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人成骨细胞在可注射羟丙基甲基纤维素水凝胶中的三维培养与分化

Three-dimensional culture and differentiation of human osteogenic cells in an injectable hydroxypropylmethylcellulose hydrogel.

作者信息

Trojani Christophe, Weiss Pierre, Michiels Jean-François, Vinatier Claire, Guicheux Jérôme, Daculsi Guy, Gaudray Patrick, Carle Georges F, Rochet Nathalie

机构信息

FRE 2720 CNRS/UNSA IFR50, Faculté de Médecine, Avenue de Valombrose, 06107 Nice Cedex 02, France.

出版信息

Biomaterials. 2005 Sep;26(27):5509-17. doi: 10.1016/j.biomaterials.2005.02.001.

Abstract

The present work evaluates a newly developed silated hydroxypropylmethylcellulose (Si-HPMC)-based hydrogel as a scaffold for 3D culture of osteogenic cells. The pH variation at room temperature catalyzes the reticulation and self-hardening of the viscous polymer solution into a gelatine state. We designed reticulation time, final consistency and pH in order to obtain an easy handling matrice, suitable for in vitro culture and in vivo injection. Three human osteogenic cell lines and normal human osteogenic (HOST) cells were cultured in 3D inside this Si-HPMC hydrogel. We show here that osteosarcoma cells proliferate as clonogenic spheroids and that HOST colonies survive for at least 3 weeks. Mineralization assay and gene expression analysis of osteoblastic markers and cytokines, indicate that all the cells cultured in 3D into this hydrogel, exhibited a more mature differentiation status than cells cultured in monolayer on plastic. This study demonstrates that this Si-HPMC hydrogel is well suited to support osteoblastic survival, proliferation and differentiation when used as a new scaffold for 3D culture and represents also a potential basis for an innovative bone repair material.

摘要

本研究评估了一种新开发的基于硅化羟丙基甲基纤维素(Si-HPMC)的水凝胶作为成骨细胞三维培养支架的性能。室温下的pH变化催化粘性聚合物溶液网状化并自硬化成凝胶状态。我们设计了网状化时间、最终稠度和pH值,以获得易于操作的基质,适用于体外培养和体内注射。三种人成骨细胞系和正常人成骨(HOST)细胞在这种Si-HPMC水凝胶内进行三维培养。我们在此表明,骨肉瘤细胞以克隆性球体形式增殖,并且HOST集落存活至少3周。成骨细胞标志物和细胞因子的矿化测定及基因表达分析表明,在这种水凝胶中进行三维培养的所有细胞,与在塑料上单层培养的细胞相比,表现出更成熟的分化状态。本研究表明,这种Si-HPMC水凝胶用作三维培养的新支架时,非常适合支持成骨细胞的存活、增殖和分化,并且也代表了一种创新骨修复材料的潜在基础。

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