1 College of Mechanical and Electrical Engineering, Hohai University, Changzhou, Jiangsu, PR China.
3 Mechanical and Aerospace Engineering Department, University of California, Los Angeles, Los Angeles, CA, USA.
SLAS Discov. 2017 Jun;22(5):626-634. doi: 10.1177/2472555217701247. Epub 2017 Mar 24.
Three-dimensional (3D) in vitro tissue models provide an approach for the systematic, repetitive, and quantitative study of drugs. In this study, we constructed an in vitro 3D acrylated hyaluronic acid (AHA) hydrogel model encapsulating fibroblasts, performed long-period 3D culture, and tested cellular topological changes and proliferation variation in the presence of herpes simplex virus-1 (HSV-1) as an infecting virus and acyclovir (ACV) as the treatment drug. The AHA hydrogels were formed by using Michael addition chemistry of bis-cysteine containing MMP-degradable cross-linker onto AHA prefunctionalized with cell adhesion peptides (RGD). Cellular structures of 3T3 fibroblasts in hydrogel presented different morphological evolution processes and proliferation rates between different groups, including HSV-1 treated alone, ACV treated alone, HSV-1 and ACV cotreated, and control samples. In AHA hydrogel, ACV blocked HSV-1 infection/replication on fibroblasts. Yet, the proliferation of ACV-treated fibroblasts was slower than that of the control group. A significantly longer period was required for cells in 3D AHA gel to regain a healthy status when compared with cells in two-dimensional (2D) culture. This hydrogel-based 3D culture model potentially lays a foundation for analyzing the response of self-organized 3D tissues to viruses and drugs in a way that is closer to nature.
三维（3D）体外组织模型为药物的系统、重复和定量研究提供了一种方法。在本研究中，我们构建了一种体外 3D 丙烯酰化透明质酸（AHA）水凝胶模型，其中包埋了成纤维细胞，进行了长期 3D 培养，并测试了单纯疱疹病毒-1（HSV-1）作为感染病毒和阿昔洛韦（ACV）作为治疗药物存在时细胞拓扑变化和增殖变化。AHA 水凝胶是通过使用含有双半胱氨酸的双官能 MMP 可降解交联剂对细胞粘附肽（RGD）预功能化的 AHA 进行迈克尔加成化学形成的。3T3 成纤维细胞在水凝胶中的细胞结构在不同组之间表现出不同的形态演变过程和增殖率，包括单独用 HSV-1 处理、单独用 ACV 处理、HSV-1 和 ACV 共同处理以及对照样品。在 AHA 水凝胶中，ACV 阻断了 HSV-1 对成纤维细胞的感染/复制。然而，ACV 处理的成纤维细胞的增殖速度比对照组慢。与二维（2D）培养中的细胞相比，3D AHA 凝胶中的细胞恢复健康状态需要更长的时间。这种基于水凝胶的 3D 培养模型为分析自我组织的 3D 组织对病毒和药物的反应奠定了基础，这种方法更接近自然。
