Legends
Fig. 1 Schematic illustration of the HIPEC procedure. Saline
solution and fluid containing cisplatin were heated up through a water
bath (37 or 42 °C) and circulated continuously into the abdominal cavity
(perfusion rate: 180 ml per hour), through an inlet catheter into the
right and an outlet catheter into the left lower abdomen. The
temperature and flow are constantly monitored, and the outflow fluid is
trapped by a drainage bag.
Fig. 2 Overview about the peritoneal carcinomatosis index
(PCI). (A) Total and PCI of the respective organs was recorded
according to the principle of Jacquet and Sugarbaker et al.(1996), which has been adapted for the animal model. PCI lesion size
score: 0 = no tumor; 1 = tumor ≤ 1 mm; 2 = tumor > 1 mm ≤ 3
mm; 3 = tumor > 3 mm. (B) Exemplary photographic
representation of the main tumor localizations. There are up to five
intra-abdominal tumor sites, which are preferentially located on the
large curvature of the stomach, peri-hepatic, peri-splenic, mesenteric
and peritoneally on the abdominal wall.
Fig. 3 In vivo detection of RH-30 tumors by MR imaging.
RH-30 tumors were detected by MR imaging of the abdominal cavity(A) without (native) and (B) with contrast agent.(C) For evaluation of the fluid distribution in the abdominal
cavity during peritoneal lavage, MR images were also taken during HIPEC
treatment. MR imaging confirmed the observed tumor dissemination
depicted in Figure 2. Tumor lesions are indicated by red asterisks.
Fig. 4 Evaluation of changes in cell morphology and
proliferation marker expression after HIPEC treatment. (A)Hematoxylin and eosin staining revealed no cell morphological changes in
RH-30 tumors. (B) Immunohistochemical analysis of the
proliferation marker Ki-67 (brown) showed no obvious changes in protein
expression through the individual treatments. (C) The detailed
examination of Ki-67 protein expression and determination of the Ki-67
proliferation index using a Ki-67 quantifier module of a pathology
software (Cognition Master Professional Suite: Ki67 Quantifier, VMscope
GmbH, Berlin, Germany) showed a slight reduction in the proliferation
capacity.
Fig. 5 Investigation of markers for apoptosis induction after
HIPEC treatment. (A) Apoptosis marker expression of the
activated variant of Caspase 3 (black) revealed no HIPEC-specific
apoptosis induction. The nuclei were counterstained with hematoxylin
(blue). (B) Terminal desoxynucleotidyl transferase
(TUNEL)-labelled DNA fragmentation as a sign of apoptosis induction
(green fluorescence signal) is visible in the marginal areas of the
HIPEC treated tumors in a dose and temperature dependent manner. The
nuclei were counterstained in blue with DAPI.