were examined by light and scanning electron microscopy. For stem cell identification, we used the antibodies anti-m-cadherin, anti-CD34 and anti-Myf-5. The markers of muscle differentiation used were: anti-vimentin, anti-alpha-actin, anti-desmin and anti-myosin. The expression profiles of the different markers of muscle differentiation and TGFbeta1 in the cell cultures were confirmed by Western blotting. Proliferation rates were determined by monitoring tritiated thymidine incorporation. The thymidine incorporation rate was substantially higher for the population of undifferentiated cells than for control fibroblasts obtained from the same animal. During the first five days of culture, most cells were negative for all the markers examined, with the exception of m-cadherin, CD34 and Myf-5, although discrete signs of vimentin expression started to emerge. After 14 days of culture, the adult stem cells showed vimentin (94.2%) and desmin (33.8%) expression yet scarce labeling for myosin (16.2%) and alpha-actin (8.3%). Control fibroblasts showed intense labeling for vimentin (99.3%) and alpha-actin (62.2%), while less than 2% of the population expressed myosin (0.9%) and desmin (1.6%). After two weeks of culture, muscle-derived stem cells show good proliferative and adhesion properties as they initiate differentiation. These conditions seem ideal for obtaining the desired construct.