Growth differentiation factor 6 and transforming growth factor-beta differentially mediate mesenchymal stem cell differentiation, composition, and micromechanical properties of nucleus pulposus constructs.
Growth differentiation factor 6 and transforming growth factor-beta differentially mediate mesenchymal stem cell differentiation, composition, and micromechanical properties of nucleus pulposus constructs.
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DOI：
10.1186/ar4505
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发表时间：
2014-03-12
影响因子：
4.9
通讯作者：
Hoyland JA
中科院分区：
文献类型：
--
作者：
Clarke LE;McConnell JC;Sherratt MJ;Derby B;Richardson SM;Hoyland JA
目前，有大量的研究集中在开发新的基于细胞的再生和组织工程疗法，用于治疗椎间盘退变和相关的背痛。骨髓源性（BM）间充质干细胞（MSC）和脂肪源性MSC（AD-MSC）都被认为是用于这种疗法的合适细胞。然而，目前还没有达成共识的最佳生长因子需要驱动分化髓核（NP）样表型。本研究的目的是调查生长分化因子-6（GDF 6），与其他转化生长因子（TGF）超家族成员相比，对骨髓间充质干细胞的椎间盘分化，基质成分和工程化NP组织构建体的微观力学的影响。将患者匹配的人AD-MSC和BM-MSC接种到I型胶原水凝胶中，并在补充有TGF-β3、GDF 5或GDF 6的分化培养基中培养。14天后，定量聚合酶链反应分析软骨形成和新的NP标记基因和硫酸化糖胺聚糖（sGAG）含量的构建体和培养基成分进行了测量。此外，通过使用扫描声学显微镜（SAM）分析结构微观力学。与TGF-β或GDF 5刺激相比，BM-MSC和AD-MSC的GDF 6刺激导致新NP标记基因表达的显著增加、更高的聚集蛋白聚糖与II型胶原基因表达比率以及更高的sGAG产生。这些作用在AD-MSCs中大于BM-MSCs。此外，通过使用SAM测量的声波速度以及因此的组织硬度在GDF 6刺激的AD-MSC构建体中最低。数据表明，AD-MSC的GDF 6刺激诱导分化为NP样表型，并导致更富含蛋白聚糖的基质。微机械分析显示，GDF 6处理的AD-MSC具有较低刚性的基质组成，表明生长因子诱导更类似于天然NP样组织的基质。因此，这种细胞和生长因子的组合可能是基于细胞的椎间盘（IVD）再生疗法的理想选择。
Currently, there is huge research focus on the development of novel cell-based regeneration and tissue-engineering therapies for the treatment of intervertebral disc degeneration and the associated back pain. Both bone marrow-derived (BM) mesenchymal stem cells (MSCs) and adipose-derived MSCs (AD-MSCs) are proposed as suitable cells for such therapies. However, currently no consensus exists as to the optimum growth factor needed to drive differentiation to a nucleus pulposus (NP)-like phenotype. The aim of this study was to investigate the effect of growth differentiation factor-6 (GDF6), compared with other transforming growth factor (TGF) superfamily members, on discogenic differentiation of MSCs, the matrix composition, and micromechanics of engineered NP tissue constructs. Patient-matched human AD-MSCs and BM-MSCs were seeded into type I collagen hydrogels and cultured in differentiating media supplemented with TGF-β3, GDF5, or GDF6. After 14 days, quantitative polymerase chain reaction analysis of chondrogenic and novel NP marker genes and sulfated glycosaminoglycan (sGAG) content of the construct and media components were measured. Additionally, construct micromechanics were analyzed by using scanning acoustic microscopy (SAM). GDF6 stimulation of BM-MSCs and AD-MSCs resulted in a significant increase in expression of novel NP marker genes, a higher aggrecan-to-type II collagen gene expression ratio, and higher sGAG production compared with TGF-β or GDF5 stimulation. These effects were greater in AD-MSCs than in BM-MSCs. Furthermore, the acoustic-wave speed measured by using SAM, and therefore tissue stiffness, was lowest in GDF6-stiumlated AD-MSC constructs. The data suggest that GDF6 stimulation of AD-MSCs induces differentiation to an NP-like phenotype and results in a more proteoglycan-rich matrix. Micromechanical analysis shows that the GDF6-treated AD-MSCs have a less-stiff matrix composition, suggesting that the growth factor is inducing a matrix that is more akin to the native NP-like tissue. Thus, this cell and growth-factor combination may be the ideal choice for cell-based intervertebral disc (IVD)-regeneration therapies.
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DOI：
10.1152/ajpheart.00355.2006
发表时间：
2007-03-01
影响因子：
4.8
作者：
Graham, H. K.;Trafford, A. W.
通讯作者：
Trafford, A. W.
DOI：
10.1016/0304-4165(86)90306-5
发表时间：
1986-09-04
期刊：
BIOCHIMICA ET BIOPHYSICA ACTA
影响因子：
--
作者：
FARNDALE, RW;BUTTLE, DJ;BARRETT, AJ
通讯作者：
BARRETT, AJ
影响因子：
14
作者：
Richardson, Stephery M.;Hughes, Nesta;Hoyland, Judith A.
通讯作者：
Hoyland, Judith A.
影响因子：
3
作者：
Roughley, PJ
通讯作者：
Roughley, PJ
影响因子：
5.3
作者：
Graham, Helen K.;Akhtar, Riaz;Sherratt, Michael J.
通讯作者：
Sherratt, Michael J.