Appleford Mark R, Oh Sunho, Cole Judith A, Carnes David L, Lee Myunghyun, Bumgardner Joel D, Haggard Warren O, Ong Joo L
University of Tennessee Health Science Center, Memphis, TN, USA.
Biomaterials. 2007 Jun;28(17):2747-53. doi: 10.1016/j.biomaterials.2007.02.018. Epub 2007 Mar 9.
The objective of this research was to investigate stress-signaling patterns in response to two-dimensional (2-D) and three-dimensional (3-D) calcium phosphate (CP) materials using human embryonic palatal mesenchyme cells (HEPM, CRL-1486, ATCC, Manassas, VA), an osteoblast precursor cell line. Control discs and scaffolds were fabricated from hydroxyapatite and beta tri-CP ceramics. Phospho-specific antibody cell-based ELISA technique was utilized on members of the mitogen-activated protein kinase cascade including; the extracellular signal-regulated kinases (ERK1/2), p38, c-Jun N-terminal kinase (JNK), and the anti-apoptosis mediator protein kinase B (AKT). Quantification of these signals was evaluated during the early attachment phase of osteoblast precursor cells. In this study, it was observed that 3-D CP scaffolds significantly activated the stress mediators p38 and JNK but not ERK1/2. This signal trend was matched with an up-regulation in AKT, suggesting the ability of cells to manage high stress signals in response to 3-D CP architecture and that 3-D CP scaffolds are necessary for studies simulating a natural trabecular bone organization. The absence of these signals in 2-D CP surfaces indicated the importance of local architecture conditions on cell stress response. It was concluded from this study that osteoblast precursor cells cultured in 3-D CP scaffolds experience greater stress-signaling patterns when compared to 2-D CP surfaces.
本研究的目的是使用人胚胎腭间充质细胞(HEPM,CRL-1486,美国典型培养物保藏中心,弗吉尼亚州马纳萨斯),一种成骨细胞前体细胞系,来研究对二维(2-D)和三维(3-D)磷酸钙(CP)材料作出反应时的应激信号模式。对照圆盘和支架由羟基磷灰石和β-磷酸三钙陶瓷制成。对丝裂原活化蛋白激酶级联反应的成员,包括细胞外信号调节激酶(ERK1/2)、p38、c-Jun氨基末端激酶(JNK)以及抗凋亡介质蛋白激酶B(AKT),采用基于磷酸特异性抗体的细胞酶联免疫吸附测定技术。在成骨细胞前体细胞的早期附着阶段对这些信号进行定量评估。在本研究中,观察到3-D CP支架显著激活应激介质p38和JNK,但未激活ERK1/2。这种信号趋势与AKT的上调相匹配,表明细胞有能力应对3-D CP结构产生的高应激信号,并且3-D CP支架对于模拟天然小梁骨组织的研究是必要的。2-D CP表面缺乏这些信号表明局部结构条件对细胞应激反应的重要性。从本研究得出的结论是,与2-D CP表面相比,在3-D CP支架中培养的成骨细胞前体细胞经历更大的应激信号模式。