Halloran Daniel R, Heubel Brian, MacMurray Connor, Root Denise, Eskander Mark, McTague Sean P, Pelkey Heather, Nohe Anja
Department of Biological Sciences, University of Delaware, Newark, DE 19716, USA.
Orthopedic Surgery, ChristianaCare Hospital, Wilmington, DE 19801, USA.
J Dev Biol. 2022 Jan 18;10(1):6. doi: 10.3390/jdb10010006.
Proper formation of the skeleton during development is crucial for the mobility of humans and the maintenance of essential organs. The production of bone is regulated by osteoblasts and osteoclasts. An imbalance of these cells can lead to a decrease in bone mineral density, which leads to fractures. While many studies are emerging to understand the role of osteoblasts, less studies are present about the role of osteoclasts. This present study utilized bone marrow cells isolated directly from the bone marrow of femoral heads obtained from osteoarthritic (OA) patients after undergoing hip replacement surgery. Here, we used tartrate resistant acid phosphatase (TRAP) staining, Cathepsin K, and nuclei to identity osteoclasts and their functionality after stimulation with macrophage-colony stimulation factor (M-CSF) and receptor activator of nuclear factor kappa-β ligand (RANKL). Our data demonstrated that isolated cells can be differentiated into functional osteoclasts, as indicated by the 92% and 83% of cells that stained positive for TRAP and Cathepsin K, respectively. Furthermore, isolated cells remain viable and terminally differentiate into osteoclasts when stimulated with RANKL. These data demonstrate that cells isolated from human femoral heads can be differentiated into osteoclasts to study bone disorders during development and adulthood.
发育过程中骨骼的正常形成对于人类的活动能力和重要器官的维持至关重要。骨的生成受成骨细胞和破骨细胞调节。这些细胞的失衡会导致骨矿物质密度降低，进而引发骨折。虽然有许多研究开始了解成骨细胞的作用，但关于破骨细胞作用的研究较少。本研究使用了直接从接受髋关节置换手术后的骨关节炎（OA）患者股骨头骨髓中分离出的骨髓细胞。在这里，我们使用抗酒石酸酸性磷酸酶（TRAP）染色、组织蛋白酶K和细胞核来鉴定破骨细胞及其在用巨噬细胞集落刺激因子（M-CSF）和核因子κ-β受体激活剂配体（RANKL）刺激后的功能。我们的数据表明，分离出的细胞可以分化为功能性破骨细胞，分别有92%和83%的细胞TRAP和组织蛋白酶K染色呈阳性。此外，分离出的细胞在受到RANKL刺激时仍保持活力并终末分化为破骨细胞。这些数据表明，从人类股骨头分离出的细胞可以分化为破骨细胞，以研究发育和成年期的骨疾病。
