Basic principles of emulsion templating and its use as an emerging manufacturing method of tissue engineering scaffolds

Tissue engineering (TE) aims to regenerate critical size defects, which cannot heal naturally, by using highly porous matrices called TE scaffolds made of biocompatible and biodegradable materials. There are various manufacturing techniques commonly used to fabricate TE scaffolds. However, in most cases, they do not provide materials with a highly interconnected pore design. Thus, emulsion templating is a promising and convenient route for the fabrication of matrices with up to 99% porosity and high interconnectivity. These matrices have been used for various application areas for decades. Although this polymer structuring technique is older than TE itself, the use of polymerised internal phase emulsions (PolyHIPEs) in TE is relatively new compared to other scaffold manufacturing techniques. It is likely because it requires a multidisciplinary background including materials science, chemistry and TE although producing emulsion templated scaffolds is practically simple. To date, a number of excellent reviews on emulsion templating have been published by the pioneers in this field in order to explain the chemistry behind this technique and potential areas of use of the emulsion templated structures. This particular review focusses on the key points of how emulsion templated scaffolds can be fabricated for different TE applications. Accordingly, we first explain the basics of emulsion templating and characteristics of PolyHIPE scaffolds. Then, we discuss the role of each ingredient in the emulsion and the impact of the compositional changes and process conditions on the characteristics of PolyHIPEs. Afterward, current fabrication methods of biocompatible PolyHIPE scaffolds and polymerisation routes are detailed, and the functionalisation strategies that can be used to improve the biological activity of PolyHIPE scaffolds are discussed. Finally, the applications of PolyHIPEs on soft and hard TE as well as in vitro models and drug delivery in the literature are summarised

Boosting the osteogenic and angiogenic performance of multiscale porous polycaprolactone scaffolds by In vitro generated extracellular matrix decoration

Tissue engineering (TE)-based bone grafts are favorable alternatives to autografts and allografts. Both biochemical properties and the architectural features of TE scaffolds are crucial in their design process. Synthetic polymers are attractive biomaterials to be used in the manufacturing of TE scaffolds, due to various advantages, such as being relatively inexpensive, enabling precise reproducibility, possessing tunable mechanical/chemical properties, and ease of processing. However, such scaffolds need modifications to improve their limited interaction with biological tissues. Structurally, multiscale porosity is advantageous over single-scale porosity; therefore, in this study, we have considered two key points in the design of a bone repair material; (i) manufacture of multiscale porous scaffolds made of photocurable polycaprolactone (PCL) by a combination of emulsion templating and three-dimensional (3D) printing and (ii) decoration of these scaffolds with the in vitro generated bone-like extracellular matrix (ECM) to create biohybrid scaffolds that have improved biological performance compared to PCL-only scaffolds. Multiscale porous scaffolds were fabricated, bone cells were cultured on them, and then they were decellularized. The biological performance of these constructs was tested in vitro and in vivo. Mesenchymal progenitors were seeded on PCL-only and biohybrid scaffolds. Cells not only showed improved attachment on biohybrid scaffolds but also exhibited a significantly higher rate of cell growth and osteogenic activity. The chick chorioallantoic membrane (CAM) assay was used to explore the angiogenic potential of the biohybrid scaffolds. The CAM assay indicated that the presence of the in vitro generated ECM on polymeric scaffolds resulted in higher angiogenic potential and a high degree of tissue infiltration. This study demonstrated that multiscale porous biohybrid scaffolds present a promising approach to improve bioactivity, encourage precursors to differentiate into mature bones, and to induce angiogenesis

Synergistic effect of type and concentration of surfactant and diluting solvent on the morphology of emulsion templated matrices developed as tissue engineering scaffolds

Emulsion templating is an advantageous route for the fabrication of tissue engineering scaffolds. Emulsions are mostly stabilised using surfactants, and the performances of the surfactants depend on various parameters such as emulsification temperature and the presence of the electrolytes. In this study, we suggest that diluting solvent type also has a dramatic impact on the efficiency of the surfactant and morphology of the polymerised emulsions. For this, morphologies of polycaprolactone methacrylate-based polymerised emulsions, which are designed for tissue engineering applications and in vitro biocompatibilities, were shown by our group, prepared using four different surfactants, and three different solvents were investigated. Results showed that the diluting solvent used in the emulsion composition has a strong impact on the performance of the surfactant and consequently on the morphology of polymerised emulsions. Increasing surfactant concentration and diluting solvent volume have an opposite impact on the characteristics of emulsions. Scaffolds with average pore sizes changing from 10 to 78 μm could be fabricated. Establishing these relations enables us to have control over the overall morphology of polymerised emulsions and precisely engineer them for specific tissue engineering applications by tuning solvent and surfactant type and concentration

Decellularised extracellular matrix decorated PCL PolyHIPE scaffolds for enhanced cellular activity, integration and angiogenesis

