The influence of nutrient supply and cell density on the growth and survival of intervertebral disc cells in 3D culture

The adult human intervertebral disc (IVD) is normally avascular. Changes to the extracellular matrix in degenerative disc disease may promote vascularisation and subsequently alter cell nutrition and disc homeostasis. This study examines the influence of cell density and the presence of glucose and serum on the proliferation and survival of IVD cells in 3D culture. Bovine nucleus pulposus (NP) cells were seeded at a range of cell densities (1.25 × 10(5)-10(6) cells/mL) and cultured in alginate beads under standard culture conditions (with 3.15 g/L glucose and 10 % serum), or without glucose and/or 20% serum. Cell proliferation, apoptosis and cell senescence were examined after 8 days in culture. Under standard culture conditions, NP cell proliferation and cluster formation was inversely related to cell seeding density, whilst the number of apoptotic cells and enucleated "ghost" cells was positively correlated to cell seeding density. Increasing serum levels from 10% to 20% was associated with increased cluster size and also an increased prevalence of apoptotic cells within clusters. Omitting glucose produced even larger clusters and also more apoptotic and senescent cells. These studies demonstrate that NP cell growth and survival are influenced both by cell density and the availability of serum or nutrients, such as glucose. The observation of clustered, senescent, apoptotic or "ghost" cells in vitro suggests that environmental factors may influence the formation of these phenotypes that have been previously reported in vivo. Hence this study has implications for both our understanding of degenerative disc disease and also cell-based therapy using cells cultured in vitro.

Thermal effects on the photoelastic coefficient of polymer optical fibers

We measure the radial profile of the photoelastic coefficient C(r) in single-mode polymer optical fibers (POFs), and we determine the evolution of C(r) after annealing the fibers at temperatures from 40°C to 80°C. We demonstrate that C(r) in the fibers drawn from a preform without specific thermal pre-treatment changes and converges to values between 1.2 and 1.6×10-12 Pa-1 following annealing at 80°C. The annealed fibers display a smoothened radial profile of C(r) and a lowered residual birefringence. In contrast, the mean value of C(r) of the fiber drawn from a preform that has been pre-annealed remains constant after our annealing process and is significantly higher, i.e., 4×10-12 Pa-1. The annealing process decreases the residual birefringence to a lower extent as well. These measurements indicate the impact of annealing on the thermal stability of the photoelastic coefficient of POFs, which is an essential characteristic in view of developing POF-based thermomechanical sensors.