Hatami Javad, Silva Sandra G, Oliveira Mariana B, Costa Rui R, Reis Rui L, Mano João F
3B's Research Group, Biomaterials, Biodegradables and Biomimetics, University of Minho, Headquarters of the European Institute of Excellence of Tissue Engineering and Regenerative Medicine, Avepark-Parque de Ciência e Tecnologia, Zona Industrial da Gandra, 4805-017 Barco GMR, Portugal.
ICVS/3B's, PT Government Associated Laboratory, Braga/Guimarães, Portugal.
Polymers (Basel). 2017 Sep 13;9(9):440. doi: 10.3390/polym9090440.
The construction of multilayered films with tunable properties could offer new routes to produce biomaterials as a platform for 3D cell cultivation. In this study, multilayered films produced with five bilayers of chitosan and alginate (CHT/ALG) were built using water-soluble modified mesyl and tosyl⁻CHT via layer-by-layer (LbL) self-assembly. NMR results demonstrated the presences of mesyl (2.83 ppm) and tosyl groups (2.39, 7.37 and 7.70 ppm) in the chemical structure of modified chitosans. The buildup of multilayered films was monitored by quartz-crystal-microbalance (QCM-D) and film thickness was estimated using the Voigt-based viscoelastic model. QCM-D results demonstrated that CHT/ALG films constructed using mesyl or tosyl modifications (mCHT/ALG) were significantly thinner in comparison to the CHT/ALG films constructed with unmodified chitosan ( < 0.05). Adhesion analysis demonstrated that human adipose stem cells (hASCs) did not adhere to the mCHT/ALG multilayered films and formed aggregates with sizes between ca. 100⁻200 µm. In vitro studies on cell metabolic activity and live/dead staining suggested that mCHT/ALG multilayered films are nontoxic toward hACSs. Multilayered films produced via LbL assembly of ALG and off-the-shelf, water-soluble modified chitosans could be used as a scaffold for the 3D aggregates formation of hASCs in vitro.
构建具有可调性质的多层膜可为生产生物材料提供新途径，作为3D细胞培养的平台。在本研究中，通过层层（LbL）自组装，使用水溶性改性甲磺酰基和对甲苯磺酰基壳聚糖（CHT）制备了含有五层壳聚糖和藻酸盐（CHT/ALG）的多层膜。核磁共振结果表明，改性壳聚糖的化学结构中存在甲磺酰基（2.83 ppm）和对甲苯磺酰基（2.39、7.37和7.70 ppm）。通过石英晶体微天平（QCM-D）监测多层膜的形成，并使用基于Voigt的粘弹性模型估算膜厚度。QCM-D结果表明，与使用未改性壳聚糖构建的CHT/ALG膜相比，使用甲磺酰基或对甲苯磺酰基改性构建的CHT/ALG膜明显更薄（P<0.05）。粘附分析表明，人脂肪干细胞（hASC）不粘附于mCHT/ALG多层膜，而是形成大小约为100-200 µm的聚集体。关于细胞代谢活性和活/死染色的体外研究表明，mCHT/ALG多层膜对hACS无毒。通过ALG与市售水溶性改性壳聚糖的LbL组装制备的多层膜可作为体外hASC 3D聚集体形成的支架。
