Investigation of the limits of nanoscale filopodial interactions

Mesenchymal stem cells are sensitive to changes in feature height, order and spacing. We had previously noted that there was an inverse relationship between osteoinductive potential and feature height on 15-, 55- and 90 nm-high titania nanopillars, with 15 nm-high pillars being the most effective substrate at inducing osteogenesis of human mesenchymal stem cells. The osteoinductive effect was somewhat diminished by decreasing the feature height to 8 nm, however, which suggested that there was a cut-off point, potentially associated with a change in cell–nanofeature interactions. To investigate this further, in this study, a scanning electron microscopy/three-dimensional scanning electron microscopy approach was used to examine the interactions between mesenchymal stem cells and the 8 and 15 nm nanopillared surfaces. As expected, the cells adopted a predominantly filopodial mode of interaction with the 15 nm-high pillars. Interestingly, fine nanoscale membrane projections, which we have termed ‘nanopodia,’ were also employed by the cells on the 8 nm pillars, and it seems that this is analogous to the cells ‘clinging on with their fingertips’ to this scale of features

In vitro analyses of the toxicity, immunological, and gene expression effects of cobalt-chromium alloy wear debris and Co ions derived from metal-on-metal hip implants

Joint replacement has proven to be an extremely successful and cost-effective means of relieving arthritic pain and improving quality of life for recipients. Wear debris-induced osteolysis is, however, a major limitation and causes orthopaedic implant aseptic loosening, and various cell types including macrophages, monocytes, osteoblasts, and osteoclasts, are involved. During the last few years, there has been increasing concern about metal-on-metal (MoM) hip replacements regarding adverse reactions to metal debris associated with the MoM articulation. Even though MoM-bearing technology was initially aimed to extend the durability of hip replacements and to reduce the requirement for revision, they have been reported to release at least three times more cobalt and chromium ions than metal-on-polyethylene (MoP) hip replacements. As a result, the toxicity of metal particles and ions produced by bearing surfaces, both locally in the periprosthetic space and systemically, became a concern. Several investigations have been carried out to understand the mechanisms responsible for the adverse response to metal wear debris. This review aims at summarising in vitro analyses of the toxicity, immunological, and gene expression effects of cobalt ions and wear debris derived from MoM hip implants

Obesity is associated with higher complication rates in revision total hip arthroplasty

We examined differences in complication rates between obese and non-obese patients undergoing revision total hip arthroplasty. Sixty-five patients with a BMI ≥30 kg/m2 and 54 patients with a BMI of <30 kg/m2 were identified. Obese patients were 2.5 times more likely to suffer a complication than non-obese (38.4% cf 14.8%, p = 0.02). Obese patients were more likely to experience dislocation, leg length discrepancy, fracture, implant loosening, infection and pulmonary embolus. The obese group had a significantly higher revision rate (12.3% cf 1.8%, p = 0.039). Obese patients should be counselled pre-operatively on their elevated risk

Elective hip arthroplasty rates and related complications in people with diabetes mellitus

Background and Aims: Diabetes mellitus (DM), poor glycaemic control and raised body mass index (BMI) have been associated with postoperative complications in arthroplasty, although the relative importance of these factors is unclear. We describe the prevalence of DM in elective hip arthroplasty in a UK centre, and evaluate the impact of these factors. Methods: We analysed retrospective data for DM patients undergoing arthroplasty over a 6-year period and compared with non-diabetic matched controls (1 DM patient: 5 controls). DM was present in 5.7% of hip arthroplasty patients (82/1443). Results: Postoperative complications occurred in 12.2% of DM patients versus 12.9% of controls (p = 1.000); surgical complications were present in 6.1% of those with DM and 2.4% of controls (p = 0.087), while medical complications occurred in 8.5% of DM patients versus 10.7% of controls (p = 0.692). Complications developed in 23.1% of DM patients with poor glycaemic control (HbA1c &gt; 53 mmol/mol) versus 9.8% with good control (p = 0.169). In DM patients and controls combined, complications occurred in 16.3% of obese patients versus 10.0% of non-obese patients (p = 0.043). In the DM cohort, 13.7% of overweight patients had complications versus 0% with a normal or low BMI (p = 0.587). Conclusions: DM rates were lower than expected, and glycaemic control was good. Overall complication rates were unrelated to the presence of DM or to glycaemic control, although surgical complications were observed more frequently in those with DM and poor glycaemic control was uncommon within our cohort. Complications were more frequent in those with a higher BMI. Whether some patients with DM but without an increased risk of complications are currently being excluded from surgery requires exploration

