Department of Industrial Chemistry, Faculty of Engineering, Tokyo University of Science, 12-1 Ichigayafunagawara-machi, Shinjuku-ku, Tokyo 162-0826, Japan; Graduate School of Chemical Sciences and Technology, Tokyo University of Science, 12-1 Ichigayafunagawara-machi, Shinjuku-ku, Tokyo 162-0826, Japan.
Graduate School of Chemical Sciences and Technology, Tokyo University of Science, 12-1 Ichigayafunagawara-machi, Shinjuku-ku, Tokyo 162-0826, Japan.
Colloids Surf B Biointerfaces. 2016 Nov 1;147:351-359. doi: 10.1016/j.colsurfb.2016.08.020. Epub 2016 Aug 18.
The surfaces of polystyrene (PS) cell culture plates were functionalized with hydroxyapatite (HAp) under body fluid conditions utilizing protein adsorption layers and a pretreatment with an alternate soaking process (ASP) using solutions containing calcium and phosphate ions. Adsorption layers of human serum albumin (HSA) formed on the surface of each well of commercial 24-well PS plates by solution processes. CaCl2 and K2HPO4 solutions were alternately added to the wells, the plates were incubated to form the precursors, and this was followed by the addition of simulated body fluid (SBF) and a further incubation for 24h. These treatments resulted in the surfaces of the PS cell culture plates being completely covered with bone-like HAp. The coating of PS plates with HAp promoted the adhesion of mesenchymal stem cells (MSCs) and maintained cell growth that was as fast as that on tissue culture-treated PS (TCPS) plates. Osteogenic differentiation was greater, whereas adipogenic and chondrogenic differentiation was less in the culture on HAp-coated PS plates than in that on TCPS plates. The present method is useful for preparing HAp-coated PS plates at clean benches without the need for any expensive apparatus. HAp coated on PS plates by this method was a bone-like apatite with high bioactivity; therefore, the present HAp-coated PS plates are promising materials for assays of bone-related cells in the bone remodeling process.
在体液条件下，利用蛋白质吸附层和采用含有钙和磷酸盐离子的交替浸泡处理（ASP）的预处理，使聚苯乙烯（PS）细胞培养板的表面功能化生成羟基磷灰石（HAp）。通过溶液处理，在商业 24 孔 PS 板的每个孔表面形成人血清白蛋白（HSA）吸附层。将 CaCl2 和 K2HPO4 溶液交替添加到孔中，将板孵育以形成前体，然后加入模拟体液（SBF）并进一步孵育 24 小时。这些处理导致 PS 细胞培养板的表面完全被类骨 HAp 覆盖。PS 板涂覆 HAp 促进间充质干细胞（MSCs）的黏附，并保持细胞生长速度与组织培养处理 PS（TCPS）板相当。在 HAp 涂覆 PS 板上的培养中，成骨分化更高，而脂肪和成软骨分化更低。该方法在清洁台上制备 HAp 涂覆 PS 板非常有用，无需任何昂贵的设备。通过该方法涂覆在 PS 板上的 HAp 是一种具有高生物活性的类骨磷灰石；因此，目前的 HAp 涂覆 PS 板是用于骨重塑过程中骨相关细胞检测的有前途的材料。
