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Utilization of a Continuous Streptococcal Surface to Measure Interbacterial Adherence in vitro and in vivo

Microbiology Program, School of Dentistry, University of Minnesota, Minneapolis, Minnesota 55455, Department of Microbiology, Medical School, University of Minnesota

C.G. Bloomquist

Microbiology Program, School of Dentistry, University of Minnesota, Minneapolis, Minnesota 55455

M.C. Coulter

Microbiology Program, School of Dentistry, University of Minnesota, Minneapolis, Minnesota 55455

L.J. Fenner

Microbiology Program, School of Dentistry, University of Minnesota, Minneapolis, Minnesota 55455

R.J. Skopek

Microbiology Program, School of Dentistry, University of Minnesota, Minneapolis, Minnesota 55455

C.F. Schachtele

Microbiology Program, School of Dentistry, University of Minnesota, Minneapolis, Minnesota 55455, Department of Microbiology, Medical School, University of Minnesota

Cell-to-cell interactions are essential for the formation of dental plaque. A continuous layer of Streptococcus sanguis SA-1 cells fixed to a solid surface has been used to evaluate interactions among this bacterium, Haemophilus parainfluenzae, and Streptococcus sobrinus. S. sanguis cells were attached to a Falcon 3001 tissue culture plates or bovine enamel chips, coated with a biological adhesive. Scanning electron microscopy of the chips showed the streptococci as a contiguous surface. Radiolabeled bacteria were used to measure a second-species interbacterial adherence to the streptococcal-coated culture plates. Strains of H. parainfluenzae known to coaggregate (strain HP-28) and not to coaggregate (strains HP-42 and HP-80), in suspension with S. sanguis strain SA-1, were studied for adherence. Ten-fold-higher numbers of coaggregating strain HP-28 adhered in vitro to the streptococcal layer than did the non-coaggregating strains. S. sobrinus strain 6715 did not show appreciable adherence to the S. sanguis surface. Saliva did not affect the adherence of coaggregating or non-coaggregating H. parainfluenzae strains to S. sanguis strain SA-1. Bovine enamel chips, coated with streptococci, mounted on modified orthodontic appliances and placed in the mouths of three volunteers, facilitated the measurement of interbacterial adherence in vivo of streptomycin-resistant strains of H. parainfluenzae (HP-28R or HP-42R). Suspensions of bacteria were placed into the mouth, distributed throughout, and expectorated. After 15 or 120 minutes, the appliance with the chips was removed, the chips sonified, and colony-forming units (CFU) of streptomycin-resistant haemophili determined per chip. Similar to the in vitro experiments, ten times as many coaggregating HP-28R than non-coaggregating HP-42R adhered in vivo to the S. sanguis SA-1 layer in two of the three subjects. The measurement per mm² in vitro and in vivo of cell-to-cell interactions facilitates accurate comparisons between the different strains and species and will facilitate studies on the interbacterial interactions involved in plaque formation.

Journal of Dental Research, Vol. 67, No. 12, 1455-1460 (1988)
DOI: 10.1177/00220345880670120301


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