Intestinal Anastomotic Healing Research Articles

Role of the serosa in intestinal anastomotic healing: insights from in-depth histological analysis of human and murine anastomoses.

Anastomotic leakage following colorectal surgery remains a significant complication despite advances in surgical techniques. Recent findings on serosal injury repair in coelomic cavities, such as the peritoneum, challenge the current understanding of the cellular origins and mechanisms underlying intestinal anastomotic healing. Understanding the contribution of each layer of the intestinal wall during anastomotic healing is needed to find new therapeutic strategies to prevent anastomotic leakage. The aim of this experimental study was to investigate the role of the serosal layer of the intestinal wall in anastomotic healing. Comprehensive histologic analysis of human and murine anastomoses was performed to elucidate histologic changes in the different intestinal layers during anastomotic healing. In vivo staining of the extracellular matrix (ECM) in the serosal layer was performed using a fluorophore-conjugated N-hydroxysuccinimide-ester before anastomosis surgery in a murine model. Histological examination of both human and murine anastomoses revealed that closure of the serosal layer occurred first during the healing process. In vivo serosal ECM staining demonstrated that a significant portion of the newly formed ECM within the anastomosis was indeed deposited onto the serosal layer. Furthermore, mesenchymal cells within the anastomotic scar were positive for mesothelial cell markers, podoplanin and Wilms tumour protein. In this experimental study, the results suggest that serosal scar formation is an important mechanism for anastomotic integrity in intestinal anastomoses. Mesothelial cells may significantly contribute to scar formation during anastomotic healing through epithelial-to-mesenchymal transition, potentially suggesting a novel therapeutic target to prevent anastomotic leakage by enhancing physiological healing processes.

Short cycles of remote ischemic preconditioning had no effect on tensile strength in small intestinal anastomoses: an experimental animal study

PurposeThis study aimed to investigate the effect of remote ischemic preconditioning (RIPC) on the healing of small intestinal anastomoses, evaluated by tensile strength and histologic wound healing on postoperative day 5. MethodsA total of 22 female pigs were randomized 1:1 into either an intervention or control group. The intervention group received 5 cycles of 3-minute ischemia followed by 3-minute reperfusion on the right forelimb. Two end-to-end anastomoses, a distal and a proximal, were created in the small intestine 30 and 60 min after RIPC, respectively. On postoperative day 5, the anastomoses were harvested and underwent a maximal anastomotic tensile strength (MATS) test (MATS 1–3) followed by histologic analyses. ResultsMATS 1, when a tear became visible in the serosa, was significantly increased in the proximal anastomoses of the RIPC group compared with the control group (4.91 N vs 3.83 N; P = .005). No other significant differences were found when comparing these 2 groups. ConclusionOur study showed no convincing results of RIPC on intestinal anastomotic healing to recommend its use in a general clinical setting. Further animal studies on RIPC’s effect after relative or absolute intestinal ischemia may be recommended.

Shenhuang plaster enhances intestinal anastomotic healing in rabbits through activation of the TGF-β and Hippo/YAP signaling pathways.

Although many efforts have been made to improve management strategies and diagnostic methods in the past several decades, the prevention of anastomotic complications, such as anastomotic leaks and strictures, remain a major clinical challenge. Therefore, new molecular pathways need to be identified that regulate anastomotic healing, and to design new treatments for patients after anastomosis to reduce the occurrence of complications. Rabbits were treated with a MST1/2 inhibitor XMU-XP-1, a Chinese medicine formula Shenhuang plaster (SHP) or a control vehicle immediately after surgery. The anastomotic burst pressure, collagen deposition, and hydroxyproline concentration were evaluated at 3 and 7 days after the surgery, and qRT-PCR and western-blot analyses were used to characterize mRNA and protein expression levels. Both XMU-XP-1 and SHP significantly increased anastomotic burst pressure, collagen deposition, and the concentration of hydroxyproline in intestinal anastomotic tissue at postoperative day 7 (POD 7). Importantly, SHP could induce TGF-β1 expression, which activated its downstream target Smad-2 to activate the TGF-β1 signaling pathway. Moreover, SHP reduced the phosphorylation level of YAP and increased its active form, and treatment with verteporfin, a YAP-TEAD complex inhibitor, significantly suppressed the effects induced by SHP during anastomotic tissue healing. This study demonstrated that activation of the Hippo-YAP pathway enhances anastomotic healing, and that SHP enhances both the TGF-β1/Smad and YAP signaling pathways to promote rabbit anastomotic healing after surgery. These results suggest that SHP could be used to treat patients who underwent anastomosis to prevent the occurrence of anastomotic complications.

