The effect of elevated temperature on the wear of stainless steel in contact with tungsten carbide

The bearings in the air motors of modern jet aircraft engines must operate dry in hostile conditions at temperatures up to 500° C, where the thrust races in the actuators operate at temperatures up to 300° C. One of the few metallurgical combinations which can function efficiently under these conditions is martensitic stainless steel on tungsten carbide. The work described was initiated to isolate the wear mechanisms of two such steels in contact with tungsten carbide at temperatures up to 500° C. Experiments were carried out on angular contact bearings similar to these used in service, where both rolling and sliding is present and also for pure sliding conditions using a pin-on-disc apparatus. Wear measurements of the bearings were obtained with wear rates, friction and surface temperatures from the pin-on-disc machine for a series of loads and speeds. Extensive X-ray diffraction analysis was carried out on the wear debris, with also S.E.M. analysis and hardness tests on the worn surfaces along with profilometry measurements of the disc. The oxidational parameters of the steel were obtained from measurements of oxide growth rates by ellipsometry. Three distinct mechanisms of wear were established and the latter two were found to be present in both configurations. These involve an oxidational-abrasive mechanism at loads below 40 N with pin surface temperatures up to about 300 °C, with the mechanism changing to severe wear for higher loads. As the temperature increases a third wear mechanism appears due to transfer of relatively soft oxide films to the steel surface reducing the wear rate. Theoretical K factors were derived and compared with experimental values which were found to be in good agreement for the severe wear mechanism. The pin-on-disc experiments may be useful as a screening test for material selection, without the considerable cost of producing the angular contact bearings.

The boundary lubricated friction and wear of low alloy steel

Pin on disc wear machines were used to study the boundary lubricated friction and wear of AISI 52100 steel sliding partners. Boundary conditions were obtained by using speed and load combinations which resulted in friction coefficients in excess of 0.1. Lubrication was achieved using zero, 15 and 1000 ppm concentrations of an organic dimeric acid additive in a hydrocarbon base stock. Experiments were performed for sliding speeds of 0.2, 0.35 and 0.5 m/s for a range of loads up to 220 N. Wear rate, frictional force and pin temperature were continually monitored throughout tests and where possible complementary methods of measurement were used to improve accuracy. A number of analytical techniques were used to examine wear surfaces, debris and lubricants, namely: Scanning Electron Microscopy (SEM), Auger Electron Spectroscopy (AES), Powder X-ray Diffraction (XRD), X-ray Photoelectron Spectroscopy (XPS), optical microscopy, Back scattered Electron Detection (BSED) and several metallographic techniques. Friction forces and wear rates were found to vary linearly with load for any given combination of speed and additive concentration. The additive itself was found to act as a surface oxidation inhibitor and as a lubricity enhancer, particularly in the case of the higher (1000 ppm) concentration. Wear was found to be due to a mild oxidational mechanism at low additive concentrations and a more severe metallic mechanism at higher concentrations with evidence of metallic delamination in the latter case. Scuffing loads were found to increase with increasing additive concentration and decrease with increasing speed as would be predicted by classical models of additive behaviour as an organo-metallic soap film. Heat flow considerations tended to suggest that surface temperature was not the overriding controlling factor in oxidational wear and a model is proposed which suggests oxygen concentration in the lubricant is the controlling factor in oxide growth and wear.

Oxidational wear of low alloy steel in gases other than air

A pin on disc wear machine has been used to study the oxidational wear of low alloy steel in a series of experiments which were carried out under dry wear sliding conditions at range of loads from 11.28 to 49.05 N and three sliding speeds of 2 m/s, 3.5 m/s and 5 m/s, in atmosphere of air, Ar, CO2, 100% O2, 20% O2-80% Ar and 2% O2-98% Ar. Also, the experiments were conducted to study frictional force, surface and contact temperatures and surface parameters of the wearing pins. The wear debris was examined using x-ray diffraction technique for the identification of compounds produced by the wear process. Scanning electron microscopy was employed to study the topographical features of worn pins and to measure the thickness of the oxide films. Microhardness tests were carried out to investigate the influence of the sub-surface microhardness in tribological conditions. Under all loads, speeds and atmospheres parabolic oxidation growth was observed on worn surfaces, although such growth is dependent on the concentration of oxygen in the atmospheres employed. These atmospheres are shown to influence wear rate and coefficient of friction with change in applied load. The nature of the atmosphere also has influence on surface and contact temperatures as determined from heat flow analysis. Unlubricated wear debris was found to be a mixture of αFe2O3, Fe3O4 and FeO oxide. A model has been proposed for tribo-oxide growth demonstrating the importance of diffusion rate and oxygen partial pressure, in the oxidation processes and thus in determination of wear rates.

