Study On The Esophageal Acellular Matrix Combined With Bone Marrow Mesenchymal Stem Cells In Construction Of Tissue Engineered Esophagus

The esophagus is the passage that connected the throat and stomach. The food isentered into the esophagus from pharyngeal swallowing and through the esophagusinto the stomach cavity. Esophageal organic disease or dynamic disorder or functionalesophageal disease can influence the function of the esophagus. Esophageal diseasesinclude esophageal abnormalities, movement disorders, inflammation, and tumor. Butsome of them are asymptomatic or mild, little influence on health. Some diseaseshave influence on eating, even threatened life and the surgery is necessary. Thetreatment of esophageal diseases mainly are surgery, radiotherapy, chemical treatmentand biological treatment. These methods are not very good to improve cure rateduring the esophageal diseases. The middle of the20th century, people started usingtheir other tissues or organs to replace the damaged esophagus,however,surgicaltrauma is serious and the postoperative is complicate. So people have been trying tofind an ideal agent for artificial esophagus. With the progress of materials science,theresearch of artificial esophagus has entered a new stage. So far, there can be used forartificial esophagus biological substitute materials mainly include syntheticbiodegradable polymeric materials and Extracellular matrix (Extracellular matrix,ECM) derived materials. Because no cell esophageal stents can be absorbedgradually,at the same time can induce remodeling of tissue specificity,thus it isbecome the object of study. Stem cell technology is one of the frontier technology inthe field of regenerative medicine research.In recent years,stem cell treatment alsoprovides a new research strategy for a variety of diseases and one of the aspects canbe used as organ tissue engineering seed cells.Method:The tissue was removed cells by the chemical detergent sodium deoxycholic acidand made from natural extracellular matrix (ECM) esophageal stent. The histologicobservation was made by immunohistochemical staining,HE staining and Massonstaining, VG staining for, etc. Then we observed the ultrastructural with scanningelectron microscopy (sem), transmission electron microscopy and optical microscope. We separated and extracted the rat bone marrow mesenchymal cells (BMSCs) and invitro cultured,amplified,identified by flow cytometry. We used a low concentration ofperacetic acid solution to sterilization ECM scaffold. The BMSCs was vaccinated inthe ECM and cultivated,then we observed and compared HE dyeing.Result:1. The treated ECM is white, soft,good elasticity,natural tubular ECM forming good.The result which was observed by HE staining shows that the ECM internal has nocomponents residue. The result which was observed by Masson staining, VG stainingand immunohistochemical staining shows that the collagen, elastin, fiber connectionprotein of ECM did not differ significantly compared with the tissue that did notremove the cells. The three-dimensional space and three dimensional networkstructure of ECM was observed by scanning electron microscopy (SEM) andtransmission electron microscopy (SEM). The ECM was used to germiculture aftersterilized with peracetic acid solution. Results show that the sterilization effect isgood.2. We separated and cultivated BMSCs by whole bone marrow adherent methodfrom bone marrow,the third generation of cells was detected by flow cytometry.The positive expression rates of CD29,CD90,CD3,CD45were98.21%,99.58%,2.43%and2.38%and that was in accordance with BMSCs antigen expression.3. After BMSCs and ECM compound cultivating,HE staining of the experimentalgroup showed numerous aizen nucleus structure,compact connection and cells growthwas visible in the pores.Conclusion:The use of whole bone marrow adherent culture among law separablemesenchymal stem cells in P3generation of more than98%purity. Sodiumdeoxycholate joint DNaseI to decellularize esophageal.It removed cell components intissue effectively and retained the extracellular matrix; The size and volume of thestent were not changed significantly, which confirmed we obtained the idealesophageal stent that has no cells. We cutured the cells mixed with tissue by themethod of quadratic precipitation, and then BMSCs grew better in the scaffold.