Is Osteogenic Differentiation of Human Nucleus Pulposus Cells a Possibility for Biological Spinal Fusion?
Is Osteogenic Differentiation of Human Nucleus Pulposus Cells a Possibility for Biological Spinal Fusion?
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DOI：
10.1177/1947603518754628
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发表时间：
2020-04-01
期刊：
影响因子：
2.8
通讯作者：
Roberts, Sally
中科院分区：
文献类型：
--
作者：
Brown, Sharon J.;Turner, Sarah A.;Roberts, Sally
本研究的目的是研究是否可以开发一种简单的、生物学上稳定的方法来诱导退变椎间盘（IVD）的钙化，从而为需要脊柱融合的患者提供替代治疗。设计将从14个人IVD分离的髓核（NP）细胞单层培养，并暴露于成骨培养基，1，25-二羟基维生素D-3（VitD（3）），甲状旁腺激素（PTH）和骨形态发生蛋白（BMP）2/7，以确定它们是否可以成为成骨细胞。同样，将11名患者的体外培养物在成骨培养基中培养，并预先暴露于或未暴露于VitD（3）和BMP-2。成骨分化通过碱性磷酸酶活性评估，钙化区域通过茜素红或von Kossa染色鉴定。在单层培养过程中，成骨基因的表达采用聚合酶链反应测定，并对BMP抑制剂的外植体组织进行评估。人骨髓间充质基质细胞（MSCs）用于comparation.ResultsStandard成骨培养基是最佳的促进矿化的人NP细胞在单层。在10 nM VitD（3）下观察到一些成骨分化，但在应用PTH或BMP后没有观察到。在11个IVD组织块中，有2个检测到钙化区域，一个在成骨培养基中培养，另一个在添加VitD（3）和BMP-2的培养基中培养。结论人NP细胞可以单层成骨，细胞外基质也可以发生钙化，但并不一致。细胞或细胞外基质中的抑制因子可能会阻碍骨生成，这表明此时稳健的生物融合需要进一步优化。
ObjectiveThe purpose of this study was to investigate whether a simple, biologically robust method for inducing calcification of degenerate intervertebral discs (IVD) could be developed to provide an alternative treatment for patients requiring spinal fusion.DesignNucleus pulposus (NP) cells isolated from 14 human IVDs were cultured in monolayer and exposed to osteogenic medium, 1,25-dihydroxyvitamin D-3 (VitD(3)), parathyroid hormone (PTH), and bone morphogenic proteins (BMPs) 2/7 to determine if they could become osteogenic. Similarly explant cultures of IVDs from 11 patients were cultured in osteogenic media with and without prior exposure to VitD(3) and BMP-2. Osteogenic differentiation was assessed by alkaline phosphatase activity and areas of calcification identified by alizarin red or von Kossa staining. Expression of osteogenic genes during monolayer culture was determined using polymerase chain reaction and explant tissues assessed for BMP inhibitors. Human bone marrow-derived mesenchymal stromal cells (MSCs) were used for comparison.ResultsStandard osteogenic media was optimum for promoting mineralization by human NP cells in monolayer. Some osteogenic differentiation was observed with 10 nM VitD(3), but none following application of PTH or BMPs. Regions of calcification were detected in 2 of the eleven IVD tissue explants, one cultured in osteogenic media and one with the addition of VitD(3) and BMP-2.ConclusionsHuman NP cells can become osteogenic in monolayer and calcification of the extracellular matrix can also occur, although not consistently. Inhibitory factors within either the cells or the extracellular matrix may hinder osteogenesis, indicating that a robust biological fusion at this time requires further optimization.