Intervertebral disc cells as competent phagocytes in vitro: implications for cell death in disc degeneration.
BACKGROUND
Apoptosis has been reported to occur in the intervertebral disc. Elsewhere in the body, apoptotic cells are cleared from the system via phagocytosis by committed phagocytes such as macrophages, reducing the chance of subsequent inflammation. These cells, however, are not normally present in the disc. We investigated whether disc cells themselves can be induced to become phagocytic and so have the ability to ingest and remove apoptotic disc cells, minimising the damage to their environment.


METHODS
Bovine nucleus pulposus cells from caudal intervertebral discs were grown in culture and exposed to both latex particles (which are ingested by committed phagocytes) and apoptotic cells. Their response was monitored via microscopy, including both fluorescent and video microscopy, and compared with that seen by cell lines of monocytes/macrophages (THP-1 and J774 cells), considered to be committed phagocytes, in addition to a nonmacrophage cell line (L929 fibroblasts). Immunostaining for the monocyte/macrophage marker, CD68, was also carried out.


RESULTS
Disc cells were able to ingest latex beads at least as efficiently, if not more so, than phagocytic THP-1 and J774 cells. Disc cells ingested a greater number of beads per cell than the committed phagocytes in a similar time scale. In addition, disc cells were able to ingest apoptotic cells when cocultured in monolayer with a UV-treated population of HeLa cells. Apoptotic disc cells, in turn, were able to stimulate phagocytosis by the committed macrophages. CD68 immunostaining was strong for THP-1 cells but negligible for disc cells, even those that had ingested beads.


CONCLUSIONS
In this study, we have shown that intervertebral disc cells are capable of behaving as competent phagocytes (that is, ingesting latex beads) and apoptotic cells. In terms of number of particles, they ingest more than the monocyte/macrophage cells, possibly due to their greater size. The fact that disc cells clearly can undergo phagocytosis has implications for the intervertebral disc in vivo. Here, where cell death is reported to be common yet there is normally no easy access to a macrophage population, the endogenous disc cells may be encouraged to undergo phagocytosis (for example, of neighbouring cells within cell clusters).
Jones
Philip
P
Centre for Spinal Studies, Robert Jones & Agnes Hunt Orthopaedic & District Hospital NHS Trust, Oswestry, Shropshire, UK.
Gardner
Lucy
L
Menage
Janis
J
Williams
Gwyn T
GT
Roberts
Sally
S
D000818:Animals; D015703:Antigens, CD / Q000378:metabolism; D015214:Antigens, Differentiation, Myelomonocytic / Q000378:metabolism; D017209:Apoptosis / Q000502:physiology*; D015415:Biomarkers / Q000378:metabolism; D002417:Cattle; D002450:Cell Communication / Q000502:physiology; D002460:Cell Line; D002478:Cells, Cultured; D005347:Fibroblasts / Q000166:cytology / Q000502:physiology; D006367:HeLa Cells; D006801:Humans; D007403:Intervertebral Disc / Q000166:cytology* / Q000502:physiology; D008264:Macrophages / Q000166:cytology / Q000502:physiology; D051379:Mice; D008863:Microspheres; D009000:Monocytes / Q000166:cytology / Q000502:physiology; D010586:Phagocytes / Q000166:cytology* / Q000502:physiology; D010587:Phagocytosis / Q000502:physiology*; D013122:Spinal Diseases / Q000503:physiopathology; D000097382:CD68 Molecule
D015703:Antigens, CD; D015214:Antigens, Differentiation, Myelomonocytic; D015415:Biomarkers; C067980:CD68 antigen, human; D000097382:CD68 Molecule