Differences in both matrix metalloproteinase 9 concentration and zymographic profile between plasma and serum with clot activators are due to the presence of amorphous silica or silicate salts in blood collection devices.
Matrix metalloproteinases (MMPs) are promising diagnostic tools, and blood sampling/handling alters MMP concentrations between plasma and serum and between serum with and without clot activators. To explain the higher MMP-9 expression in serum collected with clot accelerators relative to serum with no additives and to plasma, we analyzed the effects of increasing amounts of silica and silicates (components of clot activators) in citrate plasma, serum, and buffy coats collected in both plastic and glass tubes from 50 healthy donors, and we analyzed the effects of silica and silicate on cultured leukemia cells. The levels of MMP-2 did not show significant changes between glass and plastic tubes, between serum and plasma, between serum with and without clot accelerators, or between silica and silicate treatments. No modification of MMP-9 expression was obtained by the addition of silica or silicate to previously separated plasma and serum. Increasing the amounts of nonsoluble silica and soluble silicate added to citrate and empty tubes prior to blood collection resulted in increasing levels of MMP-9 relative to citrate plasma and serum. Silica and silicate added to buffy coats and leukemia cells significantly induced MMP-9 release/secretion, demonstrating that both silica and silicate induce the release of pro- and complexed MMP-9 forms. We recommend limiting the misuse of serum and avoiding the interfering effects of clot activators.
Mannello
Ferdinando
F
Institute of Histology and Laboratory Analysis, Faculty of Sciences and Technologies, University Carlo Bo, 61029 Urbino, Italy. ferdinando.mannello@uniurb.it
Tanus-Santos
Jose E
JE
Meschiari
Cesar A
CA
Tonti
Gaetana A
GA
D000925:Anticoagulants / Q000494:pharmacology; D001800:Blood Specimen Collection / Q000295:instrumentation / Q000379:methods*; D019343:Citric Acid / Q000494:pharmacology; D004591:Electrophoresis, Polyacrylamide Gel; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D020778:Matrix Metalloproteinase 2 / Q000097:blood; D020780:Matrix Metalloproteinase 9 / Q000097:blood*; D010949:Plasma; D044967:Serum; D017640:Silicates / Q000494:pharmacology*; D012822:Silicon Dioxide / Q000494:pharmacology*
D000925:Anticoagulants; D017640:Silicates; D019343:Citric Acid; D012822:Silicon Dioxide; D020778:Matrix Metalloproteinase 2; D020780:Matrix Metalloproteinase 9