CAJ | 학술논문

目的去细胞瓣膜上种植人主动脉瓣膜间质细胞,生物反应器内构建组织工程心脏瓣膜.方法 PEG交联去细胞瓣,GRGDSPC多肽共价修饰.RGD-PEG-去细胞瓣上种植人主动脉瓣膜间质细胞,缝制于生物反应器内.接种2、4、8h后,细胞计数观察细胞黏附情况.体外培养10d后行形态学观察和DNA含量测定.结果免疫荧光检测显示,GRGDSPC多肽与PEG-去细胞瓣有效共价接枝.接种2、4、8h后,RGD-PEG-去细胞瓣组和单纯去细胞瓣组细胞计数分别为:(4.98±0.46)×107/L比(3.35 ±0.15)×107/L,(5.71 ±0.29)× 104 个/ml比(3.55±0.18)×107/L,(5.97±0.45)×107/L比(3.62±0.18) ×107/L,P＜0.05.RGD-PEG-去细胞瓣组较单纯去细胞瓣组DNA含量明显增高,(25.98±2.45)μg/瓣叶比(19.61±1.94)μg瓣叶,差异有统计学意义(P＜0.05).结论生物反应器内,GRGDSPC接枝的PEG-去细胞瓣复合支架,可促进种子细胞的黏附,改善瓣膜支架生物学性能.
목적 거세포판막상충식인주동맥판막간질세포,생물반응기내구건조직공정심장판막.방법 PEG교련거세포판,GRGDSPC다태공개수식.RGD-PEG-거세포판상충식인주동맥판막간질세포,봉제우생물반응기내.접충2、4、8h후,세포계수관찰세포점부정황.체외배양10d후행형태학관찰화DNA함량측정.결과 면역형광검측현시,GRGDSPC다태여PEG-거세포판유효공개접지.접충2、4、8h후,RGD-PEG-거세포판조화단순거세포판조세포계수분별위:(4.98±0.46)×107/L비(3.35 ±0.15)×107/L,(5.71 ±0.29)× 104 개/ml비(3.55±0.18)×107/L,(5.97±0.45)×107/L비(3.62±0.18) ×107/L,P＜0.05.RGD-PEG-거세포판조교단순거세포판조DNA함량명현증고,(25.98±2.45)μg/판협비(19.61±1.94)μg판협,차이유통계학의의(P＜0.05).결론 생물반응기내,GRGDSPC접지적PEG-거세포판복합지가,가촉진충자세포적점부,개선판막지가생물학성능.
Objective Human aortic valve interstitial cells (HAVICs) were seeded onto decellularized valves in order to fabricate tissue engineering heart valves in a bioreactor system.Methods Decellularized valves were cross-linked by polyethylene glycol.PEGylated decellularized valves were covalently modified by GRGDSPC peptides.The modified effect of GRGDSPC peptides was determined by using immunofluorescence.HAVICs were seeded onto PEGylated decellularized valves modified by GRGDSPC peptides.Then the valves were sutured into a bioreactor system.Cell adhesion was tested respectively after 2,4 and 8 h.After 10 days of culture,hematoxylin-eosin staining,scanning electron microscopy and DNA content test were performed.Results Immunofluorescence showed that PEGylated decellularized valves could be effectively modified by GRGDSPC peptides through covalent bonding.As compared with decellularized valves,PEGylated decellularized valves modified by GRGDSPC peptides significantly promoted the adhesion of HAVICs.Number of adherent cells after 2,4 and 8 h was respectively (4.98 ±0.46) × 107/L vs.(3.35 ±0.15) ×107/L,(5.71 ±0.29)× 107/L vs.(3.55 ±0.18) ×107/L,(5.97 ±0.45) ×107/L vs.(3.62 ±0.18) × 107/L,P ＜0.05.The DNA content of PEGylated decellularized valves modified by GRGDSPC peptides was significantly higher than that of decellularized valves [(25.98 ± 2.45) μg/valve vs.(19.61 ±1.94) μg/valve,P ＜ 0.05].Conclusion Modification of PEGylated decellutarized valves by GRGDSPC peptides can promote the adhesion of cells seeded on valves in a bioreactor system.