Establishment of a syngeneic model of hepatic colorectal oligometastases

BACKGROUND: Regional and systemic therapies aimed at improving the outcome for patients with colorectal hepatic metastases have met with modest yet tangible success. Currently, liver resection remains the only curative treatment, but only a minority of patients are candidates for surgery. Animal models are an ideal way to study new treatments for patients with metastatic colorectal cancer. We propose a syngeneic animal model of hepatic colorectal metastases that simulates oligometastases, which is a clinical state considered amenable to regional therapeutic strategies. MATERIALS AND METHODS: BDIX (BD-9) rats underwent intrasplenic injection of DHD/K12/TRb (Prob/K12) cells to create hepatic metastases via the portal system. After injection of 5 x 10(6) cells, rats underwent laparotomy to determine metastatic burden. Histological analysis confirmed the presence of metastases from resected tumors. RESULTS: Fifty-three animals were prospectively treated and observed for the development of oligometastases defined as between 1 and 10 hepatic lesions. Thirty-six (68%) of the animals developed detectable metastases while 32 (60%) developed oligometastases (average = 4.40 +/- 2.67). Four animals had overwhelming metastatic liver and peritoneal disease. All animals underwent peritoneal examination and thoracotomy to ensure localized disease. Histological analysis of five hepatectomy specimens confirmed the presence of metastatic cancer. Animals with oligometastases were healthy as evidenced by normal feeding and grooming behavior. CONCLUSIONS: An animal model of oligometastatic colorectal cancer to the liver can reproducibly mimic the stage IV state in humans conducive to regional therapy and can be used reliably to test novel treatments and mechanisms of metastatic colorectal cancer.     

Instillation of taurolidine/heparin after laparotomy reduces intraperitoneal tumour growth in a colon cancer rat model

OBJECTIVE: To investigate whether irrigation of the abdominal cavity after laparotomy for caecum resection with taurolidine/heparin or adhesion prophylactic substances reduces intraperitoneal tumour growth or the local recurrence rate in a colon carcinoma rat model. METHODS: 60 BDIX rats underwent caecum resection after intraperitoneal inoculation of 1 x 10(4) colon carcinoma cells (DHD/K12/TRb). Intergel, Interceed, taurolidine/heparin or NaCl 0.9% were intraperitoneally applied after randomisation. Finally, the total number and total weight of intraperitoneal metastases were determined as well as the adhesion score according to Moreno. Metastatic tissue was examined histologically and immunohistochemically (E-cadherin, CD44, beta(1)-integrin). RESULTS: Taurolidine/heparin significantly reduced not only the total number (3 vs. 11 in the control group) but also the total weight (65 vs. 330 mg) of intraperitoneal metastases in comparison to the control group (p = 0.003 and p = 0.005). E-Cadherin expression in the metastatic tissue of animals treated with taurolidine/heparin was significantly decreased (p = 0.016). CONCLUSION: Taurolidine/heparin effectively reduces intraperitoneal tumour growth when used as an intraoperative lavage. These results represent a good rationale for intraoperative adjuvant irrigation with taurolidine/heparin during resection of colon cancer.     

Evaluation of povidone abdominal washing for prevention of peritoneal cancer cell seeding: experimental study in the rat

GOAL: The aim of the study was to evaluate the in vitro cytototoxicity of diluted povidone iodine on colon cancer cells and its in vivo antitumoral effect in a model of peritoneal carcinomatosis in the rat. METHODS: Cell cytotoxicity of a povidone iodine diluted solution was assessed, in vitro, on rat colon cancer cells (DHD/K12/PROb) and human colon cancer cells (HT29). The antitumoral effect of diluted povidone iodine washing was measured in BDIX rats after the intraperitoneal inoculation of 10(6) DHD/K12/PROb cells. Results were compared to an abdominal washing within a 9 g/l salinel solution. In one experiment, peritoneal scars and a colocolic anastomosis were performed after the injection of cancer cells. RESULTS: A short 10 min incubation of human and rat colon cancer cells with diluted povidone iodine resulted in a complete cell killing. In animals, a peritoneal washing with 1% diluted povidone iodine completely inhibited the tumor growth in parietal peritoneum. However, development of peritoneal tumor nodules was not inhibited in the omentum, in scarified peritoneum or in intestinal anastomosis. CONCLUSIONS: Despite its high in vitro efficacy, diluted povidone iodine has an incomplete effect in the prevention of peritoneal carcinomatosis, with only a partial inhibition in scarred peritoneum epiplo?c area and intestinal anastomosis. In contrary, it procures a complete inhibition of tumor growth in normal peritoneum.     

