Influence of 4% icodextrin solution on peritoneal tissue response and adhesion formation.

BACKGROUND: Postoperative peritoneal adhesion formation following abdominal surgery remains a relevant surgical problem. The application of soluble physico-chemical barriers like 4% icodextrin is one approach to protect the peritoneal surface from getting linked to adhesive scar. The aim of this study was to investigate the influence of 4% icodextrin on peritoneal tissue response both of visceral and parietal peritoneum, adhesion formation and wound healing. METHODS: 40 rats were divided into two groups. After creation of an intraabdominal defect, either 4% icodextrin (Adept®) or sodium chloride was applied. Animals were sacrificed after 7 and 21 days. Adhesions were scored by an adhesion score. Furthermore, immunohistochemical investigations were conducted to determine the discrete influence of icodextrin on the parietal and visceral peritoneal tissue responses (CD68+ macrophages, CD3+ T-lymphocytes, vimentin for mesenchymal cells, HBME-1 for mesothelial cells, and as components of wound healing COX-2, C-myc, catenin). RESULTS: Postoperative peritoneal adhesions were predominantly present in the sodium chloride group as compared to the icodextrin group (14/19 (74%) vs. 9/19 (47%); p = 0.048). The adhesion score however did not reveal any significant differences, (p = 0.614). Furthermore, the expression of vimentin in both the parietal and visceral peritoneum after 21 days was significantly lower in the icodextrin group than in the sodium chloride group (p = 0.038 and p = 0.028, respectively). No significant differences were observed for macrophages, lymphocytes, reperitonealisation or the expression of COX-2, C-myc or Catenin. CONCLUSIONS: The intraperitoneal application of 4% icodextrin reduces adhesion formation in comparison to sodium chloride. 4% icodextrin solution reduces the inflammatory and mesenchymal infiltrate in the wounded area, thus improving the ratio of mesothel cells to mesenchymal infiltrate. As demonstrated, icodextrin is able to ameliorate the local tissue response. Further experimental studies would be done to elaborate the impact on the early response of the adaptive immune system, which may then trigger the subsequent wound healing and tissue repair.

Balancing zinc deficiency leads to an improved healing of colon anastomosis in rats.

BACKGROUND: In colorectal surgery, anastomotic leakage is a relevant complication. The aim of this study was to investigate whether intraperitoneally (i.p.) administered zinc improves the healing of colon anastomosis in rats. MATERIALS AND METHODS: Male Wistar rats (66) received zinc-deficient diet for 21 days. To determine the effective dose of zinc which is necessary to compensate this deficiency, preliminary analysis in 30 rats were performed. In these rats, analysis by atom-absorption spectrophotometry revealed a dose of 1.0 mg zinc aspartate/kg body weight to be the compensatory dosage. In the remaining zinc-deficient rats (n= 36), a transverse colonic anastomosis was performed. Eighteen rats received either a zinc supplementation i.p. or 0.9% NaCl i.p. (n = 18; control group). On postoperative days 3, 5, and 14, the surface of the mucosal villi, expression of MMP 2, MMP 8, MMP 13, TIMP 1, as well as the collagen types I/III ratio were analyzed. RESULTS: Protein expression of MMP 2 and MMP 8 was significantly higher in the anastomosis of the zinc group on day 3 and on day 5. The collagen types I/III ratio was significantly increased in the zinc group on days 5 and 14. CONCLUSION: Balancing zinc deficiency benefits wound healing of colonic anastomosis qualitatively due to an increased collagen type I/III ratio. Surprisingly, these zinc supplements, however, increased the expression of MMP 2 and MMP 8 that are supposed to impair wound healing in case of an over-expression. Thus, further investigations are needed to elucidate the influence of zinc supplementation on regulation of MMPs.

Morphology, quality, and composition in mature human peritoneal adhesions.

