Directly auto-transplanted mesenchymal stem cells induce bone formation in a ceramic bone substitute in an ectopic sheep model.
Directly auto-transplanted mesenchymal stem cells induce bone formation in a ceramic bone substitute in an ectopic sheep model.
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DOI：
10.1111/j.1582-4934.2010.01131.x
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发表时间：
2011-06
影响因子：
5.3
通讯作者：
Beier JP
中科院分区：
文献类型：
--
作者：
Boos AM;Loew JS;Deschler G;Arkudas A;Bleiziffer O;Gulle H;Dragu A;Kneser U;Horch RE;Beier JP
骨髓间充质干细胞（MSC）是骨组织工程研究的热点。在大多数动物移植模型中，MSC在自体细胞移植之前被分离和扩增，这对于将来的临床应用可能是至关重要的。因此，本研究比较了直接自体移植与体外扩增MSC（含或不含骨形态发生蛋白-2（BMP-2））在绵羊模型中诱导大体积陶瓷骨替代物中骨形成的潜力。从骨髓抽吸物中分离MSC并直接自体移植或体外扩增，并在皮下植入前结合BMP-2和β-磷酸三钙/羟基磷灰石（β-TCP/HA）颗粒进行荧光激活细胞分选（FACS）和RT-PCR分析。在1至12周后将构建体固定，随后进行组织学和RT-PCR评价。经Ficoll梯度离心法筛选后，绵羊MSC为CD 29+、CD 44+和CD 166+，而直接自体移植的MSC表达较低水平的CD 29和CD 166。直接自体移植和扩增的MSC都在体内不断增殖，并且随着时间的推移凋亡减少。直接自体移植的MSC导致在使用β-TCP/HA基质的异位绵羊模型中重新骨形成，与仅应用60 μg/ml BMP-2或植入扩增的MSC相当。骨基质蛋白在直接自体移植后的构建体和扩增的MSC以及BMP-2构建体中上调。用免疫组织化学方法和RT-PCR检测上调。CD 31免疫组化染色显示三组均见密集的血管形成。直接自体移植或单独扩增MSC与β-TCP/HA颗粒可形成异位骨。因此，BMP-2的刺激可能成为未来，从而提供了一个有吸引力的，临床上可行的方法，骨组织工程。
Bone tissue engineering approaches increasingly focus on the use of mesenchymal stem cells (MSC). In most animal transplantation models MSC are isolated and expanded before auto cell transplantation which might be critical for clinical application in the future. Hence this study compares the potential of directly auto-transplanted versus in vitro expanded MSC with or without bone morphogenetic protein-2 (BMP-2) to induce bone formation in a large volume ceramic bone substitute in the sheep model. MSC were isolated from bone marrow aspirates and directly auto-transplanted or expanded in vitro and characterized using fluorescence activated cell sorting (FACS) and RT-PCR analysis before subcutaneous implantation in combination with BMP-2 and β-tricalcium phosphate/hydroxyapatite (β-TCP/HA) granules. Constructs were explanted after 1 to 12 weeks followed by histological and RT-PCR evaluation. Sheep MSC were CD29+, CD44+ and CD166+ after selection by Ficoll gradient centrifugation, while directly auto-transplanted MSC-populations expressed CD29 and CD166 at lower levels. Both, directly auto-transplanted and expanded MSC, were constantly proliferating and had a decreasing apoptosis over time in vivo. Directly auto-transplanted MSC led to de novo bone formation in a heterotopic sheep model using a β-TCP/HA matrix comparable to the application of 60 μg/ml BMP-2 only or implantation of expanded MSC. Bone matrix proteins were up-regulated in constructs following direct auto-transplantation and in expanded MSC as well as in BMP-2 constructs. Up-regulation was detected using immunohistology methods and RT-PCR. Dense vascularization was demonstrated by CD31 immunohistology staining in all three groups. Ectopic bone could be generated using directly auto-transplanted or expanded MSC with β-TCP/HA granules alone. Hence BMP-2 stimulation might become dispensable in the future, thus providing an attractive, clinically feasible approach to bone tissue engineering.
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影响因子：
5.3
作者：
Steffens L;Wenger A;Stark GB;Finkenzeller G
通讯作者：
Finkenzeller G
影响因子：
2.9
作者：
Maus, Uwe;Andereya, Stefan;Niedhart, Christopher
通讯作者：
Niedhart, Christopher
影响因子：
--
作者：
Sales, Virna L.;Mettler, Bret A.;Mayer, John E., Jr.
通讯作者：
Mayer, John E., Jr.
影响因子：
14
作者：
Niemeyer, Philipp;Fechner, Katharina;Kasten, Philip
通讯作者：
Kasten, Philip
影响因子：
3.3
作者：
Maegawa, Naoki;Kawamura, Kenji;Ohgushi, Hajime
通讯作者：
Ohgushi, Hajime