scholarly journals Ultrastructural changes of the peritoneum in a rabbit model of peritoneal dialysis

2013 ◽  
Vol 70 (11) ◽  
pp. 1023-1028 ◽  
Author(s):  
Natasa Jovanovic ◽  
Snezana Zunic-Bozinovski ◽  
Dusan Trpinac ◽  
Zeljko Lausevic ◽  
Slobodan Krstic ◽  
...  
Keyword(s):  
Peritoneal Dialysis ◽  
Basement Membrane ◽  
Rabbit Model ◽  
Mesothelial Cells ◽  
Ultrastructural Changes ◽  
Renal Diseases ◽  
Peritoneal Membrane ◽  
Tissue Samples ◽  
Peritoneal Tissue ◽  
Number Of Patients

Background/Aim. The number of patients with end-stage renal diseases treated with chronic dialysis is increasing over the last years. Long-term peritoneal dialysis is associated with progressive development of structural and functional alterations of peritoneal membrane. The aim of the study was to analyze ultrastructural alterations of mesothelial monolayer and submesothelial tissue in a modified nonuremic experimental model of peritoneal dialysis in rabbits. Methods. The study was performed on 5 healthy Chinchilla rabbits. Surgical procedures of implantation and removal of peritoneal catheter, prevention of catheter clothing, prevention of infection and dialysate instillation were performed according to previously described protocols. Peritoneal tissue samples were collected upon catheter placement and removal after a 5-week follow-up and processed for transmission electron microscopy (TEM) examination. Results. The rabbits tolerated anesthesia, surgical procedure and the applied regimen of dialysate instillations well. The animals recovered completely and no adverse effects were noted. In the animals treated with peritoneal dialysis instillations, TEM revealed alterations of the mesothelial monolayer and submesothelial tissue. The mesothelial cells in direct contact with dialysis fluid were prone to shrinking. They lost the typical cobblestone morphology and assumed a flattened shape. The mesothelial cells were often detached from the basement membrane. These cells showed euchromatic nuclei, higher number of microvilli in their apical part and very numerous vesicles. A higher quantity of collagen fibers was noticed in the peritoneal lamina propria in close relation to the basement membrane of mesothelium. The nuclei of the fibroblasts were also euchromatic. Numerous mitochondria, granules and vesicles were present in their cytoplasm. Conclusion. The used rabbit model of peritoneal dialysis is simple, practical to perform, reproducible, not expensive and not requiring advanced devices. It is suitable for obtaining peritoneal tissue samples for histological examination and can be used to analyze the effects of dialysis solutions on the rabbit peritoneal membrane.

scholarly journals Modification of peritoneal dialysis model on rabbits: A pilot study

2008 ◽  
Vol 136 (Suppl. 1) ◽  
pp. 38-43
Author(s):  
Snezana Zunic-Bozinovski ◽  
Biljana Stojimirovic ◽  
Zeljko Lausevic ◽  
Slobodan Krstic ◽  
Jasna Trbojevic-Stankovic ◽  
...  
Keyword(s):  
Peritoneal Dialysis ◽  
Pilot Study ◽  
Recovery Period ◽  
Rabbit Model ◽  
Experimental Period ◽  
Experimental Models ◽  
Peritoneal Membrane ◽  
Long Term Effects ◽  
Peritoneal Tissue

