Verónica Gómez-Gil

Early tissue incorporation and collagen deposition in lightweight polypropylene meshes: bioassay in an experimental model of ventral hernia.

BACKGROUND This study was designed to assess the early host tissue incorporation of several polypropylene lightweight (PP-LW) meshes used to repair abdominal wall defects and to correlate collagen deposition with the biomechanical response shown by PP-LW versus polypropylene heavyweight (PP-HW) meshes. METHODS Ventral hernial defects (7 x 5 cm) were created in the anterior abdominal wall of New Zealand rabbits and repaired by fixing PP-LW mesh of different pore sizes or a low porosity HW mesh to the edges of the defect. Rabbits were killed 14 days after implant, and specimens were taken from the central mesh area to examine collagen deposition by light microscopy, real time reverse transcription polymerase chain reaction, immunohistochemistry, and Western blotting. The biomechanical resistance of the biomaterials was also assessed. RESULTS All the materials showed excellent incorporation in host tissue. Relative amounts of collagen III mRNA were considerably higher than collagen I mRNA. Higher collagen I and III mRNA levels were noted for pore sizes equal to or greater than 3.45 +/- 0.19 mm(2) (Ultrapro/Optilene Elastic. These two meshes showed significantly higher levels of collagen III than Parietene and Surgipro with smaller pores. Biomechanical resistance values for Optilene were significantly higher than those recorded for Surgipro and Parietene. CONCLUSIONS (a) LW meshes of pore size larger than 3 mm(2) induced the genetic overexpression of collagen types I and III; (b) the larger pore-sized LW meshes induced more collagen type III deposition and its faster conversion to collagen I; (c) Optilene, the most porous LW mesh examined, showed the greatest tensile strength 14 days after implant.

Comparing the Behavior of Different Polypropylene Meshes (Heavy and Lightweight) in an Experimental Model of Ventral Hernia Repair

New generation prosthetic biomaterials for abdominal wall repair have been designed to be less dense, by having larger pores than that of the standard polypropylene meshes, to improve abdominal wall compliance. The aim of the present study was to analyze the functional and morphologic properties of these new meshes. For this purpose, 7 x 5 cm(2) defects were created in the anterior abdominal wall of 36 male New Zealand White rabbits and repaired using different polypropylene meshes: a heavyweight mesh (HW), Surgipro, and two lightweight meshes (LW), Parietene and Optilene. Six animals each implanted with biomaterial were sacrificed on postoperative days 14 and 90. Histological and morphometric analysis, adhesion assessment, and biomechanical resistance tests were performed. Similar behavior was shown by the LW and HW meshes in terms of the adhesions and macrophage response induced. After 14 days, the tensile strength of Optilene was greater than the strengths recorded for the other two biomaterials, probably because of its high elasticity. By 90 days, however, the tensile strengths of the three biomaterials were comparable. In conclusion, despite an initial tensile strength advantage shown by the mesh with larger pores, at 90 days postimplant, tensile strengths were similar. Compared with HW, LW prostheses have the benefit that less foreign material was implanted, preserving the elasticity of the recipient host tissue.

Postimplant behavior of lightweight polypropylene meshes in an experimental model of abdominal hernia.

