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生物活性玻璃对大鼠鼻软骨细胞培养中组织特异性基因表达的调节作用。

The modulation of tissue-specific gene expression in rat nasal chondrocyte cultures by bioactive glasses.

作者信息

Asselin Audrey, Hattar Susan, Oboeuf Martine, Greenspan David, Berdal Ariane, Sautier Jean-Michel

机构信息

Laboratoire Biologie Orofaciale et Pathologie, INSERM E0110, Institut Biomédical des Cordeliers, Université Paris 7, UFR d'Odontologie, Esc. E-2è étage, Paris Cedex 06 F-75270, France.

出版信息

Biomaterials. 2004 Nov;25(25):5621-30. doi: 10.1016/j.biomaterials.2004.01.016.

Abstract

Since bone repair may occur, following endochondral ossification, we have investigated the behaviour of chondrocytes isolated from nasal septum cartilage of foetal rats and cultured up to 21 days in the presence of a melt-derived bioactive glass (Bioglass 45S5) and a less reactive glass with 60 wt% silica content (60S). In both cultures, chondrocytes proliferate and form typical cartilaginous nodules on day 5 of cultures. However, on day 12, the nodules in contact with 45S5 granules became darker than in 60S cultures, corresponding to the emergence of matrix biomineralization. Transmission electron microscopy showed a collagen-rich matrix composed of densely packed fibres and mineralized foci formed of needle-shaped crystals in contact with an electron-dense layer located at the periphery of the material. The specific activity of alkaline phosphatase was significant higher in 45S5 cultures on day 15 than in 60S cultures. Real time RT-PCR was used to monitor gene expression levels of specific chondrogenic markers. The transcription factor Sox9 was expressed throughout the culture period, but with no significant differences between the two kinds of cultures. In contrast, Runx2 expression was higher in experiment cultures on day 12. Type II collagen mRNA and aggrecan, showed an almost similar expression pattern with a strong expression at the beginning of cultures but higher in experiment cultures. Indian hedgehog was strongly expressed between day 9 and 12 with a significant stimulation in 45S5 cultures. Similarly, type X collagen mRNA seemed to be up-regulated in 45S5 cultures on day 20. In conclusion, this study shows hat 45S5 Bioglass has the ability to support the growth of chondrocytes and to stimulate some chondrogenic molecular markers.

摘要

由于骨修复可能会在软骨内成骨后发生,我们研究了从胎鼠鼻中隔软骨分离出的软骨细胞的行为,这些软骨细胞在熔融衍生的生物活性玻璃(生物玻璃45S5)和二氧化硅含量为60 wt%的低活性玻璃(60S)存在的情况下培养长达21天。在两种培养物中,软骨细胞在培养第5天增殖并形成典型的软骨结节。然而,在第12天,与45S5颗粒接触的结节比在60S培养物中的结节颜色更深,这与基质生物矿化的出现相对应。透射电子显微镜显示,富含胶原蛋白的基质由紧密堆积的纤维组成,矿化灶由针状晶体形成,与位于材料外围的电子致密层接触。在第15天,45S5培养物中碱性磷酸酶的比活性显著高于60S培养物。实时RT-PCR用于监测特定软骨生成标志物的基因表达水平。转录因子Sox9在整个培养期均有表达,但两种培养物之间无显著差异。相比之下,Runx2在实验培养物的第12天表达较高。II型胶原蛋白mRNA和聚集蛋白聚糖显示出几乎相似的表达模式,在培养开始时表达强烈,但在实验培养物中更高。印度刺猬因子在第9天至12天强烈表达,在45S5培养物中有显著刺激。同样,X型胶原蛋白mRNA在第20天似乎在45S5培养物中上调。总之,本研究表明45S5生物玻璃有能力支持软骨细胞生长并刺激一些软骨生成分子标志物。

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