Grooved surface topography alters matrix-metalloproteinase production by human fibroblasts

Extracellular matrix (ECM) remodelling is an essential physiological process in which matrix-metalloproteinases (MMPs) have a key role. Manipulating the manner in which cells produce MMPs and ECMs may enable the creation of a desired tissue type, i.e. effect repair, or the prevention of tissue invasion (e.g. metastasis). The aim of this project was to determine if culturing fibroblasts on grooved topography altered collagen deposition or MMP production. Human fibroblasts were seeded on planar or grooved polycaprolactone substrates (grooves were 12.5 µm wide with varying depths of 240 nm, 540 nm or 2300 nm). Cell behaviour and collagen production were studied using fluorescence microscopy and the spent culture medium was assessed using gel zymography to detect MMPs. Total collagen deposition was high on the 240 nm deep grooves, but decreased as the groove depth increased, i.e. as cell contact guidance decreased. There was an increase in gelatinase on the 2300 nm deep grooved topography and there was a difference in the temporal expression of MMP-3 observed on the planar surface compared to the 540 nm and 2300 nm topographies. These results show that topography can alter collagen and MMP production. A fuller understanding of these processes may permit the design of surfaces tailored to tissue regeneration e.g. tendon repair

Nanopit-induced osteoprogenitor cell differentiation: the effect of nanopit death

We aimed to assess osteogenesis in osteoprogenitor cells by nanopits and to assess optimal feature depth. Topographies of depth 80 nm, 220 nm and 333 nm were embossed onto polycaprolactone (PCL) discs. Bone marrow-derived mesenchymal stromal cells were seeded onto PCL discs, suspended in media and incubated. Samples were fixed after three and 28 days. Cells were stained for the adhesion molecule vinculin and the osteogenic transcription factor RUNX2 after three days. Adhesion was lowest on planar controls and it was the shallowest, 80 nm deep pits that supported optimal adhesion formation. 80 nm and 220 nm deep pits induced most RUNX2 accumulation. After 28 days, osteocalcin and osteopontin expression were used as markers of osteoblastic differentiation. 220 nm deep pits produced cells with the highest concentrations of osteopontin and osteocalcin. All topographies induced higher expression levels than controls. We demonstrated stimulation of osteogenesis in a heterogeneous population of mesenchymal stromal cells. All nanopit depths gave promising results with an optimum depth of 220 nm after 28 days. Nanoscale modification of implant surfaces could optimise fracture union or osteointergration

The Tribology of Explanted Hip Resurfacings Following Early Fracture of the Femur

A recognized issue related to metal-on-metal hip resurfacings is early fracture of the femur. Most theories regarding the cause of fracture relate to clinical factors but an engineering analysis of failed hip resurfacings has not previously been reported. The objective of this work was to determine the wear volumes and surface roughness values of a cohort of retrieved hip resurfacings which were removed due to early femoral fracture, infection and avascular necrosis (AVN). Nine resurfacing femoral heads were obtained following early fracture of the femur, a further five were retrieved due to infection and AVN. All fourteen were measured for volumetric wear using a co-ordinate measuring machine. Wear rates were then calculated and regions of the articulating surface were divided into “worn” and “unworn”. Roughness values in these regions were measured using a non-contacting profilometer. The mean time to fracture was 3.7 months compared with 44.4 months for retrieval due to infection and AVN. Average wear rates in the early fracture heads were 64 times greater than those in the infection and AVN retrievals. Given the high wear rates of the early fracture components, such wear may be linked to an increased risk of femoral neck fracture

