The potential and limitations of a cell-seeded collagen/hyaluronan scaffold to engineer an intervertebral disc-like...

The use of a cell-seeded biomatrix for tissue engineering of the intervertebral disc.

To evaluate the ability of a biomatrix to support the viability of intervertebral disc cells and to accumulate the extracellular matrix that they produce.

Intervertebral disc degeneration is a common occurrence during adult life that has adverse economic consequences on the health care system. Current surgical treatments are aimed at removing or replacing the degenerate tissue, which can alter the biomechanics of the spine and result in degeneration at adjacent disc levels. The ideal treatment of the degenerate disc would involve biologic repair, and tissue-engineering techniques offer a means to achieve this goal.

Scaffolds of type I collagen and hyaluronan were seeded with bovine nucleus pulposus or anulus fibrosus cells and maintained in culture for up to 60 days in the presence of fetal calf serum or a variety of growth factors to try to generate a tissue whose properties could mimic those of the nucleus pulposus with respect to proteoglycan content.

During the culture period, various proteoglycans (aggrecan, decorin, biglycan, fibromodulin, and lumican) and collagens (types I and II) accumulated in the scaffold. Proteoglycan accumulation in the scaffold was greatest under conditions in which transforming growth factor-beta1 was present, but under all conditions, more proteoglycan was lost into the culture medium than retained in the scaffold. Both the nucleus and anulus cells behaved in a similar manner with respect to their ability to synthesize matrix macromolecules and have them retained in the scaffold. By day 60 of culture, the proteoglycan content of the scaffolds never exceeded 10% of that present in the mature nucleus pulposus, although this figure could have been considerably increased if most of the proteoglycan being synthesized could have been retained. Furthermore, proteoglycan retention was not uniform within the scaffold, but increased near its periphery.

This work demonstrates that although it is possible to maintain functional disc cells in a biomatrix, it will be necessary to optimize proteoglycan synthesis and retention if any resulting tissue is to be of value in the biologic repair of the degenerate disc. The ability of the anulus cells to replicate the matrix production of the nucleus cells, at least in the collagen/hyaluronan scaffold, suggests that repair may not be limited to the availability of authentic nucleus cells.