Mode of growth of bacterial pathogens in chronic polymicrobial human osteomyelitis

Direct examination of material from two cases of persistent (2 and 60 years) osteomyelitis by morphological and culture techniques showed that the pathogens comprised several bacterial species whose cells grew predominantly in discrete exopolysaccharide-enclosed microcolonies made up of a single bacterial morphotype. Bacterial microcolonies were seen between tissue elements in infected connective tissue, and the microcolonies adherent to bone surfaces coalesced to form extensive biofilms that occluded the surfaces of dead bone in sequestrae. Decalcification techniques were required to examine the interior of infected bone, but recognizable remnants were associated with very large amounts of fibrous, ruthenium red-stained material. All bacterial growth in these persistent infections occurred within an intercellular matrix, and some elements of this matrix, which was fibrous in transmission electron microscopy and amorphous in scanning electron microscopy, were associated with the surfaces of bacterial cells in a manner that suggested their production by these organisms. All of the implications of this microcolony mode of bacterial growth in osteomyelitis, and in other chronic bacterial diseases, have yet to be determined.