Ascites-induced compression alters the peritoneal microenvironment and promotes metastatic success in ovarian cancer.

The majority of women with recurrent ovarian cancer (OvCa) develop malignant ascites with volumes that can reach > 2 L. The resulting elevation in intraperitoneal pressure (IPP), from normal values of 5 mmHg to as high as 22 mmHg, causes striking changes in the loading environment in the peritoneal cavity. The effect of ascites-induced changes in IPP on OvCa progression is largely unknown. Herein we model the functional consequences of ascites-induced compression on ovarian tumor cells and components of the peritoneal microenvironment using a panel of in vitro, ex vivo and in vivo assays. Results show that OvCa cell adhesion to the peritoneum was increased under compression. Moreover, compressive loads stimulated remodeling of peritoneal mesothelial cell surface ultrastructure via induction of tunneling nanotubes (TNT). TNT-mediated interaction between peritoneal mesothelial cells and OvCa cells was enhanced under compression and was accompanied by transport of mitochondria from mesothelial cells to OvCa cells. Additionally, peritoneal collagen fibers adopted a more linear anisotropic alignment under compression, a collagen signature commonly correlated with enhanced invasion in solid tumors. Collectively, these findings elucidate a new role for ascites-induced compression in promoting metastatic OvCa progression.

Fibrin Deposit on the Peritoneal Surface Serves as a Niche for Cancer Expansion in Carcinomatosis Patients.

Peritoneal metastasis (PM) is a very serious complication of gastrointestinal and gynecological malignancies which is poorly documented. Modified mesothelial cell layer and their microenvironments can favor fibrin deposition for cancer cell adhesion. Scanning and transmission electron microscopy of peritoneal surface and cancer cell clusters from cancer patients was done. Ascites and its impact on mesothelial cells were assessed by cytokine array. Neprilysin, matrix metalloprotease, epithelial mesenchymal transition (EMT) related molecules (E-cadherin, Snail, Slug, Twist, Vimentin and Fibronectin), tissues factor (TF), endothelial protein C receptors (EPCR) were quantified by q-PCR. Fibrin in the simples were stained using anti fibrin F1E1 antibody. Migration ability was assessed by scratch assay. Cell viability and neprilysin activity were analyzed by bioluminescence. Cancer cells-fibrin interaction was investigated by scanning electron microscopy (SEM) and microcinematography (MCG). Mesothelial cells change their morphology after incubation with carcinomatosis peritoneal fluids in vitro. EMT associated with upregulation of neprilysin, matrix metalloproteinase-2, tissue factor and cytokines secretions such as interleukin-6, and 8, hepatocyte growth factor and granulocyte chemotactic protein-2 mRNA and protein were observed. EPCR expression as a natural anticoagulant was decreased. In parallel, carcinomatosis cell clusters extracted from peritoneal fluids were found to be associated with fibrin. Kinetic analysis of cancer cell-fibrin interaction in vitro studied by MCG showed that fiber filaments generated from clots inhibited cancer cell adhesion on fibrin clots. These results indicated that fibrin deposit on the peritoneal surface serve as niches for cancer expansion in carcinomatosis patients.

The mechanical microenvironment regulates ovarian cancer cell morphology, migration, and spheroid disaggregation.

There is growing appreciation of the importance of the mechanical properties of the tumor microenvironment on disease progression. However, the role of extracellular matrix (ECM) stiffness and cellular mechanotransduction in epithelial ovarian cancer (EOC) is largely unknown. Here, we investigated the effect of substrate rigidity on various aspects of SKOV3 human EOC cell morphology and migration. Young's modulus values of normal mouse peritoneum, a principal target tissue for EOC metastasis, were determined by atomic force microscopy (AFM) and hydrogels were fabricated to mimic these values. We find that cell spreading, focal adhesion formation, myosin light chain phosphorylation, and cellular traction forces all increase on stiffer matrices. Substrate rigidity also positively regulates random cell migration and, importantly, directional increases in matrix tension promote SKOV3 cell durotaxis. Matrix rigidity also promotes nuclear translocation of YAP1, an oncogenic transcription factor associated with aggressive metastatic EOC. Furthermore, disaggregation of multicellular EOC spheroids, a behavior associated with dissemination and metastasis, is enhanced by matrix stiffness through a mechanotransduction pathway involving ROCK, actomyosin contractility, and FAK. Finally, this pattern of mechanosensitivity is maintained in highly metastatic SKOV3ip.1 cells. These results establish that the mechanical properties of the tumor microenvironment may play a role in EOC metastasis.

