| Objective 1.To explore a simplified mothed by which we can construct a tracheoesophageal fistula(TEF)model by midtronic aortic perforator.2.To study the safety and effectiveness of stem cell therapy by injecting human adiposederived mesenchymal stem cells(AMSCs)into tissue around fistula.3.To analyse differential protein expression by collecting fistula tissues for transcriptome sequencing.Method 1.animal preparation 13 beagles were divided into 3 groups.Group 1: blank control group,1 animal(1101);Group 2: operation control group,TEF operation + PBS injection,6 animals(2101~2106);Group 3: stem cell transplant group,TEF operation + stem cell injection,6 animals(3101~3106).2.Operation After anesthesia,TEF construction was performed.The incision in the middle of the neck was taken to expose trachea and esophageal.The fistula was performed on the posterior wall of trachea and adjacent anterior wall of esophageal by midtronic perforator at the level of 3-4 tracheal cartilage ring.The diameter of fistula was 8 mm.2ml mixture including 106 stem cell and PBS was injected into tissue around fistula in group 3.The equal PBS solution was injected into same tissue in group 2.After injection,3-0 suture was used to suture the fistulas between trachea and esophageal.The incision was sutured.The TEF operation was completed.3.Postoperation 1.Nasogastric nutrition was the first nutrition option.It should be substitute by parenteral nutrition immediately if animals present intolerant to nasogastric intubation.2.The result of general conditions and fiberoptic bronchoscope was recorded every week.3.Animals was planned to be executed on 35 day.The general condition of fistula tissue,lung and Masson staining section of fistula tissues were observed.4.Fistula tissues of group 2 and 3 were collected for transcriptome sequencing.Result 1.Midtronic aortic perforator can simplify the method constructing TEF model effectively.2.In the first week after operation,3 animals in group 2 and 1 animal in group 3 died.2 animals in group 2 died on 32 day and 35 day.The rest animals survived on 35 day.3.In group 3,excepting the dead animals in the first week,comparing with the initial diameter of 8 mm,the fistulas of 2 cases closed,the diameter of other 2 cases decreased to 3 mm,1 case decreased to 4 mm.On the other hand,in group 2,excepting the dead animals in the first week,the fistulas diameter of 2 cases were 7 mm,1 case were 6.5 mm.The mean diameter of fistulas in group 3 was smaller than group 2.4.The range of inflammatory consolidation in lung of group 3 was smaller than group 2 after stem cell transplantation on 35 day.5.The result of transcriptome sequencing presented the differential expression in HIF-1α、TNF-1 and PI3K/Akt pathway.Conculsion 1.Midtronic perforator simplified the construction of TEF modeling 2.Stem cell transplantation can promote the healing of fistula without obvious side effects with good efficacy and safety.3.According to the result of transcriptome sequencing,HIF-1α 、 TNF-1 and PI3K/Akt pathway may play an important role in fistula healing process,but further specific mechanisms should be verified. |
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