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结合重组人骨形态发生蛋白(rhBMP)-9的可注射双相磷酸钙(BCP)载体系统的体外评估

In vitro evaluation of an injectable biphasic calcium phosphate (BCP) carrier system combined with recombinant human bone morphogenetic protein (rhBMP)-9.

作者信息

Fujioka-Kobayashi Masako, Schaller Benoit, Zhang Yufeng, Pippenger Benjamin E, Miron Richard J

机构信息

Department of Cranio-Maxillofacial Surgery, Inselspital, Bern University Hospital, University of Bern, Bern, Switzerland.

Department of Oral Surgery, Institute of Biomedical Sciences, Tokushima University Graduate School, Tokushima, Japan.

出版信息

Biomed Mater Eng. 2017;28(3):293-304. doi: 10.3233/BME-171675.

DOI:10.3233/BME-171675
PMID:28527192
Abstract

Bone morphogenetic protein 9 (BMP9) has previously been characterized as the strongest osteoinductive growth factor among the BMP family. The aim of the present study was to evaluate the possibility of combining rhBMP9 with an injectable biphasic calcium phosphate (I-BCP, maxresorb inject®), since I-BCP is an easy to handle biomaterial with ideal properties for bone augmentation procedures. The adsorption potential of rhBMP9 as well as the cell behavior of bone stromal ST2 cells were investigated on cell viability, adhesion, proliferation and osteogenic differentiation for I-BCP combined with/without rhBMP9 in vitro. I-BCP demonstrated excellent adsorption/retention potential of rhBMP9 with a slow and steady release over a 10 day period by ELISA. Cell attachment at 8 hours and cell proliferation at 1, 3 and 5 days was decreased on I-BCP with/without rhBMP9 when compared to control tissue-culture plastic. While I-BCP had little influence on osteoblast differentiation, its combination with rhBMP9 significantly increased ALP activity at 7 days and mRNA levels of osteoblast differentiation markers including ALP and osteocalcin at 14 days. I-BCP served as an excellent carrier for rhBMP9 clearly demonstrating its osteoinductive potential. We therefore confirm the great potential of rhBMP9 to serve as a future regenerative growth factor for bone applications.

摘要

骨形态发生蛋白9(BMP9)先前已被表征为骨形态发生蛋白家族中最强的骨诱导生长因子。本研究的目的是评估重组人骨形态发生蛋白9(rhBMP9)与可注射双相磷酸钙(I-BCP,maxresorb inject®)联合使用的可能性,因为I-BCP是一种易于处理的生物材料,具有用于骨增量手术的理想特性。在体外研究了rhBMP9与I-BCP联合/不联合rhBMP9时,rhBMP9的吸附潜力以及骨基质ST2细胞在细胞活力、黏附、增殖和成骨分化方面的细胞行为。通过酶联免疫吸附测定法(ELISA),I-BCP在10天内表现出对rhBMP9优异的吸附/保留潜力,并能缓慢稳定释放。与对照组织培养塑料相比,在有/无rhBMP9的I-BCP上,8小时时的细胞附着以及1、3和5天时的细胞增殖均有所下降。虽然I-BCP对成骨细胞分化影响不大,但其与rhBMP9联合使用时,在7天时显著增加了碱性磷酸酶(ALP)活性,在14天时显著增加了包括ALP和骨钙素在内的成骨细胞分化标志物的mRNA水平。I-BCP是rhBMP9的优良载体,清楚地证明了其骨诱导潜力。因此,我们证实了rhBMP9作为未来骨应用再生生长因子的巨大潜力。

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