Diogo Gabriela S, Marques Catarina F, Sotelo Carmen G, Pérez-Martín Ricardo I, Pirraco Rogério P, Reis Rui L, Silva Tiago H
3B's Research Group, I3Bs - Research Institute on Biomaterials, Biodegradables and Biomimetics, University of Minho, Headquarters of the European Institute of Excellence on Tissue Engineering and Regenerative Medicine, AvePark, Parque de Ciência e Tecnologia, Zona Industrial da Gandra, 4805-017 Barco, Guimarães, Portugal.
ICVS/3B's - PT Government Associate Laboratory, Braga/Guimarães, Portugal.
ACS Biomater Sci Eng. 2020 Jun 8;6(6):3664-3672. doi: 10.1021/acsbiomaterials.0c00436. Epub 2020 May 5.
Mineralization processes based on coprecipitation methods have been applied as a promising alternative to the most commonly used methods of polymer-ceramic combination, direct mixing, and incubation in simulated body fluid (SBF) or modified SBF. In the present study, for the first time, the mineralization (ideally hydroxyapatite formation) of blue shark ( (PG)) collagen to fabricate 3D printable cell-laden hydrogels is proposed. In the first part, several parameters for collagen mineralization were tested until optimization. The hydroxyapatite formation was confirmed by FT-IR, XRD, and TEM techniques. In the second part, stable bioinks combining the biomimetically mineralized collagen with alginate (AG) (1:1, 1:2, 1:3, and AG) solution were used for 3D printing of hydrogels. The addition of Ca ions into the system did present a synergistic effect: by one side, the mineralization of the collagen occurred, and at same time, they were also useful to ionically cross-link the blends with alginate, avoiding the addition of any cytotoxic chemical cross-linking agent. Mouse fibroblast cell line survival during and after printing was favored by the presence of PG collagen as exhibited by the biological performance of the hydrogels. Inspired in a concept of marine byproduct valorization, 3D bioprinting of mineralized blue shark collagen is thus proposed as a promising approach, envisioning the engineering of mineralized tissues.
基于共沉淀法的矿化过程已被用作一种有前景的替代方法，以取代聚合物 - 陶瓷结合、直接混合以及在模拟体液（SBF）或改性SBF中孵育等最常用的方法。在本研究中，首次提出了对蓝鲨（PG）胶原蛋白进行矿化（理想情况下形成羟基磷灰石）以制备可3D打印的载细胞水凝胶。在第一部分中，对胶原蛋白矿化的几个参数进行了测试，直至优化。通过傅里叶变换红外光谱（FT - IR）、X射线衍射（XRD）和透射电子显微镜（TEM）技术确认了羟基磷灰石的形成。在第二部分中，将仿生矿化的胶原蛋白与藻酸盐（AG）（1:1、1:2、1:3以及纯AG）溶液相结合的稳定生物墨水用于水凝胶的3D打印。向系统中添加钙离子确实呈现出协同效应：一方面，胶原蛋白发生矿化，同时，它们也有助于使混合物与藻酸盐进行离子交联，从而避免添加任何具有细胞毒性的化学交联剂。水凝胶的生物学性能表明，PG胶原蛋白的存在有利于小鼠成纤维细胞系在打印过程中和打印后的存活。受海洋副产品增值概念的启发，因此提出矿化蓝鲨胶原蛋白的3D生物打印是一种有前景的方法，有望实现矿化组织的工程构建。
