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The effect of DNA topoisomerase inhibitor-loaded films on postsurgical adhesion formation in the rat
oxidant are associated with side effects of inflammation, such as cell injury and tissue damage. In view of these observations the objective of this study is to determine whether there is a difference in the expression levels of MPO in fibroblasts isolated from normal peritoneum and from adhesions, and whether this expression is modulated by hypoxia. DESIGN: Comparison of fibroblast primary cultures from normal peritoneum and adhesion tissues of the same patients. MATERIALS AND METHODS: We have obtained fibroblast primary cultures from normal peritoneum and adhesion tissues of the same patients. MPO synthesis by these fibroblasts was evaluated at mRNA utilizing the real-time RT/PCR and at the protein levels utilizing MPO ELISA kit. Fibroblasts were cultured under normal (20% O2) and hypoxic (2% O2) condition for 24 h. RESULTS: The studies herein define normal fibroblasts to have higher MPO mRNA (32 %; p⬍0.001) and protein (28%; p⬍0.001) levels than adhesion fibroblasts. Hypoxia decreased MPO mRNA by (35 %; p⬍0.001) and protein levels by (40%; p⬍0.001) in normal peritoneal fibroblasts to approximately of levels observed for adhesion fibroblasts while has no effects on MPO expression in adhesion fibroblasts. CONCLUSION: Our data suggests that hypoxia drive normal fibroblasts to mimic the adhesion fibroblasts by reducing their ability to produce MPO. Induction of MPO in adhesion fibroblasts to levels seen in normal fibroblasts may be important for the elimination of the adhesion phenotype during peritoneal healing. Supported by: NIH RO1 HL 66367
Wednesday, October 20, 2004 2:30 P.M. O-271 Lactocapromer in membrane (XA-1-M)- and foil application (XA1-F) serving as “artificial peritoneum” for adhesion prevention versus established “adhesion barriers” Interceed© (ORC), Adept© (IDT) and lactated ringer’s solution (lRS) in an experimental animal study. M. Wallwiener, R. Kurek, C. Wallwiener, D. Wallwiener, H. Planck, J. Mewald. University of Tuebingen, Tuebingen, Germany; German Center of Excellence on Biomaterials and Organ (BMOZ), Denkendorf, Germany. OBJECTIVE: Peritoneal adhesions, as a consequence of healing processes, are still a significant factor in postoperative complications. About 80 –90% of abdominal adhesions can be traced back to prior surgeries. Pelvis Surgery remains on of the most frequently causes for postoperative adhaesions, leading to intestinal obstruction, chronic pelvic pain, female infertility and complicated following surgeries. The objection was to determine their benefit in adhesion prophylaxis. The novel“barriers”XA-1-M and XA-1-F were evaluated in a prospective, randomised study with four study groups of female Wistar rats (each n ⫽ 18) and a control group (lRS, n ⫽ 9) in comparison to established adhesion prophylaxis in clinical routine. DESIGN: Randomized, experimental prospective animal study with Wistar rats (80), intraindividual and interindividual comparison. MATERIALS AND METHODS: After artificial induction of the adhesions via standardized traumatization of the parietal and visceral peritoneum (using a Perlon brush technique), the alloplastic barriers were applicated intraperitoneal after randomisation. The adhesion formation after traumatization of the peritoneum was evaluated four weeks postoperatively at 2nd-look laparotomy according to an established adhesion score. The following criteria were analysed: area covered with adhesions, quality and tenacity of the adhesions at parietal and visceral peritoneum. The Chisquare test was applied for data analysis. RESULTS: The induction of adhesions could significantly be reduced with the novel “barrier” XA-1-M in membrane application and established “barriers” such as IDT and ORC likewise, showing equally promising effects, whereas the outcome in adhesion prevention and efficacy of the XA-1-F foil and artificial ascites (lRS) were marginal. Whereas in the control group, only 11% of the animals developed no adhesions, Interceed (67%), Adept (65%) and the membrane (61%) prevented the formation of adhesion in over 60% of the animals. Interceed, Adept and XA-1-M also demonstrated a significant cut back in the formation of adhesions type 3– 4 to 11% / 0% / 17% compared to 44% of the adhesions in the control group. CONCLUSION: The results are implicating the rationale for further
FERTILITY & STERILITY威
clinical trials to gain more detailed data for the implementation in daily clinical usage. Supported by: None