INTRODUCTION. Long-term peritoneal dialysis, a well-established method of depuration in end-stage renal disease patients, is assosiated with morphological and functional alterations of the peritoneal membrane due to the use of bioincompatible dialysis solutions. Studying effects of dialysate on the peritoneal tissue in humans is still a challenge due to ethical and technical limitations. There has been a variety of peritoneal dialysis experimental models but without consensus on the ideal model so far. OBJECTIVE. We aimed to develop a new, modified experimental rabbit model of peritoneal dialysis which would be practical, easy to conduct, relatively inexpensive and convenient to study long-term effects of dialysis solution on the peritoneal membrane. METHOD. This pilot study was performed on five healthy Chinchilla rabbits of both sexes. After i.v. Thiopental injection BP 1G, 0.5 ml/kg body mass, a catheter, especially made from Tro-soluset (Troge Medical GmbH, Hamburg, Germany) infusion system, was surgically tunneled from the animals? neck to the abdomen and inserted to the bottom of the peritoneal cavity. After one week recovery period, peritoneal dialysate instillations were performed for four weeks. During the whole five week experimental period a follow-up diary was kept. RESULTS. All procedures were well tolerated by the animals. The rabbits gained body weight, had normal body temperature and no complications were noted. CONCLUSION. The presented modified peritoneal dialysis model is practical, reproducible, does not require sophisticated technology and is well tolerated by the animals. That is why it is convenient for studying long-term effects of dialysate on the rabbit?s peritoneal membrane.

Histologic Change of Peritoneal Membrane in Relation to Adequacy of Dialysis in Continuous Ambulatory Peritoneal Dialysis Patients

2003 ◽  
Vol 23 (2_suppl) ◽  
pp. 26-30 ◽  
Author(s):  
Irini Savidaki ◽  
Dionisios Karavias ◽  
Florentia Sotsiou ◽  
Sotiria Alexandri ◽  
Pantelitsa Kalliakmani ◽  
...  
Keyword(s):  
Peritoneal Dialysis ◽  
Continuous Ambulatory Peritoneal Dialysis ◽  
Structural Changes ◽  
Mesothelial Cells ◽  
Peritoneal Membrane ◽  
Histologic Change ◽  
Significant Difference ◽  
Uremic Patients ◽  
Group B ◽  
Histologic Changes

Background Long-term exposure of peritoneal membrane to bioincompatible dialysis solutions leads to structural changes and loss of ultrafiltration capability. Objective We studied the possible relationship between histologic change and the transport characteristics of peritoneal membrane and adequacy of dialysis in continuous ambulatory peritoneal dialysis (CAPD) patients. Patients and Methods The study included 18 CAPD patients (11 men, 7 women) who underwent a peritoneal biopsy either at initiation of treatment (group A, n = 9) or after a mean of 4 years on CAPD (group B, n = 9). The morphologic changes in the mesothelial cells and the vascular compartment and the thickness of the submesothelial collagenous zone were estimated and compared with observations from 6 patients with normal renal function who underwent biopsy of the parietal peritoneum during abdominal surgery. The relationship of the observed changes in CAPD patients to results from a peritoneal equilibration test (PET) and to adequacy of dialysis [total weekly creatinine clearance (CCr) and Kt/V urea] were also investigated. Results The main histologic changes in both groups of patients were loss of mesothelial cells and decrease in the normal mesothelial surface, thickening of the submesothelial collagenous zone, and presence of vascular hyalinosis. The thickness of the submesothelial collagenous zone in both groups of patients was significantly greater than that found in controls (410 μm and 580 μm vs 50 μm, p < 0.05). Although no significant difference was found between morphologic change in the peritoneal membrane of uremic patients starting on CAPD and those who had been on peritoneal dialysis (PD) for a mean period of 4 years, a trend was observed toward more severe lesions in the latter patients. The PET, CCr, and Kt/V urea were not significantly different in the two groups of patients. Those parameters also showed no significant changes when examined at initiation of CAPD and after a mean of 4 years of PD in the same patients (group B). No significant correlations were observed between the histologic changes and the PET, CCr, or Kt/V in both groups of patients. Conclusions Significant structural changes are observed in the peritoneal membrane of uremic patients, and those changes become worse with CAPD treatment. Structural changes are not followed by functional changes during the first 4 years on CAPD.