Histology and transcriptome insights into the early processes of intestinal anastomotic healing: a rat model.

Understanding the early processes underlying intestinal anastomotic healing is crucial to comprehend the pathophysiology of anastomotic leakage. The aim of this study was to assess normal intestinal anastomotic healing and disturbed healing in rats to investigate morphological, cellular and intrinsic molecular changes in the anastomotic tissue. Anastomoses were created in two groups of Wistar rats, using four sutures or 12 sutures to mimic anastomotic leakage and anastomotic healing respectively. At 6, 12, 24 hours and 2, 3, 5 and 7 days, anastomotic tissue was assessed macroscopically using the anastomotic complication score and histologically using the modified Ehrlich-Hunt score. Transcriptome analysis was performed to assess differences between anastomotic leakage and anastomotic healing at the first three time points to find affected genes and biological processes. Ninety-eight rats were operated on (49 animals in the anastomotic leakage and 49 in the anastomotic healing group) and seven rats analysed at each time point. None of the animals with 12 sutures developed anastomotic leakage macroscopically, whereas 35 of the 49 animals with four sutures developed anastomotic leakage. Histological analysis showed increasing influx of inflammatory cells up to 3 days in anastomotic healing and up to 7 days in anastomotic leakage, and this increase was significantly higher in anastomotic leakage at 5 (P = 0.041) and 7 days (P = 0.003). Transcriptome analyses revealed large differences between anastomotic leakage and anastomotic healing at 6 and 24 hours, mainly driven by an overall downregulation of genes in anastomotic leakage. Transcriptomic analyses revealed large differences between normal and disturbed healing at 6 hours after surgery, which might eventually serve as early-onset biomarkers for anastomotic leakage.

Enhancing intestinal anastomotic healing using butyrate: Systematic review and meta-analysis of experimental animal studies.

Despite advancements in surgical technique and perioperative care, intestinal anastomoses still have a 10-15 per cent risk of leakage, which results in considerable morbidity and/or mortality. Recent animal studies have suggested that administration of butyrate to the anastomotic site results in enhanced anastomotic strength, which may prevent leakage. This systematic review and meta-analysis summarises current evidence concerning the effect of butyrate administration on anastomotic healing and will form a scientific basis for the development of new research into this subject. Animal studies on the effect of butyrate-based interventions in models of intestinal anastomotic healing were systematically retrieved from online databases. Bibliographical data, study characteristics and outcome data were extracted, and internal validity of the studies was assessed. Outcomes studied through meta-analysis concerned: anastomotic strength, anastomotic leakage, collagen metabolism and general histologic parameters of wound healing. A comprehensive search and selection identified 19 relevant studies containing 41 individual comparisons. Design and conduct of most experiments were poorly reported resulting in an unclear risk of bias. Meta-analyses showed that butyrate administration significantly increases anastomotic strength (SMD 1.24, 0.88 to 1.61), collagen synthesis (SMD 1.44, 0.72 to 2.15) and collagen maturation, making anastomoses less prone to leakage in the early postoperative period (OR 0.37, 0.15 to 0.93). This systematic review and meta-analysis shows that there is potential ground to investigate the use of butyrate in clinical trials to prevent anastomotic leakage in intestinal surgery. However, more research is necessary to define the best application form, dosage and administration route.

The Impact of Phase-Specific Macrophage Depletion on Intestinal Anastomotic Healing.

Intestinal anastomotic healing (AH) is critical in colorectal surgery, since disruptive AH leads to anastomotic leakage, a feared postoperative complication. Macrophages are innate immune cells and are instrumental in orchestrating intestinal wound healing, displaying a functional dichotomy as effectors of both tissue injury and repair. The aim of this study was to investigate the phase-specific function and plasticity of macrophages during intestinal AH. Transgenic CD11b diphtheria toxin receptor (CD11b-DTR) mice were used to deplete intestinal macrophages in a temporally controlled manner. Distal colonic end-to-end anastomoses were created in CD11b-DTR, and wild-type mice and macrophages were selectively depleted during either the inflammatory (day 0-3), proliferative (day 4-10), or reparative (day 11-20) phase of intestinal AH, respectively. For each time point, histological and functional analysis as well as gene set enrichment analysis (GSEA) of RNA-sequencing data were performed. Macrophage depletion during the inflammatory phase significantly reduced the associated inflammatory state without compromising microscopic AH. When intestinal macrophages were depleted during the proliferative phase, AH was improved, despite significantly reduced perianastomotic neoangiogenesis. Lastly, macrophages were depleted during the reparative phase and GSEA revealed macrophage-dependent pathways involved in collagen remodeling, cell proliferation, and extracellular matrix composition. However, AH remained comparable at this late timepoint. These results demonstrate that during intestinal AH, macrophages elicit phase-specific effects, and that therapeutic interventions must critically balance their dual and timely defined role.

