Eur Cell Mater. 2021 May 5;41:485-501. doi: 10.22203/eCM.v041a31.
The aim of the present study was to fabricate and characterise chitosan scaffolds from animal and fungal sources, with or without gelatine as a co-polymer, and cross-linked to 3-glycidyloxyproply trimethoxysilane (GPTMS) or genipin for application in dental root tissue engineering. Chitosan-based scaffolds were prepared by the emulsion freeze-drying technique. Scanning electron microscopy (SEM) and nano-focus computed tomography (nano-CT) were used to characterise scaffold microstructure. Chemical composition and cross-linking were evaluated by Fourier transform infrared-attenuated total reflectance spectroscopy. Compression tests were performed to evaluate scaffold mechanical properties. Scaffold degradation was evaluated by gravimetric method and SEM. Scaffold bioactivity immersed in simulated body fluid was evaluated by SEM, with associated electron dispersive X-ray spectroscopy, and apatite formation was examined by X-ray diffraction. Finally, human dental pulp stem cells (hDPSCs) viability was evaluated. The fabrication method used was successful in producing scaffolds with organised porosity. Chitosan source (animal vs. fungal), co-polymerisation with gelatine and cross-linking using GPTMS or genipin had a significant effect on scaffold properties and hDPSCs response. Chitosan-genipin (CS-GEN) scaffolds had the largest pore diameter, while the chitosan-gelatine-GPTMS (CS-GEL-GPTMS) scaffolds had the smallest. Animal chitosan-gelatine co-polymerisation increased scaffold compressive strength, while fungal chitosan scaffolds (fCS-GEL-GPTMS) had the fastest degradation rate, losing 80 % of their weight by day 21. Gelatine co-polymerisation and GPTMS cross-linking enhanced chitosan scaffolds bioactivity through the formation of an apatite layer as well as improved hDPSCs attachment and viability. Tailored chitosan scaffolds with tuned properties and favourable hDPSCs response can be obtained for regenerative dentistry applications.
本研究的目的是制备和表征壳聚糖支架，其来源为动物和真菌，可添加或不添加明胶作为共聚物，并通过 3-缩水甘油氧基丙基三甲氧基硅烷（GPTMS）或京尼平交联，应用于牙根管组织工程。壳聚糖支架通过乳化冷冻干燥技术制备。扫描电子显微镜（SEM）和纳米焦点计算机断层扫描（nano-CT）用于表征支架的微观结构。通过傅里叶变换红外衰减全反射光谱评估化学组成和交联。通过压缩试验评估支架的机械性能。通过重量法和 SEM 评估支架的降解。通过 SEM 评估支架的生物活性，结合电子色散 X 射线能谱，并通过 X 射线衍射检查磷灰石的形成。最后，评估人牙髓干细胞（hDPSCs）的活力。使用的制备方法成功地生产出具有组织化孔隙率的支架。壳聚糖来源（动物与真菌）、与明胶共聚以及使用 GPTMS 或京尼平交联对支架性能和 hDPSCs 反应有显著影响。壳聚糖-京尼平（CS-GEN）支架的孔径最大，而壳聚糖-明胶-GPTMS（CS-GEL-GPTMS）支架的孔径最小。动物壳聚糖-明胶共聚物增加了支架的压缩强度，而真菌壳聚糖支架（fCS-GEL-GPTMS）的降解速度最快，在第 21 天失去了 80%的重量。明胶共聚物和 GPTMS 交联通过形成磷灰石层以及提高 hDPSCs 的黏附和活力，增强了壳聚糖支架的生物活性。具有可调性质和有利 hDPSCs 反应的定制化壳聚糖支架可用于再生牙科应用。
