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[Experimental study of adhesion of gastric cancer cells to the peritoneum].

OBJECTIVE: To identify factors in favor of tumor cell adherence to the peritoneum. METHODS: Pieces of peritoneum from BALB/c mice were collected and incubated in vitro in the presence of human gastric cancer MGC-803 cells. Cells adhered to the peritoneum was assessed by MTT staining. The result was expressed as the percentage of tumor cells adhered (AD). RESULTS: If the peritondum had been previously injured, either by scraping or exposure to air, AD was significantly more than that of the control (P < 0.01). AD was significantly increased as compared to the control when the peritoneum was cultured in the presence of white cells, platelets, plasma or CEA. CONCLUSION: Adhesion of cancer cells to the peritoneum may be facilitated by peritoneal injury, and the presence of normal blood constituents. Oncofetal antigen with adhesive property such as CEA also helps tumor cells adhere to the peritoneum. The likelihood of post-operative peritoneal metastasis can be minimized if care has been taken to protect the peritoneum from injury and to thoroughly wash the peritoneal cavity before the abdomen is closed.

Animals↗

Concomitant increase in neutrophil adhesion to inflammatory peritoneum and remote organs during peritonitis.

BACKGROUND: Neutrophils contribute to the host defense mechanism, but they can cause remote organ injury in peritonitis. The purpose of this study was to examine neutrophil adhesion to the peritoneum and remote organs simultaneously in peritonitis using a fluorescence microscopic method. STUDY DESIGN: Experiment 1: Sprague-Dawley rats (n = 16) were injected intraperitoneally (ip) with saline solution or 10(5), 10(7), or 10(9) Escherichia coli. Five hours after challenge, 1 x 10(6) fluorescein-labeled neutrophils were infused. Two minutes after neutrophil injection, five peritoneal samples (the greater omentum, mesentery, parietal peritoneum, colon, and ileum), both lungs, the liver, and the right kidney were harvested for counting of labeled neutrophils under epifluorescent microscopy. Lung myeloperoxidase (MPO) activity was also determined. Experiment 2: Rats (n = 23) were given 10(9) E. coli ip. Before challenge (0 h) or at 1, 5, or 10 h after challenge, labeled neutrophils were infused. Then, the labeled neutrophil numbers in organs and lung MPO activities were assessed as described for Experiment 1. Hemodynamic and arterial blood gas data were also obtained in another set of rats before and at 1, 5, 8 and 10 h after 10(9) E. coli ip challenge. RESULTS: Experiment 1: The labeled neutrophil numbers in the peritoneum, lungs, and kidney showed significant positive correlations with the injected bacterial numbers. Lung MPO also positively correlated with E. coli number and labeled neutrophil number in the lungs. Experiment 2: Labeled neutrophil numbers in the peritoneum and kidney peaked at 5 h. The pulmonary labeled neutrophil number rose, reaching a plateau at 5 h. No remarkable change was observed in the hepatic labeled neutrophil number. There was a positive correlation between lung MPO activity and pulmonary labeled neutrophil number. Hemodynamic and blood gas data reflected a hyperdynamic state. CONCLUSIONS: Concomitant dose-dependent increases in neutrophil adhesion in the peritoneum, lungs, and kidney were observed in this peritonitis model. Increased neutrophil adhesion was transient in the peritoneum and kidney but persistent in the lungs. Strategies modulating neutrophil adhesion in organs are anticipated to be useful for the treatment of peritonitis.

Animals↗

Morphology of the rat peritoneum after carbon dioxide and helium pneumoperitoneum: a scanning electron microscopic study.

