Increasing both CoCrMo-alloy particle size and surface irregularity induces increased macrophage inflammasome activation in vitro potentially through lysosomal destabilization mechanisms

J Orthop Res. 2013 Oct;31(10):1633-42. doi: 10.1002/jor.22411. Epub 2013 Jun 21.

Abstract

Recent investigations indicate that innate immune "danger-signaling" pathways mediate metal implant debris induced-inflammatory responses, for example, NALP3 inflammasome. How the physical characteristics of particles (size, shape, and chemical composition) affect this inflammatory reactivity remains controversial. We examined the role of Cobalt-Chromium-Molybdenum (CoCrMo) alloy particle shape and size on human macrophage phagocytosis, lysosomal destabilization, and inflammasome activation. Round/smooth versus irregularly shaped/rough CoCrMo-alloy particles of ∼1 and 6-7 µm diameter were investigated for differential lysosomal damage and inflammasome activation in human monocytes/macrophages. While spherical/smooth 1 µm CoCrMo-alloy particles did not measurably affect macrophage IL-1β production, irregular 1 µm CoCrMo-alloy particles induced significant IL-1β increases over controls. Both round/smooth particles and irregular CoCrMo-alloy particles that were 6-7 µm in size induced >10-fold increases in IL-1β production compared to similarly shaped smaller particles (p < 0.05). Larger irregular particles induced a greater degree of intracellular lysosomal damage and a >3-fold increase in IL-1β versus similarly sized round/smooth particles (at an equal dose, particles/cell). CoCrMo-alloy particle-size-induced IL-1β production was dependent on the lysosomal protease Cathepsin B, further supporting lysosomal destabilization as causative in inflammation. Phagocytosable larger/irregular shaped particles (6 µm) demonstrated the greatest lysosomal destabilization (observed immunofluorescently) and inflammatory reactivity when compared on an equal dose basis (particles/cell) to smaller/spherical 1 µm particles in vitro.

Keywords: Cathepsin B; inflammasome; lysosomal destabilization; metal particles; monocytes/macrophages.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Alloys / chemistry
  • Alloys / pharmacology*
  • Carrier Proteins / metabolism
  • Caspase 1 / metabolism
  • Cathepsin B / metabolism
  • Cells, Cultured
  • Chromium / chemistry
  • Chromium / pharmacology
  • Cobalt / chemistry
  • Cobalt / pharmacology
  • Humans
  • In Vitro Techniques
  • Inflammasomes / immunology*
  • Inflammasomes / metabolism
  • Interleukin-1beta / metabolism
  • Lysosomes / immunology*
  • Lysosomes / metabolism
  • Macrophage Activation / immunology
  • Macrophages / cytology
  • Macrophages / immunology*
  • Macrophages / metabolism
  • Molybdenum / chemistry
  • Molybdenum / pharmacology
  • Monocytes / cytology
  • Monocytes / immunology
  • Monocytes / metabolism
  • NLR Family, Pyrin Domain-Containing 3 Protein
  • Particle Size
  • Phagocytosis / immunology
  • RNA, Small Interfering / genetics
  • Surface Properties

Substances

  • Alloys
  • Carrier Proteins
  • Inflammasomes
  • Interleukin-1beta
  • NLR Family, Pyrin Domain-Containing 3 Protein
  • NLRP3 protein, human
  • RNA, Small Interfering
  • Chromium
  • Cobalt
  • Molybdenum
  • CTSB protein, human
  • Cathepsin B
  • Caspase 1