An in vitro model of radiation-induced craniofacial bone growth inhibition
An in vitro model of radiation-induced craniofacial bone growth inhibition
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DOI：
10.1097/scs.0b013e31814c916f
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发表时间：
2007-09-01
影响因子：
0.9
通讯作者：
Forrest, Christopher R.
中科院分区：
文献类型：
--
作者：
Gevorgyan, Artur;La Scala, Giorgio C.;Forrest, Christopher R.
放射诱导的颅面骨生长抑制是儿童头颈癌幸存者接受放射治疗的结果。早先，幼兔眶颧骨复合体(OZC)被建立为一种可靠的动物模型。本研究的目的是建立兔OZC的细胞培养模型，以研究辐射对头面部骨骼的影响。本研究选用7周龄幼年新西兰大白兔。在无菌条件下采集两个OZC的骨膜，通过连续消化的方法引入细胞培养，并在融合培养中传代。分析细胞增殖(甲基四氮唑法)、碱性磷酸酶活性、I型胶原表达和矿化情况。用电子显微镜观察细胞的超微结构。随后，用假照射或15Gy射线照射兔，进行细胞培养并分析细胞数量。从OZC骨膜培养出的细胞具有成骨细胞样表型，在成骨环境中具有高碱性磷酸酶活性、1型胶原表达和矿化。电子显微镜证实了体外成骨的超微结构特征。最后，与对照组动物相比，接受15Gy射线照射的动物获得的细胞数量显著减少(P<；0.01)。从幼兔OZC骨膜中分离培养出具有成骨细胞样细胞表型的原代细胞。该细胞培养系统对体内辐射的反应是细胞数量显著减少。这一体外模型将被用来研究颅面部成骨样细胞的辐射和辐射防护的细胞机制。
Radiation-induced craniofacial bone growth inhibition is a consequence of therapeutic radiation in the survivors of pediatric head and neck cancer. Previously, the infant rabbit orbitozygomatic complex (OZC) was established as a reliable animal model. The purpose of this study was to develop a cell culture model from the rabbit OZC to study the effects of radiation in the craniofacial skeleton. Infant (7-week-old) New Zealand white rabbits were used in this study. Periostea from both OZC were harvested in sterile conditions, introduced into cell culture by way of sequential digestion, and subcultured at confluence. Cultures were analyzed for cellular proliferation (methylthiazoletetrazolium assay), alkaline phosphatase activity, collagen type I expression, and mineralization. Electron microscopy was performed to reveal the in vitro ultrastructure. Subsequently, rabbits were irradiated with sham or 15 Gy radiation, and cell cultures were developed and analyzed for cell numbers. Cell cultures, grown from OZC periostea, expressed osteoblast-like phenotype, with high alkaline phosphatase activity, collagen type 1 expression, and mineralization in an osteogenic environment. Electron microscopy confirmed the characteristic ultrastructural features of osteogenesis in vitro. Finally, significantly (P < 0.01) fewer cells were obtained from animals treated with 15 Gy radiation compared with those from control animals. A primary cell culture with osteoblast-like cellular phenotype was developed from infant rabbit OZC periosteum. This cell culture system responded to in vivo administered radiation by a significant decrease in cell numbers. This in vitro model will be subsequently used to study the cellular mechanisms of radiation and radioprotection in craniofacial osteoblast-like cells.