申请人:Board of Regents, The University of Texas System
公开号:US05330911A1
公开(公告)日:1994-07-19
An absorbed protein-independent cell-adhesive surface is disclosed. The treated surface comprises a chemically derivatized material to which small peptides, having less than 12 amino acid residues and including YIGSR, RGD or REDV amino acid sequence are covalently linked to. The peptides of the present invention include a terminal glycine amino acid. Tresyl chloride activation of surface hydroxyl moieties provides the active surface sites by which a terminal glycine arm of a selected peptide attaches to form covalent bonds between the substrate and peptide. Peptides high with cell adhesive properties are bound in high efficiency. By way of example, surfaces which may be used in conjunction with the present invention include polymer, metal, and ceramic surfaces. The most preferred polymer surfaces include PHEMA and PET polymer surfaces, with the most preferred glass surfaces being glycophase glass. The present methods also include a pretreatment method which provides hydroxyl moieties to surfaces devoid of readily available hydroxyl moieties. The pretreatment, by way of example, comprises immersion of the surface in a mixture of formaldehyde and acetic acid. Methods of preparing the treated surfaces are also included in the present invention. Also included are surface-treated biomedical implant devices and cell culturing devices. The treated surface promotes an enhanced rate and an enhanced amount of cell adhesion to the surface, independent of media serum concentrations or other absorbed proteins. The treated surfaces of the present invention are thermally stable, reusable, peptide efficient (attached to surface only) and resistant to cell proteolysis. The invention further concerns polymeric substrates with a surface having physically interpenetrating water-soluble polymer chains, and methods for production thereof.
本发明涉及一种吸附蛋白独立的细胞粘附表面。处理后的表面包括一种化学衍生材料,小肽通过共价键连接到其中,这些小肽具有少于12个氨基酸残基,包括YIGSR,RGD或REDV氨基酸序列。本发明的肽包括一个末端甘氨酸氨基酸。通过三氯化砜活化表面羟基官能团,使得所选肽的末端甘氨酸臂连接到基质和肽之间形成共价键的活性表面位点。具有细胞粘附性能的肽以高效率结合。例如,可以与本发明一起使用的表面包括聚合物、金属和陶瓷表面。最优选的聚合物表面包括PHEMA和PET聚合物表面,最优选的玻璃表面为糖基玻璃。本发明还包括一种预处理方法,用于为缺乏易于获取的羟基官能团的表面提供羟基官能团。例如,预处理包括将表面浸泡在甲醛和醋酸的混合物中。本发明还包括制备处理后表面的方法。还包括经处理的生物医学植入物装置和细胞培养装置。处理后的表面促进细胞与表面的粘附速率和粘附量增强,不受培养基血清浓度或其他吸附蛋白的影响。本发明的处理后表面具有热稳定性,可重复使用,肽效率高(仅附着于表面),并且抗细胞蛋白酶解。本发明还涉及具有物理相互穿插的水溶性聚合物链表面的聚合物基材料,以及其生产方法。