Engineering periodontal tissue interfaces using multiphasic scaffolds and membranes for guided bone and tissue regeneration.

Periodontal diseases are one of the greatest healthcare burdens worldwide. The periodontal tissue compartment is an anatomical tissue interface formed from the periodontal ligament, gingiva, cementum, and bone. This multifaceted composition makes tissue engineering strategies challenging to develop due to the interface of hard and soft tissues requiring multiphase scaffolds to recreate the native tissue architecture. Multilayer constructs can better mimic tissue interfaces due to the individually tuneable layers. They have different characteristics in each layer, with modulation of mechanical properties, material type, porosity, pore size, morphology, degradation properties, and drug-releasing profile all possible. The greatest challenge of multilayer constructs is to mechanically integrate consecutive layers to avoid delamination, especially when using multiple manufacturing processes. Here, we review the development of multilayer scaffolds that aim to recapitulate native periodontal tissue interfaces in terms of physical, chemical, and biological characteristics. Important properties of multiphasic biodegradable scaffolds are highlighted and summarised, with design requirements, biomaterials, and fabrication methods, as well as post-treatment and drug/growth factor incorporation discussed.

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.

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.

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.

Thiolene- and Polycaprolactone Methacrylate-Based Polymerized High Internal Phase Emulsion (PolyHIPE) Scaffolds for Tissue Engineering.

Highly porous emulsion templated polymers (PolyHIPEs) provide a number of potential advantages in the fabrication of scaffolds for tissue engineering and regenerative medicine. Porosity enables cell ingrowth and nutrient diffusion within, as well as waste removal from, the scaffold. The properties offered by emulsion templating alone include the provision of high interconnected porosity, and, in combination with additive manufacturing, the opportunity to introduce controlled multiscale porosity to complex or custom structures. However, the majority of monomer systems reported for PolyHIPE preparation are unsuitable for clinical applications as they are nondegradable. Thiol-ene chemistry is a promising route to produce biodegradable photocurable PolyHIPEs for the fabrication of scaffolds using conventional or additive manufacturing methods; however, relatively little research has been reported on this approach. This study reports the groundwork to fabricate thiol- and polycaprolactone (PCL)-based PolyHIPE materials via a photoinitiated thiolene click reaction. Two different formulations, either three-arm PCL methacrylate (3PCLMA) or four-arm PCL methacrylate (4PCLMA) moieties, were used in the PolyHIPE formulation. Biocompatibility of the PolyHIPEs was investigated using human dermal fibroblasts (HDFs) and human osteosarcoma cell line (MG-63) by DNA quantification assay, and developed PolyHIPEs were shown to be capable of supporting cell attachment and viability.

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.

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.

Landmarks in vaginal mesh development: polypropylene mesh for treatment of SUI and POP.

Vaginal meshes used in the treatment of stress urinary incontinence (SUI) and pelvic organ prolapse (POP) have produced highly variable outcomes, causing life-changing complications in some patients while providing others with effective, minimally invasive treatments. The risk:benefit ratio when using vaginal meshes is a complex issue in which a combination of several factors, including the inherent incompatibility of the mesh material with some applications in pelvic reconstructive surgeries and the lack of appropriate regulatory approval processes at the time of the premarket clearance of these products, have contributed to the occurrence of complications caused by vaginal mesh. Surgical mesh used in hernia repair has evolved over many years, from metal implants to knitted polymer meshes that were adopted for use in the pelvic floor for treatment of POP and SUI. The evolution of the material and textile properties of the surgical mesh was guided by clinical feedback from hernia repair procedures, which were also being modified to obtain the best outcomes with use of the mesh. Current evidence shows how surgical mesh fails biomechanically when used in the pelvic floor and materials with improved performance can be developed using modern material processing and tissue engineering techniques.

A Novel Bilayer Polycaprolactone Membrane for Guided Bone Regeneration: Combining Electrospinning and Emulsion Templating.

Guided bone regeneration is a common dental implant treatment where a barrier membrane (BM) is used between epithelial tissue and bone or bone graft to prevent the invasion of the fast-proliferating epithelial cells into the defect site to be able to preserve a space for infiltration of slower-growing bone cells into the periodontal defect site. In this study, a bilayer polycaprolactone (PCL) BM was developed by combining electrospinning and emulsion templating techniques. First, a 250 µm thick polymerised high internal phase emulsion (polyHIPE) made of photocurable PCL was manufactured and treated with air plasma, which was shown to enhance the cellular infiltration. Then, four solvent compositions were investigated to find the best composition for electrospinning a nanofibrous PCL barrier layer on PCL polyHIPE. The biocompatibility and the barrier properties of the electrospun layer were demonstrated over four weeks in vitro by histological staining. Following in vitro assessment of cell viability and cell migration, cell infiltration and the potential of PCL polyHIPE for supporting blood vessel ingrowth were further investigated using an ex-ovo chick chorioallantoic membrane assay. Our results demonstrated that the nanofibrous PCL electrospun layer was capable of limiting cell infiltration for at least four weeks, while PCL polyHIPE supported cell infiltration, calcium and mineral deposition of bone cells, and blood vessel ingrowth through pores.

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.

Development of a 2-dof uterine manipulator with LED illumination system as a new transvaginal uterus amputation device for gynecological surgeries.

BACKGROUND: Hysterectomy, the most common major gynecological operation worldwide, consists of removal of the uterus and can be performed abdominally, vaginally, or laparoscopically. A uterine manipulator is a key device used for uterine manipulation and cannulation in hysterectomies. The challenges of conventional manipulators are to move the uterus in two distinct planes and to identify cervical landmarks during circular cut and coagulation. MATERIAL AND METHODS: In this study, a structural synthesis of the two degrees of freedom parallel manipulator was performed considering the constraints noted by surgeons. Computer-aided design and assembly of the manipulator, the cervicovaginal cap with LEDs, and the external parts were performed before rapid prototyping. The final design of the uterine manipulator was then manufactured from stainless steel and tested on an artificial uterus model using a test chamber. RESULTS: This article presents the design, production and testing processes of an innovative manipulator with a motion capability up to 80° workspace both in the sagittal and coronal planes and an illumination system, easily detectable by the laparoscope, was successfully implemented on the manipulator's cervical cap in order to overcome the drawbacks of conventional uterine manipulators. CONCLUSIONS: Despite all the current studies and uterine manipulators on the market, no research has incorporated all the features mentioned above.