Wear and corrosion of mechanical seals

Mechanical seals are used extensively to seal machinery such as pumps, mixers and agitators in the oil, petrochemical and chemical industries. The performance of such machinery is critically dependent on these devices. Seal failures may result in the escape of dangerous chemicals, possibly causing injury or loss of life. Seal performance is limited by the choice of face materials available. These range from cast iron and stellited stainless steel to cemented and silicon carbides. The main factors that affect seal performance are the wear and corrosion of seal faces. This research investigated the feasibility of applying surface coating/treatments to seal materials, in order to provide improved seal performance. Various surface coating/treatment methods were considered; these included electroless nickel plating, ion plating, plasma nitriding, thermal spraying and high temperature diffusion processes. The best wear resistance, as evaluated by the Pin-on-Disc wear test method, was conferred by the sprayed tungsten carbide/nickel/tungsten-chromium carbide deposit, produced by the high energy plasma spraying (Jet-Kote) process. In general, no correlation was found between hardness and wear resistance or surface finish and friction. This is due primarily to the complexity of the wear and frictional oxidation, plastic deformation, ploughing, fracture and delamination. Corrosion resistance was evaluated by Tafel extrapolation, linear polarisation and anodic potentiodynamic polarisation techniques. The best corrosion performance was exhibited by an electroless nickel/titanium nitride duplex coating due to the passivity of the titanium nitride layer in the acidified salt solution. The surface coating/treatments were ranked using a systematic method, which also considered other properties such as adhesion, internal stress and resistance to thermal cracking. The sealing behaviour of surface coated/treated seals was investigated on an industrial seal testing rig. The best sealing performances were exhibited by the Jet-Kote and electroless nickel silicon carbide composite coated seals. The failure of the electroless nickel and electroless nickel/titanium nitride duplex coated seals was due to inadequate adhesion of the deposits to the substrate. Abrasion of the seal faces was the principal wear mechanism. For operation in an environment similar to the experimental system employed (acidified salt solution) the Jet-Kote deposit appears to be the best compromise.

The transition from severe to mild wear in law alloy steel

The wear rates of sliding surfaces are significantly reduced if mild oxidational wear can be encouraged. It is hence of prime importance in the interest of component life and material conservation to understand the factors necessary to promote mild, oxidational wear, The present work investigates the fundamental mechanism of the running-in wear of BS EN 31!EN 8 steel couples. under various conditions of load. speed and test duration. Unidirectional sliding experiments were carried out on a pin-on~disc wear machine where frictional force, wear rate, temperature and contact resistance were continuously monitored during each test. Physical methods of analysis (x-ray, scanning electron microscopy etc.) were used to examine the wear debris and worn samples. The wear rate versus load curves revealed mild wear transitions, which under long duration of running, categorized mild wear into four distinct regions.α-Fe20s. Fe304, FeO and an oxide mixture were the predominant oxides in four regions of oxidational wear which were identified above the Welsh T2 transition. The wear curves were strongly effected by the speed and test duration. A surface model was used to calculate the surface parameters, and the results were found to be comparable with the experimentally observed parameters. Oxidation was responsible for the transition from severe to mild wear at a load corresponding to the Welsh T2 transition. In the running-in period sufficient energy input and surface hardness enabled oxide growth rate to increase and eventually exceeded the rate of removal, where mild wear ensued. A model was developed to predict the wear volume up to the transition. Remarkable agreement was found between the theoretical prediction and the experimentally-measured values. The oxidational mechanjsm responsible for transitjon to mild wear under equilibrium conditions was related to the formation of thick homogenous oxide plateaux on subsurface hardened layers, FeO was the oxide formed initially at the onset of mild wear but oxide type changed.during the total running period to give an equilibrium oxide whose nature depended on the loads applied.

