ABSTRACT
No consensus has yet been reached to associate oral bacteria conclusively with the etio-pathogenesis of bisphosphonate-induced osteonecrosis of the jaw (BONJ). Therefore, the present study examined the effects of oral bacteria on the development of BONJ-like lesions in a mouse model. In the pamidronate (Pam)-treated mice, but not control non-drug-treated mice, tooth extraction followed by oral infection with Fusobacterium nucleatum caused BONJ-like lesions and delayed epithelial healing, both of which were completely suppressed by a broad-spectrum antibiotic cocktail. Furthermore, in both in vitro and in vivo experiments, the combination of Pam and Fusobacterium nucleatum caused the death of gingival fibroblasts (GFs) and down-regulated their production of keratinocyte growth factor (KGF), which induces epithelial cell growth and migration. Therefore, in periodontal tissues pre-exposed to bisphosphonate, bacterial infection at tooth extraction sites caused diminished KGF expression in GFs, leading to a delay in the epithelial wound-healing process that was mitigated by antibiotics.
Subject(s)
Fusobacterium nucleatum/pathogenicity , Jaw Diseases/microbiology , Osteonecrosis/microbiology , Surgical Wound Infection/microbiology , Animals , Anti-Bacterial Agents/therapeutic use , Apoptosis , Bone Density Conservation Agents/adverse effects , Cell Movement , Cell Survival , Cells, Cultured , Diphosphonates/adverse effects , Disease Models, Animal , Epithelial Cells/metabolism , Epithelial Cells/microbiology , Female , Fibroblast Growth Factor 7/biosynthesis , Fibroblasts/metabolism , Fibroblasts/microbiology , Gingiva/cytology , Gingiva/microbiology , Jaw Diseases/chemically induced , Mice , Osteonecrosis/chemically induced , Pamidronate , Surgical Wound Infection/drug therapy , Tooth Extraction/adverse effectsABSTRACT
Cyanine 5.18 (or Cy5) is a fluorochrome emitting in the long-red/far-red range, usually regarded as unsuitable for direct observation by the human eye. We describe here the optimization of a direct visualization approach to Cy5 labeling, based on a standard fluorescence microscope with mercury light excitation and applicable to both immunocytochemistry and fluorescent in situ hybridization. Crucial factors were (a) an excitation path in the microscope not absorbing light in the orange-red range, up to 640 nm, (b) a 588-640-nm excitation filter range, distinctly below the excitation optimum for Cy5, (c) a 650-700-nm emission filter range, transmitting the low-wavelength portion of Cy5 emission, and (d) high-efficiency filter set components allowing a narrow gap between excitation and emission ranges without visible cross-talk of excitation light in the emission path.
Subject(s)
Carbocyanines , Fluorescent Dyes , Immunohistochemistry/instrumentation , In Situ Hybridization/instrumentation , Animals , Cattle , Ganglia, Spinal/chemistry , Microscopy, Fluorescence , Optics and Photonics , Pituitary Gland/chemistry , SwineABSTRACT
We describe fluorescence immunostaining of four different antigens in the same section. The fluorochrome conjugates used show highest emission in the blue, green, yellow, and red regions of the visible light spectrum, respectively. Specially designed single fluorochrome filter combinations allow selective visualization of each fluorochrome label in turn, without visible crosstalk with the others.
