Healing Of Intestinal Anastomoses Research Articles

Effect of Peritoneum Graft on Intestinal Anastomosis Healing After Deserosalization in Dogs

Effect of Peritoneum Graft on Intestinal Anastomosis Healing After Deserosalization in Dogs

Mesenchymal stem cell secretome-loaded fibrin glue improves the healing of intestinal anastomosis.

Anastomotic leakage is a serious complication following gastrointestinal surgery and one of the leading causes of patient mortality. Despite the significant clinical and economic burden, there are currently no reliable treatment options to improve the healing of intestinal anastomosis and subsequently prevent anastomotic leakage. Recently, the development of regenerative medicine has shown promise for improving anastomotic healing. Recent studies have illustrated that stem cell-derived secretome can enhance tissue regeneration without the safety and ethical limitations of stem cell transplantation. Herein, we developed a fibrin glue topical delivery system loaded with mesenchymal stem cells (MSCs)-derived secretome for controlled delivery of bioactive factors, and evaluated its application potential in improving the healing of intestinal anastomosis. Under in vitro conditions, the MSCs secretome significantly promoted cell proliferation viability in a dose-dependent manner and resulted in the controlled release of growth factors via fibrin glue delivery. We established a rat surgical anastomotic model and experimentally found that MSCs secretome-loaded fibrin glue enhanced anastomotic bursting pressure, increased granulation tissue formation and collagen deposition, and significantly promoted anastomotic healing. Mechanistically, fibrin glue accelerated cell proliferation, angiogenesis, and macrophage M2 polarization at the surgical anastomotic site by releasing bioactive factors in the secretome, and it also alleviated the inflammatory response and cell apoptosis at the anastomotic site. Our results demonstrated for the first time that MSCs-derived secretome could promote the healing of intestinal anastomosis. Considering the accessibility and safety of the cell-free secretome, we believed that secretome-loaded fibrin glue would be a cell-free therapy to accelerate the healing of intestinal anastomosis with great potential for clinical translation.

Preparation of a pig experimental model of defective colon anastomosis - review.

Anastomotic leak is a serious complication in colorectal surgery. The problem of intestinal anastomosis healing disorders is dealt with by a number of experimental studies, especially in applied research. The design of animal models is diverse and the results of individual studies are difficult to compare. This paper summarizes the main issues of planning animal models of intestinal anastomoses, which are discussed individually. The main part of the text is a description of the defective intestinal anastomosis model of the pig. The anastomosis is constructed in the model with a localized mucosal protrusion. The animals are monitored for 3 weeks postoperatively, the clinical condition and changes in vital values and laboratory parameters are monitored, and CT examinations are performed at defined points. At the end of the observation period, samples are taken, macroscopic findings in the abdominal cavity are evaluated, adhesions are scored, and signs of leakage or passage disorders are assessed. The preparations are evaluated histologically both by standard methods analyzing vascularity, inflammatory infiltration and the proportion of collagen, and by methods developed de novo for the needs of the experiment, such as the analysis of the integrity of the intestinal wall at the site of the mucosal protrusion. We value the experimental model for the possibility of a systematic and detailed analysis of the healing state of the anastomosis in combination with a detailed observation protocol, which produces clinically relevant results.

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.

Nanofiber Fractionalization Stimulates Healing of Large Intestine Anastomoses in Rabbits.

