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学科主题: 临床医学
题名:
Recombinant human bone morphogenetic protein-2-induced ossification of the ligamentum flavum in rats and the associated global modification of histone H3
作者: 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
发表日期: 2014-09-01
DOI: 10.3171/2014.4.SPINE13319
卷: 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
所属项目编号: 81071505 ; 30801156 ; 200800011041
项目资助者: National Natural Science Foundation of China ; Ministry of Education of China
WOS记录号: WOS:000340688600005
Citation statistics:
内容类型: 期刊论文
URI标识: http://ir.bjmu.edu.cn/handle/400002259/53186
Appears in Collections:北京大学第三临床医学院_骨科_期刊论文

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作者单位: Peking Univ, Hosp 3, Dept Orthoped, Beijing 100191, Peoples R China

Recommended Citation:
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.
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