CAJ | 학술논문

目的制作可注射式壳聚糖/β-甘油磷酸钠/胶原蛋白支架并观察其对成肌细胞生长分化的影响.方法将2wt％的壳聚糖(C)、50wt％的β-甘油磷酸钠(β-GP)及2g/L的胶原蛋白(Co)溶液于冰上按体积比5∶1∶6混合,制成温敏性可注射凝胶支架C/GP/Co.用不同浓度的支架浸提液培养成肌细胞,通过相对增殖率评价该支架的细胞毒性；比较成肌细胞在常规培养和支架表面培养时的生长分化情况；将成肌细胞包裹于支架中进行三维培养,动态观察细胞生长情况.结果浸提液培养实验证实C/GP/Co支架无细胞毒性；成肌细胞在支架表面培养时增殖率无显著变化(P＞0.05),但融合指数及肌管体积显著增加[(39.5±5.7)比(45.3±5.5),(4.56±0.89)比(6.53±1.02),(P＜0.05)]；激光共聚焦显微镜及扫描电镜观察可见成肌细胞在三维支架内生长、增殖状态良好.结论 C/GP/Co支架有望用于成肌细胞培养及移植.
목적 제작가주사식각취당/β-감유린산납/효원단백지가병관찰기대성기세포생장분화적영향.방법 장2wt％적각취당(C)、50wt％적β-감유린산납(β-GP)급2g/L적효원단백(Co)용액우빙상안체적비5∶1∶6혼합,제성온민성가주사응효지가C/GP/Co.용불동농도적지가침제액배양성기세포,통과상대증식솔평개해지가적세포독성；비교성기세포재상규배양화지가표면배양시적생장분화정황；장성기세포포과우지가중진행삼유배양,동태관찰세포생장정황.결과 침제액배양실험증실C/GP/Co지가무세포독성；성기세포재지가표면배양시증식솔무현저변화(P＞0.05),단융합지수급기관체적현저증가[(39.5±5.7)비(45.3±5.5),(4.56±0.89)비(6.53±1.02),(P＜0.05)]；격광공취초현미경급소묘전경관찰가견성기세포재삼유지가내생장、증식상태량호.결론 C/GP/Co지가유망용우성기세포배양급이식.
Objective To prepare injectable chitosan/β-glyceral phosphate disodium/collagen scaffold and observe its effects upon the growth and differentiation of myoblasts.Methods After the preparations,2 wt％ chitosan ? solution,50 wt％ β-glyceral phosphate disodium (GP) solution and 2 mg/ml collagen ( Co ) solution were mixed on ice according to a volume ratio of 5 ∶ 1 ∶ 6 to yield a thermosensitive injectable scaffold.To assess the cytotoxicity of scaffold,the extraction fluids of different concentrations were added into the myoblast culture.Then relative growth rate (RGR) was calculated.The growth and differentiation state of myoblasts in routine culture or gel-coated plates were compared.Myoblasts were encapsulated in C/GP/Co solution before gelation to perform a 3-dimensional culture and then observed dynanically.Results No cytotoxicity was demonstrated.No significant difference of proliferation index was demonstrated (P ＞ 0.05 ) but fusion index and size of myotube increased significantly ( P ＜ 0.05 ).Thriving viability and proliferation were verified by the observations of laser confocal scanning microscope and scanning electron microscope.Conclusion C/GP/Co scaffold is a promising carrier for the culture and transplantation of myoblasts.