Gwiazda Marcin, Kumar Sudheesh, Świeszkowski Wojciech, Ivanovski Saso, Vaquette Cedryck
Faculty of Materials Science and Engineering, Warsaw University of Technology, Warsaw, Poland; Griffith Health Institute, Griffith University, Gold Coast, Australia.
Griffith Health Institute, Griffith University, Gold Coast, Australia.
J Mech Behav Biomed Mater. 2020 Apr;104:103631. doi: 10.1016/j.jmbbm.2020.103631. Epub 2020 Jan 21.
The effect of melt electrospun writing fiber arrangement on cellular behavior has not yet been thoroughly investigated. Cellular orientation is particularly important in the context of ligament tissue engineering for orthopedic applications whereby a high degree of cell alignment is present in the native tissue. The aim of this study was to investigate the response of human mesenchymal stem cells (hMSC) to three different patterned porous polycaprolactone scaffolds (aligned, crimped and random) fabricated by melt electrospinning writing, resulting in 20 μm diameter electrospun fibers. Cell orientation was investigated over 4 weeks in vitro and it was demonstrated that the aligned pattern was capable of orientating the hMSCs towards the main direction of the fibers and this feature was maintained over the entire culture period whereas the orientation was rapidly lost in the crimped pattern. In order to fabricate a functional scaffold for ligament tissue engineering, the scaffolds were rolled in three bundles, subsequently braided and combined with a bone compartment (consisting of a melt electrospun scaffold seeded with osteogenically induced hMSCs) for the development of a Bone-Ligament-Bone (BLB) construct. The mechanical properties of non-cellularized and cellularized BLB constructs were assessed under both quasi-static and cyclic conditions. This revealed that the in vitro maturation significantly softened the BLB constructs and that the mechanical properties were several fold lower than those of native tissue. The cyclic testing demonstrated that the presence of cell sheets resulted in increased resilience and elasticity, even though the global mechanical properties were decreased for the in vitro matured constructs (regardless of the pattern). In conclusion, we demonstrated that melt electrospinning writing fiber organization can induce spontaneous cell alignment and that large cellularized BLB constructs with complex geometry can achieve mechanical resilience under cyclic stretching.
熔体静电纺丝书写纤维排列对细胞行为的影响尚未得到充分研究。在用于骨科应用的韧带组织工程中，细胞取向尤为重要，因为天然组织中存在高度的细胞排列。本研究的目的是研究人间充质干细胞（hMSC）对通过熔体静电纺丝书写制备的三种不同图案化多孔聚己内酯支架（排列、卷曲和随机）的反应，所得电纺纤维直径为20μm。在体外4周内研究了细胞取向，结果表明排列图案能够使hMSC向纤维的主要方向取向，并且这一特征在整个培养期内得以维持，而在卷曲图案中取向迅速丧失。为了制备用于韧带组织工程的功能性支架，将支架卷成三束，随后编织并与骨部分（由接种有成骨诱导hMSC的熔体静电纺丝支架组成）结合，以开发骨-韧带-骨（BLB）构建体。在准静态和循环条件下评估了非细胞化和细胞化BLB构建体的力学性能。这表明体外成熟显著软化了BLB构建体，并且力学性能比天然组织低几倍。循环测试表明，即使体外成熟构建体（无论图案如何）的整体力学性能下降，细胞片层的存在仍导致弹性和韧性增加。总之，我们证明了熔体静电纺丝书写纤维组织可诱导细胞自发排列，并且具有复杂几何形状的大型细胞化BLB构建体在循环拉伸下可实现力学弹性。
