However, the authors have had no involvement that might raise the question of bias in the work reported or in the conclusions, implications, or opinions stated.. 0.7?mM MgCl2) and were then sterilized by filtration. The PDFs were prepared and adjusted to pH 5.0 just before injection every day. Rats in the chlorhexidine gluconate- (CG-) treated group intraperitoneally received 15?mL/kg/day 0.1% CG/15% ethanol/saline for 21 days. The solution was aseptically prepared. Rats in the talc-treated group intraperitoneally received 75?mL/kg/day talc suspension, which was prepared by dispersing 1?g of talc in 15?mL of saline followed by autoclave sterilization. The talc-treated group received one administration every 7 days for three weeks. The concentrations of MGO, FA, CG, and talc were decided based on previous reports [7C16]. As a control, a group treated with the PDF without adding MGO, FA, CG, or talc was also set. The control rats were given an Rabbit Polyclonal to CA14 intraperitoneal injection of 100?mL/kg/day PDF for 21 days. If solution remained in the peritoneal cavity, it was drained before the injection. When PDF had been injected for more than 3 weeks, it was difficult to inject it because of extensive peritoneal adhesion. Therefore, on the 22nd day after the start of the Z-VAD(OH)-FMK experiment, peritoneal equilibration test (PET) was performed. Subsequently, the parietal peritoneum was sampled for histological analysis from corresponding sites in each rat. We performed our experiments in accordance with the NIH Guide for the Care and Use of Laboratory Animals. The animals were housed in an air-conditioned room at a constant temperature Z-VAD(OH)-FMK of 23 2C and a relative humidity of 50 10% and kept under a 12-hour light/dark cycle with free access to sufficient pellet food and water. Adequate attention was paid to maintaining a hygienic environment and to preventing infectious peritonitis. Furthermore, a sterility test was performed using the dialysate drained for PET to check for the presence of aerobic bacteria, anaerobic bacteria, and fungi in drained dialysate, and then all rats were confirmed to be uninfected. 2.2. Peritoneal Equilibration Test (PET) In order to analyze peritoneal function, the peritoneal permeability of glucose was estimated by PET. First, intra-abdominal fluid was drained out. After 50?mL/kg PDF containing 2.5% glucose had been intraperitoneally injected, drained dialysate was collected immediately at 0 minutes and at 90 minutes. The injection volume was set based on that used in a human clinical context. Glucose levels were determined by SRL Co., Ltd. (Tokyo, Japan). The ratio of the glucose level in drained dialysate obtained 90 minutes after the injection to that obtained immediately after the injection was defined as the value of less than 0.05 was accepted as significant. 3. Results In the MGO-, FA-, and CG-treated rats, fist-like round liver edge and considerable adhesion of the bowel and belly were observed. In the MGO- or CG-treated rats, Z-VAD(OH)-FMK the bowel adhesion, called an abdominal cocoon, was observed like a mass surrounded by thickened peritoneum (Number 1). In the talc-treated rats, considerable adhesion of the peritoneum was observed, even though cocoon-like adhesion was not formed. By analysis of cells sections of parietal peritoneum, peritoneal thickening occurred in the MGO-, FA-, CG-, and talc-treated rats. The thickness of the peritoneum in each group is definitely shown in Number 2(a). In the MGO-, FA-, CG-, and talc-treated organizations, mononuclear cell infiltration, neovascularization, and fibrosis were observed in the peritoneum. In the MGO-treated rats, the peritoneal cells consisted of dense collagen fibers. However, in the surface of the peritoneum, collagen was scarce, and basophilic spindle cells with podoplanin, cytokeratin, and 0.05 compared with the control. ?? 0.01 compared with the control. Open in a separate window Number 3 Histopathological findings of parietal peritoneum. The parietal peritoneum was analyzed histologically with HE stain (a, b, d, e, g, h, j, k, m, and n) or Azan stain (c, f, i, l, and o). Control rat: (a, b, and c). MGO-treated rat: (d, e, and f). FA-treated rat: (g, h, and i). CG-treated rat: (j, k, and l). Talc-treated rat: (m, n, and o). Mesothelial cells, spindle cells, phagocytosis by macrophages, and multinucleated huge cells are indicated by open arrow mind, arrows, closed arrow mind, and asterisks, respectively. (a, c, d, f, g, i, j, l, m, and o): 200, (b, e, h, k, and n): 400. Open in a separate window Number 4 Characterization of cells at the surface of the peritoneum in the MGO-treated rats. In the parietal peritoneum, podoplanin-positive cells (a, b), cytokeratin-positive cells (c, d), and (TGF- em /em ) induces.