Demonstration of Aquaporin-Chip in Peritoneal Tissue of Uremic and Capd Patients

1996 ◽  
Vol 16 (1_suppl) ◽  
pp. 54-57 ◽  
Author(s):  
Marja M. Pannekeet ◽  
Jos B. Mulder ◽  
Jan J. Weening ◽  
Dirk G. Struijk ◽  
Machteld M. Zweers ◽  
...  
Keyword(s):  
Endothelial Cells ◽  
Peritoneal Dialysis ◽  
Sodium Transport ◽  
Water Channels ◽  
Osmotic Gradient ◽  
Peritoneal Membrane ◽  
Specific Staining ◽  
Peritoneal Tissue ◽  
Hyperosmolar Solutions ◽  
Morphological Equivalent

Aquaporin-CHIP is a 28 kD channel forming integral membrane protein. It acts as an osmotically driven, water-selective pore. The presence of aquaporin-CHIP has been demonstrated in the proximal tubule in the kidney and in the pleura, as well as in other tissues. During peritoneal dialysis a dissociation between the transport of water and sodium using hyperosmolar solutions has been reported, suggesting the presence of ultrasmall pores. Water channels, like aquaporin-CHIP, could be the morphological equivalent of these pores. We investigated the possible presence of aquaporinCHIP in cryo-sections of peritoneal tissue using affinity purified human anti-CHIP IgG (P. Agre, Baltimore, MD). Peritoneal biopsies (omenta) were obtained at catheter insertion in 2 uremic patients with end-stage renal disease, and at catheter reimplantation of 1 patient treated with continuous ambulatory peritoneal dialysis (CAPD) for two years. Peritoneal tissue obtained at autopsy from 1 patient who had been on CAPD for four years, but in whom CAPD had been discontinued for five months, was also studied. Aquaporin-CHIP antiserum specific staining was found in the endothelial cells of the peritoneal capillaries in all patients. No obvious difference in the intensity of staining was seen between uremic and CAPD patients. This demonstration of aquaporin-CHIP in human peritoneal endothelial cells supports the hypothesis of the existence of ultrasmall pores within the peritoneal membrane. These water channels facilitate the transcellular transport of water, induced by an osmotic gradient, in the absence of sodium transport. It may be the explanation for the dissociation of water and sodium transport that occurs during hyperosmolar solutions. Aquaporin-CHIP is present in human peritoneal endothelial cells in both uremic and CAPD patients. Aquaporin-CHIP may be the morphological equivalent of the ultrasmall pores within the peritoneal membrane.

Chronic Peritoneal Inflammation by Cyanate in Rats

2000 ◽  
Vol 20 (6) ◽  
pp. 699-702 ◽  
Author(s):  
Kyo-Cheol Mun ◽  
Mi-Young Yeo ◽  
Sang-Pyo Kim ◽  
Hyun-Chul Kim ◽  
Chun-Sik Kwak
Keyword(s):  
Peritoneal Dialysis ◽  
Mononuclear Cells ◽  
Morphological Changes ◽  
Sodium Bicarbonate Solution ◽  
Mesothelial Cells ◽  
Control Group ◽  
Tissue Samples ◽  
Waste Products ◽  
Visceral Peritoneum ◽  
Standard Manner

Objective During peritoneal dialysis, the peritoneum is exposed to waste products, including urea. Urea forms cyanate spontaneously at body temperature and pH, and cyanate carbamylates amino acids, peptides, and proteins. Cyanate may contribute to peritoneal injury with morphological changes in the peritoneum. To test this hypothesis, we injected cyanate into rats. Methods Experiments were performed in two groups of 7 rats each. In the cyanate group, each rat received 1 mL of 1.5 μmol/L potassium cyanate dissolved in 40 mmol/L sodium bicarbonate solution intraperitoneally each experiment day. In the control group, each rat received 1 mL of 1.5 μmol/L potassium bicarbonate instead of potassium cyanate. The rats in both groups were anesthetized and killed at the 85th day after the first injection. After formalin fixation, tissue samples from abdominal walls and livers were sliced, embedded in a standard manner, and stained with hematoxylin and eosin. Results Parietal peritoneum from rats in the cyanate group showed a mild increase in the number of fibroblasts, with collagen deposits, infiltration by mononuclear cells, vascular congestion, round-shaped transformation of mesothelial cells, widening of submesothelial spaces, and abundant denudation of mesothelial cells. The visceral peritoneum from rats in the cyanate group showed collagen deposits with fibroblastic proliferation. Conclusions Cyanate can induce chronic inflammation in the peritoneum, and exposure of the peritoneum to cyanate may contribute to peritoneal injury in patients being treated with peritoneal dialysis.

