Reyes-Peces María V, Pérez-Moreno A, de-Los-Santos Deseada María, Mesa-Díaz María Del Mar, Pinaglia-Tobaruela Gonzalo, Vilches-Pérez Jose Ignacio, Fernández-Montesinos Rafael, Salido Mercedes, de la Rosa-Fox Nicolás, Piñero Manuel
Department of Condensed Matter Physics 1, Faculty of Science, University of Cadiz, 11510 Cádiz, Spain.
Instituto de Investigación e Innovación Biomédica de Cádiz (INIBICA), 11009 Cádiz, Spain.
Polymers (Basel). 2020 Nov 17;12(11):2723. doi: 10.3390/polym12112723.
This study introduces a new synthesis route for obtaining homogeneous chitosan (CS)-silica hybrid aerogels with CS contents up to 10 wt%, using 3-glycidoxypropyl trimethoxysilane (GPTMS) as coupling agent, for tissue engineering applications. Aerogels were obtained using the sol-gel process followed by CO supercritical drying, resulting in samples with bulk densities ranging from 0.17 g/cm to 0.38 g/cm. The textural analysis by N-physisorption revealed an interconnected mesopore network with decreasing specific surface areas (1230-700 m/g) and pore sizes (11.1-8.7 nm) by increasing GPTMS content (2-4 molar ratio GPTMS:CS monomer). In addition, samples exhibited extremely fast swelling by spontaneous capillary imbibition in PBS solution, presenting swelling capacities from 1.75 to 3.75. The formation of a covalent crosslinked hybrid structure was suggested by FTIR and confirmed by an increase of four hundred fold or more in the compressive strength up to 96 MPa. Instead, samples synthesized without GPTMS fractured at only 0.10-0.26 MPa, revealing a week structure consisted in interpenetrated polymer networks. The aerogels presented bioactivity in simulated body fluid (SBF), as confirmed by the in vitro formation of hydroxyapatite (HAp) layer with crystal size of approximately 2 µm size in diameter. In vitro studies revealed also non cytotoxic effect on HOB osteoblasts and also a mechanosensitive response. Additionally, control cells grown on glass developed scarce or no stress fibers, while cells grown on hybrid samples showed a significant ( < 0.05) increase in well-developed stress fibers and mature focal adhesion complexes.
本研究引入了一种新的合成路线,以获得壳聚糖(CS)含量高达10 wt%的均匀壳聚糖-二氧化硅杂化气凝胶,使用3-缩水甘油氧基丙基三甲氧基硅烷(GPTMS)作为偶联剂,用于组织工程应用。气凝胶通过溶胶-凝胶法制备,随后进行CO2超临界干燥,得到的样品堆积密度范围为0.17 g/cm³至0.38 g/cm³。通过N-物理吸附进行的结构分析表明,随着GPTMS含量(GPTMS:CS单体的摩尔比为2-4)的增加,形成了相互连接的介孔网络,比表面积(1230-700 m²/g)和孔径(11.1-8.7 nm)减小。此外,样品在PBS溶液中通过自发毛细管吸附表现出极快的溶胀,溶胀能力为1.75至3.75。FTIR表明形成了共价交联的杂化结构,抗压强度增加了四百倍或更多,高达96 MPa,证实了这一点。相反,未使用GPTMS合成的样品在仅0.10-0.26 MPa时就会断裂,表明其结构由互穿聚合物网络组成,较为薄弱。气凝胶在模拟体液(SBF)中表现出生物活性,体外形成的羟基磷灰石(HAp)层的晶体尺寸直径约为2 µm,证实了这一点。体外研究还表明,对人成骨细胞(HOB)无细胞毒性作用,并且具有机械敏感反应。此外,在玻璃上生长的对照细胞形成的应力纤维稀少或没有,而在杂化样品上生长的细胞显示出发育良好的应力纤维和成熟的粘着斑复合物显著增加(P<0.05)。