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Recombinant human bone morphogenetic protein-2-induced ossification of the ligamentum flavum in rats and the associated global modification of histone H3
Hou, Xiao-Fei; Fan, Dong-Wei; Sun, Chui-Guo; Chen, Zhong-Qiang
关键词Ossification Of The Ligamentum Fiavum Rhbmp-2 Histone Modifications Osteogenic Transcription Factors Immunohistochemistry Lumbar Spine Degenerative Disease
刊名JOURNAL OF NEUROSURGERY-SPINE
2014-09-01
DOI10.3171/2014.4.SPINE13319
21期:3页:334-341
收录类别SCI
文章类型Article
WOS标题词Science & Technology
类目[WOS]Clinical Neurology ; Surgery
研究领域[WOS]Neurosciences & Neurology ; Surgery
关键词[WOS]STEM-CELL DIFFERENTIATION ; CHRONIC CORD COMPRESSION ; OSTEOGENIC DIFFERENTIATION ; EPIGENETIC REGULATION ; PROTEIN ; TRANSCRIPTION ; INHIBITION ; ACTIVATION ; SIGNALS ; LESION
英文摘要

Object. The primary object of this investigation was to study recombinant human bone morphogenetic protein-2 (rhBMP-2)-induced ossification of the ligamentum flavum and associated histone H3 modification in a rat model. In an additional set of studies the authors investigated spinal cord and behavioral changes in the same model.

Methods. The authors report on 2 separate sets of studies. A total of 90 rats were used for the 2 sets of studies (45 each); in each study, a lyophilized rhBMP-2 and collagen mixture (20 mu g rhBMP-2 and 200 mu l collagen) was implanted in the lumbar extradural space in 18 rats; another 18 animals were used for a sham-operation control group and underwent implantation of lyophilized collagen without rhBMP-2 at the same level; an additional 9 animals were used as untreated controls. Lumbar spinal samples were harvested from the rhBMP-2 groups and the sham-operation control groups at 1 week, 3 weeks, and 9 weeks after the operation. Samples were also obtained from untreated controls at the game time points. All samples were scanned using micro-CT and then made into paraffin-embedded sections. The sections from the first set of 45 rats were stained using elastica van Gieson and toluidine blue, and the expression of histone modifications (H3K9ac, H3K18ac, H3K4me3, and H3K36me3) and osteogenic transcription factors (osterix, Runx2) was detected by immunohistochemistry. In the second set of studies, hindlimb motor function was assessed at 1 week, 3 weeks, and 9 weeks after surgery. After behavioral evaluation, samples were harvested, scanned using micro-CT, and then made into paraffin-embedded sections. The sections were stained using Luxol fast blue. The expression of NeuN was also detected using immunohistochemistry.

Results. Ossification was seen in the rhBMP-2 group from 1 week after insertion, and the volume of ossified mass increased at land 9 weeks. There was no ossification seen in the sham-surgery and normal controls. The pathological changes of ossification involved ligament degeneration, cartilage formation, and, finally, bone replacement. Spinal cord evaluation showed a significant decrease in white matter content and number of neurons at 9 weeks after operation in the rhBMP-2 treated group (compared with findings in the sham-surgery and control groups as well as findings at the earlier time points in the rhBMP-2 group).

Using immunohistochemical staining, histone modifications (H3K9ac, H3K18ac, H3K4me3, and H3K36me3) and osteogenic transcription factors (osterix, Runx2) all were found to be expressed in the fibrocartilage area of the rat ossified ligamentum flavum samples (rhBMP2 group).

Conclusions. This rhBMP-2 induced OLF is a typical endochondral ossification, which is similar to clinical OLF. The compressed spinal cord around the ossification site showed signs of a chronic degenerative process. Histone H3 modifications (H3K9ac, H3K18ac, H3K4me3, and H3K36me3) may play an important role in OLF.

语种英语
WOS记录号WOS:000340688600005
项目编号81071505 ; 30801156 ; 200800011041
资助机构National Natural Science Foundation of China ; Ministry of Education of China
引用统计
被引频次:4[WOS]   [WOS记录]     [WOS相关记录]
文献类型期刊论文
条目标识符http://ir.bjmu.edu.cn/handle/400002259/53186
专题北京大学第三临床医学院_骨科
作者单位Peking Univ, Hosp 3, Dept Orthoped, Beijing 100191, Peoples R China
推荐引用方式
GB/T 7714
Hou, Xiao-Fei,Fan, Dong-Wei,Sun, Chui-Guo,et al. Recombinant human bone morphogenetic protein-2-induced ossification of the ligamentum flavum in rats and the associated global modification of histone H3[J]. JOURNAL OF NEUROSURGERY-SPINE,2014,21(3):334-341.
APA Hou, Xiao-Fei,Fan, Dong-Wei,Sun, Chui-Guo,&Chen, Zhong-Qiang.(2014).Recombinant human bone morphogenetic protein-2-induced ossification of the ligamentum flavum in rats and the associated global modification of histone H3.JOURNAL OF NEUROSURGERY-SPINE,21(3),334-341.
MLA Hou, Xiao-Fei,et al."Recombinant human bone morphogenetic protein-2-induced ossification of the ligamentum flavum in rats and the associated global modification of histone H3".JOURNAL OF NEUROSURGERY-SPINE 21.3(2014):334-341.
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