Shirosaki Yuki, Tsuru Kanji, Hayakawa Satoshi, Osaka Akiyoshi, Lopes Maria Ascensão, Santos José Domingos, Costa Maria Adelina, Fernandes Maria Helena
Biomaterial Laboratory, Graduate School of Natural Science and Technology, Okayama University, Tsushima, Okayama-shi 700-8530, Japan.
Acta Biomater. 2009 Jan;5(1):346-55. doi: 10.1016/j.actbio.2008.07.022. Epub 2008 Aug 5.
We attempted to prepare chitosan-silicate hybrid for use in a medical application and evaluated the physico-chemical properties and osteocompatibility of the hybrids as a function of gamma-glycidoxypropyltrimethoxysilane (GPTMS) concentration. Chitosan-silicate hybrids were synthesized using GPTMS as the reagent for cross-linking of the chitosan chains. Fourier transform infrared spectroscopy, (29)Si CP-MAS NMR spectroscopy and the ninhydrin assay were used to analyze the structures of the hybrids, and stress-strain curves were recorded to estimate their Young's modulus. The swelling ability, contact angle and cytocompatibility of the hybrids were investigated as a function of the GPTMS concentration. A certain fraction of GPTMS in each hybrid was linked at the epoxy group to the amino group of chitosan, which was associated with the change in the methoxysilane group of GPTMS due to hybridization. The cross-linking density was around 80% regardless of the volume of GPTMS. As the content of GPTMS increased, the water uptake decreased and the hydrophilicity of the hybrids increased except when the content exceeded amolar ratio of 1.5, when it caused a decrease. The values of the mechanical parameters assessed indicated that significant stiffening of the hybrids was obtained by the addition of GPTMS. The adhesion and proliferation of the MG63 osteoblast cells cultured on the chitosan-GPTMS hybrid surface were improved compared to those on the chitosan membrane, regardless of the GPTMS concentration. Moreover, human bone marrow osteoblast cells proliferated on the chitosan-GPTMS hybrid surface and formed a fibrillar extracellular matrix with numerous calcium phosphate globular structures, both in the presence and in the absence of dexamethasone. Therefore, the chitosan-GPTMS hybrids are promising candidates for basic materials that can promote bone regeneration because of their controllable composition (chitosan/GPTMS ratio).
我们试图制备用于医学应用的壳聚糖 - 硅酸盐杂化物，并评估了这些杂化物的物理化学性质以及作为γ - 缩水甘油氧基丙基三甲氧基硅烷（GPTMS）浓度函数的骨相容性。使用GPTMS作为壳聚糖链交联试剂合成壳聚糖 - 硅酸盐杂化物。采用傅里叶变换红外光谱、（29）Si CP - MAS NMR光谱和茚三酮测定法分析杂化物的结构，并记录应力 - 应变曲线以估计其杨氏模量。研究了杂化物的溶胀能力、接触角和细胞相容性作为GPTMS浓度的函数。每个杂化物中一定比例的GPTMS在环氧基团处与壳聚糖的氨基相连，这与杂交导致的GPTMS甲氧基硅烷基团的变化有关。无论GPTMS的体积如何，交联密度约为80％。随着GPTMS含量的增加，吸水率降低，杂化物的亲水性增加，但当含量超过1.5的摩尔比时除外，此时亲水性会降低。评估的力学参数值表明，通过添加GPTMS可使杂化物显著变硬。与壳聚糖膜相比，无论GPTMS浓度如何，在壳聚糖 - GPTMS杂化物表面培养的MG63成骨细胞的粘附和增殖均得到改善。此外，人骨髓成骨细胞在壳聚糖 - GPTMS杂化物表面增殖，并在存在和不存在地塞米松的情况下形成具有大量磷酸钙球状结构的纤维状细胞外基质。因此，壳聚糖 - GPTMS杂化物因其可控的组成（壳聚糖/GPTMS比例）而有望成为促进骨再生的基础材料。
