Abstract
Study Design: Experimental study to assess tissue engineered solutions to disc degeneration.
Objectives: To investigate the use of a novel biodegradable hydrogel which is capable of minimally invasive introduction into an intervertebral disc (IVD) and support of cultured nucleus pulposus (NP) cells for the purpose of developing a tissue-engineered solution to retard progression of IVD degeneration. There were 3 objectives: (1) To introduce a slowly polymerising alginate hydrogel into the NP cavity of a bovine vertebral disc model. (2) To demonstrate the viability and metabolic activity of cultured NP cells in the hydrogel in vitro. (3) To determine the effect of Synvisc (hylan G-F 20) on NP cell proliferation and extracellular matrix (ECM) production.
Summary of Background Data: The cause of intervertebral disc degeneration (IVDD) is multifactorial. One proposed mechanism is that IVDD originates in the NP and progresses radially to the annulus fibrosis (AF). There is a growing interest in tissue-engineered solutions where a biological repair is induced. By preventing the abnormality at the NP it may be possible to halt progression of IVDD. Injection of NP cells into an early degenerative IVD, where the AF is still intact, may retard the degenerative process.
Subjects/Methods: CaSO4 and CaCO3 alginates were injected into the NP cavity of a bovine tail. After 90 minutes the tail was dissected to reveal the gel. NP cells released from pooled bovine NP tissue were dispersed into the CaSO4 and CaCO3 alginate gels (10x106 cells.mL−1) with and without Synvisc and cultured for 21 days.
Results: Injectable alginate suspensions formed solid viscoelastic gels, filling the exact shape of the NP cavity. NP DNA and ECM synthesis was significantly greater in the CaCO3 alginate gel than in the CaSO4 alginate gel (p< 0.05). Synvisc significantly increased sulphated GAG (p< 0.01) and collagen (p< 0.05) production. These effects were supported histologically and immunohistologically where cells in the CaCO3 and Synvisc gels stained more intensely for proteoglycan and collagen type II.
Conclusions: Both CaCO3 alginate gel and CaSO4 alginate gel are injectable and are capable of sustaining NP cells in-vitro. Cells remain viable, maintain their phenotype, proliferate and produce ECM during the culture period. The CaCO3 alginate gel provides a three-dimensional matrix more favourable to NP cellular activity than the CaSO4 alginate gel. Synvisc behaves as a chondro-stimulant significantly enhancing NP cell metabolic activity.
These abstracts were prepared by Mr. Brian J C Freeman FRCS (Tr & Orth). Correspondence should be addressed to him at The Centre for Spinal Studies and Surgery, University Hospital, Queens Medical Centre, Nottingham NG7 2UH.