The Peritoneum: Health, Disease, and Perspectives regarding Tissue Engineering and Cell Therapies.

There are several pathologies associated with the peritoneum, such as mesothelioma and peritonitis. Moreover, the peritoneum is widely used in ultrafiltration procedures, i.e., peritoneal dialysis, presenting advantages over hemodialysis. On the other hand, ultrafiltration failure may lead to dialysis-induced fibrosis and hypervolemia. Therefore, the pathophysiological study of this tissue is of extreme biomedical importance. Studies investigating the biology of the cells dwelling in the peritoneum wall provide evidence of their plasticity and progenitor features. For instance, both mesothelial and submesothelial cells present characteristics similar to mesenchymal stem cells, including osteogenic and adipogenic differentiation potential, support of extramedullary hematopoiesis, modulation of inflammatory responses, and regulation of tumor progression. Indeed, the participation of each cell type in peritoneal pathological and physiological phenomena is still under debate, especially regarding a possible differentiation pathway connecting these peritoneal cells. The primary aim of this review is to raise this discussion. In order to do so, we will firstly provide an overview of the peritoneum anatomy, histology, and ontology, and finally we will address how a better understanding of peritoneal cell biology may contribute to future cell therapy and tissue engineering approaches.

Post-translational modification of the membrane type 1 matrix metalloproteinase (MT1-MMP) cytoplasmic tail impacts ovarian cancer multicellular aggregate dynamics.

Membrane type 1 matrix metalloproteinase (MT1-MMP, MMP-14) is a transmembrane collagenase highly expressed in metastatic ovarian cancer and correlates with poor survival. Accumulating evidence shows that the cytoplasmic tail of MT1-MMP is subjected to phosphorylation, and this post-translational modification regulates enzymatic activity at the cell surface. To investigate the potential role of MT1-MMP cytoplasmic residue Thr567 phosphorylation in regulation of metastasis-associated behaviors, ovarian cancer cells that express low endogenous levels of MT1-MMP were engineered to express wild-type MT1-MMP, a phosphomimetic mutant (T567E), or a phosphodeficient mutant (T567A). Results show that Thr567 modulation influences behavior of both individual cells and multicellular aggregates (MCAs). The acquisition of either wild-type or mutant MT1-MMP expression results in altered cohesion of epithelial sheets and the formation of more compact MCAs relative to parental cells. Cells expressing MT1-MMP-T567E phosphomimetic mutants exhibit enhanced cell migration. Furthermore, MCAs formed from MT1-MMP-T567E-expressing cells adhere avidly to both intact ex vivo peritoneal explants and three-dimensional collagen gels. Interaction of these MCAs with peritoneal mesothelium disrupts mesothelial integrity, exposing the submesothelial collagen matrix on which MT1-MMP-T567E MCAs rapidly disperse. Together, these findings suggest that post-translational regulation of the Thr567 in the MT1-MMP cytoplasmic tail may function as a regulatory mechanism to impact ovarian cancer metastatic success.

Ultrastructural Investigation of Pelvic Peritoneum in Patients With Chronic Pelvic Pain and Subtle Endometriosis in Association With Chromoendoscopy.

