Quantitative analysis and localization of mRNA transcripts of type I collagen, osteocalcin, MMP 2, MMP 8, and MMP 13 during bone healing in a rat calvarial experimental defect model
Quantitative analysis and localization of mRNA transcripts of type I collagen, osteocalcin, MMP 2, MMP 8, and MMP 13 during bone healing in a rat calvarial experimental defect model
复制标题
DOI：
10.1002/ar.20717
复制
发表时间：
2008-08-01
影响因子：
2
通讯作者：
Sasano, Yasuyuki
中科院分区：
文献类型：
--
作者：
Itagaki, Tomoko;Honma, Takahiro;Sasano, Yasuyuki
本研究检测了大鼠颅骨实验性缺损模型中骨愈合过程中基质金属蛋白酶（MMPs）、I型胶原和骨钙素的表达。使用10周龄雄性Wistar大鼠。在大鼠麻醉下，在顶骨中制作全层标准化环钻缺损。在第1天和第3天以及第1、2、3、5、8、10、12、18和24周，从填充原始骨缺损的组织中提取RNA，并通过实时聚合酶链反应处理以定量分析I型胶原、骨钙素和基质金属蛋白酶（MMPs）2、8和13的表达。或者，通过主动脉灌注固定大鼠，并对切除的颅骨进行原位杂交。I型胶原，骨钙素和MMPs 2和13的表达增加到第2周，此后下降，而MMP 8的表达是最高的第1天。I型胶原和骨钙素的mRNA转录本定位于成骨细胞和骨细胞，其中一些表达MMPs 2，8和13。成骨细胞和骨细胞可能在骨缺损愈合过程中用MMPs重塑细胞外基质中发挥作用。
The study examined the expression of matrix metalloproteinases (MMPs), type I collagen and osteocalcin during bone healing in a rat calvarial experimental defect model. Twelve-week-old male Wistar rats were used. A full-thickness standardized trephine defect was made in the parietal bone, with the rat under anesthesia. RNA was extracted from tissue that filled the original bone defect on days 1 and 3 and in weeks 1, 2, 3, 5, 8, 10, 12, 18, and 24 and processed for quantitative analysis of expression of type I collagen, osteocalcin and matrix metalloproteinases (MMPs) 2, 8, and 13 by using real-time polymerase chain reaction. Alternatively, the rats were fixed by perfusion through the aorta and resected calvaria were processed for in situ hybridization for these molecules. The expression of type I collagen, osteocalcin and MMPs 2 and 13 increased toward week 2 and decreased thereafter, whereas the expression of MMP 8 was the highest on day 1. The mRNA transcripts of type I collagen and osteocalcin were localized in osteoblasts and osteocytes, some of which expressed MMPs 2, 8, and 13. Osteoblasts and osteocytes may play a role in the remodeling of extracellular matrices with MMPs during healing of a defect in bone.