中华实验外科杂志
中華實驗外科雜誌
중화실험외과잡지
CHINESE JOURNAL OF EXPERIMENTAL SURGERY
2015年
8期
1844-1847
,共4页
周建良%聂彬恩%朱志刚%丁静丽%胡时栋%陈佳
週建良%聶彬恩%硃誌剛%丁靜麗%鬍時棟%陳佳
주건량%섭빈은%주지강%정정려%호시동%진가
组织工程心脏瓣膜%去细胞猪主动脉瓣膜%聚乙二醇%共价修饰%内皮化
組織工程心髒瓣膜%去細胞豬主動脈瓣膜%聚乙二醇%共價脩飾%內皮化
조직공정심장판막%거세포저주동맥판막%취을이순%공개수식%내피화
Tissue engineered heart valve%Decellularized porcine aortic valve%Polyethyl glycol%Covalent modification%Endothelialization
目的 观察枝状化聚乙二醇(PEG)修饰后的去细胞猪主动脉瓣(DPAV)的内皮细胞生长性能.方法 自合成枝状化聚乙二醇(4-arm-PEG-acrylate),采用核磁共振氢谱(1HNMR),对复合物进行表征检测,利用4-arm-PEG-acrylate上的丙烯酸酯与巯基化的DPAV结合对其进行PEG化,体外获取人脐静脉内皮细胞,以细胞密度为1×105接种于各组瓣膜支架材料上,8d后MTS试剂、苏木素-伊红(HE)染色、扫描电镜(SEM)评价内皮细胞的生长.结果 1HNMR(D2O):3.51～3.86 ppm处PEG信号峰,4.14(4H,CH2OAr),5.80 ～6.48 ppm处丙烯酸酯双键.SEM可见枝状化PEG修饰的DPAV胶原纤维交联增粗,人脐静脉内皮细胞在DPAV上种植8天DPAV组细胞数为(0.88 ±0.03)×105/cm2,PEG-DPAV组细胞数为(1.18 ±0.04)×105/cm2,PEG-DPAV组高于DPAV组(P＜0.05).HE染色和SEM可见两组瓣膜支架表面仅有散在的细胞存在,呈梭形或不规则形,细胞数PEG-DPAV高于DPAV组,部分可见细胞间连接紧密形成一层近似融合的细胞层,但细胞紧密连接不连续.结论 枝状化PEG修饰的去细胞猪主动脉瓣膜能够提高内皮细胞的黏附增殖,但却不能在去细胞瓣膜支架表面形成一层完整的融合层,因此PEG化的瓣膜并不能达到理想的内皮化,有必要进行进一步修饰.
目的 觀察枝狀化聚乙二醇(PEG)脩飾後的去細胞豬主動脈瓣(DPAV)的內皮細胞生長性能.方法 自閤成枝狀化聚乙二醇(4-arm-PEG-acrylate),採用覈磁共振氫譜(1HNMR),對複閤物進行錶徵檢測,利用4-arm-PEG-acrylate上的丙烯痠酯與巰基化的DPAV結閤對其進行PEG化,體外穫取人臍靜脈內皮細胞,以細胞密度為1×105接種于各組瓣膜支架材料上,8d後MTS試劑、囌木素-伊紅(HE)染色、掃描電鏡(SEM)評價內皮細胞的生長.結果 1HNMR(D2O):3.51～3.86 ppm處PEG信號峰,4.14(4H,CH2OAr),5.80 ～6.48 ppm處丙烯痠酯雙鍵.SEM可見枝狀化PEG脩飾的DPAV膠原纖維交聯增粗,人臍靜脈內皮細胞在DPAV上種植8天DPAV組細胞數為(0.88 ±0.03)×105/cm2,PEG-DPAV組細胞數為(1.18 ±0.04)×105/cm2,PEG-DPAV組高于DPAV組(P＜0.05).HE染色和SEM可見兩組瓣膜支架錶麵僅有散在的細胞存在,呈梭形或不規則形,細胞數PEG-DPAV高于DPAV組,部分可見細胞間連接緊密形成一層近似融閤的細胞層,但細胞緊密連接不連續.結論 枝狀化PEG脩飾的去細胞豬主動脈瓣膜能夠提高內皮細胞的黏附增殖,但卻不能在去細胞瓣膜支架錶麵形成一層完整的融閤層,因此PEG化的瓣膜併不能達到理想的內皮化,有必要進行進一步脩飾.
목적 관찰지상화취을이순(PEG)수식후적거세포저주동맥판(DPAV)적내피세포생장성능.방법 자합성지상화취을이순(4-arm-PEG-acrylate),채용핵자공진경보(1HNMR),대복합물진행표정검측,이용4-arm-PEG-acrylate상적병희산지여구기화적DPAV결합대기진행PEG화,체외획취인제정맥내피세포,이세포밀도위1×105접충우각조판막지가재료상,8d후MTS시제、소목소-이홍(HE)염색、소묘전경(SEM)평개내피세포적생장.결과 1HNMR(D2O):3.51～3.86 ppm처PEG신호봉,4.14(4H,CH2OAr),5.80 ～6.48 ppm처병희산지쌍건.SEM가견지상화PEG수식적DPAV효원섬유교련증조,인제정맥내피세포재DPAV상충식8천DPAV조세포수위(0.88 ±0.03)×105/cm2,PEG-DPAV조세포수위(1.18 ±0.04)×105/cm2,PEG-DPAV조고우DPAV조(P＜0.05).HE염색화SEM가견량조판막지가표면부유산재적세포존재,정사형혹불규칙형,세포수PEG-DPAV고우DPAV조,부분가견세포간련접긴밀형성일층근사융합적세포층,단세포긴밀련접불련속.결론 지상화PEG수식적거세포저주동맥판막능구제고내피세포적점부증식,단각불능재거세포판막지가표면형성일층완정적융합층,인차PEG화적판막병불능체도이상적내피화,유필요진행진일보수식.
Objective To investigate the endothelialization performance of decellularized porcine aortic valve (DPAV) modified with branched polyethylene glycol.Methods Synthesis of 4-arm-Polyethyl glycol (PEG)-acrylate,characterized by 1H nuclear magnetic resonance (NMR),then react with DPAV which had been introduced thiol through N-Succinimidyl-S-acetylthioacetate (SATA),human umbilical vein endothelial cells (HUVECs)were harvested in vitro and the cell were seeded in each group with density of 1 × 105,CellTiter 96(R) AQueous One Solution Cell Proliferation Assay、Hematoxylin and eosin (HE) stain and scanning electron microscopy (SEM) were performed for each group to detect the HUVECs growth 8 d after the seeding.Results 1 HNMR (D2O):3.51-3.86 ppm belong to PEG peak,4.14 (4H,CH2OAr),5.80-6.48 ppm belong to acrylate bond.SEM shows that DPAV modified with branched PEG become thickening.aftr HUVECs had grown for eight days,cell numbers in DPAV group was(0.88 ±0.03) × 105/cm2,while PEG-DPAV group was(1.18 ±0.04) × 105/cm2.the cell numbers were higher in group PEG-DPAV as compared to those in group DPAV (P ＜ 0.05).HE and SEM observations indicated that A confluent monolayer of endothelial cells was present on the valve surface in group PEG-DPAV at 8th d after the seeding,but not complete.Conclusion DPAV modified with branched PEG can improve the growth of endothelial cells,but cannot forming a complete fusion layer in the surface of decellularized valve scaffolds,so the PEGyalated DPAV can not achieve the desired endothelialization,and should be further modified.