Wound healing involves a complex series of events where cell–cell and cell-extracellular matrix (ECM) interactions play a key role. Wounding can be simple, such as the loss of the epithelial integrity, or deeper and more complex, reaching to subcutaneous tissues, including blood vessels, muscles and nerves. Rapid neovascularisation of the wounded area is crucial for wound healing as it has a key role in supplying oxygen and nutrients during the highly demanding proliferative phase and transmigration of inflammatory cells to the wound area. One approach to circumvent delayed neovascularisation is the exogenous use of pro-angiogenic factors, which is expensive, highly dose-dependent, and the delivery of them requires a very well-controlled system to avoid leaky, highly permeable and haemorrhagic blood vessel formation. In this study, we decorated polycaprolactone (PCL)-based polymerised high internal phase emulsion (PolyHIPE) scaffolds with fibroblast-derived ECM to assess fibroblast, endothelial cell and keratinocyte activity in vitro and angiogenesis in ex ovo chick chorioallantoic membrane (CAM) assays. Our results showed that the inclusion of ECM in the scaffolds increased the metabolic activity of three types of cells that play a key role in wound healing and stimulated angiogenesis in ex ovo CAM assays over 7 days. Herein, we demonstrated that fibroblast-ECM functionalised PCL PolyHIPE scaffolds appear to have great potential to be used as an active wound dressing to promote angiogenesis and wound healing

Assessment of the angiogenic potential of 2-deoxy-D-ribose using a novel in vitro 3D dynamic model in comparison with established in vitro assays

Angiogenesis is a highly ordered physiological process regulated by the interaction of endothelial cells with an extensive variety of growth factors, extracellular matrix components and mechanical stimuli. One of the most important challenges in tissue engineering is the rapid neovascularization of constructs to ensure their survival after transplantation. To achieve this, the use of pro-angiogenic agents is a widely accepted approach. The study of angiogenesis has gained momentum over the last two decades. Although there are various in vitro, ex vivo, and in vivo angiogenesis models that enable testing of newly discovered pro-angiogenic agents, the problem with researching angiogenesis is the choice of the most appropriate assay. In vivo assays are the most representative and reliable models, but they are expensive, time-consuming and can cause ethical concerns whereas in vitro assays are relatively inexpensive, practical, and reproducible, but they are usually lack of enabling the study of more than one aspect of angiogenesis, and they do not fully represent the complexity of physiological angiogenesis. Therefore, there is a need for the development of an angiogenesis model that allows the study of angiogenesis under physiologically more relevant, dynamic conditions without causing ethical concerns. Accordingly, in this study, we developed 3D in vitro dynamic angiogenesis model, and we tested the angiogenic potential of 2-deoxy-D-ribose (2dDR) in comparison with vascular endothelial growth factor (VEGF) using newly developed in vitro 3D dynamic model and well-established in vitro models. Our results obtained using conventional in vitro assays demonstrated that 2dDR promoted proliferation, migration and tube formation of human aortic endothelial cells (HAECs) in a dose-dependent manner. Then, the angiogenic activity of 2dDR was further assessed using the newly developed 3D in vitro model, which enabled the monitoring of cell proliferation and infiltration simultaneously under dynamic conditions. Our results showed that the administration of 2dDR and VEGF significantly enhanced the outgrowth of HAECs and the cellular density under either static or dynamic conditions

Quality characteristics and oxidative stabilityof chicken kavurma formulated with chicken abdominal fat as beef fat replacer

Kavurma is a traditional cooked ready-to-eat meat product that mainly produce in Middle East countries. In kavurma formulation, main ingredients are beef/mutton meat, beef/mutton fat and salt. In this regard, fat has high influence on product’s general characteristics. Due to increasing demand on poultry products, food industry working on novel formulations include chicken meat and chicken abdominal fat. Chicken abdominal fat is an important by-product of chicken meat industry and rich in mono and polyunsaturated fatty acids. For this reason, chicken abdominal fat could be used to improve healthier products. In this study, effects of using chicken abdominal fat (CAF) in chicken kavurma formulation as partial beef fat (BF) replacer on pH, color, textural and sensorial quality and oxidative stability during cold storage (4°C) for 3 months was studied. For this purpose, one control (C: 100% BF) sample and four modified samples, P1 (87.5% BF+12.5% CAF), P2 (75% BF+25% CAF), P3 (62.5% BF+37.5% CAF) and P4 (50% BF+50% CAF), were produced. Proximate composition and texture profile analysis of samples were determined after production whereas pH, lipid oxidation parameters, color and sensory properties of samples were performed on days 0, 30, 60 and 90. Using CAF in kavurma formulations more than 25% resulted higher pH drop during storage, and resulted lower taste and general acceptability scores compared to C samples at the end of storage. P2, P3 and P4 samples had lower TBARS value compared to C during storage period probably result of antioxidative antioxidative ingredients in chicken fed. As expected, due to the semi liquid characteristic of CAF, using this fat type resulted softer products. To sum up, using CAF as BF replacer resulted lower TBARS compared to C, but it had some negative influence on sensory and quality characteristics at high ratio

