本研究的目的是通过调整骨骼的形成状态来进一步改善骨骼的体外3D成骨细胞(OB)-破骨细胞(OC)共培养模型。再吸收,以及它们在基础研究中未来应用的平衡,药物开发和个性化医疗。这是通过改变培养基组成和单核细胞接种密度来实现的,对细胞行为影响最大的两个外部参数。将单核细胞以两种接种密度接种到由MSC衍生的OB预矿化的3D丝素蛋白构建体上,并在三种不同培养基之一中共培养(OC刺激,中性和OB刺激培养基)三周。组织学显示共培养后的矿化基质和OC培养基组中的OC标记。扫描电子显微镜在OC培养基组中显示大的OC样细胞。显微计算机断层扫描显示OB培养基组的形成增加,中性介质组的平衡和OC介质组的吸收。培养上清液样品在OC培养基组中显示出较高的早期抗酒石酸酸性磷酸酶(TRAP)释放,中性介质组中的较晚和较低的释放,在OB培养基组中几乎没有释放。单核细胞接种密度增加显示OC培养基中TRAP释放和再吸收的增加不成比例,同时在中性介质中按比例增加TRAP释放,而不影响净吸收。3DOB-OC共培养模型被有效地用于显示使用OB培养基的过量矿物质沉积,使用OC培养基吸收,或使用中性介质的平衡。适用于该模型的所有三种媒体在基础研究中可能都有自己独特的应用,药物开发,个性化医疗。
The aim of the present study was to further improve an in vitro 3D osteoblast (OB) - osteoclast (OC) co-culture model of bone by tuning it towards states of formation, resorption, and equilibrium for their future applications in fundamental research, drug development and personalized medicine. This was achieved by varying culture medium composition and monocyte seeding density, the two external parameters that affect cell behavior the most. Monocytes were seeded at two seeding densities onto 3D silk-fibroin constructs pre-mineralized by MSC-derived OBs and were co-cultured in one of three different media (OC stimulating, Neutral and OB stimulating medium) for three weeks. Histology showed mineralized matrix after co-culture and OC markers in the OC medium group. Scanning Electron Microscopy showed large OC-like cells in the OC medium group. Micro-computed tomography showed increased formation in the OB medium group, equilibrium in the Neutral medium group and resorption in the OC medium group. Culture supernatant samples showed high early tartrate resistant acid phosphatase (TRAP) release in the OC medium group, a later and lower release in the Neutral medium group, and almost no release in the OB medium group. Increased monocyte seeding density showed a less-than-proportional increase in TRAP release and resorption in OC medium, while it proportionally increased TRAP release in Neutral medium without affecting net resorption. The 3D OB-OC co-culture model was effectively used to show an excess of mineral deposition using OB medium, resorption using OC medium, or an equilibrium using Neutral medium. All three media applied to the model may have their own distinct applications in fundamental research, drug development, and personalized medicine.