Background. Over the years, reticular prostheses have undergone changes in their structure and composition to give rise to todays partially absorbable lightweight meshes. This study was designed to assess the biological and biomechanical behavior of these prostheses to establish whether they offer any advantages over nonabsorbable lightweight polypropylene prostheses. Materials and Methods. 7 × 5 cm defects were created in the anterior abdominal wall of New Zealand White rabbits and repaired by securing different prostheses to the edges of the defect with a running 4/0 polypropylene suture. The lightweight biomaterials compared were two nonabsorbable meshes: Parietene® and Optilene elastic®, and two partially absorbable prostheses: Vypro II® and Ultrapro®. At 14 and 90 days postimplant, tissue/prosthesis specimens were subjected to histological, immunohistochemical, shrinkage, and biomechanical analyses. Results. Adhesion formation on the peritoneum-facing surface of the meshes was significantly less extensive in the meshes with absorbable components at 90 days postimplant. The newly formed tissue around the prosthetic filaments was comprised of collagen fibers, fibroblasts, blood vessels, and macrophages. The partially absorbable meshes showed higher macrophage proportions (due to remnants of absorbable material and their structure) than the nonabsorbable meshes at 90 days, although differences were not significant. At 90 days postimplant, similar tensile strengths were recorded for all the implants. Conclusions. All the prosthetic materials induced good host tissue ingrowth, with no significant differences in tensile strength observed. Our findings suggest that partially absorbable lightweight prostheses could offer advantages over nonabsorbable lightweight meshes since less foreign material persists in the recipient, improving abdominal wall compliance.

TGF‐β1 overexpression in the transversalis fascia of patients with direct inguinal hernia

Background  The aetiology of inguinal hernia includes changes in collagen turnover and metalloproteinase (MMP) expression, and direct hernia has been linked to increased MMP‐2 expression. Since transforming growth factor β1 (TGFβ1) plays a role in tissue remodelling, this growth factor could directly affect metalloproteinase secretion and thus the proteolytic activity of these enzymes. We hypothesized that TGFβ1 expression could also be altered in direct inguinal hernias.

Active matrix metalloproteinase-2 upregulation in the abdominal skin of patients with direct inguinal hernia.

Eur J Clin Invest 2010; 40 (12): 1113–1121

Muscle-derived stem cells used to treat skin defects prevent wound contraction and expedite reepithelialization

Stem cells derived from adult tissues may serve as cell therapy to enhance the healing process in skin wounds. This study was designed to evaluate the use of autologous muscle‐derived stem cells in an experimental skin wound model in terms of their efficiency at promoting tissue repair/regeneration. Muscle‐derived cells obtained from the dorsal muscle of New Zealand rabbits were cultured in vitro for 2 weeks. The cell population was identified using the satellite markers CD34, m‐cadherin and Myf5, and the proliferative capacity of the adult stem cells was determined. The population was then fluorescently labeled with PKH26 and seeded onto a circular 2 cm diameter defect created on the dorsal side of the ear of the rabbit from which the cells had been harvested. Similar defects on the contra lateral ears were left untreated to form the control group. Fourteen days later, specimens were taken for light, transmission, and scanning electron microscopy, as well as for immunolabeling with antibodies against vimentin, α‐actin, desmin, myosin, fibronectin, and cytokeratin 14. Areas of wound contraction and reepithelialization were determined by image analysis. Wound contraction was significantly greater in the control than the treatment group (p<0.05); control specimens also showed more myosin expression. Reepithelialized areas were significantly greater in the treatment group (p<0.05). Control wounds showed nonepithelialized areas and inflammatory granulation tissue. Reepithelialization occurred as epidermal tongues of fusiform cells. Our findings indicate that the use of autologous stem cells on skin wounds expedites and improves the organisms natural healing process.

Peritoneal adhesion formation and reformation tracked by sequential laparoscopy: Optimizing the time point for adhesiolysis

