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Intestinal ischemia reperfusion injury (IRI) is a challenging problem and it adversely affects the healing of colonic anastomosis. Our experimental study aimed to investigate the role of mesenchymal stem cells (MSC) administration in the healing of colonic anastomosis. A total of 33 rats were grouped as Control, IRI and MSC treatment groups. Three rats were reserved for obtaining MSCs. Colonic resection and anastomosis procedure was performed in all groups. Anastomotic line was wrapped with MSCs impregnated spongostan after colonic anastomosis in the rats of the MSC treatment group. All rats were sacrificed and anastomotic line were sampled for examination on the post operative seventh day. Tissue hydroxyproline (HP) levels and anastomotic bursting pressures were statistically compared. Anastomotic bursting pressures were found to be significantly high in MSC treatment group rats. The lowest anastomotic bursting pressure was detected in IRI group rats. Hydroxyproline content of the anastomotic sites were also found to be significantly higher in the rats of the MSC treatment group when compared with the IRI group rats. Our study showed that the detrimental effects of IRI on the healing process of colonic anastomosis in an experimental model may be alleviated with the treatment of MSCs.

Abstract Background Leakage following colorectal anastomosis surgery causes various complications associated with high morbidity and mortality, especially in pediatric patients. It might be caused by the use of non-steroidal anti-inflammatory drugs (NSAIDs) as postoperative analgesics. This study aimed to compare the effect of metamizole and paracetamol on colonic anastomosis and fibroblast activities, including proliferation, migration, and collagen synthesis, in Wistar rats. Methods Rats were divided into control, paracetamol and metamizole groups. The colonic anastomosis was evaluated by determining the integrity of the muscle layers, the formation of granulation tissue, and mucosal anastomosis. Fibroblast activities were analyzed by measuring the proliferation, migration, and collagen synthesis. Results Metamizole caused more damage to muscle layer integrity, more inhibition of granulation tissue formation in the anastomosis area and lower mucosal anastomosis compared with paracetamol and control groups. Metamizole had a higher cytotoxic effect than paracetamol, which suppressed the proliferation and migration of fibroblasts. Furthermore, both drugs did not affect the synthesis of collagen. Conclusion Metamizole shows worse effects on the integrity of muscle layers, inhibition of granulation tissue formation, mucosal anastomosis, fibroblast proliferation, and migration, but not collagen synthesis, than paracetamol in Wistar rat intestines following colonic anastomosis. These findings might indicate that paracetamol is safer than metamizole as analgesic following colonic anastomosis.

Objective: Symptomatic anastomotic leakage observed after colorectal surgery is one of the major complications. One of the factors affecting the anastomosis healing is the type of suture material used. This study aims to investigate the effects of the suture material polydioxanone (PDS, Ethicon), which is late absorbable and has a monofilament structure, and the suture material polyglactin 910 (Vicryl, Ethicon), which is absorbable and has a multifilament structure, on the healing of colonic anastomosis, and to compare the traditional forms of these sutures with their antibacterial effective triclosan-impregnated forms (PDS Plus, Ethicon and Vicryl Plus, Ethicon). Materials and Methods: The rats were divided into four equal groups consisting of 10 subjects each: Group I: Vicryl; Group II: Vicryl Plus; Group III: PDS; and Group IV: PDS Plus. The presence of wound infection, whether the integrity of the abdominal wall was maintained, intra-abdominal adhesion scoring, the presence of intra-abdominal abscess, and whether the macroscopic integrity of anastomosis was maintained were evaluated. Results: According to the results of this experimental study, while the highest ABP and hydroxyproline levels were observed in the PDS Plus group, the lowest values were observed in the Vicryl group. However, the difference was not statistically significant. Conclusion: Slowly absorbable and monofilament PDS suture material causes less tissue reaction and inflammatory response compared to the Vicryl suture material that is absorbable in the colonic anastomosis line and multifilament.