A current topic of ma jor interest in regenerative medicine is the development of novel materials for accelerated healing of sutures, and nanofibers seem to be suitable materials for this purpose. As various studies have shown, nanofibers are able to partially substitute missing extracellular matrix and to stimulate cell proliferation and differentiation in sutures. Therefore, we tested nanofibrous membranes and cryogenically fractionalized nanofibers as potential materials for support of the healing of intestinal anastomoses in a rabbit model. We compared cryogenically fractionalized chitosan and PVA nanofibers with chitosan and PVA nanofiber membranes designed for intestine anastomosis healing in a rabbit animal model. The anastomoses were biomechanically and histologically tested. In strong contrast to nanofibrous membranes, the fractionalized nanofibers did show positive effects on the healing of intestinal anastomoses in rabbits. The fractionalized nanofibers were able to reach deep layers that are key to increased mechanical strength of the intestine. Moreover, fractionalized nanofibers led to the formation of collagen-rich 3D tissue significantly exceeding the healing effects of the 2D flat nanofiber membranes. In addition, the fractionalized chitosan nanofibers eliminated peritonitis, significantly stimulated anastomosis healing and led to a higher density of microvessels, in addition to a larger fraction of myofibroblasts and collagen type I and III. Biomechanical tests supported these histological findings. We concluded that the fractionalized chitosan nanofibers led to accelerated healing for rabbit colorectal anastomoses by the targeted stimulation of collagen-producing cells in the intestine, the smooth muscle cells and the fibroblasts. We believe that the collagen-producing cells were stimulated both directly due to the presence of a biocompatible scaffold providing cell adhesion, and indirectly, by a proper stimulation of immunocytes in the suture.

Effect of autologous peritoneum and platelet-rich fibrin graft on healing of intestinal anastomosis in dogs

In the present study, the effect of autogenous peritoneum graft (PG) with platelet rich fibrin (PRF) was evaluated on intestinal anastomosis healing in dogs. Eighteen local breed adult dogs were anesthetized then jejunal intestinal resection and anastomosis were created on all animals. Animals were divided into 3 equal groups (6 for each). In first group (control): An end-to-end intestinal anastomosis was performed using simple interrupted suture pattern only. In group PG: The anastomosis site was sutured as in control group and was wrapped with PG. Where as in PRF+PG group: The anastomosis site was sutured also as in first group and was wrapped with PRF and PG. Healing process was studied in all above mentioned groups clinically, grossly, histologically and radiographically at 15 and 30 days post operation. Results were revealed that the utilizing of PG caused a valuable influence on anastomosis site healing. This impact was manifested by faster re-epithelization of mucosa, increased collagen deposition, fibroblast proliferation, reduced adhesions, and decreased stenosis degree comparison with control group. Also there was less adhesions, less stenosis degree, increased in epithelization of the mucosa in the group treated with PRF+PG than the group treated with PG alone. In conclusion the current study has shown that autologous peritoneal graft application has a positive and beneficial effect on intestinal anastomosis healing. Also the results highlighted the promising effects of PRF in conjunction with PG on intestinal anastomosis healing. The application of PRF with PG at the intestinal anastomosis site is successful, safe, and highly efficient with no complications.

Effects of Oral Pirfenidone on Colon Anastomosis Healing and Adhesion Formation in Rats

Introduction: Many experimental studies have examined multiple drugs or treatments to improve the healing of intestinal anastomoses. Synthetic prostacyclin analogs, immunosuppressants, erythropoietin, growth hormone, insulin-like growth factor type 1, synthetic metalloproteinases inhibitors, and hyperbaric oxygen therapy have produced promising results in low-risk models of anastomosis dehiscence. However, in high-risk models, only hyperbaric oxygen therapy has been shown to be useful. Pirfenidone (PFD), a commonly used antifibrosing drug, has not been shown to be effective for this purpose. Our objective was to evaluate the effects of PFD on anastomosis healing and adhesion genesis in a low-risk rat model of dehiscence of colonic anastomosis. Methods: An experimental study was conducted on 40 healthy Wistar rats randomly assigned to the control group or PFD experimental group (20 rats in each group). Colon anastomosis was performed 3 cm above the peritoneal reflection using the same technique in all animals. Mechanical resistance was studied by measuring bursting pressure. Adhesions were evaluated macroscopic and histologically using common staining techniques. Animals received the first PFD dose 12 h after surgery at a dose of 500 mg/kg one a day (SID) for 5 consecutive days. On day 6, the animals were reoperated on to measure the bursting pressure in situ and to classify adhesions macroscopically, and the anastomosed colon was resected for histological analysis. Results: There were no deaths, complications, or anastomosis dehiscence in either group. The mean bursting pressure was 120.8 ± 11 mm Hg and 135.5 ± 12.4 in the control and PFD groups, respectively (p < 0.001). The adhesions were less dense and had less inflammatory cell infiltration in the PFD group (p < 0.02 and 0.002, respectively). Collagen content was slightly higher in the PFD group (p = 0.04). Conclusions: Our results revealed favorable effects of PFD in this low-risk colon anastomosis model; for example, the bursting pressure was higher, and the macroscopic adhesions were soft and exhibited less inflammatory infiltration and higher collagen content in the PFD group than in the control group. The results showing that PFD treatment was associated with better healing of minor adhesions seem to be paradoxical because the therapeutic indications for this drug are directed at treating fibrosing diseases.