Perioperative Low-Dose Prednisolone Treatment Has Beneficial Effects on Postoperative Recovery and Anastomotic Healing in a Murine Colitis Model.

High-dose glucocorticoid treatment has been identified as a risk factor for anastomotic leakage in patients with inflammatory bowel disease [IBD] undergoing bowel resection surgery. By contrast, active disease during surgery is also associated with elevated morbidity. Perioperative low-dose treatment might be beneficial regarding postoperative outcomes by controlling disease activity. The present study is the first to investigate the dose-dependent effect of perioperative prednisolone therapy in a murine IBD model combining dextran sodium sulphate [DSS] colitis with intestinal anastomosis surgery. In 84 10-week-old wild-type mice, a colorectal anastomosis was performed using a microsurgical technique. Half the animals received induction of chemical colitis with 2% DSS via drinking water prior to surgery. In both groups, one-third of the animals received daily oral administration of high-dose [0.533 mg/kg] and one-third low-dose [0.133 mg/kg] prednisolone. Evaluation was performed on postoperative days 3 and 7. While high-dose prednisolone treatment led to an increased anastomotic leakage rate in mice under colitis, low-dose prednisolone treatment limited preoperative disease activity and did not influence the leakage rate. Histological examination showed a beneficial effect of low-dose prednisolone treatment on microscopic abscess formation at the anastomotic site in DSS mice as well as an increased anastomotic healing score. We demonstrate a beneficial effect of perioperative short-term low-dose prednisolone treatment on intestinal anastomotic healing in the context of colitis. Perioperative use of short-term low-dose prednisolone treatment might be beneficial in IBD patients who need to undergo surgery during active disease.

The comparison of the suture materials on intestinal anastomotic healing: an experimental study.

This experimental comparative study was to evaluate the local effects of three different suture materials on in-testinal anastomosis healing. Ethical approval was obtained from the University of Ethical Committee (E-60758568-020-176720). A prospective, experimental comparative analysis was conducted on 24 rats. They were divided into three equal groups; Group 1 underwent colonic anastomosis with Vicryl suture material, Group 2 underwent colonic anastomosis with polypropylene suture; and Group 3 underwent colonic anastomosis with polydioxanone (PDS) suture. The second operation underwent the 7th post-operative day. Adhesion score, anastomotic leakage, anastomotic bursting pressure, hydroxyproline levels, and histopathologic examination were evaluated. All animals survived, and no leakage, intestinal obstruction, or wound infection was observed during the experiment. The adhesion score was evaluated according to the Diamond classification and same in all groups. Median anastomotic bursting pressure was 125.75 mmHg (10-241) in the Vicryl group, 159.25 mmHg (113-190) in the polypropylene group, and 154.50 mmHg (20-212) in the PDS group. Hydroxyproline tissue concentrations were in the Vicryl group 1699.92±220.8 ng/mg (range: 1509.81-2186.47), in the polypropylene group 1126.24±607.12 ng/mg (range: 53.22-1815.63), and 1547.86±335.2 ng/mg (range: 973.66-1973.2) in PDS group. There was no difference among groups regarding the inflammatory response evaluated by histopathology. There was no statistical significance in all variables evaluated. This experimental study demonstrates that suture materials did not worsen tissue healing during intestinal anastomosis. Absorbable, slowly-absorbable, and non-absorbable suture materials could be used safely in every situation.