Background. Repeated surgical interventions aimed at the colostomy closure and restoration of intestinal continuity after obstructive resections are traumatic and complex reconstructive operations on the colon. The complications of recovery operations largely feature wound abscesses (30–60 %) and anastomotic leakage (5–23 %), with the mortality levels from these complications reaching 0.4–2.8 %. Pronounced dysbiotic changes, the progression of catarrhal or erosive diversion colitis and proctosigmoiditis are the risk factors for anastomotic leakage and postoperative bowel dysfunction. This determines the need for a therapeutic algorithm of intestine sanitation and the introduction of methods for preventing the anastomotic leakage after obstructive resection of the distal colon (Hartmann’s operation). Aim. To evaluate the capability and efficiency of the intraluminal application of rifaximin-α (Alfa Normix® preparation, Alphasigma company) as a sanitising intestinal antiseptic for relieving diversion colitis and prevening postoperative complications of restorative operations after obstructive resection of the colon (Hartmann’s operation).Material and methods. A prospective comparative controlled study of the results of surgical treatment in 63 patients was conducted. The patients were divided into 2 groups, which were compared using the case-control principle. The operations included the colostomy closure and the restoration of intestinal continuity by applying a colonic anastomosis. In group I (main group, 30 patients), the original “Method of postoperative prevention of colonic anastomotic leakage” (Russian patent No. 2523822) was applied. In the control group (33 patients), routine methods for preventing anastomotic leakage and antibiotic prophylaxis were used. A comparative analysis of the number and severity of purulent-septic complications (taking into account the possibility of their relief), as well as the quality and rate of intestinal dysbiosis relief and the manifestations of diversion proctitis, was carried out.Results. The main group indicated the best results in terms of the main analysed parameters, fewer complications and anastomotic leakage. Thus, the total number of complications in groups I and II was 16.7 % and 27.3 %, respectively. The proportion of complications associated with the anastomotic leakage, inflammatory changes in this area and impaired patency in groups I and II was 6.7 % and 12.1 %, respectively. Rifaximin-α (Alpha Normix®) intolerance and pathological reactions associated with its use were not observed. The possibility for effective relief of the violations of quantitative and qualitative composition of the microflora in the colon stump was revealed. The main group showed better results in terms of eliminating local manifestations of diversion colitis and anorectal dysfunction compared to group II, where traditional schemes of postoperative therapy were used.Conclusions. Intraluminal dosed use of the Alpha Normix® suspension with the rifaximin concentration of 100 mg per 5 ml is an effective method of postoperative prevention of purulent-septic complications associated with anastomotic leakage during reconstructive operations on the distal colon. The introduction of Alpha Normix® in the scheme of postoperative treatment allows its results to be stabilised due to the sanitation of the intestinal lumen, the elimination of dysbiosis, the relief of inflammatory reaction and the elimination of factors supporting the development of diversion proctitis.

ABSTRACT Background: The use of probiotics positively modifies the composition and function of intestinal flora, improving the quality of intestinal anastomosis. Aim: To evaluate the impact of probiotic use on intestinal anastomosis of rats. Method: Thirty-six adult male Wistar rats (Rattus norvegicus albinus, Rodentia Mammalia) were used, with body weight ranging from 220-320 g. The animals were housed and acclimated individually in boxes receiving water and ration ad libitum. After initial acclimatization, the control group received perioperative ration ad libitum for 12 days (seven preoperatively and five postoperatively) associated with the maltodextrin formula at a dose of 250 mg/day in isocaloric and isovolumetric form. Likewise, the probiotic group received oral supplementation of probiotics dose of 250 mg/day, associated with isocaloric and isovolumetric diet. The probiotic chosen for this study was composed of strains (doses 1x109 CFU/g)12 Lactobacillus paracasei LPC-37, Bifidobacterium lactis HN0019, Lactobacillus rhamnosus HN001 and Lactobacillus acidophilus NCFM. Probiotics or placebo were administered orally with the aid of a dosimeter spatula. Both groups underwent two colostomies, one in the right colon and the second in rectosigmoid, followed by reanastomosis with eight separate 6-0 mononylon stitches. The sacrifice took place on the fifth day. The parameters evaluated included tensile strength, histology and collagen densitometry. Results: The rate of intestinal fistula for the control and probiotic groups were, respectively, 22.22% and 11.11% (p=0.6581).Perioperative supplementation with probiotics increased collagen deposition of types I and III (p

Background: Radiotherapy is one of the most important factors which results in negative effects on wound healing and increases anastomosis leakage. Diverting loop ileostomy has been usually performed after colorectal anastomosis in cases of colorectal cancer with a history of neoadjuvant radiotherapy to decrease the chance of leakage. Considering the side effects of diverting loop ileostomy, the objective of the present study is to investigate the effect of human amniotic membrane (HAM) on colorectal anastomosis leakage after neo-adjuvant radiotherapy. Methods: In this experimental animal study, 20 crossbreed rabbits were randomly divided into two groups (case group: 13 rabbits, control group: 7 rabbits) after receiving an equal dose of external beam radiation. Four weeks after irradiation, resection of 4 cm of colorectal segment and end-to-end single layer anastomosis were conducted. In the case group, a 2×2 cm wrap of HAM applied around the site of anastomosis. Eight weeks later, all the survived rabbits were sacrificed. A segment of anastomotic sites was resected in all expired and survived rabbits and sent for pathological evaluation. Mann-Whitney U Test (SPSS for Windows, Ver. 16, Chicago, IL) was applied to analyze healing scores between the two groups. Results: Due to anastomosis dehiscence, 5 rabbits expired in the control group, but all the 13 rabbits (case group) survived after 8 weeks and showed no leakage. In addition, pathological evaluation revealed significant epithelialization and neovascularization in the case group. Statistically, healing score was higher in the case group rather than the control group (P