Influence of different gases and intraperitoneal instillation of antiadherent or cytotoxic agents on peritoneal tumor cell growth and implantation with laparoscopic surgery in a rat model

BACKGROUND: A generally accepted approach to prevent tumor implantation with laparoscopic surgery does not exist. Alternative gases in combination with intraperitoneal instillation of different antiadherent or cytotoxic agents have not been evaluated. METHODS: The effect of taurolidine, heparin, and povidone-iodine on the growth of colon adenocarcinoma DHD/K12/TRb was measured in rats undergoing laparoscopy with carbon dioxide (n = 40), helium (n = 40), or xenon (n = 40). In the procedure, 10(4) tumor cells were administered intraperitoneally, and pneumoperitoneum was established over 30 min at 8 mmHg with the different gases. The rats additionally received intraperitoneal instillation with one of the following: 1 ml of Ringer's solution, 1 ml of 0.5% taurolidine, 1 ml 0.5% taurolidine with heparin (10 U/ml), or 1 ml 0.25% of povidone-iodine. Tumor growth was measured after 4 weeks. RESULTS: Median intraperitoneal tumor weight was lower in rats receiving taurolidine (CO(2): 10 mg; helium: 50 mg; xenon: 39.5 mg) or taurolidine with heparin (CO(2): 4 mg; helium: 4.5 mg; xenon: 46.5 mg) in all gas groups than in the control groups (CO(2): 427 mg; helium: 268 mg; xenon: 345 mg) (p < 0.001). Whereas povidone-iodine caused significantly lower tumor growth in the CO(2) group (56.5 mg) (p < 0.01), the combination of helium (145 mg) and xenon (457 mg) with povidone-iodine produced no reduction of tumor growth as compared with the control groups (helium: 268 mg; xenon: 345 mg). CONCLUSIONS: Taurolidine and taurolidine with heparin significantly inhibit intraperitoneal tumor growth, with different gases used for pneumoperitoneum. Only povidone-iodine caused significant decrease of tumor growth in combination with CO(2). The combination of xenon and povidone-iodine should not be used in patients with cancer because of increased tumor growth.     

Taurolidine Inhibits Tumor Cell Growth In Vitro and In Vivo

Background: Taurolidine, a derivative of the amino acid taurine, exhibits antiendotoxin, antibacterial, and antiadherence activity. We hypothesized that Taurolidine may inhibit tumor cell growth, both in an in vitro and in vivo setting. Our aim was to examine the effect of Taurolidine on the growth of a rat metastatic colorectal tumor cell line (DHD/K12/TRb) in vitro and in vivo.Methods: In the in vitro experiments, DHD/K12/TRb cells were incubated with 5, 10, 15, 25 g/ml of Taurolidine. Cells incubated in culture medium alone were used as controls. Cell proliferation, cell viability, cell death, and cell apoptosis were measured using commercially available techniques. In the in vivo experiment, BD IX rats were randomized into two groups (n = 10/group). Group A (control) underwent laparotomy and instillation of DHD/K12/TRb tumor cells intraperitoneally followed by phosphate buffered saline (PBS). Group B received Taurolidine (100 mg/kg) instead of PBS. Animals were killed after 24 days and tumor burden assessed by counting the number of tumor nodules in the peritoneal cavity.Results: Incubation of the tumor cells with Taurolidine resulted in a 4-fold decrease in proliferation rates (25 ± 4% vs. 100 ± 28% for controls) and a 4-fold increase in cell necrosis as demonstrated by the increase in LDH release (403 ± 28% vs. 100 ± 26% for controls), at a Taurolidine concentration of 25 g/ml. A dose-dependent decrease in cell viability was also observed. In the in vivo study, local Taurolidine administration resulted in significant decreases in tumor burden (3 ± 1 nodules in Group B animals vs. 649 ± 101 nodules in Group A animals).Conclusions: Taurolidine inhibits the growth of a rat metastatic colorectal tumor cell line in vitro and in vivo and thus may have potential in the prevention of peritoneal metastases.     