The Effect of Genistein on Anastomotic Healing in Intestinal Ischemia/Reperfusion Injury

IntroductionGenistein is a natural isoflavonoid and has several pharmacological effects, such as antioxidant, antitumor activity, and improvement of glucose metabolism. The safety of intestinal anastomosis after ischemia–reperfusion (I/R) injury is a critical issue for surgeons. This experimental study aimed to investigate the effects of genistein on anastomotic healing after intestinal I/R injury. MethodsA total of 36 male Wistar Albino rats were divided into four groups: control, I/R, genistein, and genistein + I/R. The control group received segmental ileal resection and ileoileal anastomosis. The I/R group received resection + anastomosis after intestinal I/R. The genistein group was administered subcutaneous injection of 1 mg/kg genistein 12 h and 1 h before the procedure and received ileal resection + anastomosis. The genistein + I/R group received I/R + ileal resection + anastomosis after genistein injection. Anastomotic bursting pressure, hydroxyproline, superoxide dismutase, and glutathione peroxidase levels and histopathological wound healing scores of all rats were measured on postoperative day 5. ResultsThe anastomotic bursting pressure was significantly higher in the genistein and genistein + I/R groups (P < 0.001). Genistein increased the hydroxyproline concentration and the superoxide dismutase and glutathione peroxidase levels in the intestinal anastomosis (P < 0.001). In histopathological assessment, the mean wound healing score was significantly higher in the genistein group than in the other groups (P < 0.001). ConclusionsGenistein, with its anti-inflammatory and antioxidant properties, shows protective effects against increased oxidative stress after intestinal I/R injury and contributes positively to intestinal anastomotic healing.

The effect of Ginkgo biloba EGb 761 on intestinal anastomotic healing in rats with ischemia-reperfusion induced in the lower extremities

Background/Aim: Remote ischemia-reperfusion (I-R) injury for anastomotic healing is a newly identified risk factor and there is a wide array of studies being conducted. This study aimed to reveal the negative effects of lower I-R on the colonic anastomotic healing process and examine the effects of Ginkgo biloba EGb 761 treatment, a platelet activating factor (PAF) antagonist, on anastomotic healing through the inhibition of pathological mechanisms of mediators causing these negative effects. Methods: Thirty-six Wistar-Albino rats were divided into sham, I-R and I-R+EGb 761 groups, each consisting of 12 rats. In the subjects in the sham group, an end-to-end anastomosis was performed by transecting the descending colon following midline laparotomy. In the I-R group, unilateral lower extremity ischemia was created by occluding the femoral artery and collaterals using a tourniquet from the most proximal segment of the left extremity. Then, the descending colon was transected, anastomosis was performed, and reperfusion was created by a tourniquet application at the 30th minute of ischemia. Different from the I-R (control) group, the subjects in the I-R+EGb 761 group were given two equal doses of 64 mg/kg/d Ginkgo biloba EGb 761 by the orogastric route until 10 days after surgery. After all subjects were sacrificed on the 10th day of surgery, the descending colon segment containing the anastomosis area was resected and samples were taken for bursting pressure and hydroxyproline measurements. Results: In the I-R group, anastomotic bursting pressure and perianastomotic hydroxyproline values were significantly lower compared to the sham group and the I-R+EGb 761 group. However, there was no statistically significant difference in these parameters between the sham and the I-R+EGb 761 groups. Conclusion: Colonic anastomotic bursting pressure and peri-anastomotic hydroxyproline values in the sham group were significantly decreased by lower extremity I-R, and this change was prevented with the use of EGb 761.

Intestinal anastomotic healing models during experimental colitis

BackgroundAnastomotic leakage represents a major complication following resections in colorectal surgery. Among others, intestinal inflammation such as in inflammatory bowel disease is a significant risk factor for disturbed anastomotic healing. Despite technical advancements and several decades of focused research, the underlying mechanisms remain incompletely understood. Animal experiments will remain the backbone of this research in the near future. Here, instructions on a standardized and reproducible murine model of preoperative colitis and colorectal anastomosis formation are provided to amplify research on anastomotic healing during inflammatory disease.MethodsWe demonstrate the combination of experimental colitis and colorectal anastomosis formation in a mouse model. The model allows for monitoring of anastomotic healing during inflammatory disease through functional outcomes, clinical scores, and endoscopy and histopathological examination, as well as molecular analysis.DiscussionPostoperative weight loss is used as a parameter to monitor general recovery. Functional stability can be measured by recording bursting pressure and location. Anastomotic healing can be evaluated macroscopically from the luminal side by endoscopic scoring and from the extraluminal side by assessing adhesion and abscess formation or presence of dehiscence. Histologic examination allows for detailed evaluation of the healing process.ConclusionThe murine model presented in this paper combines adjustable levels of experimental colitis with a standardized method for colorectal anastomosis formation. Extensive options for sample analysis and evaluation of clinical outcomes allow for detailed research of the mechanisms behind defective anastomotic healing.

