Blaudez Fanny, Ivanovski Saso, Ipe Deepak, Vaquette Cedryck
School of Dentistry and Oral Health, Gold Coast campus, Griffith University, QLD 4222, Australia.
The University of Queensland, School of Dentistry, Herston, Queensland, Australia.
Mater Sci Eng C Mater Biol Appl. 2020 Dec;117:111282. doi: 10.1016/j.msec.2020.111282. Epub 2020 Jul 24.
Cell seeding is challenging in the case of additively manufactured 3-dimensional scaffolds, as the open macroscopic pore network impedes the retention of the seeding solution. The present study aimed at comparing several seeding conditions (no fetal bovine serum, 10% or 100% serum) and methods (Static seeding in Tissue Culture Treated plate (CT), Static seeding of the MES in non-Culture Treated plate (nCT), Seeding in nCT plate placed on an orbital shaker at 20 rpm (nCTR), Static seeding of the MES previously incubated with 100% FBS for 1 h to allow for protein adsorption (FBS)) commonly utilised in tissue engineering using highly porous melt electrowritten scaffolds, assessing their seeding efficacy, cell distribution homogeneity and reproducibility. Firstly, we demonstrated that the incubation in 100% serum was superior to the 10% serum pre-incubation and that 1 h only was sufficient to obtain enhanced cell attachment. We further compared this technique to the other methods and demonstrated significant and beneficial impact of the 100% serum pre-incubation, which resulted in enhanced efficacy, homogeneous cell distribution and high reproducibility, leading to accelerated colonisation/maturation of the tissue engineered constructs. We further showed the superior performance of this method using 3D-printed scaffolds also made of different polymers, demonstrating its capacity for up-scaling. Therefore, the pre-incubation of the scaffold in 100% serum is a simple yet highly effective method for enhancing cell adhesion and ensuring seeding reproducibility. This is crucial for tissue engineering applications, especially when cell availability is scarce, and for product standardisation from a translational perspective.
对于通过增材制造的三维支架而言，细胞接种具有挑战性，因为开放的宏观孔隙网络会阻碍接种溶液的保留。本研究旨在比较几种接种条件（无胎牛血清、10%或100%血清）和方法（在组织培养处理板（CT）中静态接种、在非培养处理板（nCT）中静态接种间充质干细胞（MES）、将nCT板置于20转/分钟的轨道振荡器上接种（nCTR）、将预先与100%胎牛血清孵育1小时以实现蛋白质吸附的MES进行静态接种（FBS）），这些条件和方法常用于使用高度多孔的熔体电写支架的组织工程中，评估它们的接种效果、细胞分布均匀性和可重复性。首先，我们证明在100%血清中孵育优于10%血清预孵育，且仅1小时就足以增强细胞附着。我们进一步将该技术与其他方法进行比较，证明100%血清预孵育具有显著且有益的影响，这导致了接种效果增强、细胞分布均匀且可重复性高，从而加速了组织工程构建体的定植/成熟。我们还使用由不同聚合物制成的3D打印支架进一步展示了该方法的卓越性能，证明了其扩大规模的能力。因此，将支架在100%血清中预孵育是一种简单而高效的方法，可增强细胞粘附并确保接种的可重复性。这对于组织工程应用至关重要，尤其是在细胞可用性稀缺时，并且从转化的角度来看对于产品标准化也很重要。
