Homing of Mesenchymal Stem Cells in Induced Degenerative Intervertebral Discs in a Whole Organ Culture System
Homing of Mesenchymal Stem Cells in Induced Degenerative Intervertebral Discs in a Whole Organ Culture System
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DOI：
10.1097/brs.0b013e3182544a8a
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发表时间：
2012-10-15
期刊：
影响因子：
3
通讯作者：
Alini, Mauro
中科院分区：
文献类型：
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作者：
Illien-Juenger, Svenja;Pattappa, Girish;Alini, Mauro
研究设计。目的研究人骨髓间充质干细胞(BMSCs)在体外培养的牛尾状间盘(IVDS)中的归巢特性。在整个器官培养中调查代谢和机械挑战是否可以诱导骨髓间充质干细胞招募进入IVD。背景数据摘要。损伤组织中的细胞释放细胞因子和介体，使祖细胞得以募集。骨髓间充质干细胞具有在静脉内存活的能力。有或没有终板的牛IVD在模拟生理或退化条件下培养1周；分析盘细胞的细胞活力和基因表达，同时分析培养液中一氧化氮的产生和趋化作用。将PKH标记的人BMSCs接种到培养的IVD上(培养第8、10、12天，每盘1×10(6)个细胞)，观察BMSCs的归巢情况；在第14天，显微镜下观察BMSCs的归巢情况。此外，还对转导的BMSCs(用编码胰岛素样生长因子1的腺病毒转导)和未转导的BMSCs进行了比较研究。用S-35掺入法定量测定视盘蛋白多糖合成率。用酶联免疫吸附试验检测IGF-1在模拟生理盘和退变盘上的分泌情况。与模拟生理盘相比，在退变条件下培养的盘显示细胞活力降低，基质降解酶上调，一氧化氮产生增加。与带或不带终板的生理条件相比，退行性病变的归巢能力更强。退变间盘的培养液对BMSCs具有趋化活性。与未转导的BMSCs相比，IGF-1转导的BMSCs移植组的IGF-1分泌量和蛋白多糖合成率均显著增加。我们首次证明了在体外器官培养中，退行性疾病会导致促进骨髓间充质干细胞募集的因子释放。此外，转导IGF-1的BMSCs显著增加了退变椎间盘中蛋白多糖的合成速度。这一发现为骨髓间充质干细胞提供了一种新的输送系统和治疗IVD再生的策略。
Study Design. Homing of human bone marrow-derived mesenchymal stem cells (BMSCs) was studied using ex vivo cultured bovine caudal intervertebral discs (IVDs).Objective. To investigate in a whole organ culture whether metabolic and mechanical challenges can induce BMSC recruitment into the IVD.Summary of Background Data. Cells from injured tissues release cytokines and mediators that enable the recruitment of progenitor cells. BMSCs have the ability to survive within the IVD.Methods. Bovine IVDs with or without endplates were cultured for 1 week under simulated physiological or degenerative conditions; disc cells were analyzed for cell viability and gene expression, whereas media was analyzed for nitric oxide production and chemotaxis. Homing of BMSCs was investigated by supplying PKH-labeled human BMSCs onto cultured IVDs (1 x 10(6) cells/disc on d 8, 10, and 12 of culture); on day 14, the number of homed BMSCs was microscopically assessed. Moreover, a comparative study was performed between transduced BMSCs (transduced with an adenovirus encoding for insulin-like growth factor 1 [IGF-1]) and nontransduced BMSCs. Disc proteoglycan synthesis rate was quantified via S-35 incorporation. The secretion of IGF-1 was evaluated by enzyme-linked immunosorbent assay on both simulated physiological and degenerative discs.Results. Discs cultured under degenerative conditions showed reduced cell viability, upregulation of matrix degrading enzymes, and increased nitric oxide production compared with simulated physiological discs. Greater homing occurred under degenerative compared with physiological conditions with or without endplate. Media of degenerative discs demonstrated a chemoattractive activity toward BMSCs. Finally, discs homed with IGF-1 transduced BMSCs showed increased IGF-1 secretion and significantly higher proteoglycan synthesis rate than discs supplied with nontransduced BMSCs.Conclusion. We have demonstrated for the first time that degenerative conditions induce the release of factors promoting BMSC recruitment in an ex vivo organ culture. Moreover, IGF-1 transduction of BMSCs strongly increases the rate of proteoglycan synthesis within degenerative discs. This finding offers a new delivery system for BMSCs and treatment strategy for IVD regeneration.