| Purpose This experiment verifies whether there are differences between different-layer suture and the same-layer suture anastomosis of small intestine and whether there are advantages of stent anastomosis compared with traditional hand-sewn method.Methods One hundred and twenty male Sprague-Dawley rats (8-week-old,270Π30g) were randomly divided into five groups(n=6):Control group(use simple interrupted suture, the same-layer anastomosis), SM vs SM group(use interrupted inverting suture, serous membrane to serous membrane anastomosis), MM vs MM group(use interrupted everting suture, mucous membrane to mucous membrane anastomosis), SM vs MM group(use intussusception-like suture, serous membrane to mucous membrane anastomosis), and Stent group (use stent anastomosis). Time to perform the anastomosis, mortality and complications, abdominal adhesion scores, bursting pressures, hydroxyproline contents were determined at postoperative days3d,7d,14d,30d. Intestinal contrast imaging under X-rays was carried out at postoperative days30in vitro. Anastomotic tissues were histologically analyzed under the hematoxylin-eosin(HE) staining, Masson staining, the immunohistochemical staining of anti-TGF-β1, anti-b-FGF and anti-α-SMA. mRNA expressions of TGF-β1, b-FGF and α-SMA were evaluated as well via real-time reverse transcription polymerase chain reactions(RT-PCR).Results Two rats died during the experiments, one was ate off by others the next day, and another died of over dose of anesthetic drug in operation. All of the other rats had no complications during the experiments. Anastomotic time of Control group, SM vs SM group, MM vs MM group, SM vs MM group and stent group were3.96±1.803min,6.75±1.113min,6.29±1.042min,6.54±1.103min,1.75±0.794min, respectively, stent anastomosis is an easy and convienient method as a result it needs least time to perform, which has significant differences statistically(P<0.01). Abdominal adhesion scores of Control group, SM vs SM group, MM vs MM group, SM vs MM group and stent group are1.00±0.905,0.83±0.702,1.87±0.815,1.13±0.741,1.46±0.509, respectively, method of serous membrane to serous membrane suture anastomosis causes least abdominal adhesion, mucous membrane to mucous membrane suture anastomosis and stent anastomosis seem to cause more abdominal adhesion than other groups, the results are significant different statistically(P<0.01). Bursting pressure show that there were no difference among5groups in the early(POD3d) and last stage(POD30d)(P>0.01), but Control group and stent group seemed stronger than other group in the middle stage(POD7d and14d)(P<0.01). Determination of hydroxyproline contents show hydroxyproline contents go up gradually over time but there were no significant differences compared with Control group at the same period(P>0.01). Intestinal contrast imaging under X-rays show none of a group had leakage or stenosis, every group has the same results on this respect. Intestine tissue histological studies show that Control group and stent group had less inflammation and began healing earlier than other groups, but each group reached the same end at POD30d, there are no differences statistically. The immunohistochemical staining of anti-TGF-β1, anti-b-FGF and anti-α-SMA in each group show that TGF-β1and b-FGF decreased gradually over time while α-SMA increased at beginning then decreased after that, and there were no significant differences compared with Control group on each time point(P>0.01); RT-PCR assay of mRNA expressions of TGF-β1, b-FGF and α-SMA show almost the same results as the immunohistochemical staining; results of expressions of TGF-β1, b-FGF and a-SMA were more or less the same as reported in literatures.Conclusion Different-layer suture anastomosis makes an equal healing ending to the same-layer suture which begins the healing process earlier. Stent anastomosis is a safe and feasible method of intestine anastomosis which makes the anstomosis easier and faster. |
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