Wednesday, October 20, 2004 2:45 P.M. O-272 The effect of DNA topoisomerase inhibitor-loaded films on postsurgical adhesion formation in the rat. J. D. Cashman, C. M. Springate, H. M. Burt, J. K. Jackson. ARC Pharmaceutics Inc., Vancouver, BC, Canada; The University of British Columbia, Vancouver, BC, Canada. OBJECTIVE: Postsurgical adhesion formation is a leading cause of infertility in women of reproductive age. Adhesions occur when trauma to the peritoneum initiates an inflammatory cascade that eventually culminates in fibroblast proliferation, deposition of collagen and the formation of dense fibrous bands between affected organs. We have previously shown that the potent antiproliferative agent camptothecin, when loaded into hyaluronic acid (HA) film, reduces both the strength and extent of postsurgical adhesions in the rat cecal sidewall model. This drug belongs to the general class of compounds termed DNA topoisomerase inhibitors, which also includes antineoplastic agents such as etoposide, menadione, and -lapachone. The aim of this study was to examine the potential of DNA topoisomerase inhibitor-loaded HA films for the prevention of postsurgical adhesion formation. DESIGN: Prospective randomized study using the cecal sidewall model in rats. MATERIALS AND METHODS: The antiadhesion properties of the DNA topoisomerase inhibitors etoposide, menadione, -lapachone, and camptothecin were investigated by application of carbodiimide crosslinked HA films containing the drug at a concentration of 7.5 % w/w at the site of surgical injury in a total of 42 rats. Control animals received no film or a HA only film. Seven days after surgery the animals were sacrificed and an adhesion score for each animal was obtained based on the strength and extent of the adhesions. Potential toxicity was determined by changes in white blood cell (WBC) count and weight of the animals. Statistical analysis was carried out using Student’s t-test and p values less than 0.05 were considered significant. RESULTS: In this model the application of HA films alone or HA films loaded with 7.5% w/w etoposide, menadione, -lapachone, or camptothecin had a significant effect in reducing the mean adhesion score (6.1 ⫾ 2.9, 6.4 ⫾ 2.7, 3.8 ⫾ 3.3, 5.6 ⫾ 3.2, 1.9 ⫾ 1.1, respectively) when compared to those animals in which no film was placed (9.6 ⫾ 2.0). In addition, the use of camptothecin-loaded films resulted in a significant reduction in adhesion formation when compared to HA films alone. No decrease in WBC counts or weight was observed at the time of assessment and no effect on wound healing was evident in any of the treatment groups. CONCLUSION: The application of carbodiimide crosslinked HA films to the surgical site in the rat cecal sidewall model resulted in a significant decrease in adhesion formation. At the dose studied, camptothecin-loaded films had a significant antiadhesion effect over that observed for HA films alone or the other DNA topoisomersase inhibitor-loaded HA films examined in this study. These experiments confirm the potential of camptothecinloaded HA films for the prevention of postsurgical adhesion formation. Supported by: ARC Pharmaceuticals Inc.
Wednesday, October 20, 2004 3:00 P.M. O-273 Expression of macrophage stimulating protein and its receptor in peritoneal mesothelial cells during a carbon dioxide pneumoperitoneum. S. Matsuzaki, M. Canis, J.-L. Pouly, B. Rabischong, R. Botchorishvili, G. Mage. Polyclinique de l’Hotel-Dieu, CHU, Clermont-Ferrand, France. OBJECTIVE: Laparoscopic surgery is considered to be less adhesiogenic than laparotomy. However, despite the increased use of laparoscopic surgeries, the incidence of intraperitoneal adhesions and the rate of adhesiolysis have remained constant. Animal experimental studies suggest that CO2
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Wednesday, October 20, 2004 2:30 P.M. O-271 Lactocapromer in membrane (XA-1-M)- and foil application (XA1-F) serving as “artificial peritoneum” for adhesion prevention versus established “adhesion barriers” Interceed© (ORC), Adept© (IDT) and lactated ringer’s solution (lRS) in an experimental animal study. M. Wallwiener, R. Kurek, C. Wallwiener, D. Wallwiener, H. Planck, J. Mewald. University of Tuebingen, Tuebingen, Germany; German Center of Excellence on Biomaterials and Organ (BMOZ), Denkendorf, Germany. OBJECTIVE: Peritoneal adhesions, as a consequence of healing processes, are still a significant factor in postoperative complications. About 80 –90% of abdominal adhesions can be traced back to prior surgeries. Pelvis Surgery remains on of the most frequently causes for postoperative adhaesions, leading to intestinal obstruction, chronic pelvic pain, female infertility and complicated following surgeries. The objection was to determine their benefit in adhesion prophylaxis. The novel“barriers”XA-1-M and XA-1-F were evaluated in a prospective, randomised study with four study groups of female Wistar rats (each n ⫽ 18) and a control group (lRS, n ⫽ 9) in comparison to established adhesion prophylaxis in clinical routine. DESIGN: Randomized, experimental prospective animal study with Wistar rats (80), intraindividual and interindividual comparison. MATERIALS AND METHODS: After artificial induction of the adhesions via standardized traumatization of the parietal and visceral peritoneum (using a Perlon brush technique), the alloplastic barriers were applicated intraperitoneal after randomisation. The adhesion formation