Ultrastructure of the human intervertebral disc during aging and degeneration: comparison of surgical and control specimens.
METHODS
Human intervertebral disc tissue from the annulus was obtained in a prospective study investigating the ultrastructural features of disc cells and extracellular matrix. Experimental studies were approved by the authors' Human Subjects Institutional Review Board. Discs were obtained from surgical specimens and control donors.


OBJECTIVE
To compare the cellular and extracellular matrix characteristics of the annulus from control and surgical disc specimens using electron microscopy and specialized fixation that visualizes proteoglycans.


BACKGROUND
The ultrastructural features of disc cells and the disc matrix have received little attention, as compared with the literature on age- and disease-related changes in bone and cartilage.


METHODS
Ultrastructural studies investigated disc tissue obtained from control and surgical disc specimens using transmission electron microscopy. Specialized fixation with ruthenium red was used to highlight matrix proteoglycans.


RESULTS
Cellular and extracellular matrix fine structure was assessed in disc specimens from 29 control donors (newborns to 79-year-olds) and surgical disc specimens from 49 patients (16- to 77-year-olds). Control and surgical tissue showed similar ultrastructural features. Unusual matrix surrounding and encircling single cells or clusters of cells was common (48% of control and 63% of surgical specimens) and often contained fibrous long-spacing collagen (41.3% of control and 36.7% of surgical specimens). Ruthenium red greatly aided visualization of proteoglycans pooled in lacunar spaces. Variable cross-sectional diameters of collagen fibrils was present in 34% of control and 59% of surgical specimens. Regions with sparse interterritorial matrix were common. Cell morphology showed both cells with apoptotic nuclei and synthetically active cells that appeared healthy.


CONCLUSIONS
Control and surgical specimens of the annulus showed similar ultrastructural features. Heterogeneity of collagen fibril diameter is an important observation because it is believed that fibril size relates to biomechanical disc function. Fibrous long-spacing collagen may reflect extracellular matrix remodeling or the presence of previous fibril depolymerization followed by repolymerization and reassociation with proteoglycans. Synthetic activity of disc cells is reflected in active rough endoplasmic reticulum, Golgi, and pools of proteoglycans in lacunar spaces and unusual extracellular matrix components that encircle cells and cell clusters. Such components may influence biomechanical quality. Departures from normal extracellular matrix organization of the aging or degenerating disc undoubtedly contribute to decreased biomechanical function of the annulus because they disrupt the normal annulus architecture. This study underscores the need for a fuller understanding of the dynamic relation between disc cells and the surrounding extracellular matrix, which they continually produce and remodel.
Gruber
Helen E
HE
Department of Orthopaedic Surgery, Carolinas Medical Center, Charlotte, North Carolina, USA. hgruber@carolinas.org
Hanley
Edward N
EN
D000293:Adolescent; D000328:Adult; D000368:Aged; D000375:Aging / Q000473:pathology*; D002467:Cell Nucleus / Q000648:ultrastructure; D002648:Child; D002675:Child, Preschool; D003094:Collagen / Q000648:ultrastructure; D005109:Extracellular Matrix / Q000648:ultrastructure; D006801:Humans; D002479:Inclusion Bodies / Q000648:ultrastructure; D007223:Infant; D007231:Infant, Newborn; D007403:Intervertebral Disc / Q000473:pathology* / Q000648:ultrastructure*; D008875:Middle Aged
D003094:Collagen