STUDY OBJECTIVE: To evaluate the pelvic peritoneum under chromoendoscopy by scanning electron microscopy (SEM) as well as light microscopy with hematoxylin and eosin staining and immunohistochemistry (IHC) assays in patients with chronic pelvic pain (CPP) associated with subtle endometriosis. DESIGN: Case series study (Canadian Task Force classification II). SETTING: A referral academic community tertiary medical center. PATIENTS: Three women aged 29 to 37 years were referred to the obstetrics and gynecology clinic of the tertiary university hospital with CPP. They were suspicious for endometriosis, were not responding to medical treatments, and had undergone previous pelvic laparoscopy to determine the stage of endometriosis and preparation of peritoneal samples under the guidance of staining with methylene blue in 0.25% dilution. INTERVENTIONS: Comparison of stained and unstained pelvic peritoneal samples after the instillation of 0.25% methylene blue into the pelvic cavity. MEASUREMENTS AND MAIN RESULTS: In 3 patients, laparoscopic examination showed minimal endometriosis. A total of 18 samples (9 stained and 9 unstained) from the 3 patients were prepared for SEM. Ten of the samples (55.6%) showed microstructural peritoneal destruction (7 of 9 stained [77.7%] and 3 of 9 [33.4%] unstained). Eighteen samples (9 stained and 9 unstained) from the 3 patients were also prepared for IHC. Six of these samples (33.3%) were S-100-positive, including 4 of 9 (44.4%) stained samples and 2 of 9 (22.2%) unstained samples. CONCLUSIONS: In general, in the context of CPP and endometriosis, there is no established relationship between the severity of pain and stage of endometriosis. In the pathophysiology of CPP associated with endometriosis, ultrastructural changes can play a significant role. Under methylene blue staining, some destroyed areas were detected, but the stained areas do not necessarily correlate with increased microstructural peritoneal destruction.

Silk fibroin hydrogel as physical barrier for prevention of post hernia adhesion.

BACKGROUND: Adhesion formation remains a major complication following hernia repair surgery. Physical barriers though effective for adhesion prevention in clinical settings are associated with major disadvantages, therefore, needs further investigation. This study evaluates silk fibroin hydrogel as a physical barrier on polypropylene mesh for the prevention of adhesion following ventral hernia repair. STUDY DESIGN: Peritoneal explants were cultured on silk fibroin scaffold to evaluate its support for mesothelial cell growth. Full thickness uniform sized defects were created on the ventral abdominal wall of rabbits, and the defects were covered either with silk hydrogel coated polypropylene mesh or with plain polypropylene mesh as a control. The animals were killed after 1 month, and the adhesion formation was graded; healing response of peritoneum was evaluated by immunohistochemistry with calretinin, collagen staining of peritoneal sections, and expression of PCNA, collagen-I, TNFα, IL6 by real time PCR; and its adverse effect if any was determined. RESULTS: Silk fibroin scaffold showed excellent support for peritoneal cell growth in vitro and the cells expressed calretinin. A remarkable prevention of adhesion formation was observed in the animals implanted with silk hydrogel coated mesh compared to the control group; in these animals peritoneal healing was complete and predominantly by mesothelial cells with minimum fibrotic changes. Expression of inflammatory cytokines decreased compared to control animals, histology of abdominal organs, haematological and blood biochemical parameters remained normal. CONCLUSION: Therefore, silk hydrogel coating of polypropylene mesh can improve peritoneal healing, minimize adhesion formation, is safe and can augment the outcome of hernia surgery.

Application of imaging flow cytometry for characterization of acute inflammation in non-classical animal model systems.

Phagocytes display marked heterogeneity in their capacity to induce and control acute inflammation. This has a significant impact on the effectiveness of antimicrobial immune responses at different tissue sites as well as their predisposition for inflammation-associated pathology. Imaging flow cytometry provides novel opportunities for characterization of these phagocyte populations through high spatial resolution, statistical robustness, and a broad range of quantitative morphometric cell analysis tools. This study highlights an integrative approach that brings together new tools in imaging flow cytometry with conventional methodologies for characterization of phagocyte responses during acute inflammation. We focus on a comparative avian in vivo challenge model to showcase the added depth gained through these novel quantitative multiparametric approaches even in the absence of antibody-based cellular markers. Our characterization of acute inflammation in this model shows significant conservation of phagocytic capacity among avian phagocytes compared to other animal models. However, it also highlights evolutionary divergence with regards to phagocyte inflammation control mechanisms based on the internalization of apoptotic cells.

Peritoneal adhesion prevention with a biodegradable and injectable N,O-carboxymethyl chitosan-aldehyde hyaluronic acid hydrogel in a rat repeated-injury model.