Mg-6Zn alloys promote the healing of intestinal anastomosis via TGF-β/Smad signaling pathway in regulation of collagen metabolism as compared with titanium alloys.

There is a great clinical need for biodegradable materials. This study aimed to investigate the effects of Mg-6Zn and titanium alloy stapler nails on intestinal anastomosis healing mediated via the TGF-β/Smad signaling pathway, as reflected in collagen metabolism in rabbits. Side-to-side ileo-ileostomy was performed with linear stapler loaded with the two nails. The results showed that no obvious postoperative complications such as abdominal infection and anastomotic leakage were observed. General observation and scanning electron microscope showed that Mg-6Zn alloy nails remained intact in the first week, degraded significantly in the second week, and were little left in the third week, while the titanium alloy nails showed intact substrate throughout the experimental period. Immunohistochemical analysis showed that the expression of TGF-β1 in Mg-6Zn alloy group was higher than that in titanium alloy group after 1week, but it increased slowly, arrived at a lower level in the third week. Collagen I showed an increased expression in Mg-6Zn alloy group, but decreased with time in titanium alloy group. An enhanced expression of collagen III in Mg-6Zn alloy group in the first week but much lower in the third week as compared to the titanium alloy group. The expression of smad2 in Mg-6Zn alloy group maintained a steady level, while in titanium alloy group it showed a general upward trend. The expression of smad3 in both groups held steady after 2weeks, then in the third week, it showed a strong uptrend in Mg-6Zn alloy group, while decreased immediately in titanium alloy group. Our findings suggest that Mg-6Zn alloy nails degraded significantly within 3weeks and could provide stability of intestinal anastomosis in the reconstruction of intestinal tract. TGF-β/Smad signaling pathway may play a role in regulation of baseline collagen synthesis throughout the process.

The effect of therankeron D6 on the healing of intestinal anastomosis in dogs

To investigate the effects of systemic administration of Theranekron (Tarantula cubensis) extract as homeopathic remedy on anastomosis healing after surgical intestinal resection. Eight healthy stray dogs (age 1-1.5 years, weight 15-20 kgs) were randomly divided into two groups; control (4 animals) and treatment group (4 animals). Intestinal resection and anastomosis were performed in all animals in both groups. Theranekron D6 (3 ml) was administered subcutaneously one week before surgery and at operation time in the treatment group whereas animals of the control group were injected 3 ml of normal saline at the same time points of treatment group. All animals were kept for 15 days following surgery and a segment of intestine was removed for histopathological and press pressure measurement. The results showed that no post-operative complications have been observed in both groups. The gross examination of the anastomosis site revealed sever adhesions with omentum and other intestinal segment of the all animals of the control group in comparison to the treatment group where lesser adhesions with omentum without any adhesion with the other intestinal segment have been noticed. There was slight difference between control and treatment group in relation to histopathological examination. In conclusion, the findings of this study showed that Theranekron was able to improve the healing process of the resected and anastomosed intestinal segment