Overnight Mesothelial Cell Exfoliation: A Magic Tool for Predicting Future Ultrafiltration Failure in Patients on Continuous Ambulatory Peritoneal Dialysis

2008 ◽  
Vol 28 (3_suppl) ◽  
pp. 107-113
Author(s):  
Talerngsak Kanjanabuch ◽  
Monchai Siribamrungwong ◽  
Rungrote Khunprakant ◽  
Sirigul Kanjanabuch ◽  
Piyathida Jeungsmarn ◽  
...  
Keyword(s):  
Peritoneal Dialysis ◽  
Continuous Ambulatory Peritoneal Dialysis ◽  
Mesothelial Cell ◽  
Mesothelial Cells ◽  
Nutrition Status ◽  
Continuous Exposure ◽  
Peritoneal Membrane ◽  
Dialysis Solution ◽  
Ultrafiltration Failure

⋄ Background Continuous exposure of the peritoneal membrane to dialysis solutions during long-term dialysis results in mesothelial cell loss, peritoneal membrane damage, and thereby, ultrafiltration (UF) failure, a major determinant of mortality in patients on continuous ambulatory peritoneal dialysis (CAPD). Unfortunately, none of tests available today can predict long-term UF decline. Here, we propose a new tool to predict such a change. ⋄ Mesothelial cells from 8-hour overnight effluents (1.36% glucose dialysis solution) were harvested, co-stained with cytokeratin (a mesothelial marker) and TUNEL (an apoptotic marker), and were counted using flow cytometry in 48 patients recently started on CAPD. Adequacy of dialysis, UF, nutrition status, dialysate cancer antigen 125 (CA125), and a peritoneal equilibration test (3.86% glucose peritoneal dialysis solution) were simultaneously assessed and were reevaluated 1 year later. ⋄ Results The numbers of total and apoptotic mesothelial cells were 0.19 ± 0.19 million and 0.08 ± 0.12 million cells per bag, respectively. Both numbers correlated well with the levels of end dialysate–to–initial dialysate (D/D0) glucose, dialysate-to-plasma (D/P) creatinine, and sodium dipping. Notably, the counts of cells of both types in patients with diabetes or with high or high-average transport were significantly greater than the equivalent counts in nondiabetic patients or those with low or low-average transport. A cutoff of 0.06 million total mesothelial cells per bag had sensitivity of 1 and a specificity of 0.75 in predicting a further decline in D/D0 glucose and a sensitivity of 0.86 and a specificity of 0.63 to predict a further decline in UF over a 1-year period. In contrast, dialysate CA125 and other measured parameters had low predictive values. ⋄ Conclusions The greater the loss of exfoliated cells, the worse the expected decline in UF. The ability of a count of mesothelial cells to predict a future decline in UF warrants further investigation in clinical practice.

