Prometheus Division of Skeletal Tissue Engineering, KU Leuven, O&N1, Herestraat 49, PB 813, 3000 Leuven, Belgium.
Department of Development and Regeneration, Skeletal Biology and Engineering Research Center, KU Leuven, O&N1, Herestraat 49, PB 813, 3000 Leuven, Belgium.
Biofabrication. 2024 Aug 22;16(4). doi: 10.1088/1758-5090/ad6e1a.
Multicellular spheroids such as microtissues and organoids have demonstrated great potential for tissue engineering applications in recent years as these 3D cellular units enable improved cell-cell and cell-matrix interactions. Current bioprinting processes that use multicellular spheroids as building blocks have demonstrated limited control on post printing distribution of cell spheroids or moderate throughput and printing efficiency. In this work, we presented a laser-assisted bioprinting approach able to transfer multicellular spheroids as building blocks for larger tissue structures. Cartilaginous multicellular spheroids formed by human periosteum derived cells (hPDCs) were successfully bioprinted possessing high viability and the capacity to undergo chondrogenic differentiation post printing. Smaller hPDC spheroids with diameters ranging from ∼100 to 150m were successfully bioprinted through the use of laser-induced forward transfer method (LIFT) however larger spheroids constituted a challenge. For this reason a novel alternative approach was developed termed as laser induced propulsion of mesoscopic objects (LIPMO) whereby we were able to bioprint spheroids of up to 300m. Moreover, we combined the bioprinting process with computer aided image analysis demonstrating the capacity to 'target and shoot', through automated selection, multiple large spheroids in a single sequence. By taking advantage of target and shoot system, multilayered constructs containing high density cell spheroids were fabricated.
近年来，作为能够改善细胞间和细胞与基质间相互作用的 3D 细胞单位，多细胞球体（如微组织和类器官）在组织工程应用中显示出巨大的潜力。目前，使用多细胞球体作为构建块的生物打印工艺对打印后细胞球体的分布控制有限，或具有中等通量和打印效率。在这项工作中，我们提出了一种激光辅助生物打印方法，能够将多细胞球体作为更大组织结构的构建块进行转移。由人骨膜来源细胞（hPDC）形成的软骨多细胞球体成功地进行了生物打印，具有高存活率和打印后发生软骨分化的能力。通过使用激光诱导前向转移方法（LIFT），成功地打印出直径在 100-150μm 之间的较小 hPDC 球体，然而更大的球体则构成了挑战。因此，开发了一种新的替代方法，称为中观物体的激光推进（LIPMO），通过这种方法，我们能够打印出直径达 300μm 的球体。此外，我们将生物打印过程与计算机辅助图像分析相结合，通过自动化选择，在单个序列中对多个大球体进行“靶向和射击”。利用目标和射击系统，制造了包含高密度细胞球体的多层结构。
