CAJ | 학술논문

背景：临床上治疗慢性静脉功能不全的主要方法是静脉瓣膜修复及带瓣静脉段移植，但这些方法创伤较大，且带瓣静脉来源有限。组织工程学和再生医学在修复病变血管方面取得的进步，而以自体来源的内皮细胞为种子细胞的组织工程带瓣静脉也见于了报道，但存在排出反应。 　　目的：构建一个有可自我更新、修复、类似天然瓣膜结构并具有功能的带瓣静脉。方法：麻醉取 Beagle 犬的骨髓获取骨髓间充质干细胞，采用密度梯度离心和贴壁法获取骨髓间充质干细胞，并进行细胞的传代、冻存复苏、流式细胞仪检测和定向诱导分化。采用热致相分离技术，以聚(乳酸-乙醇酸)共聚物为基材，利用自制带瓣静脉模具制备三维组织工程带瓣静脉支架，制备组织工程带瓣静脉支架，并研究其形态结构。将骨髓间充质干细胞种植在支架上构建可降解的带瓣静脉，在体外培养2周。
배경：림상상치료만성정맥공능불전적주요방법시정맥판막수복급대판정맥단이식，단저사방법창상교대，차대판정맥래원유한。조직공정학화재생의학재수복병변혈관방면취득적진보，이이자체래원적내피세포위충자세포적조직공정대판정맥야견우료보도，단존재배출반응。 　　목적：구건일개유가자아경신、수복、유사천연판막결구병구유공능적대판정맥。방법：마취취 Beagle 견적골수획취골수간충질간세포，채용밀도제도리심화첩벽법획취골수간충질간세포，병진행세포적전대、동존복소、류식세포의검측화정향유도분화。채용열치상분리기술，이취(유산-을순산)공취물위기재，이용자제대판정맥모구제비삼유조직공정대판정맥지가，제비조직공정대판정맥지가，병연구기형태결구。장골수간충질간세포충식재지가상구건가강해적대판정맥，재체외배양2주。
BACKGROUND:Chronic venous insufficiency is a major health problem worldwide. Clinical treatments include venous valve repair and venous segment containing valve transplantation. However, these are invasive procedures, and the supply of vein containing valves is limited. Significant progress in the fields of tissue engineering and regenerative medicine has been made towards the creation of tissue engineered vascular grafts for the repair of damaged or malformed vessels. It has been reported that using tissue engineering, a tissue engineered vein containing valves constructed with self-derived endothelial cells and al ogeneic acellular matrices can provide the complex physiological valve structure and mechanical stability, but this elicited an immunogenic response. OBJECTIVE:To create a viable and functional vein containing valves, which has the ability to grow, repair, and imitate natural tissues. METHODS:Bone marrow mesenchymal stem cells were obtained from Beagle dogs by density gradient centrifugation and adherence methods. Bone marrow mesenchymal stem cells were cultured in vitro. Fol owing isolation and culture the cells were examined using flow cytometry and identified by direct induction towards the osteogenic and adipogenic lineages. We fabricated biodegradable venous scaffold containing valves using the method of injection molding combined with thermal y induced phase separation. Based on the self-made cast, a three-dimensional biodegradable vein scaffold containing valves was constructed from poly(lactic-co-glycolic acid). Morphological structure was tested. Bone marrow mesenchymal stem cells were used as seed cells to be seeded onto the lumen of the tissue engineered vein scaffold containing valves in vitro and then incubated for 2 weeks. RESULTS AND CONCLUSION:Scanning electron microscopy images showed that the scaffold demonstrated sufficient porosity. Cultured cells expressed mesenchymal cellmarkers, CD44 and CD29, but did not express hematopoietic cellmarkers, CD34 and CD45 at the same time point. Scaffolds were nontoxic to cells and were favorable for the growth and migration of bone marrow mesenchymal stem cells. cells attached on the surface of poly(lactic-co-glycolic acid) scaffolds formed a confluent layer after incubation. The cellular constructs were tested in vitro, and the valve leaflets were functional y capable of opening and closing when stimulated. These results suggested that the tissue engineered vein containing valves have been successful y constructed by using a three-dimensional poly(lactic-co-glycolic acid) scaffold and bone marrow mesenchymal stem cells as seed cells. Tissue engineered vein containing valves is potential y useful for the substitution and regeneration of vein valves.