BACKGROUND AND AIM: Peritoneal adhesions are caused by intra-abdominal surgery and can lead to relevant complications. Adhesions are supposed to consist of avascular scar tissue. The aim of the present study was to analyze whether mature postsurgical adhesions even after years still reveal a dynamic remodeling process. MATERIALS AND METHODS: In a prospective analysis, we investigated tissue specimen of peritoneal adhesions in 40 patients after abdominal surgery. Expression of five parameters representing wound healing and remodeling were examined (MMP-2, Ki-67, apoptosis, collagen/protein ratio, and collagen type I/III ratio). RESULTS: Gender, age, and the number of previous operations had no impact on the parameters measured. Adhesion specimens were cell rich, containing mononuclear round cells, fibroblasts, adipose cells, and vascular endothelial cells. There was a positive expression of MMP-2 and apoptosis, whereas Ki-67 was marginal irrespective of adhesion maturity or quality. Adhesions classified as dense showed a significant increase in total collagen (118.2 +/- 4.9 microg/mg) and collagen type I/III ratios (3.9 +/- 0.2), whereas there were no significant differences regarding the adhesion maturity. CONCLUSION: The distinct composition of cellular components as well as of extracellular matrix proteins may reflect an interactive cross-talk between adhesion- and stroma-derived cells even in mature adhesions. Our findings support the hypothesis that the disabilities of appropriate repair of the peritoneal surface leading to persistent adhesions are a consequence of a permanent process of disturbed remodeling.

Collagen in colon disease.

The pathophysiology of wound healing in the bowel wall suggests that collagen and matrix metalloproteinases (MMPs) have an important role in the changes of the bowel wall seen in several colonic diseases. Several recent studies suggest that disturbances of the collagen texture and the extracellular matrix (ECM) metabolism are major factors leading to the onset of diverticular disease. Changes of the ECM also play a role in the development of inflammatory bowel diseases. Regarding the permanent remodeling of the bowel wall, any imbalance of the ECM could support the onset of chronic inflammation and the development of fistula formation, such as that seen in patients with Crohn's disease. Disturbances of the ECM play a role in the pathogenesis of anastomotic leakage after large bowel surgery and suggest the presence of a genetically defined risk population with disturbed wound healing mechanisms. This concept could explain the well known situation where an anastomotic breakdown is observed, despite the absence of other known risk factors and after a technically correct anastomosis.

Changes of the extracellular matrix as a risk factor for anastomotic leakage after large bowel surgery.

BACKGROUND: Despite improved surgical techniques, anastomotic leakage remains as a serious complication in colorectal surgery, producing increased morbidity and mortality. This prospective study was initiated to test the hypothesis that preexisting disorders in the extracellular matrix (ECM) may be a factor influencing the onset of anastomotic wound healing complications. METHODS: In this prospective study of 119 patients with colorectal anastomoses, 30 clinical parameters with possible influence on anastomotic complications were evaluated. From all patients, samples of macroscopically intact colonic tissue were obtained at the index operation. Crosspolarization microscopy was performed to analyze the collagen type I/III ratio, and immunohistochemical studies were done to determine the expression of matrix metalloproteinase (MMP) 1, 2, 9, and 13. The patients with uncomplicated postoperative healing were compared with those developing anastomotic leakage. RESULTS: Patients with impaired anastomotic healing exhibited a significantly lower collagen type I/III ratio compared with the controls. Significantly higher expression of MMP-1 and MMP-2 in the mucosal layers and of MMP-2 and MMP-9 in the submucosal layers was found in the normal bowel wall of the leakage group. These findings were statistically independent from the clinical parameters. CONCLUSION: The present study confirms the hypothesis that disturbances of the ECM play a role in the pathogenesis of anastomotic leakage after large bowel surgery.

Deleterious effect of zinc in a pig model of acute endotoxemia.