BACKGROUND: Laparoscopic surgery for patients with cancer has been debated because of the susceptibility that laparoscopic incisions have shown for metastatic tumor growth. Structural damage of the mesothelial layer attributable to the pneumoperitoneum may facilitate intraabdominal tumor cell adhesion and growth. The influence of carbon dioxide (CO(2)) and helium pneumoperitoneum on the morphology of the peritoneum was examined. METHODS: A total of 50 rats received colon carcinoma (DHB/TRb) cells intraperitoneally and CO(2) (n = 25) or helium (n = 25) pneumoperitoneum at 15 mmHg for 15 min. After different periods (2, 12, 24, 48, and 96 h), the rats were killed, and the peritoneum was examined by scanning electron microscopy. Control animals (n = 5) were without pneumoperitoneum. RESULTS: The control animals and most of the rats with pneumoperitoneum showed no peritoneal alterations. In four animals of each group, inflammatory alterations of the peritoneum such as bulging and retraction of mesothelial cells were observed at different time points. Tumor cells adherent to the peritoneum were found in a total of six animals. Peritoneal carcinomatosis, tumor nodules, or infiltration of the peritoneum by tumor cells was not observed. CONCLUSIONS: The study demonstrated that the morphologic integrity of the rat peritoneum is not disturbed when CO(2) or helium is used for insufflation combined with the intraperitoneal injection of carcinoma cells. Pneumoperitoneum therefore probably is not the condition causing peritoneal changes that favor intraperitoneal tumor growth.

Animals↗

Whole explants of peritoneum and endometrium: a novel model of the early endometriosis lesion.

OBJECTIVE: To determine whether whole fragments of endometrium can adhere to peritoneum with intact mesothelium. DESIGN: Tissue culture and immunohistochemical study. SETTING: University medical center. PATIENT(S): Reproductive-age women undergoing surgery for benign conditions. INTERVENTION(S): Explants of human peritoneum from the anterior abdominal wall and the posterior surface of the uterus were cultured with whole fragments of mechanically dispersed endometrium. MAIN OUTCOME MEASURE(S): Adhesion of endometrial fragments to the surface of the peritoneum was evaluated. Adherent endometrium was identified with the use of the dissecting microscope and by the performance of serial sections of the peritoneum explants. Immunohistochemical staining of the mesothelium with antibodies to cytokeratin was used to ensure an intact layer of mesothelium beneath the endometrial implants. Transmission electron microscopy also was used to evaluate this adhesion process. RESULT(S): Endometrium was identified attached to the surface of the peritoneum. Most of the implants did not have identifiable mesothelium beneath them, but most had intact mesothelium running up to the point of attachment. Approximately 10% of the endometrial implants had intact mesothelium at the site of attachment. Endometrial stromal cells, and not epithelium, attached to the mesothelium. CONCLUSION(S): Endometrium can attach to the mesothelial surface of the peritoneum. Endometrial stromal cells are involved in this attachment. Invasion through the mesothelium seems to occur rapidly.

Adult↗

[Lymphocele and complications after pelvic/para-aortic lymph node excision in relation to closure of the peritoneum].

From January 1988 to December 1991, 343 women were operated for genital carcinoma at the 1st Gynaecological Department of the University Munich. Additionally to the corresponding cancer operations, pelvic or pelvic and paraaortic lymphadenectomy was performed. In a postoperative systematic and prospective study with ultrasound, we detected a high incidence of 48% of postoperative lymphocysts. Only a few were symptomatic and even less required therapy. In this study we were especially interested in the influence of closure of the pelvic peritoneum on the incidence of lymphocysts. Thus, we inaugurated a prospective study in 1991 with 49 patients with an open peritoneum and compared these results to 294 women who had standard closure of the peritoneum. There was a significantly higher rate of lymphocysts in patients with peritoneal closure (45.2%) in comparison to 20.4% lymphocysts in patients without peritoneal closure. In the study group (open peritoneum) 6.1% of the patients with lymphocysts showed symptoms, only 4.1% needed therapy. In the control group (closed peritoneum), 14.6% of the patients with lymphocysts were symptomatic and 8.2% needed therapy. These differences are not statistically significant. Other complications such as haemorrhage, fever or ileus were comparable in both groups. Our results do not supply sufficient proof, that leaving the peritoneum open significantly reduces symptomatic lymphocysts.

Adult↗

Migration of antigen presenting cells from periphery to the peritoneum during an inflammatory response: role of chemokines and cytokines.

We demonstrate the migration of antigen presenting cells (APCs), macrophages, and dendritic cells from the subcutaneous site to the peritoneum after they have picked up the antigen, using cell tracking dye. The migration of the APCs is more universal as it was also observed after injection of MethA tumor, DH-5alpha cells, and leishmania parasites, in addition to AK-5 tumor cells. Cellular migration is mediated by several chemokines and cytokines that also induce heavy influx of immune cells into the peritoneum. MIP-3beta secreted by the mesothelial cells is involved in the cellular influx into the peritoneum, whereas IL-12 and IFN-gamma produced by the APCs induced activation of immune cells in the peritoneum. Our results suggest an antigen presentation function for the APCs in the peritoneum as studied by lymphoproliferation assays. These studies indicate antigen presentation function of the activated migratory APCs from the distant subcutaneous site to the peritoneum, suggesting it acts as an important lymphoid organ involved in the enhancement of effector cell function.

