Top

International Journal of Colorectal Disease

Published in:

01-04-2009 | Original Article

Intraperitoneally applied gentamicin increases collagen content and mechanical stability of colon anastomosis in rats

Published in: International Journal of Colorectal Disease | Issue 4/2009

Abstract

Background

Anastomotic leakage remains a serious complication in colorectal surgery, and is being caused by a multitude of factors. Recent reports reveal changes of the extracellular matrix as risk factors as well as gentamicin as a potential agent to influence wound healing. This experimental study was initiated to investigate the influence of intraperitoneally applied gentamicin on colonic anastomotic wound healing and in particular on mechanical stability, overall collagen content and collagen type I/III ratio.

Materials and methods

Sixty Sprague Dawley rats were randomized to one of two groups. In each animal, a standard transverse colonic end-to-end anastomosis was performed. Immediately postoperative, either 5 ml gentamicin (1 ml/kg bodyweight) or NaCl 0.9% was applied intraperitoneally. On postoperative days 3, 5, and 14, ten of the animals in each group were sacrificed. Measurements of the anastomosis bursting pressure were performed on postoperative days 3 and 5. At each explantation time, the collagen per protein ratio, the collagen types I/III ratio, and both the expression of MMP-2, -9, and Ki67 were analyzed.

Results

None of the animals died. None of the rats exhibited clinical evidence of anastomotic leakage. The bursting strength in the gentamicin group was significantly elevated on postoperative day 5. Both the overall collagen content and the collagen type I/III ratio in the gentamicin group were significantly increased 3, 5, and 14 days postoperatively compared to the control group. The expression of MMP-9 was significantly elevated in the gentamicin group both 3 and 5 days postoperatively. In contrast, there were no significant differences in the expression of MMP-9 14 days postoperatively. All investigated samples demonstrated positive staining for MMP-2 and Ki67 without statistically significant differences at any term, respectively.

Conclusions

The present data confirm that intraperitoneally applied gentamicin is able to enhance healing and stability of colonic anastomosis due to an increase of both the overall collagen content and collagen type I/III ratio.

Branagan G, Finnis D (2005) Prognosis after anastomotic leakage in colorectal surgery. Dis Colon Rectum 48:1021–1026PubMedCrossRef

McArdle CS, McMillan DC, Hole DJ (2005) Impact of anastomotic leakage on long-term survival of patients undergoing curative resection for colorectal cancer. Br J Surg 92:1150–1154PubMedCrossRef

Alves A, Panis Y, Trancart D, Regimbeau JM, Pocard M, Valleur P (2002) Factors associated with clinically significant anastomotic leakage after large bowel resection: multivariate analysis of 707 patients. World J Surg 26:499–502PubMedCrossRef

Schmidt O, Merkel S, Hohenberger W (2003) Anastomotic leakage after low rectal stapler anastomosis: significance of intraoperative anastomotic testing. Eur J Surg Oncol 29:239–243PubMedCrossRef

Kasperk R, Philipps B, Vahrmeyer M, Willis S, Schumpelick V (2000) Risk factors for anastomosis dehiscence after very deep colorectal and coloanal anastomosis. Chirurg 71:1365–1369PubMedCrossRef

Stumpf M, Klinge U, Wilms A, Zabrocki R, Rosch R, Junge K et al (2005) Changes of the extracellular matrix as a risk factor for anastomotic leakage after large bowel surgery. Surgery 137:229–234PubMedCrossRef

Onodera H, Ikeuchi D, Nagayama S, Imamura M (2004) Weakness of anastomotic site in diabetic rats is caused by changes in the integrity of newly formed collagen. Dig Surg 21:146–151PubMedCrossRef

10.

Grassl GA, Valdez Y, Bergstrom KS, Vallance BA, Finlay BB (2008) Chronic enteric salmonella infection in mice leads to severe and persistent intestinal fibrosis. Gastroenterology 134:768–780PubMedCrossRef

11.

Rico RM, Ripamonti R, Burns AL, Gamelli RL, DiPietro LA (2002) The effect of sepsis on wound healing. J Surg Res 102:193–197PubMedCrossRef

12.

Hopkinson I, Evans W, Chant D, Hiscox S, Berry D, Harding K (1995) Reverse transcription-polymerase chain reaction detection of collagen transcripts in healing human wounds. Eur J Clin Invest 25:539–542PubMedCrossRef

13.