Biomimetic microtopography to enhance osteogenesis <i>in vitro</i>

Biomimicry is being used in the next generation of biomaterials. Tuning material surface features such as chemistry, stiffness and topography allow the control of cell adhesion, proliferation, growth and differentiation. Here, microtopographical features with nanoscale depths, similar in scale to osteoclast resorption pits, were used to promote &lt;i&gt;in&lt;/i&gt; vitro bone formation in basal medium. Primary human osteoblasts were used to represent an orthopaedically relevant cell type and analysis of adhesions, cytoskeleton, osteospecific proteins (phospho-Runx2 and osteopontin) and mineralisation (alizarin red) was performed. The results further demonstrate the potential for biomimicry in material design and show that the osteoblast response can be tuned from changes in feature size

Cardiac function may be compromised in patients with elevated blood cobalt levels secondary to metal-on-metal hip implants

Aims Elevated blood cobalt levels secondary to metal-on-metal (MoM) hip arthroplasties are a suggested risk factor for developing cardiovascular complications including cardiomyopathy. Clinical studies assessing patients with MoM hips using left ventricular ejection fraction (LVEF) have found conflicting evidence of cobalt-induced cardiomyopathy. Global longitudinal strain (GLS) is an echocardiography measurement known to be more sensitive than LVEF when diagnosing early cardiomyopathies. The extent of cardiovascular injury, as measured by GLS, in patients with elevated blood cobalt levels has not previously been examined. Methods A total of 16 patients with documented blood cobalt ion levels above 13 µg/l (13 ppb, 221 nmol/l) were identified from a regional arthroplasty database. They were matched with eight patients awaiting hip arthroplasty. All patients underwent echocardiography, including GLS, investigating potential signs of cardiomyopathy. Results Patients with MoM hip arthroplasties had a mean blood cobalt level of 29 µg/l (495 nmol/l) compared to 0.01 µg/l (0.2 nmol/l) in the control group. GLS readings were available for seven of the MoM cohort, and were significantly lower when compared with controls (-15.5% vs -18% (MoM vs control); p = 0.025)). Pearson correlation demonstrated that GLS significantly correlated with blood cobalt level (r = 0.8521; p &lt; 0.001). However, there were no differences or correlations for other echocardiography measurements, including LVEF (64.3% vs 63.7% (MoM vs control); p = 0.845). Conclusion This study supports the hypothesis that patients with elevated blood cobalt levels above 13 µg/l in the presence of a MoM hip implant may have impaired cardiac function compared to a control group of patients awaiting hip arthroplasty. It is the first study to use the more sensitive parameter of GLS to assess for any cardiac contractile dysfunction in patients with a MoM hip implant and a normal LVEF. Larger studies should be performed to determine the potential of GLS as a predictor of cardiac complications in patients with MoM arthroplasties

Nanotopology potentiates growth hormone signalling and osteogenesis of mesenchymal stem cells

Custom engineered materials can influence the differentiation of human mesenchymal stem cells (MSCs) towards osteoblasts, chondrocytes and adipocytes, through the control of chemistry, stiffness and nanoscale topography. Here we demonstrate that polycaprolactone growth surfaces engineered with disordered (but controlled) 120 nm diameter dots (NSQ50), but not flat surfaces, promote osteogenic conversion of MSCs in the absence of other osteogenic stimuli. Differentiating MSCs on NSQ50 were found to express growth hormone receptors (GH) and stimulation with recombinant human GH (rhGH) further enhanced NSQ50-driven osteogenic conversion of MSCs. This increased osteogenesis coincided with an enhanced ability of GH to activate ERK MAP kinase on NSQ50, but not on flat topology. The importance of ERK for MSC differentiation was demonstrated by using the inhibitor of ERK activation, U0126, which completely suppressed osteogenesis of GH-stimulated MSCs on NSQ50. The ability of GH to activate ERK in MSCs may therefore be a central control mechanism underlying bone development and growth