The influence on abdominal adhesions and inflammation in rabbits after exposure to differently charged polypeptides

Daniel Åkerberg, Carl Grunditz, Monika Posaric-Bauden, Karolin Isaksson, Roland Andersson, Bobby Tingstedt

Department of Surgery, Clinical Sciences, Sk?ne University Hospital, Lund, Sweden.

DOI: 10.4236/jbise.2012.58055                

Abstract

Background: Abdominal adhesions develop on damaged peritoneal surfaces and constitute a significant health related problem. Previous animal studies have shown promising anti-adhesive effects when administering the polycation α-poly-L-lysine (αPL) and the polyanion poly-L-glutamate (PG) together. The objective of the study was to examine the effect of these differently charged polypeptides when administered by spraying and to evaluate any possible effect on fibrinolysis, fibrosis and inflammation. Methods: Rabbits were treated with PLPG after cecal abrasive surgery and analysis from peritoneal biopsies of active tPa/PAI-1 complex and from peritoneal fluid of IL-6 and active TGFb1 at day 0, 1, 4 and 10 were measured after surgery. Histological specimens were analyzed on day 10 regarding inflammation and fibrosis. Peritoneal adhesions were evaluated by adhesion score. All values were compared to the control group (NaCl). Results: PLPG-treated rabbits had a significant diminished adhesion score on day 10 as compared to the control group (p < 0.005). Significantly reduced collagen depositions on the peritoneum were seen in the PLPG group when evaluating the histological specimens (p < 0.05). No significant differences between the experimental and control groups were seen in peritoneal fluid when analyzing for active protein levels. Conclusion: This is the first study to investigate the effect on key parameters in adhesion formation as well as the preventive effect of the PLPG complex on abdominal adhesions in rabbits and also the first study where administration by spraying the polypeptides was used. PLPG was non-toxic in this setting and without significant differences in adhesion formation parameters and a significant reduction in adhesions was observed. This was verified both macroscopically and histologically.

Keywords

Polycations; Abdominal Adhesions; Coagulation; Fibrosis; Inflammation

Conflicts of Interest

The authors declare no conflicts of interest.

