Bone Research Lab, School of Dentistry of Ribeirão Preto, University of São Paulo, Av do Café s/n, Ribeirão Preto, SP, 14040-904, Brazil.
Department of Maxillofacial Surgery, Prosthesis and Traumatology, School of Dentistry, University of São Paulo, São Paulo, SP, Brazil.
J Bone Miner Metab. 2020 Jul;38(4):481-490. doi: 10.1007/s00774-020-01090-5. Epub 2020 Feb 20.
The aim of this study was to evaluate the in vitro osteogenic potential of osteoblasts from neural crest-derived frontal bone (OB-NC) and mesoderm-derived parietal bone (OB-MS) and the bone formation induced by them when injected into calvarial defects.
Calvarial bones were collected from newborn Wistar rats (3-day old) and characterized as frontal and parietal prior to OB-NC and OB-MS harvesting. The cells were cultured, and several parameters of osteoblast differentiation were evaluated. These cells, or PBS without cells (control), were locally injected into 5-mm rat calvarial defects (5 × 10 cells/defect) and after 4 weeks bone formation was evaluated by morphometric and histological analyses.
The characterization of frontal and parietal bones assured the different embryonic origin of both cell populations, OB-NC and OB-MS. The OB-NC presented higher proliferation while the OB-MS presented higher alkaline phosphatase (ALP) activity, extracellular matrix mineralization and gene expression of runt-related transcription factor 2, Alp, bone sialoprotein and osteocalcin revealing their high osteogenic potential. µCT analysis indicated that there was higher amount of bone formation in defects injected with both OB-NC and OB-MS compared to the control. Moreover, the bone tissue formed by both cells displayed the same histological characteristics.
Despite the distinct in vitro osteogenic potential, OB-NC and OB-MS induced similar bone repair in a rat calvarial defect model. Thus, osteoblasts, irrespective of their in vitro osteogenic potential linked to embryonic origins, seem to be suitable for cell-based therapies aiming to repair bone defects.
本研究旨在评估神经嵴来源的额骨(OB-NC)和中胚层来源的顶骨(OB-MS)成骨细胞的体外成骨潜能,以及将其注射到颅骨缺损部位时诱导的骨形成。
从新生 Wistar 大鼠(3 日龄)颅骨中采集颅骨,并在采集 OB-NC 和 OB-MS 之前将颅骨分为额骨和顶骨。培养细胞,并评估成骨细胞分化的几个参数。将这些细胞或不含细胞的 PBS(对照)局部注射到 5mm 大鼠颅骨缺损(5×10 个细胞/缺损)中,4 周后通过形态计量学和组织学分析评估骨形成。
额骨和顶骨的特征保证了这两种细胞群体 OB-NC 和 OB-MS 的不同胚胎起源。OB-NC 表现出更高的增殖能力,而 OB-MS 则表现出更高的碱性磷酸酶(ALP)活性、细胞外基质矿化以及 runt 相关转录因子 2、Alp、骨涎蛋白和骨钙素的基因表达,表明其具有较高的成骨潜能。µCT 分析表明,与对照组相比,注射 OB-NC 和 OB-MS 的缺陷部位有更多的骨形成。此外,两种细胞形成的骨组织具有相同的组织学特征。
尽管体外成骨潜能不同,但 OB-NC 和 OB-MS 在大鼠颅骨缺损模型中诱导了相似的骨修复。因此,成骨细胞,无论其与胚胎起源相关的体外成骨潜能如何,似乎都适合用于旨在修复骨缺损的细胞治疗。