Postoperative peritoneal adhesion is one of the serious issues because it induces severe clinical disorders. In this study, we prepared biodegradable and injectable hydrogel composed of N,O-carboxymethyl chitosan (NOCC) and aldehyde hyaluronic acid (AHA), and assessed its anti-adhesion effect in a rigorous and severe recurrent adhesion model which is closer to clinical conditions. The flexible hydrogel, which gelated in 66 seconds at 37 °C, was cross-linked by the schiff base derived from the amino groups of NOCC and aldehyde groups in AHA. In vitro cytotoxicity test showed the hydrogel was non-toxic. In vitro and in vivo degradation examinations demonstrated the biodegradable and biocompatibility properties of the hydrogel. The hydrogel discs could prevent the invasion of fibroblasts, whereas fibroblasts encapsulated in the porous 3-dimensional hydrogels could grow and proliferate well. Furthermore, the hydrogel was applied to evaluate the anti-adhesion efficacy in a more rigorous recurrent adhesion model. Compared with normal saline group and commercial hyaluronic acid (HA) hydrogel, the NOCC-AHA hydrogel exhibited significant reduction of peritoneal adhesion. Compared to control group, the blood and abdominal lavage level of tPA was increased in NOCC-AHA hydrogel group. These findings suggested that NOCC-AHA hydrogel had a great potential to serve as an anti-adhesion candidate.

Assessment of asbestos body formation by high resolution FEG-SEM after exposure of Sprague-Dawley rats to chrysotile, crocidolite, or erionite.

This work presents a comparative FEG-SEM study of the morphological and chemical characteristics of both asbestos bodies and fibres found in the tissues of Sprague-Dawley rats subjected to intraperitoneal or intrapleural injection of UICC chrysotile, UICC crocidolite and erionite from Jersey, Nevada (USA), with monitoring up to 3 years after exposure. Due to unequal dosing based on number of fibres per mass for chrysotile with respect to crocidolite and erionite, excessive fibre burden and fibre aggregation during injection that especially for chrysotile would likely not represent what humans would be exposed to, caution must be taken in extrapolating our results based on instillation in experimental animals to human inhalation. Notwithstanding, the results of this study may help to better understand the mechanism of formation of asbestos bodies. For chrysotile and crocidolite, asbestos bodies are systematically formed on long asbestos fibres. The number of coated fibres is only 3.3% in chrysotile inoculated tissues. In UICC crocidolite, Mg, Si, and Fe are associated with the fibres whereas Fe, P and Ca are associated with the coating. Even for crocidolite, most of the observed fibres are uncoated as coated fibres are about 5.7%. Asbestos bodies do not form on erionite fibres. The crystal habit, crystallinity and chemistry of all fibre species do not change with contact time, with the exception of chrysotile which shows signs of leaching of Mg. A model for the formation of asbestos bodies from mineral fibres is postulated. Because the three fibre species show limited signs of dissolution in the tissue, they cannot act as source of elements (primarily Fe, P and Ca) promoting nucleation and growth of asbestos bodies. Hence, the limited number of coated fibres should be due to the lack of nutrients or organic nature.

Utilización del Ultrasonido como Valoración Inicial de la Indemnidad de la Pared Abdominal en Traumatismos por Herida de Arm a Blanca en Abdomen / Using Ultrasound as the Initial Assessment of the Indemnity abdominal wall Injuries stab wound in abdomen

Introducción: Los traumatismos abdominales penetrantes son debidos generalmente a heridas de arma blanca o heridas de arma de fuego. Todas deben explorarse instrumentalmente bajo anestesia local con el objetivo de determinar la integridad del peritoneo. El ultrasonido es una herramienta muy útil utilizada en trauma, además del FAST, se lo puede utilizar en la urgencia como método de gran ayuda al realzar la exploración inicial de la herida para evaluar la integridad del peritoneo. Objetivos: Destacar la importancia del conocimiento anatómico y la correlación anatomo-clínico quirúrgica y ecográfica en la interpretación de imágenes obtenidas por ultrasonografía en la evaluación de la integridad de la pared del abdomen en heridas penetrantes por arma blanca. Material y métodos: FAST y ecografía de partes blandas instrumentándose la herida abdominal bajo anestesia local evaluando la indemnidad del peritoneo mediante la observación por ultrasonografía del abdomen y de la pared antero lateral del mismo en 14 de 42 pacientes con heridas por arma blanca en abdomen en el Servicio de Emergencias del Hospital Municipal de Morón y en el Servicio de Cirugía General del Hospital Aeronáutico Central. Período entre Febrero 2014 y Marzo 2015. Resultados: 42 (100%) pacientes con heridas de arma blanca en abdomen. 28 (66,66%) fueron inicialmente intervenidos quirúrgicamente. A 14 (33,34%) se le realizó FAST en búsqueda de líquido libre y ecografía de partes blandas instrumentándose la herida abdominal bajo anestesia local evaluando la indemnidad del peritoneo. Conclusiones: El conocimiento de las estructuras anatómicas y la disposición de las mismas que componen la pared anterolateral del abdomen permiten facilitar el reconocimiento de la indemnidad o no del peritoneo en las imágenes obtenidas por ultrasonido en pacientes con heridas abiertas por arma blanca, evitando así la realización de procedimientos quirúrgicos innecesarios.(AU)

