Adipose-derived stromal cells protect intervertebral disc cells in compression: implications for stem cell regenerative disc therapy.
Adipose-derived stromal cells protect intervertebral disc cells in compression: implications for stem cell regenerative disc therapy.
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脂肪来源的基质细胞在受压时保护椎间盘细胞：对干细胞再生椎间盘治疗的影响
DOI：
10.7150/ijbs.10598
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发表时间：
2015
影响因子：
9.2
通讯作者：
Luo ZJ
中科院分区：
文献类型：
--
作者：
Sun Z;Luo B;Liu ZH;Samartzis D;Liu Z;Gao B;Huang L;Luo ZJ
引言：异常的生物力学在椎间盘退变中起作用。脂肪源性基质细胞（ADSCs）与椎间盘的完整性有关，然而，它们在椎间盘髓核（NP）的机械刺激中的作用仍然未知。因此，本研究的目的是评估的影响，脂肪干细胞对NP细胞在压缩负荷培养。方法：将人髓核细胞在3.0MPa压力下培养48 h，与ADSCs共培养或不共培养（比例为50：50）。我们使用流式细胞术，活/死染色和扫描电子显微镜（SEM），以评估细胞死亡，并确定表达的特定凋亡途径，通过表征活化的半胱天冬酶-3，8和9的表达。我们进一步使用实时（RT-）PCR和免疫染色来确定细胞外基质（ECM）、基质降解介质（例如MMP、TIMP和ADAMTSs）、促炎因子和NP细胞表型标记物的表达。结果：脂肪干细胞通过抑制活化的caspase-9和caspase-3抑制人髓核细胞凋亡。此外，ADSC通过显著上调ECM基因（SOX 9、COL 2A 1和ACAN）、金属蛋白酶组织抑制剂（TIMP-1和TIMP-2）基因和细胞角蛋白8（CK 8）蛋白的表达来保护NP细胞免受压缩负荷的降解作用。或者，ADSC通过抑制压缩负荷介导的基质金属蛋白酶（MMP-3和MMP-13）、具有血小板反应蛋白基序的去整合素和金属蛋白酶（ADAMTS; ADAMTS-1和5）以及促炎因子（IL-1 β、IL-6、TGF-β 1和TNF-α）的增加而显示出保护作用。结论：我们的研究是第一个评估非生理机械刺激培养环境中ADSC对NP细胞影响的体外研究。我们的研究指出，ADSC通过抑制活化的caspase-9和-3活性、调节ECM和调节基因、抑制促炎因子和保护CK 8来保护压缩负荷诱导的NP细胞死亡和降解。因此，本研究中发现的ADSC的保护作用提供了一个重要的理解，并扩大了我们对ADSC治疗椎间盘再生的实用性的认识。
Introduction: Abnormal biomechanics plays a role in intervertebral disc degeneration. Adipose-derived stromal cells (ADSCs) have been implicated in disc integrity; however, their role in the setting of mechanical stimuli upon the disc's nucleus pulposus (NP) remains unknown. As such, the present study aimed to evaluate the influence of ADSCs upon NP cells in compressive load culture. Methods: Human NP cells were cultured in compressive load at 3.0MPa for 48 hours with or without ADSCs co-culture (the ratio was 50:50). We used flow cytometry, live/dead staining and scanning electron microscopy (SEM) to evaluate cell death, and determined the expression of specific apoptotic pathways by characterizing the expression of activated caspases-3, -8 and -9. We further used real-time (RT-) PCR and immunostaining to determine the expression of the extracellular matrix (ECM), mediators of matrix degradation (e.g. MMPs, TIMPs and ADAMTSs), pro-inflammatory factors and NP cell phenotype markers. Results: ADSCs inhibited human NP cell apoptosis via suppression of activated caspase-9 and caspase-3. Furthermore, ADSCs protected NP cells from the degradative effects of compressive load by significantly up-regulating the expression of ECM genes (SOX9, COL2A1 and ACAN), tissue inhibitors of metalloproteinases (TIMPs) genes (TIMP-1 and TIMP-2) and cytokeratin 8 (CK8) protein expression. Alternatively, ADSCs showed protective effect by inhibiting compressive load mediated increase of matrix metalloproteinases (MMPs; MMP-3 and MMP-13), disintegrin and metalloproteinase with thrombospondin motifs (ADAMTSs; ADAMTS-1 and 5), and pro-inflammatory factors (IL-1beta, IL-6, TGF-beta1 and TNF-alpha). Conclusions: Our study is the first in vitro study assessing the impact of ADSCs on NP cells in an un-physiological mechanical stimulation culture environment. Our study noted that ADSCs protect compressive load induced NP cell death and degradation by inhibition of activated caspase-9 and -3 activity; regulating ECM and modulator genes, suppressing pro-inflammatory factors and preserving CK8. Consequently, the protective impact of ADSCs found in this study provides an essential understanding and expands our knowledge as to the utility of ADSCs therapy for intervertebral disc regeneration.
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影响因子：
3.7
作者：
Bayo J;Fiore E;Aquino JB;Malvicini M;Rizzo M;Peixoto E;Andriani O;Alaniz L;Piccioni F;Bolontrade M;Podhajcer O;Garcia MG;Mazzolini G
通讯作者：
Mazzolini G
影响因子：
3
作者：
Handa, T;Ishihara, H;Obata, K
通讯作者：
Obata, K
影响因子：
6.2
作者：
Orozco, Lluis;Soler, Robert;Garcia-Sancho, Javier
通讯作者：
Garcia-Sancho, Javier
DOI：
10.1196/annals.1397.069
发表时间：
2007-01-01
期刊：
SIGNAL TRANSDUCTION PATHWAYS, PT D
影响因子：
--
作者：
Bachmeier, Beatrice E.;Nerlich, Andreas G.;Boos, Norbert
通讯作者：
Boos, Norbert
影响因子：
3
作者：
Lotz, JC;Chin, JR
通讯作者：
Chin, JR