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Guided tissue regeneration with immobilized cytokine

用固定化细胞因子引导组织再生

基本信息

批准号：
16591996
负责人：
YAMACHIKA Eiki
金额：
$ 2.24万
依托单位：
Okayama University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2004
资助国家：
日本
起止时间：
2004 至 2005
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-16591996/
关键词：
BMP Cytokine Immobilization bone Regeneration BMP-2 I型アテロコラーゲン細胞活性細胞内シグナル伝達 Smad

项目摘要

Bone morphogenetic protein 2(BMP-2) plays an important role in bone growth and regeneration. The creation of purified recombinant human BMP-2 (rhBMP-2) has led to advances in its potential for clinical use, however this protein is soluble and disperses soon after implantation. So rhBMP-2 can't have long time stimulation to BMP signaling cascade. To overcome this problem we reported the effects of immobilization of rhBMP-2 to succinylated type I atelocollagenThe mechanism of immobilized rhBMP-2 is based on the theories of covalent bonding between rhBMP-2 and succinylated type I atelocollagen. The immobilized rhBMP-2 binds with the receptor on the cell surface without being internalized into the cell, so it is able to transduce signals into the cell over a long period of time, achieving high efficiency in promoting cell proliferation.We investigated the effect of immobilized rhBMP-2 by the way of (a) histological findings, (b) alkaline phosphatase (ALP) activity, (c) BMP intracellular signaling cascade (Smad1, Smad5 and phospho Smad1/5/8) expression, and (d) type I collagen, osteonectin, osteopontin, ostocalcin, and Smad1/5/8 mRNA expression.Our results showed that (1) nonimmobilized and immobilized rhBMP-2 also increased the alkaline phosphatase (ALP) activity in ST2, MC3T3 and C2C12 cells. However significantly higher ALP activity was observed by the immobilized rhBMP-2 treatment. (2) Phosphorylation of Smad1/5/8 after nonimmobilized rhBMP-2 stimulation increased from 15 min after stimulation and decrease to the control revel after 6hrs, however the phosphorylation was prolonged by immobilized rhBMP-2 and its effect was detectable even after 48hrs.So we conclude that because immobilized rhBMP-2 can emphasize and prolonged the effect of rhBMP-2, immobilizing rhBMP-2 is an efficient method to increase bone induction.

骨形态发生蛋白2（BMP-2）在骨生长和再生中起重要作用。纯化重组人BMP-2 （rhBMP-2）的产生在其临床应用潜力方面取得了进展，然而这种蛋白质是可溶性的，在植入后很快就会分散。因此，rhBMP-2不能对BMP信号级联产生长时间的刺激。为了克服这一问题，我们报道了固定化rhBMP-2对琥珀酰化I型间胶原的影响。固定化rhBMP-2的机制是基于rhBMP-2与琥珀酰化I型间胶原之间的共价键理论。固定化后的rhBMP-2与细胞表面的受体结合而不被内化到细胞内，因此能够在较长时间内将信号转导到细胞内，达到促进细胞增殖的高效率。我们通过(a)组织学结果，(b)碱性磷酸酶（ALP）活性，(c) BMP细胞内信号级联（Smad1， Smad5和磷酸化Smad1/5/8）表达，以及(d) I型胶原，骨连接素，骨桥蛋白，骨钙素和Smad1/5/8 mRNA表达来研究固定化rhBMP-2的影响。结果表明：(1)非固定化和固定化rhBMP-2均能提高ST2、MC3T3和C2C12细胞的碱性磷酸酶（ALP）活性。然而，固定化rhBMP-2处理显著提高了ALP活性。(2)非固定化rhBMP-2刺激后Smad1/5/8的磷酸化从刺激后15 min开始升高，6h后降至对照水平，而固定化rhBMP-2则延长了磷酸化时间，即使在48h后仍可检测到其作用。因此，由于固定化rhBMP-2可以强化和延长rhBMP-2的作用，因此固定化rhBMP-2是一种有效的增强骨诱导的方法。