Osteoinductive effect β-TCP and vitamin D3 on RUNX2 mRNA expression

Noor Abdulkareem Razouki, B. A. Ghani
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Abstract

Although the autograft is regarded as the gold standard bone graft, β-TCP artificial alloplastic is widely used in bone defect healing and has been accepted in human and animal studies to be resorbed and interchanged by vital bone. The active form of vitamin D3 is calcitriol, on mesenchymal stem cells (MSCs) it has a direct influence on encouraging their osteogenic differentiation which is identified as osteoinductive. The need for osteoinductive ability is a demanding issue regarding medical and dental care. RUNX2 regards an essential transcriptional factor for osteoblast differentiation. Adult male New Zealand rabbits (Twenty-four) with an average weight of (1.5-2 kg) were elected in this study. Four holes were made intra-bone in both tibias of each animal. The experimental groups divide as follows: Group C: Bone defect will be left to heal naturally as control; Group TCP: Bone defect will be occupied with β-TCP. Group vitamin D3: Bone defect will be filled with vitamin D3; Group TCPD: Bone defect will be filled with a combination of β-TCP and vitamin D3. Animals sacrificed at (1 and 3 weeks). Purification of total RNA from formalin fixed paraffin embedded (FFPE), The cDNA of RUNX2 and VDR amplify in RT-PCR and detected with sybr green. One-way Analysis of Variance (ANOVA) and least significant difference (LSD) was used as a tool for statistical analysis. Results at 7 days highest mean value of mRNA RUNX2 gene expression Fold Change estimated in TCP followed by TCPD experimental group while at 21 days, the highest mean value in TCPD. All measured parameters showed highly significant differences in the course of healing periods from 7 days to 21 days. In mRNA VDR gene expression parameters showed a highly significant difference in 7days and 21 days durations among experimental groups and the highest mean value of mRNA VDR gene expression Fold Change is recorded in TCPD followed by vitamin D3 groups in both durations. These findings suggest positive inductive effects of local application of vitamin D3 combined with β-TCP as well as vitamin D3 on osteoblast cells and induced bone defect healing.
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β-TCP和维生素D3对RUNX2 mRNA表达的骨诱导作用
虽然自体骨移植被认为是金标准骨移植,但β-TCP人工同种异体广泛应用于骨缺损愈合,并在人类和动物研究中被重要骨吸收和交换。维生素D3的活性形式是骨化三醇,对间充质干细胞(MSCs)有直接影响,促进其成骨分化,被确定为成骨诱导。在医疗和牙科保健方面,对骨诱导能力的需求是一个要求很高的问题。RUNX2是成骨细胞分化的重要转录因子。本研究选用平均体重(1.5-2 kg)的成年雄性新西兰兔(24只)。在每只动物的两侧胫骨骨内开4个孔。实验组分为:C组:骨缺损自行愈合为对照组;TCP组:骨缺损用β-TCP填充。维生素D3组:补骨缺损用维生素D3;TCPD组:骨缺损用β-TCP和维生素D3组合填充。动物于(1和3周)处死。从福尔马林固定石蜡包埋(FFPE)中纯化总RNA, RT-PCR扩增RUNX2和VDR cDNA,并用sybr绿色进行检测。采用单因素方差分析(ANOVA)和最小显著性差异(LSD)作为统计分析工具。结果TCP组RUNX2基因mRNA表达量均值在第7天最高,TCPD组次之,而TCPD组的均值在第21天最高。在7 ~ 21天的愈合过程中,所有测量参数均显示出高度显著的差异。mRNA VDR基因表达参数在7d和21 d期间各组间差异极显著,且mRNA VDR基因表达的平均值均以TCPD组最高,维生素D3组次之。提示局部应用维生素D3联合β-TCP和维生素D3对成骨细胞有积极的诱导作用,可诱导骨缺损愈合。
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来源期刊
Asia-pacific Journal of Molecular Biology and Biotechnology
Asia-pacific Journal of Molecular Biology and Biotechnology Biochemistry, Genetics and Molecular Biology-Biotechnology
CiteScore
0.90
自引率
0.00%
发文量
25
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