Evaluation of Gentamicin Release of PMMA Cements Using Different Methods: HPLC, Elution and Inhibition Zone Testing

Clemens Kittinger, Johannes Stadler, Klaus Dieter Kühn
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Abstract

(1) Background: There is an ongoing discussion on the elution efficacy of antibiotic-impregnated cements. Our experiments were intended to clarify if there are differences in the antibiotic elution of HPLC compared with inhibition zone testing using eluates or PMMA discs. (2) Materials and Methods: Two cement brands with different concentrations of the active ingredient were tested in antimicrobial Kirby–Bauer (disc diffusion) assays. Cement platelets were directly applied on the agar plates and their zone of inhibition was measured. In parallel, the platelets were incubated in phosphate buffered saline (PBS) and at distinct points of time transferred into new buffer. At these time points, 50 µL of the bone cement eluates was used for zone of inhibition testing. Standard gentamicin sulfate solutions served as a control in the same test setup. To verify the microbiological investigations, the antibiotic content of the eluates was also measured via high-performance liquid chromatography (HPLC). (3) Results: The experiments with cement eluates showed better differentiable results than the direct application of the cement discs. The results were also comparable to investigations with HPLC and gentamicin sulfate standard solutions. (4) Conclusions: The results of elution rates are influenced by the test system and the period of observation chosen. The microbial test systems reflect the results of HPLC to the same degree and give evidence of the efficacy of the antibiotics. The HPLC tests on eluates were more suitable in representing differences in release characteristics.
采用不同方法评估 PMMA 水泥的庆大霉素释放量:高效液相色谱法、洗脱法和抑制区测试法
(1) 背景:关于抗生素浸渍水门汀的洗脱效果的讨论一直在进行。我们的实验旨在明确,与使用洗脱液或 PMMA 盘进行抑制区测试相比,高效液相色谱法的抗生素洗脱是否存在差异。(2)材料与方法:在抗菌柯比-鲍尔(圆盘扩散)试验中测试了两种水泥品牌的不同浓度的活性成分。将水泥平板直接涂抹在琼脂平板上,测量其抑制区。同时,在磷酸盐缓冲盐水(PBS)中培养血小板,并在不同的时间点将其转移到新的缓冲液中。在这些时间点,使用 50 µL 的骨水泥洗脱液进行抑制区测试。标准硫酸庆大霉素溶液在相同的测试装置中作为对照。为了验证微生物学调查,还通过高效液相色谱法(HPLC)测量了洗脱液中的抗生素含量。(3) 结果:与直接使用水泥圆片相比,使用水泥洗脱液进行的实验显示出更好的差异化结果。其结果也可与使用高效液相色谱法和硫酸庆大霉素标准溶液进行的研究相媲美。(4) 结论:洗脱率的结果受测试系统和所选观察期的影响。微生物测试系统对高效液相色谱法结果的反映程度相同,并能证明抗生素的疗效。对洗脱液进行的高效液相色谱测试更适合反映释放特性的差异。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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