Study Design.: In vitro experiment using bone morphogenetic protein-2 (BMP-2) and cells from the nucleus pulposus (NP), transitional zone (TZ), and anulus fibrosus (AF) of the human intervertebral disc (IVD). Objective.: To demonstrate the differential effect of BMP-2 on DNA synthesis, proteoglycan synthesis, and osteocalcin mRNA expression in human IVD cells from the NP, TZ, and AF, respectively. Summary of Background Data.: BMP-2 has been proven to be effective in stimulating proteoglycan synthesis in articular chondrocytes and IVD cells from the NP. Nevertheless, the effect of BMP-2 on cells from different regions of the IVD has not yet been thoroughly elucidated. Methods.: Human IVDs were harvested from surgical disc procedures and tissue from the NP, TZ, and AF was obtained. Disc tissue was enzymatically digested, and IVD cells were cultured three-dimensionally in alginate beads. Then IVD cell cultures from the NP, TZ, and AF were exposed to BMP-2. DNA synthesis and newly synthesized proteoglycan were measured. Reverse transcription-polymerase chain reaction for mRNA expression of bone sialoprotein, DLX5, osteocalcin, and collagen type I, was performed. Results.: Cells from the AF responded to BMP-2 with mitogenesis. There was no significant increase in DNA synthesis in cultures from the NP and TZ treated with BMP-2. Only cells from the NP showed a significant increase in newly synthesized proteoglycan in response to BMP-2. IVD cells from all zones demonstrated no significant expression of bone sialoprotein, DLX5, osteocalcin mRNA after treatment with BMP-2. Conclusion.: BMP-2 clearly exerted a mitogenic effect on AF cells, and stimulated proteoglycan synthesis in NP cells. However, BMP-2 did not have an osteogenic effect in any IVD region. Taken together, these results confirm that BMP-2 can be used as an anabolic agent for mitogenesis in AF cells and NP cell matrix regeneration without the possibility of osteogenesis.
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