Gilbert AI, Felton LL (1993) Infection in inguinal hernia repair considering biomaterials and antibiotics [published erratum appears in Surg Gynecol Obstet 1993 Nov;177(5):528]. Surg-Gynecol-Obstet 177:126–130PubMed

14.

Burton RC, Hughes ES, Cuthbertson AM (1975) Prophylactic use of gentamicin in colonic and rectal surgery. Med J Aust 2:597–599PubMed

15.

Tekos A, Prodromaki E, Papadimou E, Pavlidou D, Tsambaos D, Drainas D (2003) Aminoglycosides suppress tRNA processing in human epidermal keratinocytes in vitro. Skin Pharmacol Appl Skin Physiol 16:252–258PubMed

16.

Sacha PT, Zaremba ML, Jakoniuk P (1999) The influence of antibiotics on phagocytic and bacteriocidal activity of rabbit peritoneal macrophages stimulated by filtrates of cultured t-lymphocytes. Med Dosw Mikrobiol 51:399–412PubMed

17.

Wrzesniok D, Buszman E, Karna E, Nawrat P, Palka J (2002) Melanin potentiates gentamicin-induced inhibition of collagen biosynthesis in human skin fibroblasts. Eur J Pharmacol 446:7–13PubMedCrossRef

18.

Junge K, Klinge U, Rosch R, Lynen P, Binnebosel M, Conze J et al (2007) Improved collagen type I/III ratio at the interface of gentamicin-supplemented polyvinylidenfluoride mesh materials. Langenbecks Arch Surg 392:465–471PubMedCrossRef

19.

Remmele W, Stegner HE (1987) Recommendation for uniform definition of an immunoreactive score (IRS) for immunohistochemical estrogen receptor detection (ER-ICA) in breast cancer tissue. Pathologe 8:138–140PubMed

20.

Lopez-De Leon A, Rojkind M (1985) A simple micromethod for collagen and total protein determination in formalin-fixed paraffin-embedded sections. J Histochem Cytochem 33:737–743PubMedCrossRef

21.

Junqueira LC, Cossermelli W, Brentani R (1978) Differential staining of collagens type I, II and III by Sirius Red and polarization microscopy. Arch Histol Jpn 41:267–274PubMed

22.

de HI, Siemonsma MA, de Man BM, Lomme RM, Hendriks T (2002) The matrix metalloproteinase inhibitor BB-94 improves the strength of intestinal anastomoses in the rat. Int J Colorectal Dis 17:348–354CrossRef

23.

Oxlund H, Christensen H, Seyer-Hansen M, Andreassen TT (1996) Collagen deposition and mechanical strength of colon anastomoses and skin incisional wounds of rats. J Surg Res 66:25–30PubMedCrossRef

24.

Junge K, Klinge U, Rosch R, Mertens PR, Kirch J, Klosterhalfen B et al (2004) Decreased collagen type I/III ratio in patients with recurring hernia after implantation of alloplastic prostheses. Langenbecks Arch Surg 389:17–22PubMedCrossRef

25.

Si Z, Bhardwaj R, Rosch R, Mertens PR, Klosterhalfen B, Klinge U (2002) Impaired balance of type I and type III procollagen mRNA in cultured fibroblasts of patients with incisional hernia. Surgery 131:324–331PubMedCrossRef

26.

Oxlund H, Christensen H, Seyer-Hansen M, Andreassen TT (1996) Collagen deposition and mechanical strength of colon anastomoses and skin incisional wounds of rats. J Surg Res 66:25–30PubMedCrossRef

27.

Christensen H, Oxlund H, Laurberg S (1991) Postoperative biosynthetic human growth hormone increases the strength and collagen deposition of experimental colonic anastomoses. Int J Colorectal Dis 6:133–138PubMedCrossRef

28.

Christensen H, Oxlund H (1994) Growth hormone increases the collagen deposition rate and breaking strength of left colonic anastomoses in rats. Surgery 116:550–556PubMed

29.

Cheng M, Razzaque MS, Nazneen A, Taguchi T (1998) Expression of the heat shock protein 47 in gentamicin-treated rat kidneys. Int J Exp Pathol 79:125–132PubMedCrossRef

30.

Razzaque MS, Taguchi T (1997) Collagen-binding heat shock protein (HSP) 47 expression in anti-thymocyte serum (ATS)-induced glomerulonephritis. J Pathol 183:24–29PubMedCrossRef

31.

