Inflammatory cells in full-thickness anulus injury in pigs. An experimental disc herniation animal model.
METHODS
Inflammatory cells were studied by indirect immunocytochemistry in experimental full-thickness anulus fibrosus lesions in pigs.


OBJECTIVE
First, to determine the occurrence, by immunocytochemistry, of T lymphocytes and macrophages in experimentally produced, anterolateral full-thickness disc lesions in pigs, and second, to compare the presence of inflammatory cells in 1) the injury area, 2) the adjacent noninjured part of the disc, and 3) control discs.


BACKGROUND
Previous studies on disc herniation material obtained from human disc surgeries have demonstrated inflammatory cells in a subgroup of herniations. Macrophages were most prevalent, being more numerous than lymphocytes. Macrophages have furthermore been suggested to be important in the resorption process of extruded disc tissue. No similar studies on an animal model of disc herniation, however, have so far been presented.


METHODS
A full thickness anular incision, 10 mm long, was made with a scalpel in the L3-L4 or L4-L5 intervertebral discs of 12 adult pigs. The incision was made in the anterolateral part of the disc. Nucleus material was observed outside the injury site when tissue samples were taken, suggesting a disc herniation. Tissue then was analyzed from the area of injury, from the area adjacent to the injury, and from separate control discs from three additional pigs of the same age. Thin frozen sections were studied by indirect immunocytochemistry (alkaline phosphatase anti-alkaline phosphatase method) using monoclonal anti-human antibodies applicable to porcine tissues, T lymphocytes (CD3), and macrophages (CD68). Cells were graded as: -, absent; (+), only a few scattered cells; and +, abundant cells. Disc tissue samples were taken 1 month (three discs), 2 months (four discs), and 3 months (five discs) after the operation.


RESULTS
Macrophages were present more commonly than T cells, and were abundant in seven of 12 discs (58%), with T cells abundant in four of 12 discs (33%). Only a few macrophages were present in the injured tissue from one additional disc, and scattered T cells were seen in four additional discs. Abundant macrophages were also observed in one of two discs in the adjacent noninjured area, whereas only a few T lymphocytes at the most were present in such noninjured disc tissue. In four (33%) and three (25%) injured discs, respectively, no macrophages or T lymphocytes could be found. No inflammatory cells were observed in three of 12 discs (25%). The three control discs showed no inflammatory cells.


CONCLUSIONS
Inflammatory cells, predominantly macrophages, were present in a subsample of experimental discs with full-thickness anulus defects, as has previously been observed for human disc herniations. In this animal model, macrophages may have spread to adjacent noninjured parts of the disc. The induced herniation in this animal model is, however, anterolateral and may not fully correspond to clinical disc herniations, most of which are posterolateral. However, the results from this model support a role for inflammation in disc herniation.
Habtemariam
A
A
Research Laboratory, Spine Research Unit, Clinical Research Institute, Helsinki, Finland.
Virri
J
J
Grönblad
M
M
Holm
S
S
Kaigle
A
A
Karaharju
E
E
D000818:Animals; D015415:Biomarkers; D004195:Disease Models, Animal; D006801:Humans; D007405:Intervertebral Disc Displacement / Q000276:immunology* / Q000473:pathology*; D007958:Leukocyte Count; D008264:Macrophages / Q000166:cytology / Q000276:immunology; D013552:Swine; D013601:T-Lymphocytes / Q000166:cytology / Q000276:immunology
D015415:Biomarkers