Retention and activity of BMP-2 in hyaluronic acid-based scaffolds in vitro
Retention and activity of BMP-2 in hyaluronic acid-based scaffolds in vitro
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DOI：
10.1002/jbm.10011
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发表时间：
2002-03-05
期刊：
影响因子：
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通讯作者：
Valentini, RF
中科院分区：
文献类型：
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作者：
Kim, HD;Valentini, RF
骨形态发生蛋白-2（BMP-2）在合适的可植入基质中递送，具有通过从宿主组织中的未分化多能干细胞诱导新骨形成来修复局部骨骼缺损的潜力。在这项研究中，我们在体外研究了衍生的透明质酸（Hyaff-11）支架作为重组人BMP-2（rhBMP-2）在骨和软骨修复治疗中的递送载体的潜力。Hyaff-11支架采用相转化/颗粒沥滤法制备，并浸泡加载rhBMP-2。酶联免疫吸附试验和碱性磷酸酶（ALP）试验显示rhBMP-2在体外缓慢、持续释放28 d，累积释放量为初始rhBMP-2的31.82%。当与释放上清液一起孵育时，多能细胞系C3 H10 T1/2（T1/2）的ALP诱导证明rhBMP-2以生物活性形式释放，沿着成骨细胞谱系向下。rhBMP-2在Hyaff-11支架中的保留大于胶原凝胶，胶原凝胶在14天内释放了大部分最初加载的rhBMP-2。还用T1/2细胞接种负载rhBMP-2的Hyaff-11支架，并在第3、7、14和28天评价成骨细胞表型的活力和表达。细胞在整个研究过程中保持活力，并在rhBMP-2组中表达时间和剂量依赖性ALP和骨钙素表达。基于这些观察结果，Hyaff-11支架可能是骨/软骨修复中rhBMP-2的合适递送系统，因为它们能够保留rhBMP-2，以持续的方式向局部环境释放低水平的生物活性rhBMP-2，并刺激多能干细胞的分化。(C)John Wiley & Sons，Inc. J Biomed Mater Res 59：573- 584，2002.
Bone morphogenetic protein-2 (BMP-2) delivered in a suitable implantable matrix has the potential to repair local skeletal defects by inducing new bone formation from undifferentiated plurpotent stem cells resident in host tissue. In this study, we examined in vitro the potential of a derivatized hyaluronic acid (Hyaff-11) scaffold as a delivery vehicle for recombinant human BMP-2 (rhBMP-2) in bone and cartilage repair therapies. Hyaff-11 scaffolds were fabricated using a phase inversion/particulate leaching method and soak-loaded with rhBMP-2. In vitro release kinetics of rhBMP-2, demonstrated using enzyme-linked immunosorbant assay and alkaline phosphatase (ALP) assay revealed a slow, sustained rhBMP-2 release during 28 days, with a cumulative release of 31.82% of the initial rhBMP-2 loaded. rhBMP-2 was released in bioactive form as demonstrated by ALP induction of pluripotent cell line, C3H10T1/2 (T1/2), down the osteoblast lineage when incubated with the release supernatants. rhBMP-2 retention in Hyaff-11 scaffolds was greater than that from collagen gels, which released most of the initially loaded rhBMP-2 by 14 days. rhBMP-2-loaded Hyaff-11 scaffolds were also seeded with T1/2 cells and evaluated at 3, 7, 14, and 28 days for viability and expression of osteoblast phenotype. Cells remained viable throughout the study and expressed a time- and dose-dependent ALP and osteocalcin expression in the rhBMP-2 groups. Based on these observations, Hyaff-11 scaffolds may be suitable delivery systems for rhBMP-2 in bone/cartilage repair because of their ability to retain rhBMP-2, release low levels of bioactive rhBMP-2 to the local environment in a sustained manner, and stimulate differentiation of pluripotent stem cells. (C) 2001 John Wiley & Sons, Inc. J Biomed Mater Res 59: 573-584,2002.