Thomas V L, Sanford B A, Keogh B S, Triplett R G
Department of Microbiology, University of Texas Health Science Center, San Antonio 78284-7758.
Infect Immun. 1989 Feb;57(2):404-12. doi: 10.1128/iai.57.2.404-412.1989.
A rabbit model was used to study the effect of allogeneic demineralized bone powder (DBP) implants on the persistence of Staphylococcus aureus osteomyelitis. Thirty-one rabbits with chronic osteomyelitis of the tibia established by day 21, were started on systemic antibiotics followed by either no additional treatment or debridement plus either DBP (with or without supplemental antibiotics) or supplemental antibiotics only. On day 21, cultures showed a mean of 2 x 10(4) CFU/mg of debrided osseous material. By day 70, the treatment most effective in clearing infection was found in animals treated with supplemental antibiotics only (mean of 142 +/- 116 CFU/mg). In contrast, infection persisted at a 7- to 10-fold-higher level in animals receiving DBP with and without supplemental antibiotics; these results suggest that DBP contributed to persistence of infection. Longitudinal sera were tested again staphylococcal sonic extracts by immunoblot. Detection of numerous probe-positive bands indicated complex but remarkably similar antibody responses among infected animals. Antibodies attached directly to the cell surfaces of staphylococci as shown by immunogold and blocked the binding of organisms to HEp-2 and human fetal lung cells in a radioadherence assay. Antibodies could be absorbed out by intact organisms and were unreactive by immunoblot against antigens derived from cells pretreated with pronase, proteinase K, or lysostaphin. These results indicate that the major response was directed against staphylococcal cell surface proteins. Surprisingly, only one major band (molecular weight, approximately 12,000) was detected when a homologous in vivo antigen preparation was studied by immunoblot. Antibody reactive against this peptide did not appear to react with staphylococci grown in vitro.
采用兔模型研究同种异体脱矿骨粉(DBP)植入物对金黄色葡萄球菌骨髓炎持续存在的影响。在第21天建立了31只胫骨慢性骨髓炎的兔模型,先给予全身抗生素治疗,随后分为不进行额外治疗组、清创加DBP(加或不加补充抗生素)组或仅加补充抗生素组。在第21天,培养显示清创骨材料的平均菌落形成单位(CFU)为2×10⁴CFU/mg。到第70天,发现仅接受补充抗生素治疗的动物清除感染最有效(平均为142±116 CFU/mg)。相比之下,接受DBP加或不加补充抗生素的动物感染持续水平高出7至10倍;这些结果表明DBP导致了感染的持续存在。通过免疫印迹法对纵向血清进行葡萄球菌超声提取物检测。检测到大量探针阳性条带表明感染动物之间的抗体反应复杂但非常相似。免疫金染色显示抗体直接附着在葡萄球菌细胞表面,并在放射黏附试验中阻断了细菌与HEp-2细胞和人胎儿肺细胞的结合。抗体可被完整的细菌吸收,并且通过免疫印迹法对用链霉蛋白酶、蛋白酶K或溶葡萄球菌素预处理的细胞来源的抗原无反应。这些结果表明主要反应针对葡萄球菌细胞表面蛋白。令人惊讶的是,当通过免疫印迹法研究同源体内抗体制剂时,仅检测到一条主要条带(分子量约为12,000)。针对该肽的抗体似乎不与体外培养的葡萄球菌发生反应。