Introduction: Penetrating abdominal trauma are usually due to stab wounds or gunshot wounds. All instrumentally be explored under local anesthesia in order to determine the integrity of the peritoneum. Ultrasound is a very useful tool used in trauma, in addition to FAST, I can use the urgency as a means of great help to enhance the initial exploration of the wound to evaluate the integrity of the peritoneum. Objectives: Highlighting the importance of the anatomical knowledge and surgical and anatomical clinical ultrasound in interpreting images obtained by ultrasonography in the evaluation of the integrity of the abdominal wall in penetrating stab wounds correlation. Material and methods: FAST ultrasound was performed and soft tissue abdominal wound became operational under local anesthesia indemnity evaluating the peritoneum through observation by ultrasound of the abdomen and the anterolateral wall thereof in 14 of the 42 patients with stab wounds in the abdomen Service Municipal Emergency Hospital of Moron and the Department of General Surgery of the Central Aeronautical Hospital, period between February 2014 and March 2015. Results: Of the 42 (100%) patients with stab wounds to the abdomen, 28 (66.66%) initially underwent surgery, and 14 (33.34%) were performed in FAST Search of free fluid and soft tissue ultrasound became operational abdominal wound under local anesthesia indemnity evaluating the peritoneum. Conclusions: Knowledge of the anatomical structures and arrangement thereof comprising the anterolateral wall of the abdomen allow easy recognition of indemnity or not the peritoneum in the ultrasound images in patients with stab wounds open, thus avoiding making unnecessary surgical procedures.

Comparing the host tissue response and peritoneal behavior of composite meshes used for ventral hernia repair.

BACKGROUND: The use of a prosthetic material is the best treatment option for ventral hernia repair; one of the most frequently performed abdominal surgery procedures. This preclinical study compares the behavior of a new mesh (Parietex composite ventral patch [Ptx]) with that of two existing meshes used for ventral hernia repair. MATERIALS AND METHODS: Fifty-four New Zealand White rabbits (3000 g) were used in an experimental model of umbilical hernia repair (diameter 1.5 cm). The materials tested were: Ventralex ST hernia patch (Vent) (Bard Davol Inc, Warwick, RI) (n = 18); Proceed ventral patch (Ethicon, Somerville, NJ) (PVP) (n = 18) and Ptx (Covidien, Sofradim, Trevoux, France) (n = 18). At 3, 7, 14 d, and 6 wk after implant, peritoneal behavior and adhesion formation were assessed by sequential laparoscopy. Mesh mesothelial cover was determined by scanning electron microscopy. Host tissue ingrowth (collagens I and III) and the macrophage response were assessed by immunohistochemical labeling. Animals were euthanized at 2, 6 wk, and 6 mo after surgery. Data were compared using the Mann-Whitney U test. RESULTS: Adhesion formation from 3 d-6 wk was significantly greater (P < 0.05) for PVP compared with Vent or Ptx. Three encapsulated PVP implants showed "tissue-integrated" adhesions affecting the intestinal loops. All three implant types showed similar patterns of collagen l and III deposition. The PVP mesh elicited the greater macrophage response both at 2 wk and 6 mo. CONCLUSIONS: Ptx and Vent showed excellent mesothelialization, which led to minimum adhesion formation. The appropriate tissue integration of Ptx in the parietal neoperitoneum is likely attributable to its deployment system.