In vivo bone regeneration capacity of multiscale porous polycaprolactone-based high internal phase emulsion (PolyHIPE) scaffolds in a rat calvarial defect model

Globally, one of the most common tissue transplantation procedures is bone grafting. Lately, we have reported the development of polymerized high internal phase emulsions (PolyHIPEs) made of photocurable polycaprolactone (4PCLMA) and shown their potential to be used as bone tissue engineering scaffolds in vitro. However, it is essential to evaluate the in vivo performance of these scaffolds to investigate their potential in a clinically more relevant manner. Therefore, in this study, we aimed to compare in vivo performances of macroporous (fabricated using stereolithography), microporous (fabricated using emulsion templating), and multiscale porous (fabricated using emulsion templating and perforation) scaffolds made of 4PCLMA. Also, 3D-printed macroporous scaffolds (fabricated using fused deposition modeling) made of thermoplastic polycaprolactone were used as a control. Scaffolds were implanted into a critical-sized calvarial defect, animals were sacrificed 4 or 8 weeks after implantation, and the new bone formation was assessed by micro-computed tomography, dental radiography, and histology. Multiscale porous scaffolds that include both micro- and macropores resulted in higher bone regeneration in the defect area compared to only macroporous or only microporous scaffolds. When one-grade porous scaffolds were compared, microporous scaffolds showed better performance than macroporous scaffolds in terms of mineralized bone volume and tissue regeneration. Micro-CT results revealed that while bone volume/tissue volume (Bv/Tv) values were 8 and 17% at weeks 4 and 8 for macroporous scaffolds, they were significantly higher for microporous scaffolds, with values of 26 and 33%, respectively. Taken together, the results reported in this study showed the potential application of multiscale PolyHIPE scaffolds, in particular, as a promising material for bone regeneration

Preparation of interconnected Pickering polymerized high internal phase emulsions by arrested coalescence

Emulsion templating is a method that enables the production of highly porous and interconnected polymer foams called polymerized high internal phase emulsions (PolyHIPEs). Since emulsions are inherently unstable systems, they can be stabilized either by surfactants or by particles (Pickering HIPEs). Surfactant-stabilized HIPEs form materials with an interconnected porous structure, while Pickering HIPEs typically form closed pore materials. In this study, we describe a system that uses submicrometer polymer particles to stabilize the emulsions. Polymers fabricated from these Pickering emulsions exhibit, unlike traditional Pickering emulsions, highly interconnected large pore structures, and we related these structures to arrested coalescence. We describe in detail the morphological properties of this system and their dependence on different production parameters. This production method might provide an interesting alternative to poly-surfactant-stabilized-HIPEs, in particular where the application necessitates large pore structures

The association of thinking styles with research agendas among academics in the social sciences

Research agendas are understudied, despite being key to academic knowledge creation. The literature suggests that the ways that academics determine their research agendas are conditioned by individual, organisational and environmental characteristics. This study explores the cognitive aspects of academics' research agendas in the social sciences by using a theory on thinking styles as an analytical framework. The results suggest that the research agendas of academics in the social sciences are significantly associated with their thinking styles. These findings aid understanding of how academics set their research agendas. This study also represents an important landmark in research on thinking styles, focusing on academic research work as a potential venue for further studies. The findings are relevant for policymakers, research funding agencies, university administrators and academics because they have implications for academic research development processes, outcomes, and for research and academic identity socialisation during doctoral studies.info:eu-repo/semantics/acceptedVersio

Gelatin-containing porous polycaprolactone PolyHIPEs as substrates for 3D breast cancer cell culture and vascular infiltration

Tumour survival and growth are reliant on angiogenesis, the formation of new blood vessels, to facilitate nutrient and waste exchange and, importantly, provide a route for metastasis from a primary to a secondary site. Whilst current models can ensure the transport and exchange of nutrients and waste via diffusion over distances greater than 200 μm, many lack sufficient vasculature capable of recapitulating the tumour microenvironment and, thus, metastasis. In this study, we utilise gelatin-containing polymerised high internal phase emulsion (polyHIPE) templated polycaprolactone-methacrylate (PCL-M) scaffolds to fabricate a composite material to support the 3D culture of MDA-MB-231 breast cancer cells and vascular ingrowth. Firstly, we investigated the effect of gelatin within the scaffolds on the mechanical and chemical properties using compression testing and FTIR spectroscopy, respectively. Initial in vitro assessment of cell metabolic activity and vascular endothelial growth factor expression demonstrated that gelatin-containing PCL-M polyHIPEs are capable of supporting 3D breast cancer cell growth. We then utilised the chick chorioallantoic membrane (CAM) assay to assess the angiogenic potential of cell-seeded gelatin-containing PCL-M polyHIPEs, and vascular ingrowth within cell-seeded, surfactant and gelatin-containing scaffolds was investigated via histological staining. Overall, our study proposes a promising composite material to fabricate a substrate to support the 3D culture of cancer cells and vascular ingrowth