Department of Oral and Maxillofacial Surgery, University Hospital Tübingen, 72076 Tübingen, Germany.
Department of Orthopedic Surgery, University Hospital Tübingen, 72072 Tübingen, Germany.
Int J Mol Sci. 2022 Apr 23;23(9):4682. doi: 10.3390/ijms23094682.
Perfused bioreactor systems are considered to be a promising approach for the 3D culturing of stem cells by improving the quality of the tissue-engineered grafts in terms of better cell proliferation and deeper penetration of used scaffold materials. Our study aims to establish an optimal perfusion culture system for jaw periosteal cell (JPC)-seeded scaffolds. For this purpose, we used beta-tricalcium phosphate (β-TCP) scaffolds as a three-dimensional structure for cell growth and osteogenic differentiation. Experimental set-ups of tangential and sigmoidal fluid configurations with medium flow rates of 100 and 200 µL/min were applied within the perfusion system. Cell metabolic activities of 3D-cultured JPCs under dynamic conditions with flow rates of 100 and 200 µL/min were increased in the tendency after 1, and 3 days of culture, and were significantly increased after 5 days. Significantly higher cell densities were detected under the four perfused conditions compared to the static condition at day 5. However, cell metabolic and proliferation activity under dynamic conditions showed flow rate independency in our study. In this study, dynamic conditions increased the expression of osteogenic markers (ALPL, COL1A1, RUNX2, and OCN) compared to static conditions and the tangential configuration showed a stronger osteogenic effect than the sigmoidal flow configuration.
灌流生物反应器系统被认为是一种有前途的方法,通过改善组织工程移植物的质量,促进干细胞的 3D 培养,从而实现更好的细胞增殖和更深入的使用支架材料渗透。我们的研究旨在为颌骨骨膜细胞(JPC)接种的支架建立最佳的灌流培养系统。为此,我们使用β-磷酸三钙(β-TCP)支架作为细胞生长和成骨分化的三维结构。在灌流系统中,应用了切向和正弦型流体构型的实验装置,培养基流速分别为 100 和 200µL/min。在 1 和 3 天的培养后,动态条件下培养的 JPC 细胞的代谢活性呈增加趋势,在 5 天后显著增加。与静态条件相比,在灌流的四种条件下,第 5 天检测到的细胞密度明显更高。然而,在我们的研究中,细胞代谢和增殖活性在动态条件下表现出与流速无关。在这项研究中,与静态条件相比,动态条件增加了成骨标志物(ALPL、COL1A1、RUNX2 和 OCN)的表达,切向构型比正弦流构型表现出更强的成骨作用。