Inhibition of HIF-prolyl hydroxylases improves healing of intestinal anastomoses

Anastomotic leakage (AL) accounts for a major part of in-house mortality in patients undergoing colorectal surgery. Local ischemia and abdominal sepsis are common risk factors contributing to AL and are characterized by upregulation of the hypoxia-inducible factor (HIF) pathway. The HIF pathway is critically regulated by HIF-prolyl hydroxylases (PHDs). Here, we investigated the significance of PHDs and the effects of pharmacologic PHD inhibition (PHI) during anastomotic healing. Ischemic or septic colonic anastomoses were created in mice by ligation of mesenteric vessels or lipopolysaccharide-induced abdominal sepsis, respectively. Genetic PHD deficiency (Phd1–/–, Phd2+/–, and Phd3–/–) or PHI were applied to manipulate PHD activity. Pharmacologic PHI and genetic PHD2 haplodeficiency (Phd2+/–) significantly improved healing of ischemic or septic colonic anastomoses, as indicated by increased bursting pressure and reduced AL rates. Only Phd2+/– (but not PHI or Phd1–/–) protected from sepsis-related mortality. Mechanistically, PHI and Phd2+/– induced immunomodulatory (M2) polarization of macrophages, resulting in increased collagen content and attenuated inflammation-driven immune cell recruitment. We conclude that PHI improves healing of colonic anastomoses in ischemic or septic conditions by Phd2+/–-mediated M2 polarization of macrophages, conferring a favorable microenvironment for anastomotic healing. Patients with critically perfused colorectal anastomosis or abdominal sepsis could benefit from pharmacologic PHI.

In vivo performance of a bilayer wrap to prevent abdominal adhesions

There is a high prevalence of intra-abdominal adhesions following bowel resection, which can result in chronic pain, bowel obstruction, and morbidity. Although commercial adhesion barriers have been widely utilized for colonic resections, these barriers do not prevent anastomotic leakage resulting from reduced healing of the anastomosis, which can result in long-term health problems. To address this limitation, we have developed an adhesive bilayer wrap with selective bioactivity to simultaneously prevent intra-abdominal adhesion formation and promote anastomotic healing. Reactive electrospinning was used to generate a crosslinked gelatin mesh to serve as a cell-instructive substrate to improve anastomotic healing. A coating of poly(ethylene glycol) (PEG) foam was applied to the bioactive mesh to generate an antifouling layer and prevent intra-abdominal adhesions. After in vitro confirmation of selective bioactivity, the composite wrap was compared after 2 weeks to a commercial product (InterceedⓇ) in an in vivo rat colonic abrasion model for prevention of intra-abdominal adhesions. The composite bilayer wrap was able to prevent intra-abdominal adhesions when clinical placement was maintained. The composite bilayer wrap was further modified to include tissue adhesive properties for improved efficacy. Preliminary studies indicated that the adhesive composite bilayer wrap maintained a maximum shear strength comparable to InterceedⓇ and greater than fibrin glue. Overall, this work resulted in an initial proof-of-concept device that was shown to effectively prevent intra-abdominal adhesion formation in vivo. The composite bilayer wrap studied here could lead to an improved technology for improved healing of intestinal anastomoses.

Semi-Quantitative Comparison of Infrared Thermography with Indocyanine Green Imaging in Porcine Intestinal Resection

BackgroundThe quality of intestinal blood supply is extremely important for healing of intestinal anastomoses. During the surgery the blood supply of the intestine may appear sufficient even though the microperfusion is not fully adequate. The degree of blood supply of remaining intestinal segment and the positioning of the resection margins is estimated subjectively by the surgeon's experience or objectively by means of indocyanine green fluorescence imaging. The subject of our study is the evaluation of the infrared thermal imaging as another supportive non-invasive imaging method in assessment of intestinal blood supply, and to compare surgeon's decisions of the position of resection line with information obtained by infrared thermal imaging and by indocyanine green fluorescence imaging. MethodsA pilot study on thermal imaging measurements was done in a porcine model. The infrared thermocamera Workswell WIC 640 was used in our study. The thermal imaging was correlated with the indocyanine green imaging method ICG NOVADAQ and with surgeon's subjective expert decisions of the places of resection lines. Fifteen monitored resections were performed on three experimental porcine models. ResultsBased on the data evaluated, experience of the surgical team and the post-operative conditions of the porcine models, we conclude that the thermal imaging is a useful tool for determination of the optimal intestinal resection margins, and thus contributes to lowering anastomotic complications rate in colorectal surgery. The results show relatively high consistency between the used imaging techniques. Both methods showed ischemic regions of the intestine at a comparable level. The IRT methods showed even a slightly higher match with the surgeon's rating. ConclusionBased on experiments, the infrared thermography can be considered as one of complementary imaging methods suitable for use during anastomosis surgery. The sensitivity and versatility of the infrared thermography method was demonstrated. There was confirmed agreement in obtained information about the position of area with maximal blood insufficiency obtained by IRT with the information obtained by using the ICG method.