Sodium hyaluronate enhances colorectal tumour cell metastatic potential in vitro and in vivo

BACKGROUND: Sodium hyaluronate has been used intraperitoneally to prevent postoperative adhesions. However, the effect of sodium hyaluronate on tumour growth and metastasis in vitro and in vivo is still unknown. METHODS: Human colorectal tumour cell lines SW480, SW620 and SW707 were treated with sodium hyaluronate (10-500 microg/ml) and carboxymethylcellulose (0.125-1 per cent), and tumour cell proliferation and motility were determined in vitro. For the in vivo experiments male BD IX rats were randomized to a sodium hyaluronate group (n = 11; intraperitoneal administration of 0.5 x 10(6) DHD/K12 tumour cells and 5 ml 0.4 per cent sodium hyaluronate) or a phosphate-buffered saline group (n = 11; 0.5 x 10(6) DHD/K12 tumour cells and 5 ml phosphate-buffered saline intraperitoneally). Four weeks later the intraperitoneal tumour load was visualized directly. RESULTS: In vitro sodium hyaluronate increased tumour cell proliferation and motility significantly. Sodium hyaluronate-induced tumour cell motility appeared to be CD44 receptor dependent, whereas sodium hyaluronate-induced tumour cell proliferation was CD44 receptor independent. In vivo there was a significantly higher total tumour nodule count in the peritoneal cavity of the sodium hyaluronate-treated group compared with the control (P = 0.016). CONCLUSION: Sodium hyaluronate enhances tumour metastatic potential in vitro and in vivo, which suggests that use of sodium hyaluronate to prevent adhesions in colorectal cancer surgery may also potentiate intraperitoneal tumour growth. Presented to the Patey Prize Session of the Surgical Research Society and the annual scientific meeting of the Association of Surgeons of Great Britain and Ireland, Brighton, UK, 4-7 May 1999     

Prevention of Peritoneal Carcinomatosis from Colon Cancer Cell Seeding Using a Pirarubicin Solution in Rats and Nude Mice

Free malignant cells, which are frequently detected in the washing liquid from the peritoneal cavity before and after resection of human colorectal cancer, are suspected to cause recurrent peritoneal cancer. We carried out an experimental study to compare the prophylactic efficacy of washing the peritoneum with several anticancer drugs and the antiseptic povidone-iodine against the development of peritoneal carcinomatosis from colonic origin in rats and nude mice. The in vitro anticancer activity of a short, 15-minute exposure of pirarubicin, doxorubicin, 5-fluorouracil, cisplatin, mitomycin C, and 1% povidone-iodine was first evaluated by an MTT assay on DHD/K12/PROb rat and LS174T human colon cancer cells. For the in vivo experiments, BDIX rats were inoculated intraperitoneally (IP) with 1 x 10⁶ DHD/K12/PROb cells followed by peritoneal scarring and a colocolic anastomosis. A 15-minute peritoneal washing with the anticancer drugs or povidone-iodine was then performed. Nude mice were IP-inoculated with 1 × 10⁷ LS174T human cells and treated 2 hours later with IP pirarubicin. Only pirarubicin, mitomycin C, and povidone-iodine were fully cytotoxic in vitro against DHD/K12/PROb rat colon cancer cells. In contrast to pirarubicin and povidone-iodine, mitomycin C was not completely active against LS174Tcells. In vivo, pirarubicin cured DHD/K12/PROb-inoculated rats, even at the site of the peritoneal scarring and intestinal anastomosis. IP pirarubicin prevented the development of peritoneal carcinomatosis and liver metastasis in LS174T-inoculated mice. IP washing with pirarubicin cured 2-day-old, but not 7-day-old, peritoneal carcinomatosis in rats. Short exposure to IP pirarubicin is nontoxic and more active than povidone-iodine and other anticancer drugs in preventing the development of peritoneal carcinomatosis from colonic origin in rats and mice. The prophylactic effect of preoperative peritoneal washing with pirarubicin on the development of recurrent peritoneal cancer should be evaluated in a randomized clinical trial.     

