Enhancement of osteogenesis on hydroxyapatite surface coated with synthetic peptide (EEEEEEEPRGDT) in vitro
Enhancement of osteogenesis on hydroxyapatite surface coated with synthetic peptide (EEEEEEEPRGDT) in vitro
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DOI：
10.1002/jbm.10338
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发表时间：
2002-11-01
期刊：
影响因子：
--
通讯作者：
Kasugai, S
中科院分区：
文献类型：
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作者：
Itoh, D;Yoneda, S;Kasugai, S
一些牙种植体涂有羟基磷灰石（HA），它优先与骨骼结合。几种基质蛋白具有精氨酸-甘氨酸-天冬氨酸 (RGD) 序列，细胞通过整联蛋白受体附着在该序列上。我们假设用含有 RGD 的肽涂覆 HA 表面可能会增强成骨细胞的附着和分化。 HA 盘（直径 34 mm，厚度 1 mm）用含有 pepticle EEEEEEEPRGDT 的溶液（50 mM Tris/HCl 和 150 mM NaCl，pH 7.4）处理，其中 E 重复对 HA 具有高亲和力。用磷酸盐缓冲盐水洗涤后，将KUSA/Al小鼠成骨细胞接种到HA表面并培养。 30分钟后，计算附着在表面的细胞数量。培养10天后测量DNA含量和碱性磷酸酶(ALP)活性。 7天时还通过逆转录酶-聚合酶链式反应检测骨基质蛋白的表达； 10天后还通过茜素红S染色来评估培养物的矿化面积。用肽处理可刺激细胞附着并增加 DNA 含量和 ALP 活性。此外，与未处理的表面相比，经肽处理的表面上的基质蛋白表达和矿化结节形成得到更大程度的增强。我们的结果表明，用含有 RGD 的肽涂覆 HA 表面可以增强成骨细胞的附着和分化。 HA 涂层种植体的这种肽处理可以刺激种植体的骨整合。 (C) 2002 年 Wiley 期刊公司
Some dental implants are coated with hydroxyapatite (HA), which preferentially binds to bone. Several matrix proteins have an arginine-glycine-aspartic acid (RGD) sequence where cells attach via an integrin receptor. We hypothesized that coating an HA surface with an RGD-containing peptide might enhance the attachment and differentiation of osteoblasts. The HA disks (diameter 34 mm, thickness 1 mm) were treated with a solution (50 mM Tris/HCl and 150 mM NaCl, pH 7.4) containing the pepticle EEEEEEEPRGDT, in which the E repetition exerts a high affinity to HA. After washing with phosphate-buffered saline, KUSA/Al mouse osteoblastic cells were inoculated onto the HA surface and cultured. After 30 min, the number of cells attached to the surface was counted. The DNA content and alkaline phosphatase (ALP) activity were measured after 10 days in culture. Expression of bone matrix proteins was also examined by means of reverse transcriptase-polymerase chain reaction at 7 days; the mineralized area of the culture was also evaluated by staining with Alizarin Red S after 10 days. Treatment with the peptide stimulated cell attachment and increased DNA content and ALP activity. Furthermore, matrix protein expression and mineralized nodule formation were enhanced to a greater extent on the peptide-treated surface than on the nontreated surface. Our results indicate that coating an HA surface with RGD-containing peptide enhances osteoblast attachment and differentiation. This peptide treatment of HA-coated implants may stimulate the osseointegration of the implants. (C) 2002 Wiley Periodicals, Inc.