Ac2-26-Nanoparticles Induce Resolution of Intestinal Inflammation and Anastomotic Healing via Inhibition of NF-κB Signaling in a Model of Perioperative Colitis.

Although in most patients with inflammatory bowel diseases, conservative therapy is successful, a significant proportion of patients still require surgery once in their lifetime. Development of a safe perioperative treatment to dampen colitis activity without disturbance of anastomotic healing is an urgent and unmet medical need. Annexin A1 (ANXA1) has been shown to be effective in reducing colitis activity. Herein, a nanoparticle-based perioperative treatment approach was used for analysis of the effects of ANXA1 on the resolution of inflammation after surgery for colitis. Anxa1-knockout mice were used to delineate the effects of ANXA1 on anastomotic healing. A murine model of preoperative dextran sodium sulfate colitis was performed. Collagen-IV-targeted polymeric nanoparticles, loaded with the ANXA1 biomimetic peptide Ac2-26 (Ac2-26-NPs), were synthesized and administered perioperatively during colitis induction. The effects of the Ac2-26-NPs on postoperative recovery and anastomotic healing were evaluated using the disease activity index, histological healing scores, and weight monitoring. Ultimately, whole-genome RNA sequencing of the anastomotic tissue was performed to unravel underlying molecular mechanisms. Anxa1-knockout exacerbated the inflammatory response in the healing anastomosis. Treatment with Ac2-26-NPs improved preoperative colitis activity (P < 0.045), postoperative healing scores (P < 0.018), and weight recovery (P < 0.015). Whole-genome RNA sequencing revealed that the suppression of proinflammatory cytokine and nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB) signaling was associated with the treatment effects and a phenotypic switch toward anti-inflammatory M2 macrophages. Proresolving therapy with Ac2-26-NPs promises to be a potent perioperative therapy because it improves colitis activity and even intestinal anastomotic healing by the suppression of proinflammatory signaling.

The immunologic changes during different phases of intestinal anastomotic healing.

Intestinal anatomosis is a complex and multicellular process that involving three overlapped phases: exudative phase, proliferative phase, and reparative phase. Undisturbed anastomotic healings are crucial for the recovery of patients after operations but unsuccessful healings are linked with a considerable mortality. This time, we concentrate on the immunologic changes during different phases of intestinal anastomotic healing and select several major immune cells and cytokines of each phase to get a better understanding of these immunologic changes in different phases, which will be significant for more precise therapy strategies in anastomoses.

AB053. Perioperative administration of Ac2-26 loaded nanoparticles facilitates postoperative recovery in a murine model for intestinal anastomotic healing during acute colitis

AB053. Perioperative administration of Ac2-26 loaded nanoparticles facilitates postoperative recovery in a murine model for intestinal anastomotic healing during acute colitis

Effects of autologous platelet-rich plasma coated sutures on intestinal anastomotic healing in rabbits

The objective of this study was to investigate the intestinal anastomotic healing enhancing effect of platelets-rich plasma (PRP) using PRP-coated sutures in a rabbit model. A total of 30 mature male rabbits were divided into 3 groups (10 rabbits per group). Group 1 received uncoated sutures, group 2 received sodium acetate-coated sutures, and group 3 received PRP-coated sutures. Polyglactin 910 (Vicryl, USA), size 4-0 was used in all groups. Five rabbits of each group were euthanized on day 3 following the surgery while the remaining 5 rabbits were euthanized on day 10. Gross evaluation of the anastomotic site in PRP-coated sutures group demonstrated significantly (P < 0.05) lower adhesion formation scores on both days 3 and 10 of the study while in the control groups, evidence of leakage at the anastomotic site was present along with signs of haemorrhage and local inflammation. On day 10 in the control groups, there were strands of strong adhesions between the ileum, colon and cecum with large amount of fibrin deposited at the site of the anastomosis. Tissues of the anastomotic site revealed a significant level of hydroxyproline on day 10 in PRP-coated sutures group compared with control groups. Histopathological evaluation revealed significantly (P < 0.05) less inflammatory infiltration, and more angiogenesis and collagen deposition on day 10 in PRP-coated sutures group compared to the control groups. Results of this study clearly indicate promising healing enhancing effects of using PRP-coated sutures at intestinal anastomotic site with little to no obvious disadvantages.