Validation of a new experimental model of colon cancer

BACKGROUND: Most of the published animal studies that have evaluated tumor growth and port site metastases in laparoscopy have utilized a cell suspension model and thus cannot be compared to the clinical situation. Although solid tumor models have been developed, there has been no experimental model that establishes an orthotopic tumor in the rectum, reflecting the clinical situation of a solid colonic cancer. METHODS: Tumor cells (colon adenocarcinoma DHD/K1/TRb) were administered intraperitoneally in rats, which were used as solid tumor donors. A 20-mg piece of solid tumor from the donor was placed in a submucosal blister created in the rectum wall of the study rats. The approach to the submucosal blister was made through the mucosa after contralateral enterotomy. In order to validate the model, this intervention was performed in 10 cases (group A). After 10 days of intervention, the rats were submitted to resection of the rectum and histological examination of the specimen. In another 10 rats (group B), manipulation of the tumor was performed after 10 days to cause tumor cell spillage. The likelihood of tumor dissemination was investigated in this group 20 days after this intervention. RESULTS: Group A developed solid tumors in seven of 10 cases (70%). All of the tumors were localized between the muscular and the mucosal layer, with preservation of the serosa and without affecting the enterotomy. In all of the rats in group B, macroscopic tumor was observed in the upper rectum (100%) 10 days after its induction. Twenty days after tumor manipulation, nine rats had local tumor dissemination; two of them also had general tumor dissemination in the abdominal cavity. CONCLUSIONS: We established a novel solid colonic tumor model in rats for the investigation of intraoperative tumor cell spillage during resection of the colon and the development of port site metastases.     

Abdominal insufflation does not cause hematogenous spread of colon cancer

BACKGROUND AND PURPOSE: Previous investigators have suggested that port-site recurrences are possibly a result of abdominal insufflation, forcing viable cancer cells into the circulation to metastasize and thrive in areas of trauma. Using a syngeneic animal cancer model, we tested the hypothesis that pneumoperitoneum increases the incidence of wound metastasis by a blood-borne mechanism. METHODS: Male BD IX rats (N = 150) were injected intraperitoneally with 2 x 10(5) viable syngeneic 1,2-dimethylhydralazine-induced colon cancer cells (DHD-K12). Animals were divided into three groups: A (abdominal insufflation with 3-cm incision on the back into muscle remote from the peritoneum); B (3-cm back incision alone); and C (control group with 3-cm midline abdominal incision). Three weeks after surgery, the animals were euthanized and autopsied. RESULTS: In the two groups with back wounds, the incidence of cancer growth at the incision was zero, as demonstrated grossly and by histologic sample (A: 0/47, B: 0/43). In contrast, the autopsied control group had a 42% incidence of metastasis to the wound (25/59). There seemed to be no difference in the distribution of intra-abdominal disease between those rats that underwent insufflation and those that did not. CONCLUSION: It is unlikely that pneumoperitoneum promotes hematogenous wound implantation of free intraperitoneal cancer cells.     

Effects of topical tumoricidal agents on port-site recurrence of colon cancer: an experimental study in rats.

