Bioactive glass stimulates in vitro osteoblast differentiation and creates a favorable template for bone tissue formation
Bioactive glass stimulates in vitro osteoblast differentiation and creates a favorable template for bone tissue formation
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DOI：
10.1359/jbmr.2001.16.2.231
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发表时间：
2001-02-01
影响因子：
6.2
通讯作者：
Forest, N
中科院分区：
文献类型：
--
作者：
Loty, C;Sautier, JM;Forest, N
在这项研究中，我们研究了胎鼠成骨细胞在含有55%二氧化硅含量的生物活性玻璃（55 S）和生物惰性玻璃（60 S）上培养的行为，所述生物惰性玻璃以颗粒形式或以盘形式使用。（55重量%二氧化硅含量），相差显微镜允许逐步可视化与颗粒接触的骨结节的形成。未脱钙切片的超微结构观察显示，在材料的外围存在由针状晶体组成的电子致密层，该层似乎充当生物晶体的成核表面。此外，能量色散X射线（EDX）分析和电子衍射图案表明，该界面含有钙（Ca）和磷（P），是高度结晶的。当大鼠骨细胞培养在55 S磁盘，扫描电子显微镜（SEM）观察显示，细胞附着，蔓延到所有的基质，并形成多层结节状结构的第10天的文化。碱性磷酸酶（ALP）的细胞酶定位和骨涎蛋白抗体的免疫标记显示55 S培养物中形成的骨结节呈阳性染色。此外，生化测定的ALP比活性在55 S培养物中显著高于对照组。刮除细胞层后对材料表面的SEM观察表明，矿化骨结节仍然附着在55 S表面上，但不在60 S表面上。X射线微区分析表明，在这种骨组织中的钙和磷的存在。还在横截面上分析了55 S/骨界面。界面分析表明，通过介入磷灰石层与55 S表面牢固结合，X射线图谱证实了这一点。所有这些结果表明表面组成在支持成骨细胞分化和随后的骨基质并置中的重要性，从而使生物活性材料与骨牢固结合。
In this study, we have investigated the behavior of fetal rat osteoblasts cultured on bioactive glasses with 55 wt% silica content (55S) and on a bioinert glass (60S) used either in the form of granules or in the form of disks, In the presence of Bioglass granules (55 wt% silica content), phase contrast microscopy permitted step-by-step visualization of the formation of bone nodules in contact with the particles. Ultrastructural observations of undecalcified sections revealed the presence of an electron-dense layer composed of needle-shaped crystals at the periphery of the material that seemed to act as a nucleating surface for biological crystals. Furthermore, energy dispersive X-ray (EDX) analysis and electron diffraction patterns showed that this interface contains calcium (Ca) and phosphorus (P) and was highly crystalline. When rat bone cells were cultured on 55S disks, scanning electron microscopic (SEM) observations revealed that cells attached, spread to all substrata, and formed multilayered nodular structures by day 10 in culture. Furthermore, cytoenzymatic localization of alkaline phosphatase (ALP) and immunolabeling with bone sialoprotein antibody revealed a positive staining for the bone nodules formed in cultures on 55S, In addition, the specific activity of ALP determined biochemically was significantly higher in 55S cultures than in the controls. SEM observations of the material surfaces after scraping off the cell layers showed that mineralized bone nodules remained attached on 55S surfaces but not on 60S. X-ray microanalysis indicated the presence of Ca and P in this bone tissue. The 55S/bone interfaces also were analyzed on transverse sections. The interfacial analysis showed a firm bone bonding to the 55S surface through an intervening apatite layer, confirmed by the X-ray mappings. All these results indicate the importance of the surface composition in supporting differentiation of osteogenic cells and the subsequent apposition of bone matrix allowing a strong bond of the bioactive materials to bone.