Ghosh Shreya, Wu Victoria, Pernal Sebastian, Uskoković Vuk
Advanced Materials and Nanobiotechnology Laboratory, Department of Bioengineering, University of Illinois , Chicago, Illinois 60607, United States.
ACS Appl Mater Interfaces. 2016 Mar;8(12):7691-708. doi: 10.1021/acsami.6b01160. Epub 2016 Mar 17.
Osteomyelitis, an infectious disease predominantly tied to poor sanitary conditions in underdeveloped regions of the world, is in need of inexpensive, easily in situ synthesizable and administrable materials for its treatment. The results of this study stem from the attempt to create one such affordable and minimally invasive therapeutic platform in the form of a self-setting, injectable cement with a tunable drug release profile, composed of only nanoparticulate hydroxyapatite, the synthetic version of the bone mineral. Cements comprised two separately synthesized hydroxyapatite powders, one of which, HAP2, was precipitated abruptly, retaining the amorphous nature longer, and the other one of which, HAP1, was precipitated at a slower rate, more rapidly transitioning to the crystalline structure. Cements were made with four different weight ratios of the two hydroxyapatite components: 100/0, 85/15, 50/50, and 0/100 with respect to HAP1 and HAP2. Both the setting and the release rates measured on two different antibiotics, vancomycin and ciprofloxacin, were controlled using the weight ratio of the two hydroxyapatite components. Various inorganic powder properties were formerly used to control drug release, but here we demonstrate for the first time that the kinetics of the mechanism of formation of a solid compound can be controlled to produce tunable drug release profiles. Specifically, it was found that the longer the precursor calcium phosphate component of the cement retains the amorphous nature of the primary precipitate, the more active it was in terms of speeding up the diffusional release of the adsorbed drug. The setting rate was, in contrast, inversely proportional to the release rate and to the content of this active hydroxyapatite component, HAP2. The empirical release profiles were fitted to a set of equations that could be used to tune the release rate to the therapeutic occasion. All of the cements loaded with vancomycin or ciprofloxacin inhibited the growth of Gram-positive Staphylococcus aureus and Gram-negative Escherichia coli and Pseudomonas aeruginosa in both agar diffusion assays and broth dilution tests with intensities either comparable to the antibiotic per se, as in the case of ciprofloxacin, or even larger than the antibiotic alone, as in the case of vancomycin. Interestingly, even the pure cements exhibited an antibacterial effect ranging from moderate to strong, while demonstrating high levels of biocompatibility with osteoclastic RAW264.7 cells and only slightly affecting the viability of the osteoblastic MC3T3-E1 cells, in direct proportion with the amount of the more active hydroxyapatite component in the cements. This antibacterial effect was especially noticeable against Gram-negative bacteria, where the growth inhibition by the cements was comparable to or even stronger than that of the pure antibiotics. The antibiofilm assay against P. aeruginosa biofilms reiterated the antibiotic effectiveness of pure, antibiotic-free cements. That the carrier per se, composed of a nontoxic, easily prepared, bone mineral composite, can exhibit a strong antibacterial effect even in the absence of an antibiotic drug is an insight highly relevant in view of the rising resistance of an array of pathogens to traditional antibiotic therapies and the demands for the timely development of suitable alternatives.
骨髓炎是一种主要与世界欠发达地区卫生条件差相关的传染病,需要用于治疗的廉价、易于原位合成和给药的材料。本研究的结果源于试图创建一个这样的经济实惠且微创的治疗平台,其形式为具有可调药物释放曲线的自固化、可注射水泥,该水泥仅由纳米颗粒羟基磷灰石(骨矿物质的合成形式)组成。水泥由两种单独合成的羟基磷灰石粉末组成,其中一种,即HAP2,突然沉淀,较长时间保持无定形性质,另一种,即HAP1,以较慢速率沉淀,更快地转变为晶体结构。水泥由两种羟基磷灰石成分的四种不同重量比制成:相对于HAP1和HAP2为100/0、85/15、50/50和0/100。在两种不同抗生素万古霉素和环丙沙星上测量的凝固和释放速率,通过两种羟基磷灰石成分的重量比来控制。以前曾使用各种无机粉末特性来控制药物释放,但在此我们首次证明,可以控制固体化合物形成机制的动力学以产生可调的药物释放曲线。具体而言,发现水泥的前体磷酸钙成分保持初级沉淀物无定形性质的时间越长,其在加速吸附药物的扩散释放方面就越活跃。相比之下,凝固速率与释放速率以及这种活性羟基磷灰石成分HAP2的含量成反比。经验释放曲线拟合到一组方程,这些方程可用于根据治疗情况调整释放速率。所有负载万古霉素或环丙沙星的水泥在琼脂扩散试验和肉汤稀释试验中均抑制革兰氏阳性金黄色葡萄球菌以及革兰氏阴性大肠杆菌和铜绿假单胞菌的生长,其强度要么与抗生素本身相当,如环丙沙星的情况,要么甚至大于单独使用抗生素,如万古霉素的情况。有趣的是,即使是纯水泥也表现出从中度到强的抗菌作用,同时与破骨细胞RAW264.7细胞具有高度生物相容性,并且仅轻微影响成骨细胞MC3T3 - E1细胞的活力,这与水泥中活性更高的羟基磷灰石成分的量成正比。这种抗菌作用对革兰氏阴性细菌尤其明显,其中水泥的生长抑制作用与纯抗生素相当甚至更强。针对铜绿假单胞菌生物膜的抗生物膜试验重申了纯的、不含抗生素的水泥的抗生素有效性。鉴于一系列病原体对传统抗生素疗法的耐药性不断上升以及及时开发合适替代方案的需求,由无毒、易于制备的骨矿物质复合材料组成的载体本身即使在没有抗生素药物的情况下也能表现出强大的抗菌作用,这一见解具有高度相关性。