Electrospun nanofibers as support for the healing of intestinal anastomoses.

The breakdown of intestinal anastomosis is a serious postsurgical complication. The worst complication is anastomotic leakage, resulting in contaminated peritoneal cavity, sepsis, multi-organ failure and even death. In problematic locations like the rectum, the leakage rate has not yet fallen below 10 %. Such a life-threatening condition is the result of impaired healing in the anastomotic wound. It is still vital to find innovative strategies and techniques in order to support regeneration of the anastomotic wound. This paper reviews the surgical techniques and biomaterials used, tested or published. Electrospun nanofibers are introduced as a novel and potential material in gastrointestinal surgery. Nanofibers possess several, unique, physical and chemical properties, that may effectively stimulate cell proliferation and collagen production; a key requirement for the healed intestinal wound.

Long-term anti-inflammatory efficacy in intestinal anastomosis in mice using silver nanoparticle-coated suture

BackgroundIn our previous study, we coated silver nanoparticles (AgNPs) onto the surface of absorbable braided suture using layer-by-layer deposition and demonstrated significant anti-inflammatory property during the early phase of intestinal anastomosis healing in mice. The present study aimed to further investigate the long-term anti-inflammatory efficacy. MethodsAgNP-coated suture, antibiotic coated suture, and normal suture were respectively used for single layered, interrupted intestinal anastomosis. The anastomotic segments in each group were harvested on day 14, day 21, and day 28 postoperation and investigated for the degree of inflammation by cell infiltration and expression of cytokines as well as collagen deposition. ResultsWhen compared with the control groups, the AgNP-coated suture group showed better histological appearance in the intestinal anastomotic segments at each time point. Immunohistochemistry staining and quantitative evaluation further indicated less macrophage infiltration and decreased production of IL-6, IL-10, and TNF-α (p<0.05). Masson staining showed normal collagen deposition and remodeling at intestinal anastomotic tissue in the AgNP-coated suture group. ConclusionOur study shows that AgNP-coated suture provides better long-term anti-inflammatory efficacy and ideal tissue remodeling in intestinal anastomosis. Despite these findings, clinical trials are still needed for evaluation before medical application.

Comparison of the effects of platelet-rich or growth factor-rich plasma on intestinal anastomosis healing in pigs

BackgroundThe use of autologous platelet-rich plasma (PRP) and plasma rich in growth factors (PRGF) has been proposed for the treatment of several acute and chronic syndromes, such as corneal epithelial defects and dry eye syndrome, gum bleeding during oral surgery, and in orthopaedic surgery. We hypothesized that PRGF, rather than PRP, could be more effective because of its intrinsic characteristics in promoting the healing of intestinal anastomosis. The purpose of the present study was to evaluate and compare the effects of PRP and PRGF on various parameters of anastomotic healing in a swine model.MethodsEight female pigs were randomly assigned to two groups and subjected to hand sewn jeujuno-jejunal appositional extramucosal anastomoses. For each animal, a total of six anastomoses were performed: two were considered controls and received no treatment, while the remaining four anastomoses were treated with PRP or PRGF of which both were prepared at a platelet concentration that was respectively 3.4-fold and 2.81-fold higher than the original platelet count. In each animal, either PRP or PRGF was used as a treatment, to avoid interference among products. Animals were euthanized after 8 days and the anastomoses were evaluated and compared for the presence of adhesions, anastomotic leakage, bursting pressure, and histological appearance.ResultsThe concentration of platelets in PRP was 3.41-fold higher (range, 3.20–4.24) that the concentration in whole blood, while the concentration in PRGF was 2.81-fold higher (range, 2.89–4.88).The results obtained from the present study highlighted that there are no differences between anastomotic samples treated with either PRP or PRGF preparations, except for a significant increase in epithelization of the intestinal mucosa at the anastomotic site in the PRGF group.ConclusionsBoth PRP and PRGF suspensions should be considered a safe strategy and represent a relatively low-cost technology that is flexible enough to be applied in several therapeutic fields. No true benefit could be proven in our study compared to the no treatment following anastomoses formation, with the exception of enhanced epithelization of the mucosa in the PRGF group.

