Zhang Qinghao, Gerlach Jörg C, Schmelzer Eva, Nettleship Ian
Department of Mechanical Engineering and Materials Science, McGowan Institute for Regenerative Medicine, University of Pittsburgh, Pittsburgh, PA, USA.
J Vasc Res. 2017;54(6):376-385. doi: 10.1159/000481778. Epub 2017 Nov 23.
Foamed hydroxyapatite offers a three-dimensional scaffold for the development of bone constructs, mimicking perfectly the in vivo bone structure. In vivo, calcium release at the surface is assumed to provide a locally increased gradient supporting the maintenance of the hematopoietic stem cells niche. We fabricated hydroxyapatite scaffolds with high surface calcium concentration by infiltration, and used human umbilical vein endothelial cells (HUVECs) as a model to study the effects on hematopoietic lineage direction. HUVECs are umbilical vein-derived and thus possess progenitor characteristics, with a prospective potential to give rise to hematopoietic lineages. HUVECs were cultured for long term on three-dimensional porous hydroxyapatite scaffolds, which were either infiltrated biphasic foams or untreated. Controls were cultured in two-dimensional dishes. The release of calcium into culture medium was determined, and cells were analyzed for typical hematopoietic and endothelial gene expressions, surface markers by flow cytometry, and hematopoietic potential using colony-forming unit assays. Our results indicate that the biphasic foams promoted a hematopoietic lineage direction of HUVECs, suggesting an improved in vivo-like scaffold for hematopoietic bone tissue engineering.
泡沫状羟基磷灰石为骨构建体的发育提供了三维支架,完美模拟了体内骨结构。在体内,表面钙的释放被认为可提供局部增加的梯度,支持造血干细胞龛的维持。我们通过渗透法制备了具有高表面钙浓度的羟基磷灰石支架,并使用人脐静脉内皮细胞(HUVECs)作为模型来研究其对造血谱系方向的影响。HUVECs来源于脐静脉,因此具有祖细胞特征,具有产生造血谱系的潜在可能性。将HUVECs长期培养在三维多孔羟基磷灰石支架上,这些支架要么是渗透双相泡沫,要么未处理。对照组在二维培养皿中培养。测定钙向培养基中的释放,并通过流式细胞术分析细胞的典型造血和内皮基因表达、表面标志物,并使用集落形成单位测定法分析造血潜能。我们的结果表明,双相泡沫促进了HUVECs的造血谱系方向,表明其为造血骨组织工程提供了一种改进的类似体内的支架。