Clearance of Tracer Albumin from Peritoneal Cavity to Plasma at Low Intraperitoneal Volumes and Hydrostatic Pressures

1998 ◽  
Vol 18 (5) ◽  
pp. 497-504 ◽  
Author(s):  
Qing Zhu ◽  
Ola Carlsson ◽  
Bengt Rippe
Keyword(s):  
Peritoneal Dialysis ◽  
Peritoneal Cavity ◽  
Fluid Volume ◽  
Free Fluid ◽  
Annual Conference ◽  
Lymphatic Absorption ◽  
Tissue Samples ◽  
Essentially Normal ◽  
Peritoneal Tissue ◽  
Tracer Distribution

Objective To assess the clearance of radiolabeled tracer albumin (RISA) from peritoneal cavity to plasma (CI → P) in rats under essentially “normal” conditions, that is, when intraperitoneal hydrostatic pressure (IPP) is subatmospheric and the intraperitoneal (IP) “free” fluid volume (IPV) is low. Methods A volume of 0.3 mL of RISA was injected IP into anesthetized Wistar rats (wt = 300 g) when the IPV was approximately 2 mL (normal) or the IPV was approximately 10 mL, and IPP was either -1.8 mmHg (normal) or +1.5 mmHg (produced by an external cuff). Plasma samples (25 μL) were obtained repeatedly during the dwell, which lasted 30 300 min, after which the peritoneal cavity was opened to recover the IPV and residuallP RISA activity. The CI → P was assessed as the mass transfer of RISA into plasma, occurring per unit time,-divided by the calculated mean IP RISA concentration (CD). The interstitial RISA space was measured as the mass of RISA accumulated, per unit tissue weight, in peritoneal tissue samples divided by the CD. Results A markedly lower CI → P (2.47 ± 0.67 μL/min), as well as total RISA clearance out of the peritoneal cavity (CI), was found under “normal” conditions (an IPV of approximately 2 mL and an IPP of approximately -1.8 mmHg) compared to the situation during peritoneal dialysis (an IPV of approximately 20 mL and an IPP of +1 mmHg). Furthermore, the interstitial RISA space increased linearly over time even at negative IPPs and at an unchanging peritoneal interstitial fluid volume. At a low (normal) IPV the CI → P did not increase significantly with elevating IPP, and increased only marginally when tracer distribution was improved by artificial vibration of the rats. However the CI → P increased when larger volumes were infused to increase the totallPV. Conclusions It is concluded that the CI → P and CI at low IPPs and IPVs are not as high as during peritoneal dialysis. Increases in CI → P were, however, coupled to increases in IPV. This highlights the importance of the IPV per se and of a sufficient IP tracer distribution for direct lymphatic absorption to be efficient. This study was presented in part at the XVIth Annual Conference on Peritoneal Dialysis, Denver, Colorado, U.S.A., 1997 (33).

Technical Aspects in Studying Peritoneal Morphology in Animal Models of Peritoneal Dialysis

2009 ◽  
Vol 29 (2_suppl) ◽  
pp. 40-44 ◽  
Author(s):  
Soner Duman ◽  
Sait Şen
Keyword(s):  
Peritoneal Dialysis ◽  
Abdominal Wall ◽  
Histological Examination ◽  
Anterior Abdominal Wall ◽  
Anterior Wall ◽  
Abdominal Muscle ◽  
Tissue Samples ◽  
Peritoneal Tissue ◽  
Significant Difference ◽  
Peritonitis Model