BACKGROUND: Reports of metastatic spread of colon and rectal cancer to port sites after laparoscopic resection of potentially curable lesions has raised doubt regarding the efficacy and safety of laparoscopic technology in cancer surgery. Experimental study in animals has led us to believe that the mode of spread of these metastases is via the direct route. We hypothesized, therefore, that we could decrease the rate of trocar-site recurrences by treating the individual port sites with a topical tumoricidal agent. MATERIALS AND METHODS: Male BD-IX rats weighing 240 to 360 g were injected with syngeneic colon cancer to simulate free intraperitoneal cancer spread to trocar sites. All rats were subjected to a sham laparoscopic operation after 2 x 10(5) viable cancer cells had been injected into their peritoneal cavities. Five-millimeter trocars were inserted into each rat after abdominal insufflation to 10 mm Hg. Pneumoperitoneum was maintained for 10 minutes before the trocars were removed simultaneously. Trocar sites were then subjected to one of three treatments, with each animal receiving a maximum of two different treatments. Sites were treated with 70% ethanol (N = 42), povidine/ iodine (N = 40), or no topical treatment (N = 46). Three weeks later, the animals were euthanized and autopsied. Subcutaneous tumors at trocar sites or tumors with >50% volume within the wound were considered implants. RESULTS: Control sites revealed a metastasis rate of 41% (19/46). The tumor implant rate was 36% (15/42) at alcohol-treated sites and 20% (8/40) at sites treated with povidone-iodine (P < 0.05). CONCLUSION: Topical administration of povidone-iodine to trocar wounds after laparoscopic surgery can significantly reduce the incidence of port-site metastasis in a syngeneic animal model of colon cancer.     

The effect of intra-abdominal temperature on the tissue and tumor diffusion of intraperitoneal cisplatin in a model of peritoneal carcinomatosis in rats]

OBJECTIVE: To evaluate the effects of hyperthermia and hypothermia on the peritoneal and on tumor penetration by intraperitoneal cisplatin. MATERIAL AND METHODS: Twenty day old peritoneal carcinomatosis was obtained after intraperitoneal injection of 1 x 10(6) DHD/K12/PROb cells into BD IX rats. Animals were treated by intraperitoneal infusion of cisplatin (25 micrograms/mL) using hyperthermic (16.6 degrees C), normothermic (37.6 degrees C) or hyperthermic (41.8 degrees C) intraperitoneal chemotherapy. RESULTS: Hyperthermia increased cisplatin concentration in tumoral and diaphragmatic tissues compared to normothermic treatment, while renal concentrations were lower. Hypothermia produced lower cisplatin concentrations in both cancer and peritoneal tissues compared to normothermic treatment. CONCLUSION: These experiments confirmed the pharmacological advantage produced by hyperthermia in cisplatin intraperitoneal chemotherapy.     

Influence of intraperitoneal application of taurolidine/heparin on expression of adhesion molecules and colon cancer in rats undergoing laparoscopy

BACKGROUND: Recent experimental studies have shown that intraperitoneal administration of taurolidine/heparin causes a reduction of local tumor growth after laparoscopy in rat models. It might be that the anti-adherent activities of these agents are responsible for this effect. In this study we investigated the adhesion molecules E-cadherin, beta1-integrin, and CD44. MATERIALS AND METHODS: Following a 10,000 colon adenocarcinoma cells' (DHD/K12/TRb) intraperitoneal application a cecum resection and a partial parietal peritoneum resection (1 x 1 cm) were performed using a three trocar technique in 30 BD IX rats. After randomization in two groups, the cecum suture line and the parietal peritoneal defect were either lavaged with 1 mL of 0.5% taurolidine/10 IU heparin or with equal amounts of 0.9% normal saline solution. Rats were sacrificed four weeks after operation and total tumor growth was determined. E-cadherin, beta1-integrin, and CD44 were assessed immunohistochemically on the tumor tissue. RESULTS: The expression of E-cadherin was significantly reduced to 46.7% (complete loss of staining) in the taurolidine/heparin group. Although no significant difference was detected concerning the beta1-integrin and CD44 expression, a slightly reduced expression level with 26.7% of negative staining in metastases of the taurolidine/heparin group was observed. The total tumor weight (171.1 +/- 181.2 mg) as well as the total number of tumor lesions was also reduced by the substances compared to the control group (283.2 +/- 91.4 mg). CONCLUSIONS: Taurolidine/heparin led to a significant reduction of local tumor growth. Additionally a reduction of the expression of E-cadherin was observed. However, the biological behavior of this molecule is multivariant, controversial and still unclear. Further studies should elucidate its role in the epithelial tumor genesis.     