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.

Ultrastructure of lymphatic stomata in the tunica vaginalis of humans.

Lymphatic stomata are small openings of lymphatic capillaries on the surface of the mesothelium that lines the serous cavity and have the function of active absorption. They play an important role in physiological and pathological conditions. The cavity of the tunica vaginalis is a typical serous cavity of the testis, but the lymphatic stomata of the tunica vaginalis of humans have never been reported. Here, we studied their ultrastructure by scanning and transmission electron microscopy. The submesothelial connective tissue with foramina was investigated after the mesothelial cells were digested using NaOH solution. We found the lymphatic stomata in cuboidal mesothelial cell regions of the parietal layer of the tunica vaginalis of humans with a diameter of about 1-2 µm. Sometimes, closed lymphatic stomata could be observed. Our study is the first to report the existence of lymphatic stomata of the tunica vaginalis of humans. We found that the tunica vaginalis cavity is connected with the lymphatic system through the stomata, which might provide a morphological basis for the drainage of hydrocele and tumor metastasis of the tunica vaginalis of humans.

Structural deteriorations of the human peritoneum during laparoscopic cholecystectomy. A transmission electron microscopic study.

BACKGROUND: In previous studies, changes in the surface of the peritoneum during laparoscopic surgery are well defined. Nevertheless, almost all of these studies were performed on rodents via scanning electron microscopy. In the present study, structural alterations of the mesothelial cells of peritoneum were examined during laparoscopic cholecystectomy using transmission electron microscopy. METHODS: Twenty patients with symptomatic cholelithiasis were included in the study. Peritoneal biopsy was performed immediately after CO2 pneumoperitoneum creation and at the end of surgery just before gallbladder removal. Biopsies were taken from the right upper quadrant, i.e., apart from operative manipulation. Peritoneal sample cross-sections were compared using transmission electron microscopy. RESULTS: The carbon dioxide pneumoperitoneum during laparoscopic cholecystectomy caused deteriorations of the peritoneal mesothelium. Apoptosis were developed in mesothelial cells. Bulging of mesothelial cells, irregular cell junctions, focal intercellular clefts, apical cell membrane degeneration, deep nuclear invaginations, and lipid droplets in the cytoplasm of the mesothelial cells were other remarkable findings. Mesothelial edema also was determined. DISCUSSION: As seen in previous studies, basement membrane nudity appeared after carbon dioxide pneumoperitoneum could be attributable to mesothelial cell apoptosis, deterioration of the cell structure, and cell organelles.

Human prostate DU145 carcinoma cells implanted in nude mice remove the peritoneal mesothelium to invade and grow as carcinomas.

Implanted human, androgen-independent prostatic carcinoma cells (DU145) into athymic (NCr nu/nu) mice produce diverse tumors on the peritoneal surfaces of many organs. Light and ultrastructural observations show that the mesothelial covering these surfaces are typically microvilli-coated, squamous cells or secretory cuboidal cells. The peritoneal regions colonized by tumors lack mesothelial cells and are covered by actively replicating carcinoma cells that grow as poorly differentiated cell clusters made of cell aggregates to somewhat compact spheroids covered with pleiomorphic microvilli and containing an undifferentiated vascular supply. These xenografts clusters invade the diaphragm and develop into tumors with both a basal solid aspect and an upper region of cribriform morphology. Furthermore, each tumor contains two cell types: (1) a poorly differentiated clear cell type, which grows into intraperitoneal tumors and (2) a large, basophilic cell type, which invades the peritoneal stroma of organs, including of the diaphragm.

Pathological study for the effects of in utero and postnatal exposure to diesel exhaust on a rat endometriosis model.

