Department of Biomaterials, Institute of Clinical Dentistry, University of Oslo, NO-0317 P.O. Box 1109 Blindern Oslo, Norway.
Cells. 2022 Jun 22;11(13):1995. doi: 10.3390/cells11131995.
Combining biomaterial scaffolds with cells serves as a promising strategy for engineering critical size defects; however, homogenous cellular growth within large scaffolds is challenging. Mechanical stimuli can enhance bone regeneration by modulating cellular growth and differentiation. Here, we compare dynamic seeding in a perfusion flow bioreactor with static seeding for a synthetic bone scaffold for up to 21 days using the cell line MC3T3-E1 and primary human osteoblast, confocal laser scanning microscopy, and real-time reverse transcriptase-polymerase chain reaction. The secretion of bone-related proteins was quantified using multiplex immunoassays. Dynamic culture improved cellular distribution through the TiO scaffold and induced a five-fold increase in cell number after 21 days. The relative mRNA expression of osteopontin of MC3T3-E1 was 40-fold enhanced after 7 and 21 days at a flow rate of 0.08 mL/min, and that of collagen type I alpha I expression was 18-fold after 21 days. A flow rate of 0.16 mL/min was 10-fold less effective. Dynamic culture increased the levels of dickkopf-related protein 1 (60-fold), osteoprotegrin (29-fold), interleukin-6 (23-fold), interleukin-8 (36-fold), monocyte chemoattractant protein 1 (28-fold) and vascular endothelial growth factor (6-fold) in the medium of primary human osteoblasts after 21 days compared to static seeding. The proposed method may have clinical potential for bone tissue engineering.
将生物材料支架与细胞结合是工程化临界尺寸缺陷的一种有前途的策略；然而，在大支架内实现同质细胞生长是具有挑战性的。机械刺激可以通过调节细胞生长和分化来增强骨再生。在这里，我们使用 MC3T3-E1 细胞系和原代人成骨细胞，通过比较灌注流生物反应器中的动态接种和静态接种，在合成骨支架上进行长达 21 天的研究，使用共聚焦激光扫描显微镜和实时逆转录-聚合酶链反应。使用多重免疫测定法来定量骨相关蛋白的分泌。动态培养通过 TiO2 支架改善了细胞分布，并在 21 天后将细胞数量增加了五倍。在流速为 0.08 mL/min 时，MC3T3-E1 的骨桥蛋白相对 mRNA 表达在 7 和 21 天后增强了 40 倍，I 型胶原α I 表达在 21 天后增强了 18 倍。流速为 0.16 mL/min 时的效果降低了 10 倍。与静态接种相比，动态培养在第 21 天使原代人成骨细胞培养基中 dickkopf 相关蛋白 1（60 倍）、骨保护素（29 倍）、白细胞介素-6（23 倍）、白细胞介素-8（36 倍）、单核细胞趋化蛋白 1（28 倍）和血管内皮生长因子（6 倍）的水平增加。该方法可能具有临床潜力用于骨组织工程。