BACKGROUND In a high proportion of patients, operatively lysed adhesions reform. Using a rabbit adhesiogenesis model, this study assessed the efficacy of adhesiolysis and examined how this relates to the tissue composition of adhesions at the time of lysis. METHODS Polypropylene meshes (5 x 3.5 cm) were implanted on the parietal peritoneum of New Zealand white rabbits. Some animals were killed 3, 7, 14, and 90 days postimplantation to obtain adhesion tissue. Adhesion formation/reformation was monitored by sequential laparoscopy in other animals kept for 90 days and in a separate experimental group subjected to adhesiolysis at 3 days postimplantation. Immune and inflammatory response markers were determined by immunohistochemical, Western blotting, and real-time reverse transcriptase polymerase chain reaction procedures in adhesion tissue; areas occupied by adhesions were quantified in meshes. RESULTS In animals undergoing adhesiolysis, mesh areas covered by adhesions were significantly decreased at each follow-up time and affected areas became mesothelialized. Increased transforming growth factor (TGF)-beta1 expression was detected in adhesions at 3 days. Greatest TGF-beta1 and vascular endothelial growth factor (VEGF) protein expressions were observed at 7 days, whereas genetic overexpression was noted at 14 days. Active inflammatory cells peaked at the 7-day time point. CONCLUSION Adhesions formed at 3 days; at this critical time, an adhesiolysis was effective in preventing reformation of future adhesions. TGF-beta1 gene and protein expression were increased in 3-day adhesions with respect to the omentum. Levels of active TGF-beta1 and VEGF were increased at 7 days, along with the inflammatory response at this time point related to tissue remodeling, which led to stabilization of adhesions.

Characterizing omental adhesions by culturing cells isolated from a novel in vivo adhesion model.

Although it has been established that postoperative adhesions in the peritoneal cavity are the consequence of injury to the peritoneum, there is much controversy over the nature of the cells giving rise to this neotissue. Here, we establish a novel adhesiogenic model in the rabbit to analyze the phenotype and proliferation in vitro of cells comprising adhesion tissue seven days postsurgery. Adhesion‐free omentum tissue was used as control. Cells derived from adhesions and from the control omentum were subcultured and characterized through immunofluorescence and Western blotting procedures to determine markers of cell differentiation and pluripotential, and viability and proliferation assays. Our findings indicate the existence of a mesenchymal population in the omentum revealed by markers of pluripotent cells with high angiogenic capacity. This population seems to be responsible for the adhesions formed in response to mesothelial damage. Depending on the local environment, mesenchymal cells are capable of in vivo differentiation towards at least two different cell phenotypes rendering two types of adhesions with clearly differentiated characteristics. One type of adhesion shows a highly vascularized adipose morphology containing cells differentiating into a vascular lineage. The other adhesions are fibrous with large amounts of collagen and comprised mainly of myofibroblasts conferring less compliance to this tissue.

Involvement of transforming growth factor-β3 and betaglycan in the cytoarchitecture of postoperative omental adhesions.

BACKGROUND Adhesions commonly appear in patients after abdominal surgery, with considerable individual variation in adhesion composition and severity of the repair process. Here, we address the influence of transforming growth factor (TGF)-β3 and betaglycan in this response, in relation to TGF-β1, in an adhesiogenic rabbit model. MATERIALS AND METHODS Omental adhesions were recovered 3, 7, 14, and 90 d after the implantation of a polypropylene mesh on the parietal peritoneum in New Zealand White rabbits. Omentum from nonoperated animals served as control. Tissue specimens were examined for TGF-β3 and TGF-β1 (Western blotting, reverse transcription-polymerase chain reaction), and TGF-β1:TGF-β3 messenger RNA and protein expression ratios were analyzed. Immunohistochemical detection of TGF-β3 and betaglycan was performed. RESULTS Injury to the omentum led to mobilization of TGF-β3 and betaglycan-expressing cells from milky spots. Fibrous zones in adhesions were simultaneous to the presence of TGF-β1 and the membrane-bound form of betaglycan (7-d adhesions), whereas soluble betaglycan appeared in TGF-β1-positive areas showing limited fibrosis (3-d adhesions). The elevated expression of TGF-β3 concurrent with the presence of membrane-bound form of betaglycan was observed in zones of adipose regeneration (14-d adhesions), whereas zones of fibrous consistency were negative for TGF-β3. CONCLUSIONS Milky spots on the omentum contain inflammatory/immune cells positive for TGF-β3, TGF-β1, and betaglycan, playing a role in the damaged omentum repair. Our observations support the contribution of TGF-β3 to tissue repair through adipose tissue regeneration and the profibrotic role of TGF-β1 and suggest that these effects on the local wound repair response could be driven by the expression of betaglycan in its soluble or membrane-bound form.