In vivo evaluation of Mg–6Zn and titanium alloys on collagen metabolism in the healing of intestinal anastomosis

There is a great clinical need for biodegradable materials, which were used as pins of circular staplers, for gastrointestinal reconstruction in medicine. In this work we compared the effects of the Mg–6Zn and the titanium alloys on collagen metabolism in the healing of the intestinal tract in vivo. The study included Sprague-Dawley rats and their effect was compared on rat’s intestinal tract, using serum magnesium, radiology, and immunohistochemistry in vivo. Radiographic and scanning electron microscope evaluation confirmed the degradation by Mg–6Zn alloy during the implantation period. Biochemical measurements including serum magnesium, creatinine, blood urea nitrogen and glutamic–pyruvic–transaminase proved that degradation of Mg–6Zn alloy showed no impact on serum magnesium and the function of other important organs. Superior to titanium alloy, Mg–6Zn alloy enhanced the expression of collagen I/III and relatively suppressed the expression of MMP-1/-13 in the healing tissues, leading to more mature collagen formation at the site of anastomosis. In conclusion, Mg–6Zn alloy performed better than titanium alloy on collagen metabolism and promoted the healing of intestinal anastomosis. Hence, Mg-6Zn may be a promising candidate for use of stapler pins for intestinal reconstruction in the clinically.

Preoperative vitamin C supplementation improves colorectal anastomotic healing and biochemical parameters in malnourished rats.

The objective of this study was to evaluate the effect of supplementation with vitamin C on intestinal anastomosis healing in malnourished rats. Male Wistar rats were divided into three groups: (1) sham, well-nourished rats that received vehicle; (2) FR+Veh, rats that were subjected to food restriction and received vehicle; and (3) FR+VC, rats that were subjected to food restriction and received vitamin C. Four days before surgery, the animals received vitamin C (100mg/kg/day) via gavage and underwent colon resection with anastomosis in a single plane. The survival rate of rats was monitored until day 7 after surgery. Regarding anastomosis tissues, we examined intra-abdominal adhesion index, hydroxyproline content, collagen density, inflammatory parameters, and oxidative damage to proteins and lipids. Malnutrition decreases body weight and increases mortality; the survival rate was 90% in group 1, 60% in group 2, and 80% in group 3. Vitamin C was able to increase hydroxyproline concentration and density of collagen and decrease the intra-abdominal adhesion index, as well as the infiltration of neutrophils and oxidative damage to proteins in malnourished rats compared to group treated with vehicle. Preoperative vitamin C supplementation can improve the intestinal anastomosis healing, biochemical alterations, and prolong survival in rats subjected to food restriction.

Evaluating the effect of infliximab on the healing of left colonic anastomosis in the presence of intra-abdominal sepsis

Background and study aimsInfliximab (IFX) is a chimeric anti-TNF-α body which is effectively used in the treatment of inflammatory bowel diseases and a variety of autoimmune diseases. The effect of IFX on the healing of intestinal anastomosis has been evaluated in several studies, however with conflicting results. Furthermore, the effect of IFX on colonic anastomosis in sepsis has not been evaluated to date. In this study, we aimed to investigate whether IFX has an adverse effect on the healing process of colonic anastomosis either under normal or septic condition. Material and methodThe efficiency of IFX was assessed with respect to anastomotic bursting pressure (ABP), tissue hydroxyproline levels (THL) and histopathological examination of left colonic anastomosis in 40 male rats. The rats were randomly allocated into four groups of 10 rats each as control (C), septic control (SC), control IFX (C-IFX) and septic IFX (S-IFX). ResultsThe anastomotic bursting pressure was measured at 182±19.1, 158±15.4, 161±26.8 and 100±10.3mm/Hg, in C, SC, C-IFX and S-IFX; respectively. IFX administration did not influence the anastomotic strength under normal condition whereas in sepsis significantly induced the reduction of APB. The mean THL was almost similar in both control groups (p=0.87), whilst IFX reduced the level of TH in sepsis comparing with control groups (p=0.01). IFX significantly impaired immune response in sepsis resulting in poor anastomotic healing in S-IFX group. ConclusionOur study demonstrated that IFX had no detrimental effect on the healing of colonic anastomosis under normal condition whilst significantly impaired the healing process in sepsis.