The influence of a pneumoperitoneum on the peritoneal implantation of free intraperitoneal colon cancer cells

Background: In order to test the influence of a pneumoperitoneum on the peritoneal implantation of free intraperitoneal colon cancer cells, 40 male syngeneic WAG rats were at random divided into four groups. Methods: Group 1 (n=10) animals underwent a midline laparotomy and 10⁴ CC531 colon cancer cells were injected intraperitoneally (IP); in group 2 (n=10) 10⁴ CC531 cells were injected IP without further manipulation; in group 3 (n=10) a pneumoperitoneum up to 10 mmHg was created after the IP injection of the same amount of CC531 cells. The pneumoperitoneum was maintained for 15 min. Finally in group 4 (n=10) after the IP injection of 10⁴ CC531 cells and after the creation of a pneumoperitoneum, two 14-G IV catheters simulating trocars were introduced in each flank. A follow-up period of 8 weeks was used. Tumor implantation was scored according to the peritoneal cancer index of Eggermont and the index of Chauffert. Results: Tumor nodules were found varying from 60% in groups 1–3 to 50% in group 4. There was no statistical difference between the implantation rate in the four groups. A port-site recurrence was seen in group 4; all the other tumor implants were located in the mesenterium, omentum, internal genitals, or parietal peritoneum. Conclusions: The presence of a pneumoperitoneum does not enhance the implantation of free intraperitoneal malignant colon cancer cells in the rat, but the presence of a port may lead to abdominal-wall metastases.     

Hepatic metastasis alters the immune function of murine liver nonparenchymal cells.

To examine the effect of a single hepatic focus of metastatic colon tumor on the immune function of liver non-parenchymal cells (NPCs) from C57Bl/6 mice, we injected 2.5 x 10(5) liver-derived murine colon adenocarcinoma (LD-MCA-38) cells beneath the liver capsule. Three weeks following injection of the tumor cells, the immune function of the NPCs was studied. The NPCs from tumor-bearing mice exhibited increased cytotoxic and proliferative activity. The NPCs from tumor-bearing mice also contained a greater percentage of CD8+ and T-cell receptor gamma/delta+ liver-associated T lymphocytes. Levels of interleukin 6 and tumor necrosis factor were increased in the NPC supernatant, and interleukin 6 levels were increased in serum from tumor-bearing mice. We conclude that the presence of a single hepatic focus of metastatic tumor results in augmented immune function of murine liver NPCs.     

Human cord blood cells and myocardial infarction: effect of dose and route of administration on infarct size

There is no consensus regarding the optimal dose of stem cells or the optimal route of administration for the treatment of acute myocardial infarction. Bone marrow cells, containing hematopoietic and mesenchymal stem cells, in doses of 0.5 x 10(6) to >30 x 10(6) have been directly injected into the myocardium or into coronary arteries or infused intravenously in subjects with myocardial infarctions to reduce infarct size and improve heart function. Therefore, we determined the specific effects of different doses of human umbilical cord blood mononuclear cells (HUCBC), which contain hematopoietic and mesenchymal stem cells, on infarct size. In order to determine the optimal technique for stem cell administration, HUCBC were injected directly into the myocardium (IM), or into the LV cavity with the ascending aorta transiently clamped to facilitate coronary artery perfusion (IA), or injected intravenously (IV) in rats 1-2 h after the left anterior coronary artery was permanently ligated. Immune suppressive therapy was not given to any rat. One month later, the infarct size in control rat hearts treated with only Isolyte averaged 23.7 +/- 1.7% of the LV muscle area. Intramyocardial injection of HUCBC reduced the infarct size by 71% with 0.5 x 10(6) HUCBC and by 93% with 4 x 10(6) HUCBC in comparison with the controls (p < 0.001). Intracoronary injection reduced the infarction size by 47% with 0.5 x 10(6) HUCBC and by 80% with 4 x 10(6) HUCBC (p < 0.001), and IV HUCBC reduced infarct size by 51% with 0.5 x 10(6) and by 75-77% with 16-32 million HUCBC (p < 0.001) in comparison with control hearts. With 4 x 10(6) HUCBC, infarction size was 65% smaller with IM HUCBC than with IA HUCBC and 78% smaller than with IV HUCBC (p < 0.05). Nevertheless, IM, IA, and IV HUCBC all produced significant reductions in infarct size in comparison with Isolyte-treated infarcted hearts without requirements for host immune suppression. The present experiments demonstrate that the optimal dose of HUCBC for reduction of infarct size in the rat is 4 x 10(6) IM, 4 x 10(6) IA, and 16 x 10(6) IV, and that the IM injection of HUCBC is the most effective technique for reduction in infarct size.     