Background: The use of long-chain polyunsaturated fatty acids has been studied in the context of healing and tissue regeneration mainly due to its anti-inflammatory, immunoregulatory and antioncogenic properties. Previous studies have demonstrated beneficial effects with the use of enteral immunonutrition containing various farmaconutrients such as L-arginine, omega-3, trace elements, but the individual action of each component in the healing of colonic anastomosis remains unclear. Aim: To evaluate the influence of preoperative supplementation with omega-3 fatty acids on the healing of colonic anastomoses of well-nourished rats. Methods: Forty Wistar adult male rats, weighing 234.4±22.3 g were used. The animals were divided into two groups: the control group received for seven days olive oil rich in omega-9 oil through an orogastric tube, while the study group received isocaloric and isovolumetric omega-3 emulsion at a dose of 100 mg/kg/day, also for seven days. Both groups were submitted to two colotomies followed by anastomosis, in the right and left colon, respectively. Parameters evaluated included changes in body weight, anastomotic complications and mortality, as well as maximum tensile strength by using a tensiometer and collagen densitometry at the anastomotic site. Results: There were no differences in body weight or mortality and morbidity between groups. The value of the maximum tensile strength of the control group was 1.9±0.3 N and the study group 1.7±0.2, p=0.357. There was, however, a larger amount of type I collagen deposition in the study group (p=0.0126). The collagen maturation índex was 1.74±0.71 in the control group and 1.67±0.5 in the study group; p=0,719). Conclusions: Preoperative supplementation of omega-3 fatty acid in rats is associated with increased collagen deposition of type I fibers in colonic anastomoses on the 5th postoperative day. No differences were observed in the tensile strength or collagen maturation index.

OBJECTIVE: Fibrin glues have not been consistently successful in preventing the dehiscence of high-risk colonic anastomoses. Fibrinogen and thrombin concentrations in glues determine their ability to function as sealants, healers, and/or adhesives. The objective of the current study was to compare the effects of different concentrations of fibrinogen and thrombin on bursting pressure, leaks, dehiscence, and morphology of high-risk ischemic colonic anastomoses using fibrin glue in rats. METHODS: Colonic anastomoses in adult female Sprague-Dawley rats (weight, 250-350 g) treated with fibrin glue containing different concentrations of fibrinogen and thrombin were evaluated at post-operative day 5. The interventions were low-risk (normal) or high-risk (ischemic) end-to-end colonic anastomoses using polypropylene sutures and topical application of fibrinogen at high (120 mg/mL) or low (40 mg/mL) concentrations and thrombin at high (1000 IU/mL) or low (500 IU/mL) concentrations. RESULTS: Ischemia alone, anastomosis alone, or both together reduced the bursting pressure. Glues containing a low fibrinogen concentration improved this parameter in all cases. High thrombin in combination with low fibrinogen also improved adherence exclusively in low-risk anastomoses. No differences were detected with respect to macroscopic parameters, histopathology, or hydroxyproline content at 5 days post-anastomosis. CONCLUSIONS: Fibrin glue with a low fibrinogen content normalizes the bursting pressure of high-risk ischemic left-colon anastomoses in rats at day 5 after surgery.

OBJECTIVES: In addition to its roles in the stimulation of growth hormone secretion and the regulation of appetite and metabolism, ghrelin exerts immunomodulatory, anti-inflammatory and antioxidant actions in several organ systems. In this study, we investigated the effects of ghrelin on the healing of experimental colonic anastomoses. METHODS: Wistar rats were randomly divided into two groups (n = 10 in each). A segment of colon was excised, and an end-to-end anastomosis was performed in the distal colon. The Ghrelin Group received 10 ng/kg/day IP ghrelin for seven days postoperatively, whereas the Control Group received an identical volume of saline. On the seventh postoperative day, the anastomotic bursting pressures and hydroxyproline levels were measured, and adhesion formation around the anastomoses was examined. Histopathological analyses were performed to evaluate inflammatory cell infiltration, fibroblast infiltration, collagen density and neovascularization. RESULTS: In the Ghrelin Group, the bursting pressure and hydroxyproline levels were significantly higher than in the Control Group. The adhesion formation scores were lower in the Ghrelin Group than in the Control Group. Although the inflammatory cell infiltration was diminished in the Ghrelin Group, the degrees of fibroblast infiltration, collagen density and neovascularization were not significantly different between the groups. CONCLUSION: Our results indicate that ghrelin improves the healing of colonic anastomoses in rats.