Cheng M, Razzaque MS, Nazneen A, Taguchi T (1998) Expression of the heat shock protein 47 in gentamicin-treated rat kidneys. Int J Exp Pathol 79:125–132PubMedCrossRef

32.

Razzaque MS, Taguchi T (1997) Collagen-binding heat shock protein (HSP) 47 expression in anti-thymocyte serum (ATS)-induced glomerulonephritis. J Pathol 183:24–29PubMedCrossRef

33.

Vaneerdeweg W, Hendriks JM, Lauwers PR, Ieven M, Eyskens EJ (2000) Effect of gentamicin-containing sponges on the healing of colonic anastomoses in a rat model of peritonitis. Eur J Surg 166:959–962PubMedCrossRef

34.

Asch HL, Farnham PJ (1978) Effects of gentamicin on trypsin, chymotrypsin, and collagenase. J Infect Dis 138:257–259PubMedCrossRef

35.

36.

Jansen PL, Rosch R, Jansen M, Binnebosel M, Junge K, Alfonso-Jaume A et al (2007) Regulation of MMP-2 gene transcription in dermal wounds. J Invest Dermatol 127:1762–1767PubMed

37.

Inkinen K, Turakainen H, Wolff H, Ravanti L, Kahari VM, Ahonen J (2000) Expression and activity of matrix metalloproteinase-2 and -9 in experimental granulation tissue. APMIS 108:318–328PubMedCrossRef

38.

Ravanti L, Kahari VM (2000) Matrix metalloproteinases in wound repair (review). Int J Mol Med 6:391–407PubMed

39.

de Hingh HI, Siemonsma MA, de Man BM, Lomme RM, Hendriks T (2002) The matrix metalloproteinase inhibitor BB-94 improves the strength of intestinal anastomoses in the rat. Int J Colorectal Dis 17:348–354PubMedCrossRef

40.

Savage FJ, Lacombe DL, Hembry RM, Boulos PB (1998) Effect of colonic obstruction on the distribution of matrix metalloproteinases during anastomotic healing. Br J Surg 85:72–75PubMedCrossRef

41.

Savage FJ, Lacombe DL, Boulos PB, Hembry RM (1997) Role of matrix metalloproteinases in healing of colonic anastomosis. Dis Colon Rectum 40:962–970PubMedCrossRef

42.

Syk I, Agren MS, Adawi D, Jeppsson B (2001) Inhibition of matrix metalloproteinases enhances breaking strength of colonic anastomoses in an experimental model. Br J Surg 88:228–234PubMedCrossRef

43.

Somerville RP, Oblander SA, Apte SS (2003) Matrix metalloproteinases: old dogs with new tricks. Genome Biol 4:216PubMedCrossRef

44.

Seifert WF, Wobbes T, Hendriks T (1996) Divergent patterns of matrix metalloproteinase activity during wound healing in ileum and colon of rats. Gut 39:114–119PubMedCrossRef

45.

46.

Zhu X, Hu C, Zhang Y, Li L, Wang Z (2001) Expression of cyclin-dependent kinase inhibitors, p21cip1 and p27kip1, during wound healing in rats. Wound Repair Regen 9:205–212PubMedCrossRef

47.

Title: Intraperitoneally applied gentamicin increases collagen content and mechanical stability of colon anastomosis in rats
Publication date: 01-04-2009
Published in: International Journal of Colorectal Disease / Issue 4/2009
Print ISSN: 0179-1958
Electronic ISSN: 1432-1262
DOI: https://doi.org/10.1007/s00384-008-0614-x

Keynote webinar | Spotlight on medication adherence

Springer Medicine

Intraperitoneally applied gentamicin increases collagen content and mechanical stability of colon anastomosis in rats

Abstract

Background

Materials and methods

Results

Conclusions

Keynote webinar | Spotlight on medication adherence

Springer Medicine

Abstract

Background

Materials and methods

Results

Conclusions

Please log in to get access to this content

Other articles of this Issue 4/2009

Enterocolic lymphocytic phlebitis and lymphocytic colitis: drug-related coexistent pathology

Aberrant crypt foci: endoscopic assessment and cell kinetics characterization

Hepatectomy for liver metastasis of colorectal cancer

Acute pancolitis after colonoscopy in liver cirrhosis patient

Recurrent volvulus of the transverse colon after sigmoid resection

Occurrence and prognostic value of circumferential resection margin involvement for patients with rectal cancer