The influence of adhesion prophylactic substances and taurolidine/heparin on local recurrence and intraperitoneal tumor growth after laparoscopic-assisted bowel resection of colon carcinoma in a rat model

Backgroud: The goal of the study was to investigate the influence of adhesion prophylactic substances (Interceed/lntergel) as well as taurolidine/heparin on intraperitoneal tumor growth and the local recurrence rate after laparoscopic cecum resection in a rat tumor model. Methods: Sixty BDIX rats were randomized in three therapy groups and one control group. A laparoscopic-assisted cecum resection was performed via three-trocar method after intraperitoneal tumor cell application (10,000 cells) of a colon carcinoma cell line (DHD/K1/TRb) in all animals. According to the randomization, the cecum suture and a 1 × 1-cm peritoneal defect were either covered with Intergel/Interceed or 1 ml of 0.5% taurolidine 10 IU heparin. The control group underwent instillation of 1 ml 0.9% NaCl solution. After 4 weeks the animals were euthanized and intraperitoneal tumor growth, local recurrence rate, and the number of intraperitoneal adhesions were determined. Results: The local recurrence rate was not significantly affected by any of the substances. Nevertheless, taurolidine/heparin significantly reduced the total number and weight of intraperitoneal metastases. The formation of adhesions was not significantly influenced by adhesion prophylaxis substances or by taurolidine/heparin. Conclusions: Taurolidine/heparin led to a significant reduction of intraperitoneal tumor growth after intraperitoneal application, whereas local tumor recurrence was not significantly influenced. This might be due to the number of injected tumor cells in this cell suspension model. Interceed and Intergel did not reduce intraperitoneal tumor growth. Furthermore, adhesion formation was not reduced by any of the substances.     

Topical treatment with oxaliplatin for the prevention of port-site metastases in laparoscopic surgery for colorectal cancer.

BACKGROUND: The development of port-site metastases following laparoscopic resection of various malignancies continues to be a disturbing issue for laparoscopic surgeons. Previous studies revealed promising results with oxaliplatin, a third-generation platinum compound, as a first-line treatment in advanced colorectal cancer. This study evaluates the effect of topical application of oxaliplatin on the development of port-site metastases in an experimental murine model. METHODS: Nineteen female BDIX rats (immunocompetent, 6 weeks old) underwent a sham laparoscopic operation after 1 x 10(7) viable rat colon carcinoma viable cells (LMCR) had been injected into their peritoneal cavities. Three trocars (1 central camera port and 2 additional lateral ports) were introduced into the abdomen, and a pneumoperitoneum was created with carbon dioxide. Ten minutes after LMCR, cells were injected into the peritoneal cavity, the 2 lateral trocars were removed and carbon dioxide insufflation was maintained for an additional 5 minutes to allow for tumor cell seeding. Oxaliplatin (0.198 mg/kg) was then topically applied to 1 trocar site intramuscularly, while the other site was left untreated. One week later, the animals were euthanized, and the port sites were histologically examined for evidence of metastases. RESULTS: The rate of tumor implantation at the muscle layer in control sites was 68% (13/19) compared with 37% (7/19) at oxaliplatin-treated sites (P = 0.1). Also, no significant differences were detected in port-site metastasis rates in other untreated layers of the abdominal wall. CONCLUSION: Intramuscular topical application of oxaliplatin may not decrease the incidence of port-site metastasis in a syngeneic animal model of colon cancer. Nevertheless, we can see the tendency of declination. Further studies are needed to better determine its possible therapeutic role in high-risk humans undergoing laparoscopic resection of colorectal malignancies.     

Does the ultrasonically activated scalpel release viable airborne cancer cells?

