31P nuclear magnetic resonance spectroscopic evidence that canine nucleus pulposus and anulus fibrosus cells differ in principal organophosphorus biomolecules.

Abstract

31P Nuclear magnetic resonance spectroscopy was used to compare amounts and types of principal organophosphorus biomolecules in canine intervertebral disc tissues ex vivo. The goal of this study was to compare principal organophosphorus biomolecules in nucleus pulposus, anulus fibrosus, and in isolated nucleus pulposus and anulus fibrosus cells. There is no published information on principal organophosphorus biomolecules in disc tissues for any animal. Canine nucleus pulposus and anulus fibrosus were shown by hematoxylin-eosin staining to be healthy tissues characteristic of adult dogs. Viable cells liberated from these disc tissues by sequential protease digestion were directly visualized by light microscopy of wet mounts. 31P nuclear magnetic resonance spectra were recorded at room temperature for 12 hours according to conventional published methods. No resonances were detected for intact nucleus pulposus and anulus fibrosus tissue. 31P nuclear magnetic resonance spectra of viable chondrocyte-like cells from anulus fibrosus featured two distinctive resonance peaks consistent with phosphomonoesters and phosphodiesters. After undigested anulus fibrosus was removed, no resonances were detected. 31P nuclear magnetic resonance spectra of viable chondrocyte-like cells from nucleus pulposus before and after removal of undigested tissue featured only one resonance peak consistent with phosphodiesters. A phosphomonoester-containing biomolecule is present in healthy canine anulus fibrosus tissue but not in nucleus pulposus tissue, nor in nucleus pulposus cells or anulus fibrosus cells. A phosphodiester-containing biomolecule is present principally in nucleus pulposus cells. This study demonstrated that canine chondrocyte-like cells from nucleus pulposus and anulus fibrosus are phenotypically distinctive in principal organophosphorus biomolecules.