Adult bone marrow stromal cell-based tissue-engineered aggrecan exhibits ultrastructure and nanomechanical properties superior to native cartilage.
Adult bone marrow stromal cell-based tissue-engineered aggrecan exhibits ultrastructure and nanomechanical properties superior to native cartilage.
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DOI：
10.1016/j.joca.2010.07.015
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发表时间：
2010-11
影响因子：
7
通讯作者：
Ortiz C
中科院分区：
文献类型：
--
作者：
Lee HY;Kopesky PW;Plaas A;Sandy J;Kisiday J;Frisbie D;Grodzinsky AJ;Ortiz C
定量研究成人骨髓基质细胞(BMSCs)在多肽水凝胶支架中合成的聚集素的结构特征和纳米力学性能，并与成人关节软骨聚集素进行比较。将成年马骨髓间充质干细胞包裹在3D-多肽水凝胶中，在转化生长因子-β-1的作用下培养21d，诱导其向软骨方向分化。从年龄匹配的成年马关节软骨中提取BMSC-aggrecan，并与aggrecan进行比较。通过原子力显微镜成像显示聚集素的单分子，并通过高分辨率的力显微镜定量聚集素的纳米机械硬度。通过大小排除层析、Western分析和荧光辅助碳水化合物电泳(FACE)测定了聚集素的水动力大小、核心蛋白结构和CS硫酸盐组成的群体平均测量值。BMSC-aggrecan主要是全长的，而软骨-aggrecan有许多碎片。单分子测量表明，BMSC-aggrecan的核心蛋白和GAG链明显长于软骨-aggrecan。比较两个物种的全长aggrecan，BMSC-aggrecan具有更长的Gag链，而核心蛋白的踪迹长度相似。Face分析检测到骨髓间充质干细胞-GAG中4-硫酸软骨素和6-硫酸软骨素的∼比例为1：1，这一表型与来自骨骼未成熟软骨的聚集素一致。在相同的总固定电荷密度下，BMSC-aggrecan的纳米机械硬度明显大于软骨-aggrecan。BMSC-aggrecan具有更高的全长单体比例、更长的Gag链和更大的硬度，使其在生物力学方面优于成人软骨-aggrecan。聚集素的刚性不仅依赖于固定的电荷密度，而且还依赖于GAG分子的超微结构。这些结果支持将成人骨髓间充质干细胞用于基于细胞的软骨修复。
To quantify the structural characteristics and nanomechanical properties of aggrecan produced by adult bone marrow stromal cells (BMSCs) in peptide hydrogel scaffolds and compare to aggrecan from adult articular cartilage. Adult equine BMSCs were encapsulated in 3D-peptide hydrogels and cultured for 21 days with TGF-β1 to induce chondrogenic differentiation. BMSC-aggrecan was extracted and compared with aggrecan from age-matched adult equine articular cartilage. Single molecules of aggrecan were visualized by atomic force microcopy-based imaging and aggrecan nanomechanical stiffness was quantified by high resolution force microscopy. Population-averaged measures of aggrecan hydrodynamic size, core protein structures and CS sulfation compositions were determined by size-exclusion chromatography, Western analysis, and fluorescence-assisted carbohydrate electrophoresis (FACE). BMSC-aggrecan was primarily full-length while cartilage-aggrecan had many fragments. Single molecule measurements showed that core protein and GAG chains of BMSC-aggrecan were markedly longer than those of cartilage-aggrecan. Comparing full-length aggrecan of both species, BMSC-aggrecan had longer GAG chains, while the core protein trace lengths were similar. FACE analysis detected a ∼1:1 ratio of chondroitin-4-sulfate to chondroitin-6-sulfate in BMSC-GAG, a phenotype consistent with aggrecan from skeletally-immature cartilage. The nanomechanical stiffness of BMSC-aggrecan was demonstrably greater than that of cartilage-aggrecan at the same total sGAG (fixed charge) density. The higher proportion of full-length monomers, longer GAG chains and greater stiffness of the BMSC-aggrecan makes it biomechanically superior to adult cartilage-aggrecan. Aggrecan stiffness was not solely dependent on fixed charge density, but also on GAG molecular ultrastructure. These results support the use of adult BMSCs for cell-based cartilage repair.
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影响因子：
4.1
作者：
INEROT, S;HEINEGARD, D;OLSSON, SE
通讯作者：
OLSSON, SE
影响因子：
5.6
作者：
Bos, KJ;Holmes, DF;Bishop, PN
通讯作者：
Bishop, PN
影响因子：
3
作者：
Langer, RS;Vacanti, JP
通讯作者：
Vacanti, JP
影响因子：
2.4
作者：
Dean, Delphine;Han, Lin;Ortiz, Christine
通讯作者：
Ortiz, Christine
影响因子：
7
作者：
Brown, M. P.;Trumble, T. N.;Merritt, K. A.
通讯作者：
Merritt, K. A.