Bacterial cellulose membrane (BCM) has been recently recognized as a new generation of carbohydrate-based nanomaterial that possesses a great potential in tissue engineering applications. This research aims to develop an active non-resorbable guided tissue regeneration (GTR) membrane from BCM by conjugating with plant-derived recombinant human osteopontin (p-rhOPN), an economically produced and RGD-containing biomolecule. The BCM was initially grafted with poly(acrylic acid) (PAA) brushes to form poly(acrylic acid)-grafted BCM. Multiple carboxyl groups introduced to the BCM by PAA can serve as active anchoring points for p-rhOPN conjugation and yielded p-rhOPN-BCM. All chemically modified BCMs were characterized by attenuated total reflectance Fourier transform infrared spectroscopy and X-ray photoelectron spectroscopy, while their surface morphology was evaluated by field emission-scanning electron microscopy and atomic force microscopy analyses. The amount of p-rhOPN adhered on the membrane was quantified by enzyme-linked immunosorbent assay. The immunocytochemistry, two-stage quantitative real-time reverse transcriptase polymerase chain reaction and in vitro mineralization analyses strongly suggested that p-rhOPN-BCM could elicit biological functions leading to the enhancement of osteogenic differentiation of human periodontal ligament stem cells as effective as BCM conjugated with commercially available rhOPN from mammalian cells (rhOPN-BCM), suggesting its potential to be used as GTR membrane to promote bone tissue regeneration.

细菌纤维素膜（BCM）最近被公认为是新一代的基于碳水化合物的纳米材料，在组织工程应用中具有巨大的潜力。这项研究的目的是通过与植物来源的重组人骨桥蛋白（p-rhOPN）结合，从BCM开发出一种活性不可吸收的引导组织再生（GTR）膜，该重组人骨桥蛋白是一种经济生产且包含RGD的生物分子。首先将BCM接枝聚丙烯酸（PAA）刷子，以形成聚丙烯酸接枝的BCM。通过PAA引入BCM的多个羧基可用作p-rhOPN共轭的活性锚定点，并产生p-rhOPN-BCM。所有化学修饰的BCM均通过衰减全反射傅里叶变换红外光谱和X射线光电子能谱进行表征，同时通过场发射扫描电子显微镜和原子力显微镜分析评估了它们的表面形态。通过酶联免疫吸附测定来定量粘附在膜上的p-rhOPN的量。免疫细胞化学，两阶段实时定量逆转录酶聚合酶链反应和体外矿化分析强烈表明，p-rhOPN-BCM可以引起生物学功能，从而导致人牙周膜干细胞的成骨分化增强，与结合BCM一样有效。与可从哺乳动物细胞（rhOPN-BCM）购得的rhOPN一起使用，表明其潜力可作为GTR膜来促进骨组织再生。通过酶联免疫吸附测定来定量粘附在膜上的p-rhOPN的量。免疫细胞化学，两阶段实时定量逆转录酶聚合酶链反应和体外矿化分析强烈表明，p-rhOPN-BCM可以引起生物学功能，从而导致人牙周膜干细胞的成骨分化增强，与结合BCM一样有效。与可从哺乳动物细胞（rhOPN-BCM）购得的rhOPN一起使用，表明其潜力可作为GTR膜来促进骨组织再生。通过酶联免疫吸附测定来定量粘附在膜上的p-rhOPN的量。免疫细胞化学，两阶段实时定量逆转录酶聚合酶链反应和体外矿化分析强烈表明，p-rhOPN-BCM可以引起生物学功能，从而导致人牙周膜干细胞的成骨分化增强，与结合BCM一样有效。与可从哺乳动物细胞（rhOPN-BCM）购得的rhOPN一起使用，表明其潜力可作为GTR膜来促进骨组织再生。