Guizzardi Stefano, Galli Carlo, Martini Desiree, Belletti Silvana, Tinti Anna, Raspanti Mario, Taddei Paola, Ruggeri Alessandro, Scandroglio Renato
Department of Experimental Medicine, Section of Histology, University of Parma, Parma, Italy.
J Periodontol. 2004 Feb;75(2):273-82. doi: 10.1902/jop.2004.75.2.273.
Six titanium disks with six different surface treatments were examined: SS: smooth (polished) surface; TPS: plasma spray; C100: sand blasting by aluminum oxide (Al2O3) diameter 100 microm and acid etching; C150: sand blasting by Al2O3 diameter 150 microm and acid etching; B60: sand blasting by zirconium oxide (ZrO2) diameter 60 microm and acid etching; and B120: sand blasting by ZrO2 diameter 120 microm and acid etching.
The surface characteristics were determined by scanning electron microscopy (SEM) observation and a roughness tester. Raman spectroscopy was used to determine the presence of residual substances on the samples. Cells were seeded onto the disk and after 24 hours, 6 days, and 12 days were observed under SEM and growth curves generated with a cell counter. Some samples were used to determine alkaline phosphatase activity (ALP), using a colorimetric assay.
SEM observation revealed drastic differences in surface microtopography, with a higher cell density on sand-blasted and acid-etched (SLA) samples than SS and TPS, and more regularly aligned cells on B60 and B120 surfaces than on the others. The growth curves showed a greater adhesion of cells on the etched/blasted surfaces compared to the SS and TPS surfaces. The number of cells increased on all the SLA samples, especially B60, throughout the experiment. At the same time, there was considerable ALP activity on the B60 sample, while it remained at extremely low levels on SS and TPS surfaces. Raman analyses revealed Al2O3 debris on C100 and C150, partly explaining the poorer performances of these two surface treatments, since this substance was shown to be toxic for cultured osteoblasts.
Surface treatments influence the growth and the metabolic activity of cultured osteoblasts, and B60 seems to be the most favorable surface inducing a more pronounced proliferation of cells together with a high differentiation degree.
对六个经过六种不同表面处理的钛盘进行了检查:SS:光滑(抛光)表面;TPS:等离子喷涂;C100:用直径100微米的氧化铝(Al2O3)喷砂并酸蚀;C150:用直径150微米的Al2O3喷砂并酸蚀;B60:用直径60微米的氧化锆(ZrO2)喷砂并酸蚀;B120:用直径120微米的ZrO2喷砂并酸蚀。
通过扫描电子显微镜(SEM)观察和粗糙度测试仪确定表面特征。使用拉曼光谱法确定样品上残留物质的存在。将细胞接种到盘上,在24小时、6天和12天后,在SEM下观察并用细胞计数器生成生长曲线。一些样品用于通过比色法测定碱性磷酸酶活性(ALP)。
SEM观察显示表面微观形貌存在巨大差异,喷砂和酸蚀(SLA)样品上的细胞密度高于SS和TPS,并且B60和B120表面上的细胞排列比其他表面更规则。生长曲线显示,与SS和TPS表面相比,蚀刻/喷砂表面上的细胞粘附性更强。在整个实验过程中,所有SLA样品上的细胞数量都增加了,尤其是B60。同时,B60样品上有相当高的ALP活性,而在SS和TPS表面上则保持在极低水平。拉曼分析在C100和C150上发现了Al2O3碎片,部分解释了这两种表面处理性能较差的原因,因为这种物质对培养的成骨细胞有毒性。
表面处理会影响培养的成骨细胞的生长和代谢活性,B60似乎是最有利的表面,能诱导细胞更明显的增殖以及高分化程度。