Antigens, Neoplasm↗

Secretion of IL-6, monocyte chemoattractant protein-1, macrophage inflammatory protein-1alpha, and TNFalpha by cultured intact human peritoneum.

The peritoneum is an important site of host defence. The mesothelial cells, lining the peritoneum, and the fibroblasts found in the layers below are potent sources of a variety of mediators. Furthermore, granulocytes, mast cells, and macrophages, either resident or attracted by inflammatory processes, are interspersed within the tissue. We investigated the production of mediators by samples of fresh human peritoneum. The method described here has the advantage that the cellular composition of the human peritoneum remains intact. Samples of peritoneum were excised at the beginning of elective abdominal operations in infection-free patients. The tissue was placed across the wells of a microtitre plate, fixed in place by the plate cover and incubated with culture medium with or without lipopolysaccharide (LPS) for up to 5 h. The accumulation of IL-6, monocyte chemoattractant protein-1 (MCP-1), macrophage inflammatory protein-1alpha (MIP-1alpha) and TNFalpha in culture supernatants was measured by ELISA. Production of MCP-1 and IL-6 occurred spontaneously during incubation and was enhanced by as much as 4-fold in the presence of different concentrations of LPS (0. 5-500 ng/ml) in a dose-dependent manner. MIP-1alpha and TNFalpha were detected in culture supernatants of LPS-stimulated samples with concentrations about 8 times as high as those of samples cultured with no such stimulus. The addition of IL-1beta resulted in an increase in the release of IL-6 and MCP-1, similar to that observed with LPS stimulation, but failed to increase the production of TNFalpha. MIP-1alpha production was only marginally enhanced by IL-1beta. In conclusion, our experimental system is suitable for the investigation of chemokine and cytokine production by the human peritoneum, with the aim of assessing aspects of local immunocompetence.

Cell Survival↗

Construction of a transplantable tissue-engineered artificial peritoneum.

BACKGROUND: Peritoneal defects lead to serious postoperative problems. Thus the development of physiological material to cover peritoneal defects is very desirable. AIM: The aim of this study was to develop a transplantable artificial peritoneum. METHOD: The artificial peritoneum consisted of collagen gel, fibroblasts, and mesothelial cells, and histological features were analyzed. The artificial peritoneum at the site of a peritoneal defect in the rat was transplanted to the abdominal wall. RESULTS: Histological examination revealed that the artificial peritoneum consisted of a flat mesothelial monolayer upon a stromal matrix. All transplanted artificial peritoneums adapted well to the host and prevented severe adhesion. CONCLUSION: Our artificial peritoneum may be a useful transplantable bioengineered material for repair of surgical peritoneal defects.

Abdominal Wall↗

Expression pattern and regulation of genes differ between fibroblasts of adhesion and normal human peritoneum.

BACKGROUND: Injury to the peritoneum during surgery is followed by a healing process that frequently results in the attachment of adjacent organs by a fibrous mass, referred commonly as adhesions. Because injuries to the peritoneum during surgery are inevitable, it is imperative that we understand the mechanisms of adhesion formation to prevent its occurrence. This requires thorough understanding of the molecular sequence that results in the attachment of injured peritoneum and the development of fibrous tissue. Recent data show that fibroblasts from the injured peritoneum may play a critical role in the formation of adhesion tissues. Therefore, identifying changes in gene expression pattern in the peritoneal fibroblasts during the process may provide clues to the mechanisms by which adhesion develop. METHODS: In this study, we compared expression patterns of larger number of genes in the fibroblasts isolated from adhesion and normal human peritoneum using gene filters. Contributions of TGF-beta1 and hypoxia in the altered expression of specific genes were also examined using a semiquantitative RT-PCR technique. RESULTS: Results show that several genes are differentially expressed between fibroblasts of normal and adhesion peritoneum and that the peritoneal fibroblast may acquire a different phenotype during adhesion formation. Genes that are differentially expressed between normal and adhesion fibroblasts encode molecules involved in cell adhesion, proliferation, differentiation, migration and factors regulating cytokines, transcription, translation and protein/vesicle trafficking. CONCLUSIONS: Our data substantiate that adhesion formation is a multigenic phenomenon and not all changes in gene expression pattern between normal and adhesion fibroblasts are the function of TGF-beta1 and hypoxia that are known to influence adhesion formation. Analysis of the gene expression data in the perspective of known functions of genes connote to additional targets that may be manipulated to inhibit adhesion development.

