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Expression and inhibition of matrix metalloproteinase (MMP)-8, MMP-9 and MMP-12 in early colonic anastomotic repair

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Abstract

Purpose

Submucosal collagen is paramount for colonic anastomotic integrity. Matrix metalloproteinases (MMPs) mediate collagen degradation that increases the risk of wound dehiscence. Although broad-spectrum MMP inhibitors are beneficial for anastomotic strength, they can cause adverse reactions. Knowledge of specific MMPs responsible for the weakening of anastomoses can be used to optimise MMP inhibition therapy. We aimed to quantify transcript and protein levels of multiple MMPs in colonic anastomoses and evaluate the effect of inhibiting the MMPs that displayed the highest expression levels on anastomotic repair.

Methods

Left-sided colonic anastomoses were made in male Sprague-Dawley rats. After 3 days when biomechanical strength is lowest, MMP mRNA and protein levels were measured by quantitative real-time polymerase chain reaction, enzyme-linked immunosorbent assays and gelatin zymography. The effects of the MMP-8, MMP-9 and MMP-12 synthetic inhibitor AZD3342 was also studied.

Results

MMP-8, MMP-9 and MMP-12 gene and protein expression increased profoundly (p < 0.01), and MMP-13 mRNA and MMP-2 mRNA and protein modestly (p < 0.001) in the anastomoses. MMP-3 mRNA levels were not up-regulated significantly compared with adjacent uninjured colon. Increased anastomotic MMP-12 levels paralleled macrophage infiltration by immunohistochemical analyses. AZD3342 (50 mg/kg) treatment increased the anastomotic breaking strength by 29 % (p = 0.015) day 3 compared with vehicle. Improved anastomotic strength was not accompanied with alterations of type I or type III procollagen mRNA but was possibly due to inhibition of the concerted digestive action on the existent submucosal collagens by the targeted MMPs.

Conclusion

The present findings justify the concept of selective MMP inhibition to enhance anastomotic strength in colon.

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Acknowledgments

We thank Peter Schjerling for his advice on the mRNA measurements and Karen Bonde Larsen for help with microphotography. All materials and utilities were funded by the I.M. Dæhnfeldt Foundation, The A.P. Møller Foundation, The Danish Medical Association and AstraZeneca, Sweden. The authors have no disclosers to declare, except M. B. H. who is currently employed by AstraZeneca.

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Authors and Affiliations

  1. Department of Surgery K, Bispebjerg Hospital, Bispebjerg Bakke 23, DK 2400, Copenhagen NV, Denmark

    Peter-Martin Krarup, Lars Nannestad Jorgensen, Mark Berner Hansen & Magnus S. Ågren

  2. Department of Pathology, Aalborg Hospital, Aalborg, Denmark

    Mikkel Eld

  3. Institute of Sports Medicine, Department of Orthopedic Surgery M81, Bispebjerg Hospital and Centre of Healthy Ageing, Faculty of Health and Medical Sciences, University of Copenhagen, Copenhagen, Denmark

    Katja Heinemeier

  4. AstraZeneca Research and Development, Mölndal, Sweden

    Mark Berner Hansen

  5. Copenhagen Wound Healing Center, Bispebjerg Hospital, Copenhagen, Denmark

    Magnus S. Ågren

  6. Department of Diagnostic Sciences, Faculty of Health and Medical Sciences, University of Copenhagen, Copenhagen, Denmark

    Magnus S. Ågren

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  1. Peter-Martin Krarup
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Correspondence to Peter-Martin Krarup.

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Krarup, PM., Eld, M., Heinemeier, K. et al. Expression and inhibition of matrix metalloproteinase (MMP)-8, MMP-9 and MMP-12 in early colonic anastomotic repair. Int J Colorectal Dis 28, 1151–1159 (2013). https://doi.org/10.1007/s00384-013-1697-6

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