Stem cells within epithelial tissues reside in anatomical structures known as crypts that are known to contribute to the mechanical and chemical milieu important for function. To date, epithelial stem cell therapies have largely ignored the niche and focussed solely on the cell population to be transplanted. Our aim was to recreate the precise geometry of the epithelial stem cell niche using two photon polymerisation and to determine the influence of this structure alone on stem cell phenotype. We were able to recreate crypt structures and following cell seeding, a zonation in cell phenotype along the z-axis emerged. This illustrates that geometry alone, without the use of exogenous signalling molecules, influences cell response. Understanding the role of geometry in the regulation of the stem cell niche will enable significant advances in our ability to influence stem cell behaviour to expedite cellular therapies to the clinic.
Corneal infection, inflammation and injury can result in scarring and loss of transparency of the cornea, which accounts for one to two million cases of monocular blindness worldwide. Modulating and promoting wound healing of the cornea has proved to be a major challenge. The amniotic membrane is currently used as a biological bandage to achieve this; however, many limitations have been shown to exist with this technique. Hydrogels are large polymeric networks that have a high fluid content. Such polymers can be manipulated to switch between liquid and solid states depending on light, temperature and ionic changes and can be either biodegradable or nonbiodegradable. Thermosensitive hydrogels can be manipulated to form gels at body temperature and may hold potential as synthetic scaffolds for ocular surface reconstruction, wound healing and ophthalmic drug delivery.
Orthogonal wettability and topographical gradients in a combinatorial sample format are fabricated using plasma-polymer-coated microgrooved surfaces. Preferred cell proliferation is found on specific combinations of topography and chemistry. This proof-of-concept study demonstrates the potential applications of this sample format for investigating the relationship between multiple surface properties on cellular response in a high-throughput manner.
The fabrication and application of a biocompatible peptide conjugated thermo-responsive fibrous scaffolds for cellular phenotype support and enzymatic-free passaging of mammalian cells.
Using phantom samples, we investigated the feasibility of spatially-offset Raman spectroscopy (SORS) as a tool for monitoring non-invasively the mineralization of bone tissue engineering scaffold in-vivo. The phantom samples consisted of 3D-printed scaffolds of poly-caprolactone (PCL) and hydroxyapatite (HA) blends, with varying concentrations of HA, to mimic the mineralisation process. The scaffolds were covered by a 4 mm layer of skin to simulate the real in-vivo measurement conditions. At a concentration of HA approximately 1/3 that of bone (~0.6 g/cm3), the characteristic Raman band of HA (960 cm-1) was detectable when the PCL:HA layer was located at 4 mm depth within the scaffold (i.e. 8 mm below the skin surface). For the layers of the PCL:HA immediately under the skin (i.e. top of the scaffold), the detection limit of HA was 0.18 g/cm3, which is approximately one order of magnitude lower than that of bone. Similar results were also found for the phantoms simulating uniform and inward gradual mineralisation of the scaffold, indicating the suitability of SORS to detect early stages of mineralisation. Nevertheless, the results also show that the contribution of the materials surrounding the scaffold can be significant and methods for subtraction need to be investigated in the future. In conclusion, these results indicate that spatially-offset Raman spectroscopy is a promising technique for in-vivo longitudinal monitoring scaffold mineralization and bone re-growth.
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