The differentiation of progenitor cells is dependent on more than biochemical signalling. Topographical cues in natural bone extracellular matrix guide cellular differentiation through the formation of focal adhesions, contact guidance, cytoskeletal rearrangement and ultimately gene expression. Osteoarthritis and a number of bone disorders present as growing challenges for our society. Hence, there is a need for next generation implantable devices to substitute for, or guide, bone repair in vivo. Cellular responses to nanometric topographical cues need to be better understood in vitro in order to ensure the effective and efficient integration and performance of these orthopedic devices. In this study, the FDA-approved plastic polycaprolactone was embossed with nanometric grooves and the response of primary and immortalized osteoprogenitor cells observed. Nanometric groove dimensions were 240 nm or 540 nm deep and 12.5 μm wide. Cells cultured on test surfaces followed contact guidance along the length of groove edges, elongated along their major axis and showed nuclear distortion; they formed more focal complexes and lower proportions of mature adhesions relative to planar controls. Down-regulation of the osteoblast marker genes RUNX2 and BMPR2 in primary and immortalized cells was observed on grooved substrates. Down-regulation appeared to directly correlate with focal adhesion maturation, indicating the involvement of ERK 1/2 negative feedback pathways following integrin-mediated FAK activation.

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Disseminated breast cancer cells have the capacity to metastasise to the bone marrow and reside in a dormant state within the mesenchymal stem cell niche. Research has focussed on paracrine signalling factors, such as soluble proteins, within the microenvironment. However, it is now clear extracellular vesicles secreted by resident mesenchymal stem cells into this microenvironment also play a key role in the initiation of dormancy. Dormancy encourages reduced cell proliferation and migration, while upregulating cell adhesion, thus retaining the cancer cells within the bone marrow microenvironment. Here, MCF7 breast cancer cells were treated with mesenchymal stem cell–derived extracellular vesicles, resulting in reduced migration in two-dimensional and three-dimensional culture, with reduced cell proliferation and enhanced adhesion, collectively supporting cancer cell dormancy.

The aim of this work is to investigate the use of microtopographies in providing physical cues to modulate the cellular response of human mesenchymal stem cells on ceramics. Two microgrooved patterns (100 μm/50 μm, 10 μm/10 μm groove/pitch) were transcribed reversely onto alumina green ceramic tapes via an embossing technique followed by sintering. Characterization of the micropatterned alumina surfaces and their cellular response was carried out. Spread and polygonal cell morphologies were observed on the wider groove (50 μm/100 μm) surface. Cells seeded onto the narrow groove (10 μm/10 μm) surface aligned themselves alongside the grooves, resulting in more elongated cell morphology. More osteoid matrix nodules shown by osteopontin and osteocalcin biomarkers were detected on the larger grooved surfaces after cell culture of 21 days, indicating a greater level of osteogenicity. This study has shown that micropatterned wider groove (50 μm) topographies are more suitable surfaces for improving osseointegration of ceramic implants.

Nanometric movements of the substrate on which endothelial cells are growing, driven by periodic sinusoidal vibration from 1 Hz to 50 Hz applied by piezo actuators, upregulate endothelin-1 and Kruppel-like factor 2 expression, and increase cell adhesion. These movements are in the z (vertical) axis and ranges from 5 to 50 nm and are similar in vertical extent to protrusions from the cells themselves already reported in the literature. White noise vibrations do not to produce these effects. Vibrational sweeps, if suitably confined within a narrow frequency range, produce similar stimulatory effects but not at wider sweeps. These effects suggest that coherent vibration is crucial for driving these cellular responses. In addition to this, the applied stimulations are observed to be close to or below the random seismic noise of the surroundings, which may suggest stochastic resonance is being employed. The stimulations also interact with the effects of nanometric patterning of the substrates on cell adhesion and Kruppel-like factor 2 and endothelin-1 expression thus linking cell reactions to nanotopographically patterned surfaces with those to mechanical stimulation.

The ability to influence stem cell differentiation is highly desirable as it would help us improve clinical outcomes for patients in various aspects. Many different techniques to achieve this have previously been investigated. This concise study, however, has focused on the topography on which cells grow. Current uncemented orthopaedic implants can fail if the implant fails to bind to the surrounding bone and, typically, forms a soft tissue interface which reduces direct bone contact. Here, we look at the effect of a previously reported nanotopography that utilises nanodisorder to influence mesenchymal stromal cell (as may be found in the bone marrow) differentiation towards bone and to also exert this effect on mature osteoblasts (as may be found in the bone). As topography is a physical technique, it can be envisaged for use in a range of materials such as polymers and metals used in the manufacture of orthopaedic implants.

Graphical abstract

Adult stem cells, such as mesenchymal stem cells, are a multipotent cell source able to differentiate towards multiple cell types. While used widely in tissue engineering and biomaterials research, they present inherent donor variability and functionalities. In addition, their potential to form multiple tissues is rarely exploited. Here, we combine an osteogenic nanotopography and a chondrogenic hyaluronan hydrogel with the hypothesis that we can make a complex tissue from a single multipotent cell source with the exemplar of creating a three-dimensional bone–cartilage boundary environment. Marrow stromal cells were seeded onto the topographical surface and the temperature gelling hydrogel laid on top. Cells that remained on the nanotopography spread and formed osteoblast-like cells, while those that were seeded into or migrated into the gel remained rounded and expressed chondrogenic markers. This novel, simple interfacial environment provides a platform for anisotropic differentiation of cells from a single source, which could ultimately be exploited to sort osteogenic and chondrogenic progenitor cells from a marrow stromal cell population and to develop a tissue engineered interface.