Microarray gene expression during early healing of GBR-treated calvarial critical size defects
Microarray gene expression during early healing of GBR-treated calvarial critical size defects
复制标题
DOI：
10.1111/clr.12949
复制
发表时间：
2017-10-01
影响因子：
4.3
通讯作者：
Donos, N.
中科院分区：
文献类型：
--
作者：
Al-Kattan, R.;Retzepi, M.;Donos, N.
目的探讨引导骨再生（guided bone regeneration， GBR）后骨愈合过程调控的基因表达和分子途径。材料与方法选用6只6月龄Wistar雄性大鼠。根据GBR原理，在每只动物的顶骨上制造标准化的5毫米临界尺寸缺陷，并使用颅外和颅内ePTFE膜进行治疗。在7天和15天后，随机处死3只动物。从每个样品中提取总RNA，准备用于基因表达分析。评估RNA的质量和数量，然后将cRNA与Affymetrix GeneChip大鼠基因组230 2.0阵列杂交。对Affymetrix数据进行处理，进行一阶分析、质量控制和统计分析。通过pathway和Gene Ontology （GO）分析进行生物学解释。结果在7天和15天的样本中，538个基因受到不同的调控。第7天，炎症和免疫反应明显上调。此外，与血管生成和细胞周期调节相关的氧化石墨烯术语也过表达。在第15天，细胞活性和细胞代谢明显增加。骨形成过程明显过表达，一些编码生长因子、酶活性和细胞外基质形成的基因被发现上调。值得注意的是，15d时观察到Wnt信号通路的负调控。参与骨形成的细胞的基因表达谱随愈合阶段的不同而变化。一些候选基因似乎在膜内骨再生的早期阶段差异表达被提出。
ObjectivesTo investigate the gene expression and molecular pathways implicated in the regulation of the osseous healing process following guided bone regeneration (GBR).Material and methodsSix 6-month-old Wistar male rats were used. Standardized 5-mm critical size defects were created in the parietal bones of each animal and treated with an extracranial and intracranial ePTFE membrane, according to the GBR principle. Three animals were randomly sacrificed after 7 and 15days of healing. Total RNA was extracted from each sample and prepared for gene expression analysis. RNA quality and quantity were assessed, followed by hybridization of the cRNA to Affymetrix GeneChip Rat Genome 230 2.0 Arrays. The Affymetrix data were processed, and first-order analysis, quality control and statistical analysis were performed. Biological interpretation was performed via pathway and Gene Ontology (GO) analysis.ResultsBetween the 7- and 15-day samples, 538 genes were differently regulated. At day 7, inflammatory and immune responses were clearly upregulated. In addition, GO terms related to angiogenesis and cell cycle regulation were overexpressed. At day 15, a more complex cellular activity and cell metabolism were evident. The bone formation processes were significantly overexpressed, with several genes encoding growth factors, enzyme activity, and extracellular matrix formation found as upregulated. Remarkably, a negative regulation of Wnt signalling pathway was observed at 15days.DiscussionThe gene expression profile of the cells participating in osseous formation varied depending on the healing stage. A number of candidate genes that seem differentially expressed during early stages of intramembranous bone regeneration was suggested.