Induction of matrix metalloproteinase expression in human macrophages by orthopaedic particulate debris in vitro.
We exposed human macrophages isolated from the peripheral blood of healthy donors to metal and bone-cement particles from 0.2 to 10 microm in size. Zymography showed that macrophages exposed to titanium alloy and polymethylmethacrylate (PMMA) particles released a 92- and 72-kDa gelatinase in a dose- and time-dependent manner. Western immunoblotting confirmed that the 92- and 72-kDa gelatinolytic activities corresponded to matrix metalloproteinase-9 and matrix metalloproteinase-2 (MMP-9, MMP-2), respectively. Western immunoblotting also indicated that titanium alloy and PMMA particles increased the release of MMP-1. Northern blotting showed elevated mRNA signal levels for MMP-1, MMP-2, and MMP-9 after exposure to both types of particle. Collagenolytic activity also increased in the macrophage culture medium in response to both types of particle. Our findings support the hypothesis that macrophages release MMPs in proportion to the amount of particulate debris within periprosthetic tissues.
Division of Orthopaedic Surgery, Stanford University School of Medicine, California 94305-5326, USA.
Y
Y
Nakashima
D H
DH
Sun
W J
WJ
Maloney
S B
SB
Goodman
D J
DJ
Schurman
R L
RL
Smith
D000497:Alloys / Q000008:administration & dosage / Q000494:pharmacology*; D015152:Blotting, Northern; D015153:Blotting, Western; D001843:Bone Cements / Q000494:pharmacology*; D002478:Cells, Cultured; D003094:Collagen / Q000187:drug effects / Q000378:metabolism; D017364:Collagenases / Q000032:analysis / Q000187:drug effects / Q000235:genetics; D004305:Dose-Response Relationship, Drug; D004591:Electrophoresis, Polyacrylamide Gel; D018093:Gelatinases / Q000032:analysis / Q000187:drug effects / Q000235:genetics; D015971:Gene Expression Regulation, Enzymologic; D006801:Humans; D008264:Macrophages / Q000187:drug effects / Q000201:enzymology*; D020781:Matrix Metalloproteinase 1; D020778:Matrix Metalloproteinase 2; D020780:Matrix Metalloproteinase 9; D008666:Metalloendopeptidases / Q000032:analysis* / Q000187:drug effects / Q000235:genetics; D010316:Particle Size; D019904:Polymethyl Methacrylate / Q000008:administration & dosage / Q000494:pharmacology*; D012333:RNA, Messenger / Q000032:analysis / Q000235:genetics; D013997:Time Factors; D014025:Titanium / Q000008:administration & dosage / Q000494:pharmacology*
D000497:Alloys; D001843:Bone Cements; D012333:RNA, Messenger; C031462:titanium alloy (TiAl6V4); D003094:Collagen; D019904:Polymethyl Methacrylate; D014025:Titanium; D017364:Collagenases; D018093:Gelatinases; D008666:Metalloendopeptidases; D020778:Matrix Metalloproteinase 2; D020780:Matrix Metalloproteinase 9; D020781:Matrix Metalloproteinase 1