Nitrogen implantation of tool steels and engineering coatings

Ion implantation modifies the surface composition and properties of materials by bombardment with high energy ions. The low temperature of the process ensures the avoidance of distortion and degradation of the surface or bulk mechanical properties of components. In the present work nitrogen ion implantation at 90 keV and doses above 1017 ions/cm2 has been carried out on AISI M2, D2 and 420 steels and engineering coatings such as hard chromium, electroless Ni-P and a brush plated Co-W alloy. Evaluation of wear and frictional properties of these materials was performed with a lubricated Falex wear test at high loads up to 900 N and a dry pin-on-disc apparatus at loads up to 40 N. It was found that nitrogen implantation reduced the wear of AISI 420 stainless steel by a factor of 2.5 under high load lubricated conditions and by a factor of 5.5 in low load dry testing. Lower but significant reductions in wear were achieved for AISI M2 and D2 steels. Wear resistance of coating materials was improved by up to 4 times in lubricated wear of hard Cr coatings implanted at the optimum dose but lower improvements were obtained for the Co-W alloy coating. However, hardened electroless Ni-P coatings showed no enhancement in wear properties. The benefits obtained in wear behaviour for the above materials were generally accompanied by a significant decrease in the running-in friction. Nitrogen implantation hardened the surface of steels and Cr and Co-W coatings. An ultra-microhardness technique showed that the true hardness of implanted layers was greater than the values obtained by conventional micro-hardness methods, which often result in penetration below the implanted depth. Scanning electron microscopy revealed that implantation reduced the ploughing effect during wear and a change in wear mechanism from an abrasive-adhesive type to a mild oxidative mode was evident. Retention of nitrogen after implantation was studied by Nuclear Reaction Analysis and Auger Electron Spectroscopy. It was shown that maximum nitrogen retention occurs in hard Cr coatings and AISI 420 stainless steel, which explains the improvements obtained in wear resistance and hardness. X-ray photoelectron spectroscopy on these materials revealed that nitrogen is almost entirely bound to Cr, forming chromium nitrides. It was concluded that nitrogen implantation at 90 keV and doses above 3x1017 ions/cm2 produced the most significant improvements in mechanical properties in materials containing nitride formers by precipitation strengthening, improving the load bearing capacity of the surface and changing the wear mechanism from adhesive-abrasive to oxidative.

Wear of high aluminium zinc-based alloys in plain bearing applications

Two zinc-based alloys of high aluminium content, Super Cosmal alloy containing 60% Al, 6% Si, 1% Cu, 0.3% Mn and HAZCA alloy containing 60% Al, 8% Si, 2% Cu, 0.06% Mg were produced by sand casting. Foundry characteristics in particular, fluidity, mode of solidification and feeding ability were examined. Metallographic analysis of structures was carried out using optical and scanning electron microscopy and their mechanical properties were determined using standard techniques. Dry wear characteristics were determined using a pin-on-disc test, and boundary-lubricated wear was studied using full bearing tests. Results from casting experiments were evaluated and compared with the behaviour of a standard ZA-27 alloy and those from tribological tests with both ZA-27 alloy and a leaded tin-bronze (SAE660) under the same testing conditions. The presence of silicon was beneficial, reducing the temperature range of solidification, improving feeding efficiency and reducing gravity segregation of phases. Use of chills and melt degassing was found necessary to achieve soundness and enhanced mechanical properties. Dry wear tests were performed against a steel counterface for sliding speeds of 0.25, 0.5, 1.0 and 2 m/s and for a range of loads up to 15 kgf. The high aluminium alloys showed wear rates as low as those of ZA-27 at speeds of 0.25 and 0.5 m/s for the whole range of applied loads. ZA-27 performed better at higher speeds. The build up of a surface film on the wearing surface of the test pins was found to be responsible for the mild type of wear of the zinc based alloys. The constitution of the surface film was determined as a complex mixture of aluminium, zinc and iron oxides and metallic elements derived from both sliding materials. For full bearing tests, bushes were machined from sand cast bars and were tested against a steel shaft in the presence of a light spindle oil as the lubricant. Results showed that all zinc based alloys run-in more rapidly than bronze, and that wear in Super Cosmal and HAZCA alloys after prolonged running were similar to those in ZA-27 bearings and significantly smaller than those of the bronze.

