Department of Mechanics, Politecnico di Torino, Italy.
Biomacromolecules. 2010 Feb 8;11(2):309-15. doi: 10.1021/bm901169v.
The work was focused on the synthesis and characterization of the chitosan-g-fluorescein (CHFL) conjugate polymer as a biocompatible amphiphilic water-soluble photosensitizer, able to stimulate hydroxyapatite deposition upon visible light irradiation. Fluorescein (FL) grafting to chitosan (CH) chains was confirmed by UV-vis analysis of water solutions of FL and CHFL and by Fourier transform infrared spectroscopy (FTIR-ATR) analysis of CHFL and CH. Smooth CHFL cast films with 4 microm thickness were obtained by solvent casting. Continuous exposure to visible light for 7 days was found to activate the deposition of calcium phosphate crystals from a conventional simulated body fluid (SBF 1.0x) on the surface of CHFL cast films. EDX and FTIR-ATR analyses confirmed the apatite nature of the deposited calcium phosphate crystals. CHFL films preincubated in SBF (1.0x) solution under visible light irradiation and in the dark for 7 days were found to support the in vitro adhesion and proliferation of MG63 osteoblast-like cells (MTT viability test; 1-3 days culture time). On the other hand, the mineralization ability of MG63 osteoblast-like cells was significantly improved on CHFL films preincubated under visible light exposure (alkaline phosphatase activity (ALP) test for 1, 3, 7, and 14 days). The use of photoactive biocompatible conjugate polymer, such as CHFL, may lead to new therapeutic options in the field of bone/dental repair, exploiting the photoexcitation mechanism as a tool for biomineralization.
这项工作集中于壳聚糖接枝荧光素(CHFL)的共轭聚合物的合成与表征,该聚合物是一种具有生物相容性的两亲性水溶性光敏剂,能够在可见光照射下刺激羟基磷灰石的沉积。通过对 FL 和 CHFL 水溶液的 UV-vis 分析,以及对 CHFL 和 CH 的傅里叶变换红外光谱(FTIR-ATR)分析,证实了荧光素(FL)接枝到壳聚糖(CH)链上。通过溶剂浇铸获得了厚度为 4 微米的光滑 CHFL 铸膜。连续暴露在可见光下 7 天,发现能够在 CHFL 铸膜表面激活从传统模拟体液(SBF 1.0x)中钙磷晶体的沉积。EDX 和 FTIR-ATR 分析证实了沉积钙磷晶体的磷灰石性质。在可见光照射下和黑暗中预孵育在 SBF(1.0x)溶液中的 CHFL 膜 7 天,发现能够支持 MG63 成骨样细胞(MTT 活力测试;1-3 天培养时间)在体外的黏附和增殖。另一方面,MG63 成骨样细胞的矿化能力在预孵育在可见光下的 CHFL 膜中显著提高(碱性磷酸酶活性(ALP)测试 1、3、7 和 14 天)。使用光活性生物相容的共轭聚合物,如 CHFL,可能会在骨/牙科修复领域开辟新的治疗选择,利用光激发机制作为生物矿化的工具。