By antiapoptotic effects and the induction of the heat-shock response, zinc is supposed to be a promising means of therapy during sepsis. As zinc also stimulates the expression of proinflammatory cytokines, its administration during the proinflammatory stage of septic shock might have adverse effects. Therefore, this study analyzes the influence of zinc during the acute phase of endotoxemia. In a pig model of acute endotoxemia, animals were divided into two groups: group I (n = 5) with saline treatment and group II (n = 5) with zinc treatment in close succession to lipopolysaccharide (LPS) (1.0 mu g/kg Escherichia coli endotoxin WO 111:B4). Hemodynamic and pulmonary monitoring was followed by combined reflection photometry, pulse oxymetry, blood gas samples, and temperature measurement. Plasma concentrations of tumor necrosis factor (TNF)alpha and interleukin (IL)-6 were analyzed by enzyme-linked immunosorbent assay (ELISA). Morphology included the weight of the lungs, the width of the alveolar septae, and the paracentral necrosis rate of the liver. After LPS infusion, group II (zinc) showed an impressive and significant deterioration of all pulmonary and most of the hemodynamical parameters compared to group I (saline). Levels of TNFalpha and IL-6 measured were significantly higher after zinc treatment. In accordance, we found significant more morphologic damages in group II (zinc). The almost simultaneous infusion of zinc and LPS complementary induced proinflammatory effects with a deleterious outcome. The same potentials characterizing zinc as a promising tool of prophylactic therapy in sepsis seem to ban its use during the acute phase.

Suture-free small bowel anastomoses using collagen fleece covered with fibrin glue in pigs.

BACKGROUND: Several studies investigating anastomotic healing could objectify that the regularly used suture material leads to an impairment of wound healing due to ischemia at the anastomotic line. This study was initiated to test a hypothesis that a reduction of suture material leading to suture-free glued intestinal anastomoses is feasible and enables an improved wound healing. MATERIALS AND METHODS: Three different types of anastomoses were carried out at the small bowel of 16 pigs. Standard hand-sewn anastomoses, anastomoses with loose-fitting skin staples, and suture-free glued anastomoses using a fibrin covered collagen fleece. When the animals were killed, both gross inspection of the parietes, bursting pressure and tissue for histological study became the basis for evaluation. Analyses were also made regarding the collagen I/III ratio and the expression of MMP 1 and 13. RESULTS: Four leakages at the stapled, one at the sutured, and one at the glued anastomoses occurred. All other anastomoses healed without complications. The bursting pressure did not differ significantly between the groups. The macroscopic inspection and the microscopic examination both showed an improved healing pattern for the material reduced techniques without onset of a deep ulcer at the anastomotic line as seen at the conventional sutured anastomoses. These findings were supported by the immunohistochemical studies. CONCLUSIONS: These observations suggest that a suture-free bowel anastomoses using collagen fleece covered with fibrin glue is technically feasible. Obviously, a reduction of foreign body material at the anastomotic line avoids unnecessary ischemia and thus supports a physiological improved wound healing process.

Macrophage and T-lymphocyte infiltrates in human peritoneal adhesions indicate a chronic inflammatory disease.

BACKGROUND: Peritoneal adhesions are common and lead to significant clinical morbidity and mortality. Besides various individual factors, notably the inflammatory response to peritoneal defects affects adhesion formation. The aim of this study was to investigate whether there is inflammatory activity even in persistent adhesions. METHODS: Tissue specimens of 40 patients suffering peritoneal adhesions were prospectively collected. Expression profiles of seven parameters as potential mediators in cellular immune response, cell differentiation, and wound healing were analyzed (macrophages [CD68], B-lymphocytes [CD20] and T-lymphocytes [CD45], cyclo-oxygenase-2 [COX-2], Notch-3, beta-catenin, and c-myc). Furthermore, clinical details and co-morbidities were recorded. RESULTS: Infiltrates of mononuclear round cells were found in all adhesion specimens irrespective of the maturity. Immunohistochemical analysis identified mononuclear round cells as macrophages (CD68) and as T-lymphocytes (CD45). Expression of CD68 was significantly elevated in adhesion tissue with an age<12 months. Positive expression of CD45, COX-2, Notch-3, beta-catenin, and c-myc, was observed even in long-lasting adhesions. CONCLUSIONS: A persistent inflammatory process has to be considered, even in mature adhesions. Macrophages may play an important role in triggering adhesions, whereas T-cells and the Notch-3/beta-catenin complex signaling pathway may play a crucial role in maintaining adhesions. These findings indicate that adhesions should not be regarded simply as an adynamic result of an operative trauma but rather may be grasped as a permanent process in remodeled tissue.