Previous studies have shown that prenatal and postnatal exposure to diesel exhaust (DE), which is known to be one of the main constituents of air pollution, enhances the persistence of endometriosis in a rat model. The aim of this study is to investigate the pathological changes induced by DE exposure in a rat model of endometriosis. Pregnant Sprague-Dawley rats were exposed to DE or clean air beginning on gestational day 2 and neonatal rats were persistently exposed to DE or clean air. Endometriosis was induced by autotransplantation of endometrium onto the peritoneum of eight-week-old female offspring. Endometriotic lesions were examined at 7 and 14 days post-transplantation. As a result, infiltration of activated mast cells remained in deeper area of peritoneal tissue around the endometriosis model compared to the control group at 14 days post-autotransplantation. In the DE exposure group, 14 days post-transplant, the remaining lesions contained fibroblasts and activated mast cells, which were surrounded by collagen fibers. The data showed that prenatal and postnatal DE exposure enhances the activation of mast cells and prolongs the persistence of collagen fibers in the induced rat model of endometriosis.

Application of stereology to study the effects of pneumoperitoneum on peritoneum.

BACKGROUND: Scanning electron microscopy is unable to provide sufficient data to obtain definitive results for research into the morphologic effect of pneumoperitoneum on peritoneum. To overcome this difficulty, we adopted stereology to examine the effect of the type of gas insufflated, pressure, duration, and gas flow on morphologic alterations of peritoneum. METHODS: Fifty SD rats were divided into ten groups. One group served as control. Pneumoperitoneum was established at 5 mmHg and 1.0 l/min gas flow for 1, 2 or 3 h with CO2 (in groups C1h, C2h, and C3h, respectively) or with He (in groups H1h, H2h, and H3h, respectively). CO2 pneumoperitoneum was further established at 8 mmHg and 1.0 l/min gas flow for 1 h (group C8p), at 5 mmHg and 2.0 l/min gas flow for 1 h (group C2f), and at 5 mmHg and 3.0 l/min gas flow for 1 h (group C3f). After the procedures, five specimens were sampled from anterior peritoneum and measured by stereological and electron-microscopic techniques. RESULTS: Groups H1h and C1h, H2h and C2h, and H3h and C3h, respectively, were the same in terms of area fraction of basal lamina exposed and diameter of mesothelial cells (P>0.05). The magnitudes of peritoneal trauma in groups C2h, C3h, C8p, C2f, and C3f were significantly higher than that in group C1h (P<0.01), and the same result was observed in groups H2h and H3h against group H1h (P<0.01), and in group C3f against group C2f (P<0.01). Furthermore, the area fractions of basal lamina exposed in groups C3h and H3h were remarkably higher than those in groups C2h and H2h, respectively (P<0.01). The mechanism of basal lamina exposure comprises mesothelial cell desquamation and plasmatorrhexis. CONCLUSIONS: Peritoneal morphologic trauma during pneumoperitoneum can be attributed to the pressure, duration, and gas flow instead of the type of gas insufflated.

Investigation of tumor-peritoneal interactions in the pathogenesis of peritoneal metastases using a novel ex vivo peritoneal model.

BACKGROUND: Peritoneal metastasis occurs in up to 30% of patients with gastric cancer. The aim of this experimental study is to develop and validate a novel ex vivo model of the human peritoneum to better identify factors involved in the development of peritoneal metastasis in order to improve its management and prognosis. METHODS: Peritoneal discs harvested from hernia sacs obtained at inguinal hernia surgery were suspended in media using Teflon rings. Viability of the tissue was investigated using MTS assay, light and scanning electron microscopy (LM and SEM) over 72 h. To assess validity of the model, phenotypic changes in tumor cells were investigated. Changes in matrix metalloproteinases (MMP)-2 and -9 activities in HGC and AGS gastric adenocarcinoma cells after co-culture were investigated using zymography. Modulation of tumor cell adhesion to peritoneum after exposure to heparin was assessed using a fluorometric adhesion assay. Analysis was performed using Kruskal-Wallis for multiple comparisons and Mann-Witney U for comparisons between each group. RESULTS: MTS assay showed reduced viability after 72 h (P = 0.047, compared with 24 h). Mesothelial cell loss at 48 h was demonstrated by LM and SEM, confirming peritoneal viability for at least 24 h after tissue harvesting. Zymography confirmed increased MMP2 and -9 activities in tumor cells and peritoneal tissue during co-culture compared with controls, and heparin significantly reduced tumor cell adherence (P = 0.04), as observed in published in vivo models. CONCLUSION: A validated complete model of peritoneum was developed that has shown potential to determine realistic mechanisms of peritoneal metastasis.