BACKGROUND: Leakage from colonic anastomosis is a major complication causing increased mortality and morbidity. Ischemia is a well-known cause of this event. This study was designed to investigate the effects of adrenomedullin on the healing of ischemic colon anastomosis in a rat model. METHODS: Standardized left colon resection 3 cm above the peritoneal reflection and colonic anastomosis were performed in 40 Wistar rats that were divided into four groups. To mimic ischemia, the mesocolon was ligated 2 cm from either side of the anastomosis in all of the groups. The control groups (1 and 2) received no further treatment. The experimental groups (3 and 4) received adrenomedullin treatment. Adrenomedullin therapy was started in the perioperative period in group 3 and 4 rats (the therapeutic groups). Group 1 and group 3 rats were sacrificed on postoperative day 3. Group 2 and group 4 rats were sacrificed on postoperative day 7. After careful relaparotomy, bursting pressure, hydroxyproline, malondialdehyde, interleukin 6, nitric oxide, vascular endothelial growth factor, and tumor necrosis factor alpha levels were measured. Histopathological characteristics of the anastomosis were analyzed. RESULTS: The group 3 animals had a significantly higher bursting pressure than group 1 (p

BACKGROUND: Failure of anastomotic healing is one of the major complications in colorectal surgery. Because histamine plays an important role in immune and inflammatory reactions, we demonstrate the effects of famotidine on the healing of colonic anastomosis in rats. METHODS: Twenty-eight Sprague-Dawley rats were used in the study. Excision and end-to-end anastomosis was performed in the distal colon of the rat. The Famotidine Group received 2 mg/kg/day famotidine; the Control Group received the same amount of saline. Bursting pressure of anastomoses and hydroxyproline content of perianastomotic tissues were evaluated on the third and seventh days following surgery. RESULTS: Bursting pressures and hydroxyproline contents for the Famotidine Group were significantly lower than the equivalent parameters for the Control Group on both the third and seventh days post-surgery. CONCLUSIONS: According to our findings, famotidine exerts detrimental effects on the anastomotic bursting pressure and hydroxyproline content of perianastomotic tissues in the colon of rats.

PURPOSE: Thalidomide, because of its anti-inflammatory properties, as re-emerged as an option for the treatment of Crohn's disease refractory to standard therapy. We studied the effect of thalidomide on the healing of colonic anastomosis. METHODS: Sixty male rats (Rattus norvegicus), were divided into 3 groups of 20 animals each, respectively receiving 0.5 or 1.0 mg/kg thalidomide by the oral route for 7 days, or saline solution (control). All animals were submitted to continuous end-to-end anastomosis with 6-0 Prolene sutures. After sacrifice the anastomoses were analyzed macroscopically and submitted to determination of hydroxyproline, to histology and to immunohistochemistry for metalloproteinase 1, metalloproteinase 1 inhibitor and vascular endothelial growth factor (VEGF). RESULTS: Statistical analysis of the data showed no significant difference in macroscopic aspect or hydroxyproline determination (p= 0.5403). In the immunohistochemical analysis, the following p values were obtained: p = 0.5817 for VEGF, p = 0.1854 for metalloproteinase 1, and p = 0.0023 for metalloproteinase 1 inhibitor, with this last value being considered statistically significant. CONCLUSION: We conclude that thalidomide influenced collagen maturation. There was a stronger action of metalloproteinases, possibly indicating a negative tendency for the healing process.OBJETIVO: Sabe-se que agentes farmacológicos podem influenciar no processo de cicatrização. A talidomida, devido às suas propriedades antiinflamatórias, tem ressurgido como uma opção no tratamento da doença de Cröhn refratária à terapêutica convencional. Neste trabalho, estudamos o efeito da talidomida na cicatrização de anastomoses colônicas no rato. MÉTODOS: Foram utilizados 60 animais Rattus norvegius, com peso médio de 300g. Organizou-se 3 grupos de 20 animais, sendo um grupo controle (AC), um grupo (BD), com administração de talidomida 0,5 mg/kg por 7 dias e um grupo (AD) com administração de talidomida 1,0 mg/kg por 7 dias. Foi realizada anastomose término-terminal contínua de prolene 6-0. O sacrifício foi no 7º. dia pós operatório e as anastomoses foram analisadas quanto a aspecto macroscópico, dosagem de hidroxiprolina, histologia em hematoxilina-eosina e imuno-histoquímica para metaloproteinase 1, inibidor de metaloproteinase 1 e VEGF. RESULTADOS: Não houve diferença estatisticamente significante para a observação macroscópica e para dosagem de hidroxiprolina (p=0,5403). Na análise imunohistoquímica, para VEGF houve p=0,5817, para metaloproteinase 1, p=0,1854 e para inibidor de metaloproteinase, p=0,0023, considerado estatisticamente significante. CONCLUSÃO: Concluímos que a talidomida influenciou na maturação do colágeno. Notou-se maior ação das metaloproteinases, que pode significar uma tendência negativa para o processo cicatricial.