The effect of pasireotide on intestinal anastomotic healing with and without whole-body irradiation in a rat model

ObjectiveTo examine pasireotide’s effect on intestinal anastomotic healing under physiological conditions and following preoperative whole-body irradiation.Material and methodsForty-five male Wistar rats received an ileoileal end-to-end anastomosis. Group 1 (Co, n = 9) served as control. Group 2 (SOM, n = 10) received pasireotide (60 mg/kg) 6 days preoperatively. Group 3 (R-Co, n = 13) was subjected to 8 Gy whole-body irradiation 4 days preoperatively. Finally, group 4 (R-SOM, n = 13) received pasireotide 6 days preoperatively and whole-body irradiation 4 days preoperatively. On postoperative day 4, anastomotic bursting pressure, histology, IGF-1 staining, and collagen density were examined.ResultsMortality was higher in irradiated animals (30.8% vs. 5.3%, p = 0.021), and anastomotic bursting pressure was significantly lower (median, R-Co = 83 mmHg; R-SOM = 101 mmHg; Co = 149.5 mmHg; SOM = 169 mmHg). Inflammation measured by leukocyte infiltration following irradiation was reduced (p = 0.023), and less collagen was observed, though this was not statistically significant. Bursting pressure did not significantly differ between Co and SOM and between R-Co and R-SOM animals respectively. Semi-quantitative scoring of IGF-1, fibroblast bridging, or collagen density did not reveal significant differences among the groups.ConclusionWhole-body irradiation decreases the quality of intestinal anastomotic wound healing and increases mortality. Pasireotide does not significantly lessen this detrimental effect.

Role of platelet-rich fibrin on intestinal anastomosis wound healing in a rat

Objectives. The aim of the present study is to investigate the role of platelet-rich fibrin (PRF) on the intestinal anastomotic wound healing in a mesenteric ischemia/reperfusion (I/R) rat model. Methods. Forty male Sprague Dawley rats were allocated into four groups: Group I (n = 10): anastomosis to normal bowel; Group II (n = 10): anastomosis after I/R injury; Group III (n = 10): anastomosis and PRF; Group IV: anastomosis after I/R and PRF. Animals were followed up for 7 days, then sacrificed. Anastomotic complications, anastomosis bursting pressures and histopathologic evaluations of the anastomoses were the study parameters. Results. The I/R injury caused more anastomotic adhesion and very low anastomotic bursting pressure when compared with the other groups (p < 0.01). Application of PRF onto the bowel, however, dramatically decreased the local complications and significantly increased the anastomosis bursting pressures (p < 0.01). Histologic evaluation of the anastomoses showed almost complete healing in all animals. The mean histologic scores of the animals were not different between the groups. Conclusion. PRF has a beneficial effect on the intestinal healing process by prevention of local complications and increases the tensile strain of the anastomosis. This effect was more prominent, particularly when healing is disrupted. Therefore, we conclude that PRF could be an alternative treatment option to prevent anastomotic complications for elderly, co-morbid and emergency patients.

Tissular growth factors profile after teduglutide administration on an animal model of intestinal anastomosis.

Teduglutide is an enterotrophic analogue of glucagon-like peptide-2, with an indirect and poorly understood mechanism of action, approved for the rehabilitation of short-bowel syndrome. This study aims to analyze the response of tissue growth factors to surgical injury and teduglutide administration on an animal model of intestinal anastomosis. Wistar rats (n = 59) were distributed into four groups: "ileal resection" or "laparotomy", each one subdivided into "postoperative teduglutide administration" or "no treatment"; and sacrificed at the third or the seventh day, with ileal sample harvesting. Gene expression of insulin-like growth factor 1 (Igf1), vascular endothelial growth factor a (Vegfa), transforming growth factor β1 (Tgfβ1), connective tissue growth factor (Ctgf), fibroblast growth factor 2 (Fgf2), fibroblast growth factor 7 (Fgf7), epidermal growth factor (Egf), heparin-binding epidermal-like growth factor (Hbegf), platelet-derived growth factor b (Pdgfb) and glucagon-like peptide 2 receptor (Glp2r)was studied by real-time polymerase chain reaction. Upregulation of Fgf7, Fgf2, Egf, Vegfaand Glp2rat the third day and of Pdgfat the seventh day was verified in the perianastomotic segment. Teduglutide administration was associated with higher fold-change of relative gene expression of Vegfa(3.6 ± 1.3 vs.1.9 ± 2.0, p = 0.0001), Hbegf(2.2 ± 2.3 vs. 1.1 ± 0.9, p = 0.001), Igf1(1.6 ± 7.6 vs. 0.9 ± 0.7, p = 0.002) and Ctgf(1.1 ± 2.1 vs. 0.6 ± 2.0, p = 0.013); and lower fold-change of Tgfβ1, Fgf7and Glp2r. Those results underscore the recognized role of Igf1and Hbegfas molecular mediators of the effects of teduglutide and suggest that other humoral factors, like Vegfand Ctgf, may also be relevant in the perioperative context. Induction of Vegfa, Igf1and Ctgfgene expressions might indicate a favorable influence of teduglutide on the intestinal anastomotic healing.