The mechanical and wear behaviour of B(SiC) fibre-reinforced composite materials

The mechanical properties and wear behaviour of B(SiC) fibre-reinforced metal matrix composites (MMCs) and aluminium alloy (2014) produced by metal infiltration technique were determined. Tensile tests were peliormed at different conditions on both the alloy matrix and its composite, and the tensile fracture surfaces were also examined by Scanning Electron Microscopy (SEM). Dry wear of the composite materials sliding on hardened steel was studied using a pin-on-disc type machine. The effect of fibre orientation on wear rate was studied to provide wear resistance engineering data on the MMCs. Tests were carried out with the wear surface sliding direction set normal, parallel and anti-parallel to the fibre axis. Experiments were perfonned for sliding speeds of 0.6, 1.0 and 1.6 m/s for a load range from 12 N to 60 N. A number of sensitive techniques were used to examine worn surface and debris, i.e: Scanning Electron Microscopy (SEM), Backscattered Electron Microscopy (BSEM) and X-ray Photoelectron Spectroscopy (XPS). Finally, the effect of fibre orientation on the wear rate of the Borsic-reinforced plastic matrix composites (PMCs) produced by hot pressing technique was also investigated under identical test conditions. It was found that the composite had a markedly increased tensile strength compared with the matrix. The wear results also showed that the composite exhibited extremely low wear rates compared to the matrix material and the wear rate increased with increasing sliding speed and normal load. The effect of fibre orientation was marked, the lowest wear rates were obtained by arranging the fibre perpendicular to the sliding surface, while the highest wear was obtained for the parallel orientation. The coefficient of friction was found to be lowest in the parallel orientation than the others. Wear of PMCs were influenced to the greatest extent by these test parameters although similar findings were obtained for both composites. Based on the results of analyses using SEM, BSED and XPS, possible wear mechanisms are suggested to explain the wear of these materials.

Mild wear in dry and lubricated sliding systems

A vertical pin on horizontal disc machine has been used to conduct a series of experiments in air under dry and lubricating sliding conditions. For dry sliding low load and speed combinations were chosen to correspond to the mild wear region below the Welsh T1 transition. Lubricated tests were conducted under flooded conditions using Esso Technical White Oil alone and with a 0.1% stearic acid additive, for load and speed ranges that produced substantial amounts of asperity contact and thus a boundary lubricated regime of wear. The test material in all cases was AISI 52100 steel, for unlubricated sliding subjected to loads from 5 to 50 N and a range of speeds from 10-3 to 1.0 ms-1, and for lubricated sliding loads of 50 to 123 N and for speeds of 10-2 to 1.0 ms-1. Unlubricated wear debris was found to be a mixture of -Fe_2O_3 and -Fe. Unlubricated wear was found to occur via a thin film logarithmic oxide growth followed by agglomeration into thicker oxide plateaux 2 to 10 m in thickness. Lubricated wear occurred via thick film diffusion controlled oxide growth producing homogeneous oxide plateaux 0.1 to 0.2 m in thickness. X-ray photoelectron spectroscopy identified the presence of a surface film on pins worn in White Oil with stearic acid, which is thought to be iron stearate. A model has been developed for unlubricated wear based upon the postulated growth of thin film oxides by a logarithmic rate law. The importance of sliding geometry and environment to the dominant wear mechanism has been illustrated.

