Abstract：
Three-dimensional bioprinting is a potent biofabrication technique in tissue engineering but is limited by inadequate bioink availability. Plant-derived proteins are increasingly recognized as highly promising yet underutilized materials for biomedical product development and hold potential for use in bioink formulations. Herein, we report the development of a biocompatible plant protein bioink from pea protein isolate. Through pH shifting, ethanol precipitation, and lyophilization, the pea protein isolate (PPI) transformed from an insoluble to a soluble form. Next, it was modified with glycidyl methacrylate to obtain methacrylate-modified PPI (PPIGMA), which is photocurable and was used as the precursor of bioink. The mechanical and microstructural studies of the hydrogel containing 16% PPIGMA revealed a suitable compress modulus and a porous network with a pore size over 100 μm, which can facilitate nutrient and waste transportation. The PPIGMA bioink exhibited good 3D bioprinting performance in creating complex patterns and good biocompatibility as plenty of viable cells were observed in the printed samples after 3 days of incubation in the cell culture medium. No immunogenicity of the PPIGMA bioink was identified as no inflammation was observed for 4 weeks after implantation in Sprague Dawley rats. Compared with methacrylate-modified gelatin, the PPIGMA bioink significantly enhanced cartilage regeneration in vitro and in vivo, suggesting that it can be used in tissue engineering applications. In summary, the PPIGMA bioink can be potentially used for tissue engineering applications.
摘要：
三维生物打印是组织工程中一种有效的生物制造技术，但受到生物墨水可用性不足的限制。植物衍生的蛋白质越来越被认为是生物医学产品开发的非常有前途但未充分利用的材料，并且具有用于生物墨水制剂的潜力。在这里，我们报道了从豌豆蛋白分离物中开发出生物相容性植物蛋白生物墨水。通过pH值变化，乙醇沉淀，和冻干，豌豆分离蛋白（PPI）从不溶性转化为可溶性形式。接下来,用甲基丙烯酸缩水甘油酯改性得到甲基丙烯酸酯改性PPI(PPIGMA),它是可光固化的，被用作生物墨水的前体。含有16％PPIGMA的水凝胶的机械和微观结构研究表明，合适的压缩模量和孔径超过100μm的多孔网络，这可以促进营养和废物的运输。PPIGMA生物墨水在创建复杂图案和良好的生物相容性方面表现出良好的3D生物打印性能,因为在细胞培养基中孵育3天后在打印的样品中观察到大量的活细胞。没有鉴定PPIGMA生物墨水的免疫原性,因为在SpragueDawley大鼠中植入后4周内没有观察到炎症。与甲基丙烯酸酯改性明胶相比，PPIGMA生物墨水在体外和体内显着增强软骨再生，表明它可以用于组织工程应用。总之,PPIGMA生物墨水可能用于组织工程应用。