Objective Peritoneal biopsies are considered useful for gaining a better understanding of the pathophysiology of the peritoneum during experimental peritoneal dialysis (PD). Different peritoneal tissue samples (i.e., abdominal wall, liver, diaphragm, intestine, and omentum) may be used, but there can be artifacts due to peritoneal tissue processing. Aim To investigate differences in peritoneal membranes from different parts of the peritoneum, and also 2 different fixatives, in experimental PD and a peritonitis model in rats. Methods Peritoneal tissues from the anterior abdominal wall, liver, omentum, and intestine were taken from each of 3 groups of animals: sham, experimental PD, and peritonitis model. Tissue samples were immediately fixed with 4% formaldehyde and routinely processed for histological examination. Two parietal peritoneal tissue samples according to longitudinal and horizontal sections of anterior wall inner abdominal muscle were also taken. All samples were immediately fixed with 4% formaldehyde and B5 fixative (B5), and then routinely processed for histological examination. Results In all groups, histopathological findings were more commonly seen in the abdominal wall samples. There were no changes observed in peritoneal membranes other than those of anterior abdominal wall samples from both sham and PD model rats. However, there was a significant difference between anterior and posterior facets of liver in the peritonitis model. Furthermore, the antimesenteric site of intestinal peritoneum was less affected than the mesenteric site. There were no significant histopathological differences between B5 and 4% formaldehyde fixation ( p > 0.05). Conclusion Our results suggest that peritoneum obtained from the anterior abdominal wall is the most affected area and therefore the most suitable site to investigate peritoneal changes in the experimental rat PD model. There were no significant differences between fixation with 4% formaldehyde and B5 solution. Abdominal wall samples should be of the same direction of inner abdominal muscle, that is, horizontal sectioning should be used for measurements of the submesothelial area.

Factors Contributing to Peritoneal Tissue Remodeling in Peritoneal Dialysis

2009 ◽  
Vol 29 (6) ◽  
pp. 605-617 ◽  
Author(s):  
Margot N. Schilte ◽  
Johanna W.A.M Celie ◽  
Piet M. ter Wee ◽  
Robert H.J. Beelen ◽  
Jacob van den Born
Keyword(s):  
Peritoneal Dialysis ◽  
Structural Changes ◽  
Tissue Remodeling ◽  
Therapeutic Strategies ◽  
Molecular Pathways ◽  
Peritoneal Membrane ◽  
Volume Loading ◽  
Peritoneal Cells ◽  
Peritoneal Tissue

Peritoneal dialysis (PD) is associated with functional and structural changes of the peritoneal membrane. In this review we describe factors contributing to peritoneal tissue remodeling, including uremia, peritonitis, volume loading, the presence of a catheter, and the PD fluid itself. These factors initiate recruitment and activation of peritoneal cells such as macrophages and mast cells, as well as activation of peritoneal cells, including mesothelial cells, fibroblasts, and endothelial cells. We provide an overview of cytokines, growth factors, and other mediators involved in PD-associated changes. Activation of downstream pathways of cellular modulators can induce peritoneal tissue remodeling, leading to ultrafiltration loss. Identification of molecular pathways, cells, and cytokines involved in the development of angiogenesis, fibrosis, and membrane failure may lead to innovative therapeutic strategies that can protect the peritoneal membrane from the consequences of long-term PD.

scholarly journals Surgical Technique Using An Improvised Peritoneal Catheter In An Experimental Non-Uremic Rabbit Model Of Peritoneal Dialysis

2015 ◽  
Vol 65 (3) ◽  
pp. 319-327 ◽  
Author(s):  
Krstić Slobodan ◽  
Trbojević-Stanković Jasna ◽  
Žunić Snežana ◽  
Jovanović Nataša ◽  
Stojimirović Biljana
Keyword(s):  
Peritoneal Dialysis ◽  
Surgical Technique ◽  
Light And Electron Microscopy ◽  
Rabbit Model ◽  
Experimental Models ◽  
Peritoneal Catheter ◽  
Tissue Samples ◽  
Custom Made ◽  
Catheter Implantation ◽  
Rabbit Animal Model

AbstractExperimental models have strongly contributed to the comprehension of the processes of peritoneal damage that take place during peritoneal dialysis treatment in human patients. A variety of peritoneal dialysis models have been developed, mostly using rats and rabbits.In this study we present the successful development of a custom-made improvised peritoneal catheter for an experimental non-uremic rabbit model of peritoneal dialysis.A detailed description of the surgical technique of peritoneal catheter implantation, care and removal is provided.This innovative approach to constructing a peritoneal catheter in rabbit animal model of peritoneal dialysis is easy, reproducible and inexpensive. The surgical technique applied provided adequate tissue samples for both light and electron microscopy.

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