上转换纳米粒子介导的光动力疗法杀伤脊髓星形胶质细胞的实验研究
상전환납미입자개도적광동력요법살상척수성형효질세포적실험연구
The photodynamic effects mediated by upconversion nanoparticles on rat astrocytes in vitro
  • 万方数据
    中华骨科杂志 중화골과잡지
    CHINESE JOURNAL OF ORTHOPAEDICS
    2015年 4期 450-455 ,共6页

    马超%冯世庆%魏金栋%张治国%张佳音%班德翔%刘洋

    마초%풍세경%위금동%장치국%장가음%반덕상%류양

    大鼠%纳米技术%光化学疗法%脊髓损伤 大鼠%納米技術%光化學療法%脊髓損傷
    대서%납미기술%광화학요법%척수손상
    Rats%Nanotechnology%Photochemotherapy%Spinal cord injuries
    目的 探讨上转换纳米粒子(upconversion nanoparticles,UCNPs)介导的光动力疗法对大鼠脊髓星形胶质细胞的体外杀伤效应.方法 体外无菌条件下取新生SD大鼠脊髓组织,采用机械吹打及胰酶消化法制成细胞悬液,通过差速贴壁和恒温摇床震荡去除杂细胞,利用GFAP免疫荧光染色鉴定,成功分离培养大鼠脊髓星形胶质细胞.采用MTT法检测不同浓度UCNPs-MC540[以上转换纳米为载体和能量转换器,并使用偶联光敏剂部花青540(Merocyanine 540)构建的复合物]、不同能量980 nm激光照射对脊髓星形胶质细胞活力的影响以及在980 nm近红外激光照射下UCNPs-MC540介导的光动力疗法的细胞杀伤效应;应用浓度为200 μg/ml的UCNPs-MC540培养液与星形胶质细胞共培养12h后,给予2 000 J/cm2的980 nm近红外激光进行照射,以同样条件培养而未作激光照射光动力治疗的细胞作为对照组;分别制作透射电镜标本,透射电子显微镜下观察细胞形态改变.结果 不同浓度UCNPs-MC540及不同照射剂量980 nm激光照射处理后细胞存活率的差异无统计学意义.在浓度100 μg/ml的UCNPs-MC540作用细胞12h后分别给予不同能量激光照射,细胞存活率随着激光剂量的增加而下降,差异有统计学意义.不同浓度的UCNPs-MC540作用于细胞12h后予2000 J/cm2能量激光照射,随着UCNPs-540浓度的增加,细胞的存活率降低,差异有统计学意义.与未作激光照射光动力治疗细胞的对照组比较,细胞与浓度为200 μg/ml的UCNPs-MC540共培养,能量为2 000 J/cm2激光照射后,透射电镜检查显示细胞呈凋亡表现,形成具有特征性的凋亡小体.结论 由上转换荧光纳米NaYF∶Er,Yb与光敏剂MC540连接的纳米复合物UC-NPs-MC540介导的光动力疗法对星形胶质细胞有良好的杀伤效果,其机制可能为诱导细胞发生凋亡,为脊髓损伤的治疗提供新的思路,扩大光动力疗法的研究应用范围.
    목적 탐토상전환납미입자(upconversion nanoparticles,UCNPs)개도적광동력요법대대서척수성형효질세포적체외살상효응.방법 체외무균조건하취신생SD대서척수조직,채용궤계취타급이매소화법제성세포현액,통과차속첩벽화항온요상진탕거제잡세포,이용GFAP면역형광염색감정,성공분리배양대서척수성형효질세포.채용MTT법검측불동농도UCNPs-MC540[이상전환납미위재체화능량전환기,병사용우련광민제부화청540(Merocyanine 540)구건적복합물]、불동능량980 nm격광조사대척수성형효질세포활력적영향이급재980 nm근홍외격광조사하UCNPs-MC540개도적광동력요법적세포살상효응;응용농도위200 μg/ml적UCNPs-MC540배양액여성형효질세포공배양12h후,급여2 000 J/cm2적980 nm근홍외격광진행조사,이동양조건배양이미작격광조사광동력치료적세포작위대조조;분별제작투사전경표본,투사전자현미경하관찰세포형태개변.결과 불동농도UCNPs-MC540급불동조사제량980 nm격광조사처리후세포존활솔적차이무통계학의의.재농도100 μg/ml적UCNPs-MC540작용세포12h후분별급여불동능량격광조사,세포존활솔수착격광제량적증가이하강,차이유통계학의의.불동농도적UCNPs-MC540작용우세포12h후여2000 J/cm2능량격광조사,수착UCNPs-540농도적증가,세포적존활솔강저,차이유통계학의의.여미작격광조사광동력치료세포적대조조비교,세포여농도위200 μg/ml적UCNPs-MC540공배양,능량위2 000 J/cm2격광조사후,투사전경검사현시세포정조망표현,형성구유특정성적조망소체.결론 유상전환형광납미NaYF∶Er,Yb여광민제MC540련접적납미복합물UC-NPs-MC540개도적광동력요법대성형효질세포유량호적살상효과,기궤제가능위유도세포발생조망,위척수손상적치료제공신적사로,확대광동력요법적연구응용범위.
    Objective To investigate the phototoxicity effects of the nanocompound of upconversion nanoparticles (UCNPs) on rat astrocytes in vitro.Methods The spinal astrocytes cells were cultured successfully in vitro and then incubated with the UCNPs-MC540 of various concentrations and exposured 980 nm infrared laser irradiation of different energy densities.The cell survival rates of each group were detected by MTT assay.The cellular morphology was observed via transmission electron microscope after photodynamic therapy.Results UCNPs-MC540 of different concentrations without laser irradiation or laser of different energy had no significant effects on cell survival rates.when cells incubated with 100 μg/ml UCNPs-MC540 for 12 h underwent laser irradiation of different energy,the cellular survival rates significantly decreased with the increased energy densities.when the cells incubated with UCNPs-MC540 of various concentrations for 12 h underwent laser irradiation of 2 000 J/cm2,the cellular survival rates significantly decreased with the increased concentrations.Compared with controls,the TEM show the apoptosis sign in the cells incubated with 200 μg/ml UCNPs-MC540 after laser irradiation of 2 000 J/cm2.Conclusion The UCNPs-MC540 mediated photodynamic therapy have effective killing effect on astrocytes by the mechanism of induction the apoptosis.
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