Dutta Sayan Deb, Bin Jin, Ganguly Keya, Patel Dinesh K, Lim Ki-Taek
Department of Biosystems Engineering, Institute of Forest Science, Kangwon National University Chuncheon-24341 Republic of Korea
School of Stomatology, Affiliated Hospital of Yanbian University Yanji-136200 Beijing China.
RSC Adv. 2021 Jun 7;11(33):20342-20354. doi: 10.1039/d1ra01143j. eCollection 2021 Jun 3.
3D bioprinted hydrogel has gained enormous attention, especially in tissue engineering, owing to its attractive structure and excellent biocompatibility. In this study, we demonstrated that 3D bioprinted cell-laden 'thermoresponsive' poloxamer-407 (P407) gels have the potential to stimulate osteogenic differentiation of apical papilla stem cells (SCAPs) under the influence of low voltage-frequency (5 V-1 Hz, 0.62 mT) electromagnetic fields (EMFs). SCAPs were initially used for cell-laden 3D printing to biomimic the apical papilla of human teeth. The developed hydrogel exhibited higher mechanical strength as well as good printability, showing high-quality micro-architecture. Moreover, the as-printed hydrogels (5 mm × 5 mm) were loaded with plasminogen activator inhibitor-1 (PAI-1) for testing the combined effect of PAI-1 and EMFs on SCAP differentiation. Interestingly, the 3D hydrogels showed improved viability and differentiation of SCAPs under EMFs' influence as examined by live/dead assay and alizarin Red-S staining, respectively. Therefore, our results confirmed that P407 hydrogels are non-toxic for encapsulation of SCAPs, yielding high cell viability and accelerate the cell migration potential. The 3D hydrogels with PAI-1 exhibited high mRNA expression levels for osteogenic/odontogenic gene markers (, , , and ) control after 14 days of culture. Our findings suggest that 3D bioprinted P407 hydrogels are biocompatible for SCAP encapsulation, and the applied low voltage-frequency EMFs could effectively improve dental tissue regeneration, particularly for oral applications.
由于其吸引人的结构和出色的生物相容性,3D生物打印水凝胶受到了极大关注,尤其是在组织工程领域。在本研究中,我们证明了3D生物打印的载有细胞的“热响应性”泊洛沙姆-407(P407)凝胶在低电压频率(5V - 1Hz,0.62mT)电磁场(EMF)影响下具有刺激根尖乳头干细胞(SCAPs)成骨分化的潜力。SCAPs最初被用于载有细胞的3D打印,以模拟人类牙齿的根尖乳头。所开发的水凝胶表现出更高的机械强度以及良好的可打印性,呈现出高质量的微观结构。此外,将打印好的水凝胶(5mm×5mm)加载纤溶酶原激活物抑制剂-1(PAI-1),以测试PAI-1和EMF对SCAP分化的联合作用。有趣的是,通过活/死检测和茜素红S染色分别检测发现,3D水凝胶在EMF影响下显示出SCAPs活力和分化的改善。因此,我们的结果证实P407水凝胶对封装SCAPs无毒,产生高细胞活力并加速细胞迁移潜力。含有PAI-1的3D水凝胶在培养14天后,与对照组相比,成骨/牙源性基因标志物( 、 、 和 )的mRNA表达水平较高。我们的研究结果表明,3D生物打印的P407水凝胶对封装SCAPs具有生物相容性,并且施加的低电压频率EMF可以有效地改善牙组织再生,特别是在口腔应用方面。
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