Teduglutide effects on gene regulation of fibrogenesis on an animal model of intestinal anastomosis

BackgroundTeduglutide is an enterotrophic analog of glucagon-like peptide 2 approved for the rehabilitation of short-bowel syndrome. This study aims to analyze the effects of teduglutide administration on the gene regulation of fibrogenesis during the intestinal anastomotic healing on an animal model. MethodsWistar rats (n = 62) were assigned into four groups: “Ileal Resection and Anastomosis” or “Laparotomy,” each one subdivided into “Postoperative Teduglutide Administration” or “No Treatment,” and sacrificed at the third or at the seventh days, with ileal sample harvesting. Gene expression of matrix components and remodeling factors (matrix metalloproteinases [Mmp] and tissue inhibitors of metalloproteinases [Timp]) and growth factors was studied by real-time polymerase chain reaction. Net collagen deposition was assessed through the Collagen-to-Mmp-to-Timp ratio of fold change of relative gene expression. ResultsGene expression profiles revealed a balance toward net degradation of collagen at the third day of the intestinal anastomotic healing. Teduglutide appeared to be associated with an overall accumulation of collagen at the third day of the anastomotic repair, attributable to the upregulation of Collagen type 1 alpha 1, Collagen type 3 alpha 1, and Collagen type 4 alpha 1, Timp1, and Timp2 and downregulation of Mmp13 and to a net degradation of collagen at the seventh day, derived from repression of Collagen type 3 alpha 1, Collagen type 5 alpha 1 and Timp1 expression. ConclusionsTeduglutide appeared to be associated with a favorable influence on fibrogenesis at the third day of the intestinal anastomotic repair and to a trend to fibrolysis at the seventh day.

Effects of teduglutide on histological parameters of intestinal anastomotic healing

Intestinal anastomotic failure continues to be one of the most relevant surgical incidents. Teduglutide is an enterotrophic long-acting equivalent of glucagon-like peptide-2 (GLP-2) recommended for the treatment of adult patients with short bowel syndrome. This study evaluates the effects of short-term teduglutide administration on histological parameters of intestinal anastomotic healing in an animal model. Wistar rats (n = 31) were submitted to ileal resection and standard anastomosis, with or without postoperative teduglutide ministration, and sacrificed at the third or seventh day with tissue and blood sampling. Intestinal anastomotic healing was assessed with the modified Houdart–Hutschenreiter classification after hematoxylin and eosin staining. Collagen content and distribution were evaluated with the Gordon and Sweet technique; neoangiogenesis and epithelial proliferation indexes with anti-CD31 and anti-Ki67 immunostaining, respectively. GLP-2 plasma levels were determined by quantitative enzyme immunoassay. Teduglutide administration was associated with higher levels of reepithelialization (P = 0.022) and a higher neoangiogenesis index (neovessel density = 16.0 ± 10.8/mm2 vs. 5.3 ± 3.4/mm2; P = 0.0001) at the seventh postoperative day. Lower expression of type I collagen (P = 0.015) and higher content of type III collagen (P = 0.007) in the submucosa were also documented at the seventh day in teduglutide-treated animals. Drug administration was not associated with statistically significant differences of GLP-2 plasma levels. These findings indicate a positive impact of teduglutide on the reepithelialization and neoangiogenesis events of the proliferative phase of intestinal anastomotic repair. Additional studies are necessary to clarify its effects on the fibrogenesis process.