Application of wear resistant coatings by the brush plating technique

The development of cobalt molybdenum and cobalt tungsten brush plating electrolytes is described. Their optimum compositions and operating conditions for commercial applications have been determined. The effects of composition, pH, applied voltage, stylus speed and pressure upon deposit composition and efficiency have been investigated. Transmission and Scanning Electron Microscopy have been employed to study the cobalt alloy deposits produced. Evaluation of the wear resistant properties of the cobalt alloys developed in this work was carried out in the laboratory using a pin and disc technique and a simulated hot forging test, and by industrial trials involving the "on site" plating of hot forging dies and cold pressing tools. It was concluded that the electrolytes developed in tl1is work enabled cobalt alloys containing 6% Mo or 8% W to be deposited at 17-20V. Brush plated cobalt deposits possessed a mixed CPU and FCC crystallographic structure at room temperature. The application of 13µm of either of the cobalt alloys resulted in improved wear performance in both pin and disc and simulated hot forging tests. The results of the industrial trials indicated that by the use of these alloys, the life of hot forging dies may be increased by 20-100%. A commercial forging organisation is using electrolytes developed in this work to coat dies prior to forging nimonic alloys. Reductions in forging temperature and improved forging qualities have been reported. Cold pressing tools coated with the alloys showed a reduced tendency to "pick-up" and scoring of the pressed panels. Reports of a reduced need for lubrication of panels before pressing have also been received.

Topographical guidance of intervertebral disc cell growth in vitro:towards the development of tissue repair strategies for the anulus fibrosus

The anulus fibrosus (AF) of the intervertebral disc consists of concentric sheets of collagenous matrix that is synthesised during embryogenesis by aligned disc cells. This highly organised structure may be severely disrupted during disc degeneration and/or herniation. Cell scaffolds that incorporate topographical cues as contact guidance have been used successfully to promote the healing of injured tendons. Therefore, we have investigated the effects of topography on disc cell growth. We show that disc cells from the AF and nucleus pulposus (NP) behaved differently in monolayer culture on micro-grooved membranes of polycaprolactone (PCL). Both cell types aligned to and migrated along the membrane's micro-grooves and ridges, but AF cells were smaller (or less spread), more bipolar and better aligned to the micro-grooves than NP cells. In addition, AF cells were markedly more immunopositive for type I collagen, but less immunopositive for chondroitin-6-sulphated proteoglycans than NP cells. There was no evidence of extracellular matrix (ECM) deposition. Disc cells cultured on non-grooved PCL did not show any preferential alignment at sub-confluence and did not differ in their pattern of immunopositivity to those on grooved PCL. We conclude that substratum topography is effective in aligning disc cell growth and may be useful in tissue engineering for the AF. However, there is a need to optimise cell sources and/or environmental conditions (e.g. mechanical influences) to promote the synthesis of an aligned ECM.

Human intervertebral disc cells promote nerve growth over substrata of human intervertebral disc aggrecan

Study Design. Coculture assays of the migration and interaction of human intervertebral disc cells and chick sensory nerves on alternate substrata of collagen and aggrecan. Objective. To examine the effects of aggrecan on disc cell migration, how disc cells and sensory nerves interact, and whether disc cells affect previously reported inhibitory effects of aggrecan on sensory nerve growth. Summary of Background Data. Human intervertebral disc aggrecan is inhibitory to sensory nerve growth in vitro, suggesting that a loss of aggrecan from the disc may have a role in the increased innervation seen in disc degeneration. Endothelial cells that appear to co-migrate with nerves into degenerated intervertebral disc express neurotrophic factors, but the effects of disc cells on nerve growth are not known. Methods. Human disc cells were seeded onto tissue culture plates that had been coated with type I collagen and human intervertebral disc aggrecan. Explants of chick dorsal root ganglions (DRGs) were subsequently added to the plates and sensory neurite outgrowth stimulated by the addition of nerve growth factor. Time-lapse video and fluorescence microscopy were used to examine the migration and interaction of the disc cells and sensory neurites, in the context of the different matrix substrata. The effects of disc cell conditioned medium on nerve growth were also examined. Results. Disc cells spread and migrated on collagen until they encountered the aggrecan substrata, where some cells, but not all, were repelled. In coculture, DRG neurites extended onto the collagen/disc cells until they encountered the aggrecan, where, like the disc cells, many were repelled. However, in the presence of disc cells, some neurites were able to cross onto this normally inhibitory substratum. The number of neurite crossings onto aggrecan correlated significantly with the number of disc cells present on the aggrecan. In control experiments using DRG alone, all extending neurites were repelled at the collagen/aggrecan border. Conditioned medium from disc cell cultures stimulated DRG neurite outgrowth on collagen but did not increase neurite crossing onto aggrecan substrata. Conclusions. Human disc cells migrate across aggrecan substrata that are repellent to sensory DRG neurites. Disc cells synthesize neurotrophic factors in vitro that promote neurite outgrowth. Furthermore, the presence of disc cells in coculture with DRG partially abrogates the inhibitory effects of aggrecan on nerve growth. These findings have important implications for the regulation of nerve growth into the intervertebral disc, but whether disc cells promote nerve growth in vivo remains to be determined.

