Expression of Type I, III, and VI Collagen mRNAs in Experimentally Injured Porcine Intervertebral Disc

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Abstract
Young domestic pigs were incised with a scalpel blade into the anterior part of annulus fibrosus of lumbar discs in order to study the reparative processes in the annulus fibrosus and the secondary reactions in the nucleus pulposus. Northern and slot-blot hybridizations were used to investigate type I, III, and VI collagen gene expression in the disc tissue. For this purpose a method for RNA isolation was modified so as to be applicable to the intervertebral disc, which has a low cell density and a high proteoglycan content in its extracellular matrix. The amount of total RNA was found to be very low, particularly in the nucleus pulposus. Intact RNA could be isolated from most parts of the injured discs, but only from the outer annulus of the control discs. Hybridizations showed that healing of the wound in the annulus fibrosus involves an increase in the synthesis of particularly type I and type III collagens. However, no changes in the collagen gene expression were detectable in the nucleus pulposus two weeks after the injury.