after traumatization of the peritoneum was evaluated four weeks postoperatively at 2nd-look laparotomy according to an established adhesion score. The following criteria were analysed: area covered with adhesions, quality and tenacity of the adhesions at parietal and visceral peritoneum. The Chisquare test was applied for data analysis. RESULTS: The induction of adhesions could significantly be reduced with the novel “barrier” XA-1-M in membrane application and established “barriers” such as IDT and ORC likewise, showing equally promising effects, whereas the outcome in adhesion prevention and efficacy of the XA-1-F foil and artificial ascites (lRS) were marginal. Whereas in the control group, only 11% of the animals developed no adhesions, Interceed (67%), Adept (65%) and the membrane (61%) prevented the formation of adhesion in over 60% of the animals. Interceed, Adept and XA-1-M also demonstrated a significant cut back in the formation of adhesions type 3– 4 to 11% / 0% / 17% compared to 44% of the adhesions in the control group. CONCLUSION: The results are implicating the rationale for further
FERTILITY & STERILITY威
clinical trials to gain more detailed data for the implementation in daily clinical usage. Supported by: None
Wednesday, October 20, 2004 2:45 P.M. O-272 The effect of DNA topoisomerase inhibitor-loaded films on postsurgical adhesion formation in the rat. J. D. Cashman, C. M. Springate, H. M. Burt, J. K. Jackson. ARC Pharmaceutics Inc., Vancouver, BC, Canada; The University of British Columbia, Vancouver, BC, Canada. OBJECTIVE: Postsurgical adhesion formation is a leading cause of infertility in women of reproductive age. Adhesions occur when trauma to the peritoneum initiates an inflammatory cascade that eventually culminates in fibroblast proliferation, deposition of collagen and the formation of dense fibrous bands between affected organs. We have previously shown that the potent antiproliferative agent camptothecin, when loaded into hyaluronic acid (HA) film, reduces both the strength and extent of postsurgical adhesions in the rat cecal sidewall model. This drug belongs to the general class of compounds termed DNA topoisomerase inhibitors, which also includes antineoplastic agents such as etoposide, menadione, and -lapachone. The aim of this study was to examine the potential of DNA topoisomerase inhibitor-loaded HA films for the prevention of postsurgical adhesion formation. DESIGN: Prospective randomized study using the cecal sidewall model in rats. MATERIALS AND METHODS: The antiadhesion properties of the DNA topoisomerase inhibitors etoposide, menadione, -lapachone, and camptothecin were investigated by application of carbodiimide crosslinked HA films containing the drug at a concentration of 7.5 % w/w at the site of surgical injury in a total of 42 rats. Control animals received no film or a HA only film. Seven days after surgery the animals were sacrificed and an adhesion score for each animal was obtained based on the strength and extent of the adhesions. Potential toxicity was determined by changes in white blood cell (WBC) count and weight of the animals. Statistical analysis was carried out using Student’s t-test and p values less than 0.05 were considered significant. RESULTS: In this model the application of HA films alone or HA films loaded with 7.5% w/w etoposide, menadione, -lapachone, or camptothecin had a significant effect in reducing the mean adhesion score (6.1 ⫾ 2.9, 6.4 ⫾ 2.7, 3.8 ⫾ 3.3, 5.6 ⫾ 3.2, 1.9 ⫾ 1.1, respectively) when compared to those animals in which no film was placed (9.6 ⫾ 2.0). In addition, the use of camptothecin-loaded films resulted in a significant reduction in adhesion formation when compared to HA films alone. No decrease in WBC counts or weight was observed at the time of assessment and no effect on wound healing was evident in any of the treatment groups. CONCLUSION: The application of carbodiimide crosslinked HA films to the surgical site in the rat cecal sidewall model resulted in a significant decrease in adhesion formation. At the dose studied, camptothecin-loaded films had a significant antiadhesion effect over that observed for HA films alone or the other DNA topoisomersase inhibitor-loaded HA films examined in this study. These experiments confirm the potential of camptothecinloaded HA films for the prevention of postsurgical adhesion formation. Supported by: ARC Pharmaceuticals Inc.
Wednesday, October 20, 2004 3:00 P.M. O-273 Expression of macrophage stimulating protein and its receptor in peritoneal mesothelial cells during a carbon dioxide pneumoperitoneum. S. Matsuzaki, M. Canis, J.-L. Pouly, B. Rabischong, R. Botchorishvili, G. Mage. Polyclinique de l’Hotel-Dieu, CHU, Clermont-Ferrand, France. OBJECTIVE: Laparoscopic surgery is considered to be less adhesiogenic than laparotomy. However, despite the increased use of laparoscopic surgeries, the incidence of intraperitoneal adhesions and the rate of adhesiolysis have remained constant. Animal experimental studies suggest that CO2
S109