References

[1]	Tingstedt B, Andersson E, Isaksson K, Andersson R. (2008) Clinical impact of abdominal adhesions: what is the magnitude of the problem? Scand J Gastroenterol, 43, 255-61.
[2]	Tingstedt B, Isaksson J, Andersson R. (2007) Long-term follow-up and cost analysis following surgery for small bowel obstruction caused by intra-abdominal adhesions. Br J Surg, 94, 743-8.
[3]	Sulaiman H, Dawson L, Laurent GJ, Bellingan GJ, Herrick SE. (2002) Role of plasminogen activators in peritoneal adhesion formation. Biochem Soc Trans, 30, 126-31.
[4]	Liakakos T, Thomakos N, Fine PM, Dervenis C, Young RL. (2001) Peritoneal adhesions: etiology, pathophysiology, and clinical significance. Recent advances in prevention and management. Dig Surg, 18, 260-73.
[5]	Chegini N. (2022) Peritoneal molecular environment, adhesion formation and clinical implication. Front Biosci, 7, e91-115.
[6]	Di Filippo C, Falsetto A, De Pascale V, Tufariello E, De Lucia D, Rossi F. (2006) Plasma levels of t-PA and PAI-1 correlate with the formation of experimental post-surgical peritoneal adhesions. Mediators Inflamm, 4, 13901.
[7]	Thompson J. Pathogenesis and prevention of adhesion formation.(1998) Dig Surg, 15, 153-7.
[8]	Prushik SG, Stucchi AF, Matteotti R, Aarons CB, Reed KL, Gower AC. (2010) Open adhesiolysis is more effective in reducing adhesion reformation than laparoscopic adhesiolysis in an experimental model. Br J Surg., 97, 420-7.
[9]	Neudecker J, Junghans T, Raue W, Ziemer S, Schwenk W. (2005) Fibrinolytic capacity in peritoneal fluid after laparoscopic and conventional colorectal resection: data from a randomized controlled trial. Langenbecks Arch Surg, 390, 523-7.
[10]	Lucas PA, Warejcka DJ, Young HE, Lee BY. (1996) Formation of abdominal adhesions is inhibited by antibodies to transforming growth factor-beta1. J Surg Res, 65, 135-8.
[11]	Cheong YC, Laird SM, Shelton JB, Ledger WL, Li TC, Cooke ID. (2002) The correlation of adhesions and peritoneal fluid cytokine concentrations: a pilot study. Hum Reprod, 17, 1039-45.
[12]	Nehez L, Vodros D, Axelsson J, Tingstedt B, Lindman B, Andersson R. (2005) Prevention of postoperative peritoneal adhesions: effects of lysozyme, polylysine and polyglutamate versus hyaluronic acid. Scand J Gastroenterol, 40, 1118-23.
[13]	http://www.avma.org/issues/animal_welfare/euthanasia.pdf
[14]	Lang RA, Weisgerber C, Gruntzig PM, Weis C, Odermatt EK, Kirschner MH. (2009) Polyvinyl alcohol gel prevents adhesion reformation after adhesiolysis in a rabbit model. J Surg Res, 153, 12-6.
[15]	Hooker GD, Taylor BM, Driman DK. (1999) Prevention of adhesion formation with use of sodium hyaluronate-based bioresorbable membrane in a rat model of ventral hernia repair with polypropylene mesh-a ran-domized, controlled study. Surgery, 125, 211-6.
[16]	Nehez L, Tingstedt B, Vodros D, Axelsson J, Lindman B, Andersson R. (2006) Novel treatment in peritoneal adhesion prevention: protection by polypeptides. Scand J Gastroenterol, 41, 1110-7.
[17]	Nehez L, Tingstedt B, Axelsson J, Andersson R. (2007) Differently charged polypeptides in the prevention of post-surgical peritoneal adhesions. Scand J Gastroenterol, 42, 519-23.
[18]	Tingstedt B, Nehez L, Axelsson J, Lindman B, Andersson R. (2006) Increasing anastomosis safety and preventing abdominal adhesion formation by the use of polypeptides in the rat. Int J Colorectal Dis, 21, 566-72.
[19]	Tingstedt B, Nehez L, Lindman B, Andersson R. (2007) Effect of bioactive polypeptides on leaking large bowel anastomosis and intestines in the rat. J Invest Surg, 20, 229-35.
[20]	Tingstedt B, Nehez L, Lindman B, Andersson R. (2007) Efficacy of bioactive polypeptides on bleeding and intra-abdominal adhesions. Eur Surg Res, 39, 35-40.
[21]	Ar'Rajab A, Mileski W, Sentementes JT, Sikes P, Harris RB, Dawidson IJ. (1996) The role of neutrophils in peritoneal adhesion formation. J Surg Res, 61, 143-6.
[22]	Isaksson K, Akerberg D, Andersson R, Tingstedt B. (2010) Toxicity and dose response of intra-abdominally administered poly-L-alpha-lysine and poly-L-glutamate for postoperative adhesion protection. Eur Surg Res, 44, 17-22.
[23]	Renvall S, Lehto M, Penttinen R. (1987) Development of peritoneal fibrosis occurs under the mesothelial cell layer. J Surg Res, 43, 407-12.
[24]	Miyamoto T, Tamura M, Kabashima N, Serino R, Shibata T, Furuno Y. (2010) An integrin-activating peptide, PHSRN, ameliorates inhibitory effects of conventional peritoneal dialysis fluids on peritoneal wound healing. Nephrol Dial Transplant, 25, 1109-19.
[25]	van der Wal JB, Jeekel J. (2007) Biology of the peritoneum in normal homeostasis and after surgical trauma. Colorectal Dis, 9 Suppl 2, 9-13.
[26]	Imudia AN, Kumar S, Saed GM, Diamond MP. (2008) Pathogenesis of Intra-abdominal and pelvic adhesion development. Semin Reprod Med, 26, 289-97.
[27]	Mutsaers SE. (2008) The mesothelial cell. Int J Biochem Cell Biol, 36, 9-16.
[28]	Hellebrekers BW, Trimbos-Kemper GC, Bakkum EA, Trimbos JB, Declerck PJ, Kooistra T. (2000) Short-term effect of surgical trauma on rat peritoneal fibrinolytic activity and its role in adhesion formation. Thromb Haemost, 84, 876-81.
[29]	Ghellai AM, Stucchi AF, Chegini N, Ma C, Andry CD, Kaseta JM. (2004) Role of transforming growth factor beta-1 in peritonitis-induced adhesions. J Gastrointest Surg, 4, 316-23.
[30]	Saba AA, Kaidi AA, Godziachvili V, Dombi GW, Dawe EJ, Libcke JH. (1996) Effects of interleukin-6 and its neutralizing antibodies on peritoneal adhesion formation and wound healing. Am Surg, 62, 569-72.
[31]	Rougier JP, Guia S, Hagege J, Nguyen G, Ronco PM. (1998) PAI-1 secretion and matrix deposition in human peritoneal mesothelial cell cultures: transcriptional regulation by TGF-beta 1. Kidney Int, 54, 87-98.
[32]	Topley N, Jorres A, Luttmann W, Petersen MM, Lang MJ, Thierauch KH. (1993) Human peritoneal mesothelial cells synthesize interleukin-6: induction by IL-1 beta and TNF alpha. Kidney Int, 43, 226-33.
[33]	van Hinsbergh VW, Kooistra T, Scheffer MA, Hajo van Bockel J, van Muijen GN. (1990) Characterization and fibrinolytic properties of human omental tissue mesothelial cells. Comparison with endothelial cells. Blood, 75, 1490-7.
[34]	Freyria AM, Paul J, Belleville J, Broyer P, Eloy R. (1991) Rat peritoneal macrophage procoagulant and fibrinolytic activities. An expression of the local inflammatory response. Comp Biochem Physiol A Comp Physiol, 99, 517-24.
[35]	Ivarsson ML, Holmdahl L, Falk P, Molne J, Risberg B. (1998) Characterization and fibrinolytic properties of mesothelial cells isolated from peritoneal lavage. Scand J Clin Lab Invest, 58, 195-203.