A study of the micro-mechanics of granular media

The development of more realistic constitutive models for granular media, such as sand, requires ingredients which take into account the internal micro-mechanical response to deformation. Unfortunately, at present, very little is known about these mechanisms and therefore it is instructive to find out more about the internal nature of granular samples by conducting suitable tests. In contrast to physical testing the method of investigation used in this study employs the Distinct Element Method. This is a computer based, iterative, time-dependent technique that allows the deformation of granular assemblies to be numerically simulated. By making assumptions regarding contact stiffnesses each individual contact force can be measured and by resolution particle centroid forces can be calculated. Then by dividing particle forces by their respective mass, particle centroid velocities and displacements are obtained by numerical integration. The Distinct Element Method is incorporated into a computer program 'Ball'. This program is effectively a numerical apparatus which forms a logical housing for this method and allows data input and output, and also provides testing control. By using this numerical apparatus tests have been carried out on disc assemblies and many new interesting observations regarding the micromechanical behaviour are revealed. In order to relate the observed microscopic mechanisms of deformation to the flow of the granular system two separate approaches have been used. Firstly a constitutive model has been developed which describes the yield function, flow rule and translation rule for regular assemblies of spheres and discs when subjected to coaxial deformation. Secondly statistical analyses have been carried out using data which was extracted from the simulation tests. These analyses define and quantify granular structure and then show how the force and velocity distributions use the structure to produce the corresponding stress and strain-rate tensors.

Study of the deposition, properties and applications of electroless deposits containing particles

The deposition efficiencies of a number of electroless nickel and cobalt plating solutions were studied and in the case of nickel compared with a commercial plating solution Nifoss 80. At the optimum plating conditions (92ºC and pH 4.5) Nifoss 80 produced nickel layers most efficiently, the alkaline cobalt solution operated most efficiently at 90ºC and pH 9. The methods of producing compostte layers containing 2-3 µm carbide particles and chromium powder is described. Nickel and cobalt layers containing approximately 27% carbide particles, or 40% (Ni) and 30% (Co) chromium particles by volume were obtained. This value is independent of the particle concentration in the plating solution within the range (20~200 g/l). Hardness of the nickel. as deposited was 515 Hv, this was increased to a maximum of 1155 Hv by heat treatment at 200ºC for 5 hours in vacuum. Incorporation. of .chromium carbide particles resulted in a maximum hardness of 1225 Hv after heating at 500ºC for 5 hours in vacuum and chromium particles resulted in a maximum hardness of 16S0 Hv after heat treatment at 400ºC for 2 hours in vacuum. Similarly the hardness of cobalt as deposited was 600 Hv, this was increased to a maximum of 1300 Hv after heat treatment at 400ºC for 1 hour. Incorporation of chromium carbide particles resulted jn a maximum hardness of 1405 Hv after heating at 400ºC for 5 hours in vacuum and chromium particles resulted in a maximum hardness of 1440 Hv after. heat treating for 2 hours at 400ºC in vacuum. The structure of the deposits was studied by optical and scanning electron microscopy. The wear rate and coefficient of friction was determined by a pin and disc method. Wear rate and coefficient of friction decreased with increase in hardness. The wear resistance of the materials was also determined using a simulated forging test. Dies made of standard die steel were coated and the wear rates of the layers as deposited and after heat treatment were compared with those of uncoated tools. The wear resistance generally increased with hardness, it was 50-75% more than the uncoated die steel. Acetic acid salt spray test and outdoor exposure for six months was used to study the corrosion behaviour of the deposits and potentiodynamic curves plotted to find their corrosion potential. Nickel deposit exhibited less staining than carbide composite deposits and nickel-chromium deposits had the most noble corrosion potential.

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.