Fujioka-Kobayashi Masako, Schaller Benoit, Saulacic Nikola, Zhang Yufeng, Miron Richard J
Department of Periodontology, College of Dental Medicine, Nova Southeastern University, Fort Lauderdale, Florida.
Department of Cranio-Maxillofacial Surgery, Bern University Hospital, Inselspital, Switzerland.
J Biomed Mater Res A. 2017 Feb;105(2):408-418. doi: 10.1002/jbm.a.35921. Epub 2016 Oct 18.
Within the past years, BMP9 has been characterized as one of the most osteogenic bone-inducers among the BMP family, however up until recently, BMP9 has only been available through adenovirus transfection experiments (gene therapy) not approved for clinical use. The aim of this study was to investigate recombinant rhBMP9 versus rhBMP2 at 2 concentrations (10 and 100 ng/mL) in combination with 2 bone grafts: (1) a natural bone mineral (NBM) without collagen versus (2) a novel NBM integrated with atelo-collagen type I (NBM-Col). Scanning electron microscopy revealed that while NBM demonstrated a mineralized roughened surface morphology, NBM-Col particles contained many more visible collagen fibrils throughout the scaffold surface significantly increasing rhBMP adsorption from 8 h to 10 days (as quantified by ELISA). Thereafter, ST2 preosteoblasts were used to investigate cell attachment, proliferation, and differentiation. While little change was observed for cell attachment/proliferation, osteoblast differentiation demonstrated a significant increase in alkaline phosphatase (ALP) activity when scaffolds were loaded with rhBMP9 when compared to rhBMP2. Furthermore, a 2-3 fold increase in alizarin red staining, and in mRNA levels of osteoblast differentiation markers Runx2, Collagen1α2, ALP, and osteocalcin was observed when rhBMP9 was combined with NBM-Col when compared to NBM without collagen at equivalent doses and when compared to rhBMP2. The results from this study demonstrate that (1) the use of rhBMP9 significantly and markedly induced osteoblast differentiation when compared to rhBMP2 and (2) the incorporation of atelo-collagen type I into NBM bone grafts markedly improved these findings by serving as a scaffold capable of improving growth factor adsorption and osteoblast behavior. © 2016 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 105A: 408-418, 2017.
在过去几年中，骨形态发生蛋白9（BMP9）已被确定为BMP家族中最具成骨诱导性的因子之一。然而直到最近，BMP9仅通过腺病毒转染实验（基因治疗）获得，而该方法尚未获批用于临床。本研究的目的是在两种浓度（10和100 ng/mL）下，将重组人BMP9（rhBMP9）与rhBMP2，分别与两种骨移植材料联合使用进行对比研究：（1）不含胶原蛋白的天然骨矿物质（NBM），以及（2）整合了I型去端胶原蛋白的新型NBM（NBM-Col）。扫描电子显微镜显示，NBM呈现出矿化粗糙的表面形态，而NBM-Col颗粒在整个支架表面含有更多可见的胶原纤维，从8小时到10天显著增加了rhBMP的吸附（通过酶联免疫吸附测定法量化）。此后，使用ST2前成骨细胞研究细胞黏附、增殖和分化。虽然细胞黏附/增殖变化不大，但与rhBMP2相比，当支架负载rhBMP9时，成骨细胞分化显示碱性磷酸酶（ALP）活性显著增加。此外，与等量剂量下不含胶原蛋白的NBM以及与rhBMP2相比，当rhBMP9与NBM-Col联合使用时，茜素红染色以及成骨细胞分化标志物Runx2、胶原蛋白1α2、ALP和骨钙素的mRNA水平增加了2 - 3倍。本研究结果表明：（1）与rhBMP2相比，rhBMP9的使用显著且明显地诱导了成骨细胞分化；（2）将I型去端胶原蛋白整合到NBM骨移植材料中，通过作为一种能够改善生长因子吸附和成骨细胞行为的支架，显著改善了这些结果。© 2016威利期刊公司。《生物医学材料研究杂志》A部分：105A：408 - 418，2017年。
