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, Guimarães, Portugal.
ICVS/3B's-PT Government Associate Laboratory, Guimarães, Portugal.
J Biomed Mater Res A. 2022 Oct;110(10):1655-1668. doi: 10.1002/jbm.a.37415. Epub 2022 Jun 9.
The treatment of skeletal muscle defects is still a topic of noteworthy concern since surgical intervention is not capable of recovering muscle function. Herein, we propose myoblasts laden in laminin-inspired biofunctionalized gellan gum hydrogels as promising tissue-engineered skeletal muscle surrogates. Gellan gum-based hydrogels were developed by combining native gellan gum (GG) and GG tethered with laminin-derived peptides (CIKVAVS (V), KNRLTIELEVRTC (T) or RKRLQVQLSIRTC (Q)), using different polymer content (0.75%-1.875%). Hydrogels were characterized in terms of compressive modulus, molecules trafficking, and C2C12 adhesion. Hydrogels with higher polymeric content (1.125%-1.875%) showed higher stiffness whereas hydrogels with lower polymer content (0.75%-1.125%) showed higher fluorescein isothiocyanate-dextran molecules diffusion. Cell spreading was achieved regardless of the laminin-derived peptide but preferred in hydrogels with higher polymer content (1.125%-1.875%). Taken together, hydrogels with 1.125% of polymer content were selected for printability analysis. GG-based inks showed a non-newtonian, shear-thinning, and thixotropic behavior suitable for printing. Accordingly, all inks were printable, but inks tethered with T and Q peptides presented some signs of clogging. Cell viability was affected after printing but increased after 7 days of culture. After 7 days, cells were spreading but not showing significant signs of cell-cell communications. Therefore, cell density was increased, thus, myocytes loaded in V-tethered GG-based inks showed higher cell-cell communication, spreading morphology, and alignment 7, 14 days post-printing. Overall, myoblasts laden in laminin-inspired biofunctionalized GG-based hydrogels are a promising skeletal muscle surrogate with the potential to be used as in vitro model or explored for further in vivo applications.
骨骼肌缺损的治疗仍然是一个值得关注的话题，因为手术干预无法恢复肌肉功能。在此，我们提出负载成肌细胞的层粘连蛋白仿生功能化凝胶多糖水凝胶作为有前途的组织工程骨骼肌替代物。通过将天然凝胶多糖（GG）与层粘连蛋白衍生肽（CIKVAVS（V）、KNRLTIELEVRTC（T）或 RKRLQVQLSIRTC（Q））键合的 GG 结合使用，开发了基于 GG 的水凝胶，使用不同的聚合物含量（0.75%-1.875%）。从压缩模量、分子迁移和 C2C12 黏附等方面对水凝胶进行了表征。聚合物含量较高的水凝胶（1.125%-1.875%）表现出较高的刚性，而聚合物含量较低的水凝胶（0.75%-1.125%）则表现出较高的异硫氰酸荧光素-葡聚糖分子扩散。无论层粘连蛋白衍生肽如何，细胞都能展开，但在聚合物含量较高的水凝胶（1.125%-1.875%）中更为明显。综上所述，选择聚合物含量为 1.125%的水凝胶进行可打印性分析。基于 GG 的墨水表现出非牛顿、剪切变稀和触变行为，适合打印。因此，所有的墨水都可以打印，但与 T 和 Q 肽键合的墨水有一些堵塞的迹象。打印后细胞活力受到影响，但培养 7 天后活力增加。培养 7 天后，细胞开始展开，但没有明显的细胞间通讯迹象。因此，细胞密度增加，负载在 V 键合 GG 基墨水中的成肌细胞在打印后 7、14 天表现出更高的细胞间通讯、展开形态和排列。总之，负载在层粘连蛋白仿生功能化 GG 基水凝胶中的成肌细胞是一种有前途的骨骼肌替代物，具有作为体外模型的潜力，或可进一步探索其在体内的应用。
