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生物硅对人成骨样细胞(SaOS-2)的成骨潜能。

Osteogenic potential of biosilica on human osteoblast-like (SaOS-2) cells.

机构信息

Institute for Physiological Chemistry, Medical Center of the Johannes Gutenberg-University, Duesbergweg 6, 55128, Mainz, Germany.

出版信息

Calcif Tissue Int. 2010 Dec;87(6):513-24. doi: 10.1007/s00223-010-9408-6. Epub 2010 Aug 20.

Abstract

Biosilica is a natural polymer, synthesized by the poriferan enzyme silicatein from monomeric silicate substrates. Biosilica stimulates mineralizing activity and gene expression of SaOS-2 cells. To study its effect on the formation of hydroxyapatite (HA), SaOS-2 cells were grown on different silicatein/biosilica-modified substrates (bone slices, Ca-P-coated coverslips, glass coverslips). Growth on these substrates induced the formation of HA nodules, organized in longitudinal arrays or spherical spots. Nodules of sizes above 1 μm were composed of irregularly arranged HA prism-like nanorods, formed by aggregates of three to eight SaOS-2 cells. Moreover, growth on silicatein/biosilica-modified substrates elicited increased [(3)H]dT incorporation into DNA, indicative of enhanced cell proliferation. Consequently, an in vitro-based bioassay was established to determine the ratio between [(3)H]dT incorporation and HA formation. This ratio was significantly higher for cells that grew on silicatein/biosilica-modified substrates than for cells on Ca-P-coated coverslips or plain glass slips. Hence, we propose that this ratio of in vitro-determined parameters reflects the osteogenic effect of different substrates on bone-forming cells. Finally, qRT-PCR analyses demonstrated that growth of SaOS-2 cells on a silicatein/biosilica matrix upregulated BMP2 (bone morphogenetic protein 2, inducer of bone formation) expression. In contrast, TRAP (tartrate-resistant acid phosphatase, modulator of bone resorption) expression remained unaffected. We conclude that biosilica shows pronounced osteogenicity in vitro, qualifying this material for studies of bone replacement also in vivo.

摘要

生物硅是一种天然聚合物,由海绵酶硅酸合成酶从单体硅酸基质合成。生物硅刺激 SaOS-2 细胞的矿化活性和基因表达。为了研究其对羟基磷灰石(HA)形成的影响,将 SaOS-2 细胞生长在不同的硅酸合成酶/生物硅修饰的基底(骨片、Ca-P 涂层盖玻片、玻璃盖玻片)上。在这些基底上生长诱导 HA 结节的形成,以纵向排列或球形斑点的形式组织。尺寸大于 1μm 的结节由不规则排列的 HA 棱柱纳米棒组成,由三到八个 SaOS-2 细胞的聚集形成。此外,在硅酸合成酶/生物硅修饰的基底上生长会导致 [(3)H]dT 掺入 DNA 增加,表明细胞增殖增强。因此,建立了一种基于体外的生物测定法来确定 [(3)H]dT 掺入和 HA 形成之间的比率。在硅酸合成酶/生物硅修饰的基底上生长的细胞的该比率明显高于在 Ca-P 涂层盖玻片或普通玻璃载玻片上生长的细胞的比率。因此,我们提出,体外测定参数的这种比率反映了不同基底对成骨细胞的成骨作用。最后,qRT-PCR 分析表明,SaOS-2 细胞在硅酸合成酶/生物硅基质上生长会上调 BMP2(骨形态发生蛋白 2,骨形成诱导物)的表达。相比之下,TRAP(抗酒石酸酸性磷酸酶,骨吸收调节剂)的表达不受影响。我们得出结论,生物硅在体外具有明显的成骨作用,使该材料有资格在体内进行骨替代研究。

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