Department of Prosthodontics, West China College of Stomatology, Sichuan University, Renmin Nanlu, Chengdu, China.
J Mater Sci Mater Med. 2012 Nov;23(11):2717-26. doi: 10.1007/s10856-012-4724-1. Epub 2012 Aug 19.
The aim of the present work was to design a bio-interactive implant surface by coating recombinant human dentin matrix protein 1 (hDMP1) onto titanium and to investigate the biological function of this material. Firstly, the plasmid containing the hDMP1 cDNA was constructed and hDMP1 was expressed, purified and characterized. Then, hDMP1 was coated onto the surface of Ti substrates via a biochemical technique and the procedure was divided into three steps: in the beginning, titanium was treated by regular polishing and denoted as Cp-Ti; then, Cp-Ti received alkaline and water treatment and was nominated as AW-Ti; finally, AW-Ti was coated with hDMP1 and referred to as hDMP1-Ti. The inserts of hDMP1 genes were detected by enzyme digestion as well as gel electrophoresis, and the complete nucleotide sequence of hDMP1 was tested. The purified recombinant hDMP1 was electrophoresed on a 10 % SDS-PAGE gel. Cp-Ti, AW-Ti and hDMP1-Ti were characterized by X-ray photoelectron spectroscope and water contact angles tests. The biological activity of MG63 cells cultured in the three groups was investigated by the cell attachment, proliferation and alkaline phosphatase activity assays. The results show that hDMP1 was successfully constructed and coated onto the titanium surface, and hDMP1-Ti had higher hydrophilicity than Cp-Ti. Compared with Cp-Ti and AW-Ti, hDMP1-Ti showed better in vitro bioactivity.
本工作旨在通过将重组人牙本质基质蛋白 1(hDMP1)涂覆到钛上设计一种生物相互作用的植入物表面,并研究该材料的生物学功能。首先构建了含有 hDMP1 cDNA 的质粒,并表达、纯化和鉴定了 hDMP1。然后,通过生化技术将 hDMP1 涂覆到 Ti 基底表面上,该过程分为三步:首先,钛经过常规抛光处理,记为 Cp-Ti;然后,Cp-Ti 经过碱处理和水热处理,记为 AW-Ti;最后,AW-Ti 涂覆 hDMP1,记为 hDMP1-Ti。通过酶消化和凝胶电泳检测 hDMP1 基因的插入片段,并测试 hDMP1 的完整核苷酸序列。纯化的重组 hDMP1 在 10%SDS-PAGE 凝胶上电泳。通过 X 射线光电子能谱仪和水接触角测试对 Cp-Ti、AW-Ti 和 hDMP1-Ti 进行了表征。通过细胞附着、增殖和碱性磷酸酶活性测定研究了在这三组中培养的 MG63 细胞的生物活性。结果表明,成功构建并将 hDMP1 涂覆到钛表面上,并且 hDMP1-Ti 比 Cp-Ti 具有更高的亲水性。与 Cp-Ti 和 AW-Ti 相比,hDMP1-Ti 表现出更好的体外生物活性。