Cell Hypoxia↗

[Production of chemokines by the human peritoneum].

AIM OF THE STUDY: Peritonitis is characterised by a continued infiltration of the peritoneal cavity with leukocytes, attracted by chemotactic mediators. The aim of this study was to demonstrate the capacity of the human peritoneum to secrete chemokines and to show a therapeutic option by impairing the proinflammatory function of the peritoneum. METHODS: Peritoneum was obtained from 12 consenting patients undergoing abdominal surgery for noninflammatory diseases. After opening the peritoneal cavity a piece of the parietal peritoneum was excised and subsequently incubated with lipopolysaccharide (LPS, 50 ng/ml) +/- interleukin-10 (IL-10, 100 U/ml) for five hours in vitro. The culture supernatants were assayed for concentrations of interleukin-8 (IL-8), monocyte chemoattractant protein-1 (MCP-1) and monocyte inflammatory protein-1 alpha (MIP-1 alpha) by using the ELISA. RESULTS: The cultured peritoneum secreted MCP-1 (mean (SEM): 3416 (659) pg/ml) and IL-8 (2946 (894) pg/ml). The presence of LPS resulted in a fourfold enhancement of this secretion (MCP-1: 13563 (1613), IL-8: 9854 (1305) pg/ml) and led to the production of MIP-1 alpha (1476 (240) pg/ml). The LPS-stimulated production of all of these chemokines was significantly diminished by the presence of IL-10. CONCLUSION: The reaction of the peritoneum to LPS indicates its proinflammatory function in the context of peritonitis caused by gram-negative bacteria. This inflammatory reaction might be diminished by application of IL-10.

Chemokine CCL2↗

Heme oxygenase-1 regulates vascular tone of the peritoneum undergoing peritoneal dialysis.

The present study investigated the role of heme oxygenase-1 (HO-1) in regulating the vascular tone of the peritoneum undergoing peritoneal dialysis. First, we used reverse-transcriptase polymerase chain reaction (RT-PCR) to measure changes in the expression of HO-1 mRNA in the peritoneum of dehydrated Wistar Kyoto (WKY) rats. Second, we used a charge-coupled-device (CCD) camera to measure changes in the diameter and blood flow in the small artery of the peritoneum after intravenous administration of saline, hemin (4-12 mumol/kg), and tin-protoporphyrin (4-12 mumol/kg) in dehydrated WKY rats. In non dehydrated rats (control), no expression of HO-1 mRNA in the peritoneum was seen. In dehydrated and 10% glucose-treated dehydrated rats, HO-1 expression was significantly enhanced as compared with the control rats. Intravenous administration of saline induced no significant changes in the diameter of the small artery in dehydrated WKY rats. Intravenous administration of hemin induced a significant increase in the diameter of the small artery in dehydrated WKY rats. Intravenous administration of tin-protoporphyrin induced a significant reduction in the diameter of the small artery in dehydrated WKY rats. Our results suggest that 10% glucose injection into the peritoneal cavity induces overexpression of HO-1 in the peritoneum of dehydrated rats. In addition, HO-1 regulates peritoneal function by the regulation of vascular tone in the peritoneum.

Animals↗

Change in the transport of uric acid under the influence of pyrazinoic acid: in vitro studies with human and rabbit peritoneum.

