Department of Orthopedics and Traumatology, Okmeydani Training and Research Hospital, Istanbul, Turkey.
Department of Histology & Embryology, Faculty of Medicine, Mugla Sitki Kocman University, Mugla, Turkey.
Biotech Histochem. 2022 Nov;97(8):593-603. doi: 10.1080/10520295.2022.2049876. Epub 2022 Apr 27.
Bone healing deficiencies are challenging for orthopedic practice. The use of stem cells with scaffolds to treat bone tissue losses currently is popular for promoting regeneration of tissue. Programmable cells of monocytic origin (PCMO) may differentiate into three germ layers and may be a promising alternative treatment due to their stem cell-like properties. Parathyroid hormone (PTH) participates in bone metabolism. Intermittent administration of PTH promotes osteogenic activity of mesenchymal stem cdells (MSC). We investigated the osteogenic effects of continuous and intermittent administration of PTH on PCMO. Mononuclear cells were harvested from the peripheral blood of healthy donors. Isolated cells were cultured for six days in a de-differentiation medium. Indirect immunocytochemistry using anti-CD14, anti-CD45 and anti-CD90 primary antibodies, as well as electron microscopy were used to detect PCMO. PCMO then were cultured in an osteogenic differentiation medium supplemented with continuous or intermittent 50 ng/ml PTH. The PTH-free control group (CG), intermittent PTH treated group (IPG) and continuous PTH treated group (CPG) were cultured and assessed for their differentiation into osteogenic lineage cells by indirect immunocytochemistry using anti-collagen I, anti-osteonectin and anti-osteocalcin primary antibodies. Osteoblast-like cells obtained by continuous or intermittent PTH administration exhibited increased levels of collagen I, osteonectin and osteocalcin immunoreactivity. We found that continuous and intermittent PTH administration to PCMO enhanced their differentiation to osteogenic lineage cells and increased osteoblastic activity.
骨愈合缺陷是骨科实践中的挑战。目前，使用干细胞与支架治疗骨组织缺失物，以促进组织再生，这种方法很受欢迎。单核细胞来源的可编程细胞（PCMO）可以分化为三个胚层，由于其干细胞样特性，可能成为一种有前途的替代治疗方法。甲状旁腺激素（PTH）参与骨代谢。PTH 的间歇性给药可促进间充质干细胞（MSC）的成骨活性。我们研究了 PTH 的连续和间歇性给药对 PCMO 的成骨作用。从健康供者的外周血中采集单核细胞。分离的细胞在去分化培养基中培养六天。使用抗 CD14、抗 CD45 和抗 CD90 一抗的间接免疫细胞化学以及电子显微镜来检测 PCMO。然后，将 PCMO 在补充有连续或间歇性 50ng/ml PTH 的成骨分化培养基中培养。PTH 无对照组（CG）、间歇性 PTH 处理组（IPG）和连续 PTH 处理组（CPG）进行培养，并通过间接免疫细胞化学使用抗胶原 I、抗骨粘连蛋白和抗骨钙素一抗评估其向成骨谱系细胞的分化。通过连续或间歇性 PTH 给药获得的成骨样细胞表现出胶原 I、骨粘连蛋白和骨钙素免疫反应性增加。我们发现，连续和间歇性 PTH 给药可增强 PCMO 向成骨谱系细胞的分化，并增加成骨细胞活性。
