Identity of limulus amoebocyte lysate-active root surface materials from periodontally involved teeth.
A new approach has been developed to circumvent the problems of false positive reactions in the Limulus Amoebocyte Lysate (LAL) assay for lipopolysaccharide (LPS) in root surface materials. These LAL-reactive materials include thrombin, thromboplastin, ribonuclease, ribonucleic acid, lipoteichoic acid and peptidoglycan fragments. In the present study, hot phenol/water extraction of these substances followed by ultracentrifugation of the resulting aqueous phases reduced their concentrations to very low levels. Furthermore, the application of Polymyxin B/Sepharose 4B affinity chromatography to these extracts enabled their intrinsic LAL-activity to be determined. Use of these techniques to assay root surface materials has identified LPS as being the major LAL-reactive material present. The mean LPS yield for the periodontally involved teeth was 4.13 micrograms/tooth, representing 2.82 micrograms/root. In contrast, the mean yield of LPS for the periodontally uninvolved teeth was 3.12 ng/tooth.
Wilson
M
M
Moore
J
J
Kieser
J B
JB
D000328:Adult; D000368:Aged; D002846:Chromatography, Affinity; D005189:False Positive Reactions; D006801:Humans; D008033:Limulus Test; D008070:Lipopolysaccharides / Q000032:analysis*; D008875:Middle Aged; D010510:Periodontal Diseases / Q000378:metabolism*; D013499:Surface Properties; D014020:Tissue Extracts; D014092:Tooth Root / Q000032:analysis*; D014461:Ultracentrifugation
D008070:Lipopolysaccharides; D014020:Tissue Extracts