Bidirectional transport of uric acid (UA) through human and rabbit peritoneum, and its change under the influence of pyrazinoic acid (PA) was investigated in vitro. Fragments of parietal peritoneum were taken from the region of anterior abdominal wall from rabbits (the animal series) and from patients undergoing abdominal operations (the human series). Under control conditions, higher values of the UA transport were found in the rabbit peritoneum in comparison with the human peritoneum, but in both series it was maintained at a constant level throughout the experiment lasting 120 min. In rabbit as well as in human peritoneum it was found that PA depressed about 50% of the UA transport from the interstitial to the mesothelial side, leaving the opposite direction intact. The results obtained in vitro are in agreement with previous observations in a patient undergoing peritoneal dialysis that showed decreased peritoneal UA excretion evoked by pyrazinamide. The comparative analysis proves the rabbit peritoneum to be a good experimental model for further investigations of peritoneal urate transport in man.

Adult↗

Complete omentectomy and extensive lymphadenectomy with gastrectomy improves the survival of gastric cancer patients with metastases in the adjacent peritoneum.

BACKGROUND/AIMS: The omentum is the site where peritoneal metastases occur most frequently. It has not been shown whether complete resection of the omenta during gastrectomy improves the survival of gastric cancer patients with macroscopic peritoneal metastases. METHODOLOGY: We retrospectively analyzed 126 patients who underwent gastrectomies for gastric cancer with peritoneal metastases but without hematogenous metastases. The 126 patients were stratified according to their grade of peritoneal metastases into three groups: the P1 patients (patients with peritoneal metastases in the adjacent peritoneum but not in the distant peritoneum); the P2 patients (patients with a few peritoneal metastases in the distant peritoneum); and the P3 patients (patients with many metastases in the distant peritoneum). In each group, the survival and clinicopathological characteristics were compared between patients treated by complete resection of the greater omentum and the lesser omentum plus extensive lymphadenectomy during gastrectomy, versus patients treated by incomplete resection of the omenta and non-extensive lymphadenectomy during gastrectomy. RESULTS: Complete omentectomy and extensive lymphadenectomy during gastrectomy improved survival significantly only in the P1 patients. Other clinicopathological characteristics did not differ between them. CONCLUSION: Complete omentectomy and extensive lymphadenectomy is recommended in patients with peritoneal metastases in the adjacent peritoneum but not in the distant peritoneum.

Adenocarcinoma↗

Identification of the cadherin subtypes present in the human peritoneum and endometriotic lesions: potential role for P-cadherin in the development of endometriosis.

Endometriosis is defined as endometrial tissue outside of the uterine cavity. The pathogenesis of this common disease remains poorly understood. However, the implantation and invasion of the viable cells from retrograde menstruation into the peritoneum is a widely accepted theory. To date, the mechanisms by which cell adhesion molecules mediate the development of human endometriosis remain unclear. Cadherins are a family of cell adhesion molecules that mediate cell-cell adhesion in a homophilic manner. In this study, the cadherins present in the peritoneum and endometriotic lesions were identified by RT-PCR using degenerate primers. In addition, differences in the levels of the cadherin mRNA transcripts present in eutopic endometrium and endometriotic lesions of the same patients were then compared by semiquantitative RT-PCR. Multiple cadherins were detected in the peritoneum and endometriotic lesions. Of these, P-cadherin appears to be the predominant cadherin subtype present in the peritoneum. Similarly, P-cadherin mRNA levels in endometriotic lesions were significantly greater than those observed in the corresponding eutopic endometrium. The expression of P-cadherin in both the human peritoneum and endometriotic lesions suggests that this cell adhesion molecule may play a central role in the development of endometriosis by mediating endometrial-peritoneal cell interactions in a homophilic manner.

Adult↗

A study of non-closure of the peritoneum at vaginal hysterectomy.

The aim of this case-controlled study was to determine whether non-closure of the peritoneum is detrimental in vaginal hysterectomy. 233 patients who underwent total vaginal hysterectomy (TVH) or laparoscopically assisted vaginal hysterectomy (LAVH) at the University of Vienna/Austria were analyzed. Cohorts of patients were formed according to their peritonealization status (open, n=117, vs closed peritoneum, n=116) and further stratified according to the type of surgical procedure: simple TVH ( n=115), TVH with concurrent vaginal repair and/or urinary incontinence surgery ( n=91) and LAVH ( n=27). No significant differences could be observed in analyzed surgical outcome (operation time, blood loss and analgesia). Complications (fever, infection, hemorrhage or revision) were similar whether the peritoneum was closed or not. After simple TVH, resumption of bowel function took place earlier in patients with open peritoneum than in those where it had been sutured (1.9 vs 2.4 days, P=0.001). No readmission for prolapse of the vaginal vault was recorded. Non-closure of the peritoneum at vaginal hysterectomy appears to be safe. Omission of peritoneal closure reduces the potential risk of injury and has a beneficial effect on bowel function.