The Effect of Infliximab on Intestinal Anastomosis Healing in Rats

Intestinal anastomosis healing is a complex physiological process in which many local and systemic factors play a role. One of the significant cytokines in this process is TNF-α. Infliximab is a chimeric monoclonal antibody which binds to TNF-α with high affinity. Although this agent is used in ulcerative colitis and Crohn's disease, intestinal surgery may be required in these patients. In this study it was aimed to determine whether or not there was any negative effect of preoperative single dose infliximab treatment on intestinal anastomosis healing. Two groups of 10 rats were formed. One of these groups was administered with a single dose of infliximab 8 mg/kg as a 20-minute intravenous infusion from the femoral vein. Four days after the infusion, a full layer incision was made to the colon and anastomosis was applied to all the rats. At 7 days after anastomosis, the subjects were sacrificed. The anastomosis segment was removed and the bursting pressure was measured. Tissue samples were taken from this segment for hydroxyproline concentration and histopathological examination. A blood sample was taken to measure TNF-α values. No statistically significant difference was determined between the groups in terms of bursting pressure, tissue hydroxyproline concentration or histopathological scoring. A single dose of 8 mg/kg infliximab administered 4 days preoperatively was not found to have any negative effect on intestinal anastomosis healing in rats.

Esophagogastric anastomosis in rats: Improved healing by BPC 157 and L-arginine, aggravated by L-NAME.

AIMTo cure typically life-threatening esophagogastric anastomosis in rats, lacking anastomosis healing and sphincter function rescue, in particular.METHODSBecause we assume esophagogastric fistulas represent a particular NO-system disability, we attempt to identify the benefits of anti-ulcer stable gastric pentadecapeptide BPC 157, which was in trials for ulcerative colitis and currently for multiple sclerosis, in rats with esophagocutaneous fistulas. Previously, BPC 157 therapies have promoted the healing of intestinal anastomosis and fistulas, and esophagitis and gastric lesions, along with rescued sphincter function. Additionally, BPC 157 particularly interacts with the NO-system. In the 4 d after esophagogastric anastomosis creation, rats received medication (/kg intraperitoneally once daily: BPC 157 (10 μg, 10 ng), L-NAME (5 mg), or L-arginine (100 mg) alone and/or combined or BPC 157 (10 μg, 10 ng) in drinking water). For rats underwent esophagogastric anastomosis, daily assessment included progressive stomach damage (sum of the longest diameters, mm), esophagitis (scored 0-5), weak anastomosis (mL H2O before leak), low pressure in esophagus at anastomosis and in the pyloric sphincter (cm H2O), progressive weight loss (g) and mortality. Immediate effect assessed blood vessels disappearance (scored 0-5) at the stomach surface immediately after anastomosis creation.RESULTSBPC 157 (all regimens) fully counteracted the perilous disease course from the very beginning (i.e., with the BPC 157 bath, blood vessels remained present at the gastric surface after anastomosis creation) and eliminated mortality. Additionally, BPC 157 treatment in combination with L-NAME nullified any effect of L-NAME that otherwise intensified the regular course. Consistently, with worsening (with L-NAME administration) and amelioration (with L-arginine), either L-arginine amelioration prevails (attenuated esophageal and gastric lesions) or they counteract each other (L-NAME + L-arginine); with the addition of BPC 157 (L-NAME + L-arginine + BPC 157), there was a marked beneficial effect. BPC 157 treatment for esophagogastric anastomosis, along with NOS-blocker L-NAME and/or NOS substrate L-arginine, demonstrated an innate NO-system disability (as observed with L-arginine effectiveness). BPC 157 distinctively affected corresponding events: worsening (obtained with L-NAME administration that was counteracted); or amelioration (L-arginine + BPC 157-rats correspond to BPC 157-rats).CONCLUSIONInnate NO-system disability for esophagogastric anastomoses, including L-NAME-worsening, suggests that these effects could be corrected by L-arginine and almost completely eliminated by BPC 157 therapy.