低强度间断负压诱导人骨髓间充质干细胞向成骨细胞的分化

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杨　治1 ,刘　淼1 ,张银刚1 ,郭　雄2 ,许　鹏3
' T) e3 g0 o. @+ f4 H(1. 西安交通大学医学院第一附属医院骨科,陕西西安　710061 ;2. 西安交通大学环境与疾病相关基因
3 M/ ?" v  P, ?+ g9 Z. I9 E教育部重点实验室,环境与地方病研究室,陕西西安　710061 ;3. 西安交通大学材料学院,陕西西安　710049)
8 ^* |; Q9 z3 Y8 k+ Q摘要:目的　利用低强度间断负压诱导人骨髓间充质干细胞(MSCs) 向成骨细胞定向分化。方法　分离人骨髓,利用/ H1 x2 {1 Y9 D2 J$ S
梯度离心法进行MSCs 的培养,取第三代细胞利用低强度间断负压诱导分化;倒置显微镜下观察细胞的形态,检测碱/ W/ K; @. e1 H. R- l2 o
性磷酸酶(AL P) 活性,茜素红染色观察钙结节形成,免疫组织化学检测Ⅰ型胶原和骨保护素的表达。结果　低强度间- O, b, J) E, C0 I' o
断负压诱导2 周后,细胞呈现出成骨细胞形态,AL P 活性明显增强,茜素红染色可见钙结节形成, Ⅰ型胶原和骨保护
5 m9 d* W6 j2 e7 A素表达阳性。结论　利用低强度间断负压可以成功的诱导MSCs 向成骨细胞定向分化,诱导的细胞具有典型的成骨' Q. r  E: S$ f) M2 ?
细胞特性。
' v% w+ P6 @9 j( |( a关键词:负压;骨髓间充质干细胞;成骨细胞;诱导& m; Q8 h7 _; _) ?% K( \
中图分类号:R687 　　　文献标识码:A 　　　文章编号:167128259 (2008) 0520527203
5 _5 e$ q7 y% o% r* Z% xLow2intensity interrupted negative pressure induces human marrow
9 G, _* l0 Q, z1 Hmesenchymal stem cells differentiating into osteoblasts
; u! \& C( t, j2 M5 E& yYang Zhi1 , Liu Miao1 , Zhang Yingang1 , Guo Xiong2 , Xu Peng36 V% f4 |0 o& o; K+ Z; L
(1. Department of Ort hopaedics , t he First Affiliated Hospital , Medical School of
2 i9 f* f: g4 E* Z  y# q1 e4 |Xipan J iaotong Univer sity , Xipan 710061 ; 2. Key Laboratory of Environment and Genes Related to
2 M3 J( |: i# o) [; [: z" ^Diseases of Minist ry of Education , Instit ute of Environmental and Endemic Diseases ,
; V  s6 H) v* D; f  G7 z6 b/ |Xipan J iaotong Univer sity , Xipan 710061 ; 3. School of Material s Science and Engineering ,
6 N# ?& u' x3 @; X; s5 @; ^Xipan J iaotong University , Xipan 710049 , China)
9 p  d2 n/ w1 eABSTRACT: Objective 　To investigate t he ef f ect of low2intensity inter rup ted negative p ressure on t he. y2 T- y+ @  l
dif f e rentiation of human ma r row mesenchymal stem cells (MSCs) int o osteoblast in vit ro. Methods 　MSCs we re! v+ `9 V+ c- ^% r' m! P
isolated f rom adult ma r row using density gradient sep aration met hod , p assage 3 cells we re chosen t o induce# k$ k! h  M% M6 B4 h
dif f e rentiation ; t he dif f e rentiation of MSCs was examined by p hase2cont rast microscop y , t he dete rmination of( L4 R2 d8 [: p5 j2 m
alkaline p hosp hatase (AL P) activities , alizarin2red staining and t he immunohist ochemist ry of type Ⅰcollagen and
" |0 K$ L* x3 i) u* ]osteop rotege rin. Re sult s 　MSCs showed a typical app ea rance of osteoblasts af te r 2 weeksp induction by low2# u0 o  `: m# i' v- N) O
intensity inte r rup ted negative p ressure , t he activities of AL P we re increased signif icantly , calcium nodes we re seen
% p9 i3 X$ `7 t5 ^$ W( dby alizarin2red staining , t he exp ressions of collagen type Ⅰ and osteop rotege rin were p ositive . Conclusion 　L ow2
1 D) Q0 `# o8 [0 @& \% Yintensity inte r rup ted negative p ressure could successf ully induce human MSCs int o osteoblasts and t he induced cells
- g; V1 @. v0 v6 Pshow cha racte ristics of osteoblasts .
1 h5 r2 ^, S( G! R: p1 P: cKEY WORDS : negative p ressure ; ma r row mesenchymal stem cell ; osteoblast ; induction
& B# B9 G1 W. X3 Y5 V; e7 C) r
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