Austria↗

Effect of carbon dioxide pneumoperitoneum on tissue blood flow in the peritoneum, rectus abdominis, and diaphragm muscles.

BACKGROUND: Changes in local blood flow may play a role in the pathogenesis of port-site metastasis. This study aimed to investigate the effect of pneumoperitoneum induced by carbon dioxide (CO2) on the blood flow in the peritoneum and abdominal wall muscle layers, which are target structures for this phenomenon. METHODS: The study was performed on domestic farm swine of both genders weighing 20 to 25 kg. Intraabdominal pressures (IAP) of 0, 5, and 10 mmHg were produced by either CO2 ( n = 9) or helium (He) ( n = 6) insufflations. The colored microsphere technique was used to measure blood flow distributions in the parietal peritoneum, rectus abdominis, and diaphragm muscles. RESULTS: Insufflation of CO2 was associated with a threefold increase in blood flow of the parietal peritoneum at both 5 and 10 mmHg IAP ( p < 0.001 for both pressure levels). In contrast, insufflation of He caused a significant decrease in blood flow in the parietal peritoneum at both 5 and 10 mmHg ( p < 0.05). In the rectus abdominis and diaphragm muscles, blood flow remained unchanged after insufflation of CO2 at both 5 and 10 mmHg IAP. However, after insufflation of He, there was a substantial decrease in blood flow both in the rectus abdominis and diaphragm muscles at both 5 mmHg ( p < 0.01 and p < 0.05, respectively) and 10 mmHg ( p < 0.001 and p < 0.01, respectively). CONCLUSIONS: Despite high intraabdominal pressure, tissues surrounding the abdominal cavity, particularly the peritoneum, respond to insufflation of CO2 with increased blood flow, which may favor the growth of tumor cells.

Animals↗

Closure vs. nonclosure of the peritoneum at cesarean delivery.

OBJECTIVE: To compare the frequency and severity of celiac adhesions following cesarean sections performed with and without closure of the parietal and visceral peritoneum. METHODS: A retrospective cohort study involved 612 women who underwent their second lower-segment transverse cesarean section. RESULTS: Nonclosure of the parietal peritoneum at the time of the first cesarean section was associated with significantly more visceral adhesions than closure (16.2% vs. 8.1%; P=0.003), and closure of the visceral peritoneum had a similar effect (16.1% vs. 6.7%; P=0.02). However, the difference in rates of severe adhesions was not statistically significant (12.9% vs. 17.6%; P=0.12). After controlling for confounding variables, it was found that closure of the parietal peritoneum reduced the rate of visceral adhesions 2.7-fold. Trial of labor before and fever after surgery increased the risk of severe adhesions 6.1-fold and 5.6-fold, respectively. CONCLUSION: Nonclosure of the peritoneum at primary cesarean section is associated with a significantly increased risk of visceral adhesions.

Cesarean Section↗

Closure or nonclosure of the visceral peritoneum at cesarean delivery.

OBJECTIVE: Our purpose was to determine whether nonclosure of the visceral peritoneum at low transverse cesarean delivery has advantages over suture peritonization with regard to postoperative morbidity. STUDY DESIGN: A prospective randomized trial of 549 women undergoing cesarean section was carried out; 262 were randomized to nonclosure and 287 to closure of the visceral peritoneum. Perioperative, intraoperative, and postoperative management decisions were made without reference to treatment groups. Statistical analysis compared intraoperative and postoperative outcome between the two groups. RESULTS: Operating and anesthesia times were significantly shorter in patients receiving nonclosure. The incidence of febrile morbidity and cystitis and the need for antibiotics and narcotics were all significantly greater when the peritoneum was closed. Hospital stay was significantly shorter after nonclosure. CONCLUSION: Nonclosure of the visceral peritoneum is associated with lower febrile and infectious morbidity. Routine closure of the visceral peritoneum should be abandoned at cesarean delivery.

Adolescent↗