Background: Viable cancer cells may implant at distant sites and cause tumor recurrence. One possible mechanism is the inadvertent exfoliation of viable tumor cells during dissection. The ultrasonically activated scalpel (UAS) uses ultrasonic energy to disrupt tissues by cavitation and produces a dense cloud of cellular debris that may contain viable cells. This study aimed to investigate the viability of airborne cells released during cancer dissection using the UAS and electrosurgery. Methods: Flank tumors (n= 8) measuring 1 cm³ were induced in male WAG rats by subcutaneous injection of 2 × 10⁶ CC531s colon cancer cells. Dissection was performed in cutting mode using the maximum power output of the respective devices. Electrosurgery was performed using a standard monopolar electrosurgical unit and a needle probe, and ultrasonic dissection was performed with the Harmonic Scalpel™ utilising the open surgical handset and the hooked spatula tip. The smoke plume was aspirated by a vacuum pump and bubbled under Hank's balanced salt solution to trap particulate matter. The viability of the cellular material was blindly assessed with the trypan blue test and by in vitro culture. The morphology of the cellular debris was studied by examination of cytospin preparations. Results: Large quantities of cellular debris was trapped in the plume from both devices. However, no viable cells were isolated, nor did in vitro cell growth occur with either device. Examination of the debris from the UAS demonstrated a characteristic mixture of amorphous forms and very few morphologically intact cells. The cauterized tumor produced charred cells and tissue fragments. Conclusions: In conclusion, this study demonstrates that viable airborne cancer cells are not released after tumor ablation with the UAS or electrosurgery. Received: 6 June 1997/Accepted: 20 November 1997     

In vitro study of multicellular hepatocyte spheroids formed in microcapsules

To make highly differentiated encapsulated hepatocytes, we formed hepatocyte spheroids in microcapsules in cultures. Hepatocytes isolated from rats were encapsulated in alginate-poly-L-lysine artificial cells and cultured under different medium conditions. When high molecular weight poly-L-lysine was used to make the capsule membrane, the hepatocytes aggregated and formed spheroids inside microcapsules within 48 hs. We studied the viability and function of encapsulated hepatocyte spheroids; free hepatocyte spheroids; nonaggregated encapsulated hepatocytes; and free hepatocyte monolayers. Cell viability and protein-producing ability were monitored for up to 30 days. Hepatocyte spheroids in the encapsulated or free form remained viable and continued to secrete proteins throughout the 30 days of observation. The viability and function of nonaggregated hepatocytes in the encapsulated or free form declined rapidly. These results suggest that the tridimensional structure of the spheroids may be important in maintaining the viability and function of encapsulated hepatocytes.     

High doses of taurolidine inhibit advanced intraperitoneal tumor growth in rats

BACKGROUND: The antitumor agent taurolidine (TRD) affects tumor growth in animals. Thus far, no animal studies have been published concerning the systemic or local toxicity and the effectiveness of long-term intraperitoneal (i.p.) and intravenous (i.v.) administration on advanced tumor growths. MATERIALS AND METHODS: In a first experiment (A) the systemic toxicity of the liver and kidneys was examined only after i.v. treatment in 40 rats (BD IX). For local toxicity the superior vena cava (SVC) was histologically analyzed. In a second study (B) 20,000 colon adenocarcinoma cells (DHD/K12/TRb) were initially applied i.p. after laparotomy in 80 rats (BD IX). After 28 days a port catheter system was placed in the SVC and left for 1 week. The animals were randomized into eight groups (n = 10) and received a 7-day treatment (eight hourly, 1 ml): 1, 2, 3% TRD or Ringer's solution (control group) either i.p. or i.v. Total i.p. tumor weight was measured 4 weeks after the end of the therapy. Side effects on differential blood counts and animal weight changes were examined. RESULTS: No organ lesions were detected in liver, kidneys, and SVC in experiment A. The i.v. administration of 2% TRD (P = 0.034) and 3% TRD (P = 0.05) as well the i.p. application of 2% TRD (P = 0.05) decreased the development of advanced i.p. tumor lesions. No changes of differential blood count nor relevant animal weight changes resulted. Three port catheter-related infections were examined. CONCLUSIONS: TRD does not impair the liver tissue, kidneys, SVC, and leucopoiesis. The intravenous therapy of 2% TRD is safe and anti-tumorigenic in advanced local tumor growth in rats.