Alpha-1-antitrypsin improves anastomotic healing in intestinal epithelial cells model
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Published on: January 17, 2019
Nagoud Schukfeh1, Kokilavani Sivaraman2,3, Aileen Schmidt3,4
1Department of Pediatric Surgery, Hannover Medical School, Hannover, Germany. schukfeh.nagoud@mh-hannover.de.
Insights
Human alpha1-antitrypsin (AAT) significantly accelerates intestinal cell healing and proliferation in vitro. This suggests AAT may be a potential therapeutic agent to improve anastomotic healing in pediatric surgery.
Area of Science:
- Pediatric Surgery
- Gastroenterology
- Wound Healing Research
Background:
- Intestinal anastomosis is a common pediatric surgical procedure.
- Anastomotic leakage is a significant complication.
- Human alpha1-antitrypsin (AAT) promotes wound healing and cell proliferation.
Purpose of the Study:
- To investigate the potential of AAT in enhancing intestinal anastomotic healing.
- To test the hypothesis that AAT improves anastomotic healing.
Main Methods:
- Utilized HIEC-6 intestinal epithelial cells in a monolayer.
- Created a scratch assay and treated cells with AAT (0.5 and 1 mg/ml).
- Assessed cell proliferation and gene expression (qRT-PCR) using a Life-Cell Imaging System.
Main Results:
- AAT significantly accelerated scratch closure in HIEC-6 cells.
- Cells treated with 1 mg/ml AAT showed 97% repopulation after 18 hours vs. 60% for controls.
- AAT increased proliferation and upregulated CDKN1A, CDKN2A, ANGPTL4, WNT3, and COL3A1 mRNA levels.
Conclusions:
- AAT at physiological concentrations accelerates intestinal cell confluence and proliferation.
- Local AAT administration shows therapeutic potential for improving anastomotic healing.
Purpose:
Intestinal anastomosis is a routine procedure in pediatric surgery, with leakage being a significant complication. Human alpha1-antitrypsin (AAT), whose physiological serum concentrations range from 0.9-2.0 mg/ml, is known to accelerate wound healing and stimulate the expression of cell proliferation-related genes. We hypothesized that AAT might enhance anastomotic healing.
Methods:
In a monolayer of non-tumorigenic HIEC-6 epithelial cells derived from fetal intestine a scratch was created. Standard medium without (control) or with AAT (0.5 and 1 mg/ml) was added. Cells were observed using a Life-Cell Imaging System. Cell proliferation was assessed, and the expression of proliferation-related genes was measured by qRT-PCR.
Results:
In the presence of AAT, the scratch closed significantly faster. Cells treated with 1 mg/ml AAT showed 53% repopulation after 8 h and 97% after 18 h, while control cells showed 24% and 60% repopulation, respectively (p < 0.02). The treatment with AAT induced HIEC-6-cell proliferation and significantly increased the mRNA-expression of CDKN1A, CDKN2A, ANGPTL4, WNT3 and COL3A1 genes. AAT did not change the mRNA-expression of CXCL8 but decreased levels of IL-8 as compared to controls.
Conclusion:
At physiological concentrations AAT accelerates the confluence of intestinal cells and increases cell proliferation. The local administration of AAT may bear therapeutic potential to improve anastomotic healing.
