CAJ | 학술논문

目的探讨人骨髓间充质干细胞(BM-MSCs)经诱导转化为人髓核细胞(NPs)并构建组织工程椎间盘的价值.方法体外培养胎儿NPs及BM-MSCs并种植在聚乳酸-聚羟基乙酸共聚物(PLGA)支架上,倒置显微镜及扫描电镜进行形态学观察.将载有BM-MSCs和NPs的PLGA支架及BM-MSCs和NPs的细胞悬液植入新西兰大白兔椎间盘中,12周后对椎间盘信号强度按Thompson分级进行评定.分光光度法检测蛋白聚糖并免疫组化检测Ⅱ型胶原的表达.结果 BM-MSCs在与NPs共培养后由梭形、多角形变为成纤维细胞样,且两种细胞在PLGA支架表面贴附,形态正常,生长良好;载有BM-MSCs和NPs的PLGA支架在椎间MRI信号维持、蛋白多糖[PLGA支架组含量为(3.93±0.31)mg/100 mg,对照组为(3.52±0.26)mg/100 mg]及Ⅱ型胶原表达上较对照组差异均有统计学意义(P＜0.05).结论共培养可促使BM-MSCs向NPs转化,PLGA支架为细胞提供良好的生长环境,其力学性能维持和空间结构保障可以满足BM-MSCs与NPs构建组织工程椎间盘的需要,有效延缓了椎间盘的退变.
목적 탐토인골수간충질간세포(BM-MSCs)경유도전화위인수핵세포(NPs)병구건조직공정추간반적개치.방법 체외배양태인NPs급BM-MSCs병충식재취유산-취간기을산공취물(PLGA)지가상,도치현미경급소묘전경진행형태학관찰.장재유BM-MSCs화NPs적PLGA지가급BM-MSCs화NPs적세포현액식입신서란대백토추간반중,12주후대추간반신호강도안Thompson분급진행평정.분광광도법검측단백취당병면역조화검측Ⅱ형효원적표체.결과 BM-MSCs재여NPs공배양후유사형、다각형변위성섬유세포양,차량충세포재PLGA지가표면첩부,형태정상,생장량호;재유BM-MSCs화NPs적PLGA지가재추간MRI신호유지、단백다당[PLGA지가조함량위(3.93±0.31)mg/100 mg,대조조위(3.52±0.26)mg/100 mg]급Ⅱ형효원표체상교대조조차이균유통계학의의(P＜0.05).결론 공배양가촉사BM-MSCs향NPs전화,PLGA지가위세포제공량호적생장배경,기역학성능유지화공간결구보장가이만족BM-MSCs여NPs구건조직공정추간반적수요,유효연완료추간반적퇴변.
Objective To investigate the value of bone marrow-mesenchymal stem cells(BMMSCs)transformed by nucleus pulposus(NPs)for construction of tissue engineering disc.Methods BMMSCs and fetal NPs were cultured in vitro,planted on polylactic acid-polyglycolic acid copolymer(PLGA),and observed with inverted microscope and scanning electronic microscope.PLGA scaffolds with adherent BM-MSCs and NPs,as well as BM-MSCs and NPs suspension were implanted into intervertebral discs of New Zealand white rabbits,respectively.Intervertebral signal intensity was evaluated by Thompson grading 12 weeks later.Proteoglycan and type Ⅱ collagen were determined by spectrophotometric method and immunohistochemistry,respectively.Results Spindle or multi-angular BM-MSCs turned into fibro-like phenotype coculture of BM-MSCs and NPs,which grew well with normal morphology when they attached on PLGA scaffolds.There was statistical difference in intervertebral signal intensity,and the expression of proteoglycan and type Ⅱ collagen between PLGA scaffolds group and control group(P＜0.05),the content of proteoglycan was(3.93 ± 0.31)mg/100 mg in the PLGA scaffolds group whereas(3.52 ± 0.26)mg/100 mg in the control group.Conclusions BM-MSCs can be induced into NPs by cocultivation,and PLGA scaffolds can provide good growing conditions,and maintain high mechanical properties and spacial structure which meet the requirement of tissue engineering disc to prevent degeneration.