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使用电化学固定核酸对钛基合金进行生物活性分子表面改性。

Surface modification of titanium-based alloys with bioactive molecules using electrochemically fixed nucleic acids.

作者信息

Michael J, Beutner R, Hempel U, Scharnweber D, Worch H, Schwenzer B

机构信息

Institut für Biochemie, Technische Universität Dresden, Bergstr. 66, 01069 Dresden, Germany.

出版信息

J Biomed Mater Res B Appl Biomater. 2007 Jan;80(1):146-55. doi: 10.1002/jbm.b.30579.

Abstract

A new method of surface modification for titanium (alloys) with bioactive molecules was developed with the intention of providing a new basis of implant adaptation for particular requirements of certain medical indications. Nucleic acid single strands are fixed electrochemically via their termini (regiospecifically) by growing an oxide layer on Ti6Al7Nb anodically. It could be shown that they are accessible to subsequent hybridization with complementary strands at physiological pH. Amount of nucleic acids immobilized and hybridized were determined radioanalytically using 32P-labelled nucleic acids. Stable fixation was attained at and above potentials of 4 V(SCE). Up to 4 pmol/cm2 of nucleic acid single strands could be immobilized and hybridization efficiencies up to 1.0 were reached. Hybridization efficiency was found to depend on surface density of immobilized oligonucleotides, while hybridization rates increased when MgCl2 was added. A conjugate consisting of an oligonucleotide complementary to the immobilized strand and the hexapeptide GRGDSP with RGD as an integrin recognition site was synthesized. This conjugate was able to bind to integrins on osteoblasts. It was shown that this conjugate binds to the anchor strand fixed on Ti6Al7Nb to an extent comparable with the unconjugated complementary strand.

摘要

开发了一种用生物活性分子对钛(合金)进行表面改性的新方法,旨在为特定医学适应症的特殊要求提供植入物适配的新基础。通过在Ti6Al7Nb上阳极生长氧化层,核酸单链通过其末端(区域特异性地)进行电化学固定。结果表明,在生理pH值下,它们可与互补链进行后续杂交。使用32P标记的核酸通过放射分析确定固定和杂交的核酸量。在4 V(SCE)及以上的电位下可实现稳定固定。每平方厘米可固定高达4 pmol的核酸单链,杂交效率可达1.0。发现杂交效率取决于固定寡核苷酸的表面密度,而添加MgCl2时杂交速率会增加。合成了一种由与固定链互补的寡核苷酸和具有RGD作为整合素识别位点的六肽GRGDSP组成的共轭物。这种共轭物能够与成骨细胞上的整合素结合。结果表明,这种共轭物与固定在Ti6Al7Nb上的锚定链的结合程度与未共轭的互补链相当。

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