Immobilization of alkaline phosphatase on microporous nanofibrous fibrin scaffolds for bone tissue engineering.
Immobilization of alkaline phosphatase on microporous nanofibrous fibrin scaffolds for bone tissue engineering.
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DOI：
10.1016/j.biomaterials.2009.05.022
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发表时间：
2009-09
期刊：
影响因子：
14
通讯作者：
Somerman, Martha J.
中科院分区：
文献类型：
--
作者：
Osathanon, Thanaphum;Giachelli, Cecilia M.;Somerman, Martha J.
碱性磷酸酶(ALP)通过降解抑制羟基磷灰石形成的无机焦磷酸盐(PPI)和产生诱导羟基磷灰石形成的无机磷酸盐(PI)来促进骨形成。PI是成骨细胞分化和矿化的关键分子。本研究开发了一种将碱性磷酸酶固定在孔径和孔连接可控的纤维蛋白支架上的方法，并对这些支架的体外和体内生物学特性进行了表征。采用球形模板法制备了微孔纳米纤维蛋白支架(FS)。用1-乙基-3-(二甲氨基丙基)碳二亚胺盐酸盐(EDC)将碱性磷酸酶共价固定在纤维蛋白支架上。扫描电子显微镜观察表明，在添加β-甘油磷酸盐的条件下，固定化碱性磷酸酶纤维蛋白支架(固定化碱性磷酸酶/纤维蛋白支架)上有矿物质沉积，表明固定化碱性磷酸酶具有活性。原代颅骨细胞附着、铺展并在支架表面形成多层。当颅骨细胞接种于固定化的ALP/FS上时，也观察到矿物质沉积。此外，种植在固定化ALP/FS上的细胞显示出比对照FS更高的成骨细胞标志基因表达。植入小鼠颅骨缺损区后，固定化ALP/FS组和单纯FS组缺损区的骨体积均高于空白对照组。此外，与单纯FS组相比，固定化ALP/FS组的骨形成有统计学意义。然而，反响并不强烈。
Alkaline phosphatase (ALP) promotes bone formation by degrading inorganic pyrophosphate (PPi), an inhibitor of hydroxyapatite formation, and generating inorganic phosphate (Pi), an inducer of hydroxyapatite formation. Pi is a crucial molecule in differentiation and mineralization of osteoblasts. In this study, a method to immobilize ALP on fibrin scaffolds with tightly controllable pore size and pore interconnection was developed, and the biological properties of these scaffolds were characterized both in vitro and in vivo. Microporous, nanofibrous fibrin scaffolds (FS) were fabricated using a sphere-templating method. ALP was covalently immobilized on the fibrin scaffolds using 1-ethyl-3-(dimethylaminopropyl)carbodiimide hydrochloride (EDC). Scanning electron microscopic observation (SEM) showed that mineral was deposited on immobilized alkaline phosphatase fibrin scaffolds (immobilized ALP/FS) when incubated in medium supplemented with β-glycerophosphate, suggesting that the immobilized ALP was active. Primary calvarial cells attached, spread and formed multiple layers on the surface of the scaffolds. Mineral deposition was also observed when calvarial cells were seeded on immobilized ALP/FS. Furthermore, cells seeded on immobilized ALP/FS exhibited higher osteoblast marker gene expression compared to control FS. Upon implantation in mouse calvarial defects, both the immobilized ALP/FS and FS alone treated group had higher bone volume in the defect compared to the empty defect control. Furthermore, bone formation in the immobilized ALP/FS treated group was statistically significant compared to FS alone group. However, the response was not robust.
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影响因子：
20.1
作者：
Li, XW;Yang, HY;Giachelli, CM
通讯作者：
Giachelli, CM
碱性磷酸酶在成骨的作用。
影响因子：
15.3
作者：
SIFFERT, R S
通讯作者：
SIFFERT, R S
影响因子：
14
作者：
Osathanon, Thanaphum;Linnes, Michael L.;Rajachar, Rupak M.;Ratner, Buddy D.;Somerman, Martha J.;Giachelli, Cecilia M.
通讯作者：
Giachelli, Cecilia M.
影响因子：
7.6
作者：
Hacchou, Y.;Uematsu, T.;Furusawa, K.
通讯作者：
Furusawa, K.
影响因子：
4.8
作者：
Addison, William N.;Azari, Fereshteh;McKee, Marc D.
通讯作者：
McKee, Marc D.