Department of Biomedical and Specialty Surgical Sciences, University of Ferrara, Italy.
Department of Biomedical and Specialty Surgical Sciences, University of Ferrara, Italy.
Life Sci. 2016 May 1;152:82-93. doi: 10.1016/j.lfs.2016.03.035. Epub 2016 Mar 22.
We aimed to establish a 3D osteoblasts/osteoclasts co-culture system requiring limited amounts of human primary cells and useful as platform to 1. recapitulate an "oral bone microenvironment" in healthy or pathological condition, and 2. produce potential implantable cell constructs for regeneration of jawbone which can be negatively affected by bisphosphonates (BPs).
Osteoblasts from normal bone chips (hOBs) or from jawbone of patients taking BPs (hnOBs) were co-cultured with monocytes (hMCs) either in static (3D-C) or dynamic (3D-DyC) condition using the RCCS-4™ bioreactor for 3weeks. Cell aggregates were characterized for viability, histological features and specific osteoclastic and osteogenic markers.
In all tested conditions hOBs supported the formation of mature osteoclasts (hOCs), without differentiating agents or exogenous scaffolds. 3D-DyC condition associated with a ground based condition (Xg) rather than modeled microgravity (μXg) produced aggregates with high level of osteogenic markers including Osteopontin (OPN), Osterix (OSX), Runx2 and appreciable bone mineral matrix. hnOBs co-cultured with hMCs in 3D-Dyc/Xg condition generated OPN and mineral matrix positive aggregates.
We optimized a 3D co-culture system with a limited amount of cells preserving viability and functionality of bone cellular components and generating bone-like aggregates also by using cells from jawbone necrotic tissue. The feasibility to obtain from poor-quality bone sites viable osteoblasts able to form aggregates when co-cultured with hMCs, allows to study the development of autologous implantable constructs to overcome jawbone deficiency in patients affected by MRONJ (Medication-Related Osteonecrosis of the Jaws).
我们旨在建立一个需要少量人原代细胞的 3D 成骨细胞/破骨细胞共培养系统，该系统可用作平台：1. 重现健康或病理条件下的“口腔骨微环境”，2. 生成可用于颌骨再生的潜在可植入细胞构建体，这些构建体可能受到双膦酸盐（BPs）的负面影响。
正常骨片（hOBs）或服用 BP 的患者颌骨中的成骨细胞（hnOBs）与单核细胞（hMCs）在 RCCS-4™生物反应器中分别在静态（3D-C）或动态（3D-DyC）条件下共培养 3 周。通过细胞聚集物的活力、组织学特征和特定的破骨细胞和成骨细胞标志物对其进行了表征。
在所有测试条件下，hOBs 支持成熟破骨细胞（hOCs）的形成，而无需分化剂或外源性支架。与地面条件（Xg）相关的 3D-DyC 条件而非模拟微重力（μXg）产生了高水平成骨标志物的聚集物，包括骨桥蛋白（OPN）、骨钙素（OSX）、Runx2 和可观的骨矿物质基质。在 3D-Dyc/Xg 条件下与 hMCs 共培养的 hnOBs 生成了 OPN 和矿化基质阳性聚集物。
我们优化了一个 3D 共培养系统，该系统使用少量细胞保留了骨细胞成分的活力和功能，并通过使用来自坏死颌骨组织的细胞生成了类骨聚集物。从质量较差的骨部位获得具有活力的成骨细胞的可行性，当与 hMCs 共培养时，这些细胞能够形成聚集物，这允许研究开发自体可植入构建体以克服受 MRONJ（药物相关性颌骨坏死）影响的患者的颌骨缺陷。
