Richert Ludovic, Boulmedais Fouzia, Lavalle Philippe, Mutterer Jérôme, Ferreux Emmanuelle, Decher Gero, Schaaf Pierre, Voegel Jean-Claude, Picart Catherine
Institut National de la Santé et de la Recherche Médicale, Unité 595, Faculté de Chirurgie Dentaire, Université Louis Pasteur, 11 rue Humann, 67085 Strasbourg Cedex, France.
Biomacromolecules. 2004 Mar-Apr;5(2):284-94. doi: 10.1021/bm0342281.
Poly(L-lysine)/hyaluronan (PLL/HA) films were chemically cross-linked with a water soluble carbodiimide (EDC) in combination with a N-hydroxysulfo-succinimide (NHS) to induce amide formation. Fourier transform infrared spectroscopy confirms the conversion of carboxylate and ammonium groups into amide bonds. Quartz crystal microbalance-dissipation reveals that the cross linking reaction is accompanied by a change in the viscoelastic properties of the films leading to more rigid films. After the cross-linking reaction, both positively and negatively ending films exhibit a negative zeta potential. It is shown by fluorescence recovery after photobleaching measured by confocal laser scanning microscopy that cross-linking dramatically reduces the diffusion of the PLL chains in the network. Cross linking also renders the films highly resistant to hyaluronidase, an enzyme that naturally degrades hyaluronan. Finally, the adhesion of chondrosarcoma cells on the films terminating either with PLL or HA is also investigated. Whereas the non cross-linked films are highly resistant to cell adhesion, the cells adhere and spread well on the cross-linked films.
聚(L-赖氨酸)/透明质酸(PLL/HA)薄膜与水溶性碳二亚胺(EDC)结合N-羟基硫代琥珀酰亚胺(NHS)进行化学交联,以诱导酰胺形成。傅里叶变换红外光谱证实羧酸盐和铵基团转化为酰胺键。石英晶体微天平耗散显示交联反应伴随着薄膜粘弹性性质的变化,导致薄膜更坚硬。交联反应后,带正电和带负电的薄膜均呈现负ζ电位。通过共聚焦激光扫描显微镜测量的光漂白后荧光恢复表明,交联显著降低了PLL链在网络中的扩散。交联还使薄膜对透明质酸酶具有高度抗性,透明质酸酶是一种天然降解透明质酸的酶。最后,还研究了软骨肉瘤细胞在以PLL或HA终止的薄膜上的粘附情况。未交联的薄膜对细胞粘附具有高度抗性,而细胞在交联薄膜上能很好地粘附和铺展。