Methacrylated gelatin (GelMA) has been intensively studied as a 3D printable scaffold material in tissue regeneration fields, which can be attributed to its well-known biological functions. However, the long-term stability of photo-crosslinked GelMA scaffolds is hampered by a combination of its fast degradation in the presence of collagenase and the loss of physical crosslinks at higher temperatures. To increase the longer-term shape stability of printed scaffolds, a mixture of GelMA and tyramine-conjugated 8-arm PEG (8PEGTA) was used to create filaments composed of an interpenetrating network (IPN). Photo-crosslinking during filament deposition of the GelMA and subsequent enzymatic crosslinking of the 8PEGTA were applied to the printed 3D scaffolds. Although both crosslinking mechanisms are radical based, they operate without interference of each other. Rheological data of bulk hydrogels showed that the IPN was an elastic hydrogel, having a storage modulus of 6 kPa, independent of temperature in the range of 10 - 40°C. Tensile and compression moduli were 110 kPa and 80 kPa, respectively. On enzymatic degradation in the presence of collagenase, the gelatin content of the IPN fully degraded in 7 days, leaving a stable secondary crosslinked 8PEGTA network. Using a BioMaker bioprinter, hydrogels without and with human osteosarcoma cells (hMG-63) were printed. On culturing for 21 days, hMG-63 in the GelMA/8PEGTA IPN showed a high cell viability (>90%). Thus, the presence of the photoinitiator, incubation with H2O2, and mechanical forces during printing did not hamper cell viability. This study shows that the GelMA/8PEGTA ink is a good candidate to generate cell-laden bioinks for extrusion-based printing of constructs for tissue engineering applications.

This review on hybrid hydrogels prepared from methacrylated gelatin (GelMA) and synthetic polymers presents their advantageous properties and potential applications in biomedical engineering. It describes the preparation of the different types of frequently used hybrid hydrogel networks: co-networks, interpenetrating networks (IPNs), and semi-interpenetrating networks (semi-IPNs) and gives an overview of the different methods of preparing biomedical devices from these materials.

Endothelial cell seeding to improve the performance of small-diameter vascular grafts requires a suitable substrate, such as crosslinked collagen. In addition to providing a suitable substrate for adhesion and growth of endothelial cells, proliferation of seeded endothelial cells can be enhanced by local, sustained release of basic fibroblast growth factor (bFGF, a heparin-binding growth factor for endothelial cells). We have previously shown that collagen crosslinked using N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide (EDC) and N-hydroxysuccinimide (NHS) supports adhesion and proliferation of human umbilical vein endothelial cells (HUVECs). In the present study, HUVECs were seeded on (heparinized) EDC/NHS-crosslinked collagen, pre-loaded with bFGF. Proliferation of HUVECs on (heparinized) crosslinked collagen increased with increasing amounts of pre-loaded bFGF. The minimal cell-seeding density required for proliferation proved to be very low after pre-loading the substrates with bFGF, and was 4-fold lower for heparinized crosslinked collagen compared to crosslinked collagen (250 versus 1000 cells/cm(2)). Pro-coagulant properties (von Willebrand factor secretion and tissue factor expression) of HUVECs seeded on (heparinized) crosslinked collagen, with or without pre-loading of bFGF, were comparable to those of HUVECs on TCPS. It is concluded that heparinized, EDC/NHS-crosslinked collagen pre-loaded with bFGF is a candidate matrix for in vivo endothelial cell seeding of synthetic vascular graft materials.

In recent years, organs-on-chips (OOCs) have been developed to meet the desire for more realistic in vitro cell culture models. These systems introduce microfluidics, mechanical stretch and other physiological stimuli to in vitro models, thereby significantly enhancing their descriptive power. In most OOCs, porous polymeric membranes are used as substrates for cell culture. The polymeric material, morphology and shape of these membranes are often suboptimal, despite their importance for achieving ideal cell functionality such as cell–cell interaction and differentiation. The currently used membranes are flat and thus do not account for the shape and surface morphology of a tissue. Moreover, the polymers used for fabrication of these membranes often lack relevant characteristics, such as mechanical properties matching the tissue to be developed and/or cytocompatibility. Recently, innovative techniques have been reported for fabrication of porous membranes with suitable porosity, shape and surface morphology matching the requirements of OOCs. In this paper, we review the state of the art for developing these membranes and discuss their application in OOCs.

Vascular grafts with an inner diameter less than 6 mm are urgently needed due to the increasing prevalence of vascular disease. In this study, tubular scaffolds for vascular tissue engineering were fabricated by photo‐crosslinking of acrylate‐functionalized poly(trimethylene carbonate) (PTMC) macromers of different molecular weights in a glass mold. Porous structures were prepared by means of salt leaching. Tubular scaffolds were obtained with an inner diameter of 3 mm, a wall thickness of 1 mm, and a length of 4.5 cm. Pore sizes ranged from 0 to 290 µm, and the porosity was around 70%. The pores were homogeneously distributed and interconnected. PTMC macromers with a molecular weight of 4, 8, 13, 17, and 22 kg/mol were used. With increasing PTMC macromer molecular weight from 4 to 22 kg/mol, the E‐modulus and maximum tensile strength of the scaffolds in the radial direction increased from 0.56 to 1.12 MPa and 0.12 to 0.55 MPa, respectively. Stress–strain curves for scaffolds made of 13, 17, and 22 kg/mol PTMC macromers showed a “toe” region characteristic for native arteries, followed by a linear increase until the maximum stress was reached. The E‐moduli of the latter scaffolds are comparable to those of native arteries, whereas the maximum tensile strengths are approximately fourfold lower. This can be improved, however, by cell seeding in the porous scaffolds and subsequent mechanical stimulation in a bioreactor. It is concluded that the porous tubular scaffolds made of 13, 17, and 22 kg/mol PTMC macromers are suitable scaffolds for vascular tissue engineering. Copyright © 2016 John Wiley & Sons, Ltd.

The intervertebral disc (IVD) is the most avascular and acellular tissue in the body and therefore prone to degeneration. During IVD degeneration, the balance between anabolic and catabolic processes in the disc is deregulated, amongst others leading to alteration of extracellular matrix production, abnormal enzyme activities and production of pro-inflammatory substances like cytokines. The established treatment strategy for IVD degeneration consists of physiotherapy, pain medication by drug therapy and if necessary surgery. This approach, however, has shown limited success. Alternative strategies to increase and prolong the effects of bioactive agents and to reverse the process of IVD degeneration include the use of delivery systems for drugs, proteins, cells and genes. In view of the specific anatomy and physiology of the IVD and depending on the strategy of the therapy, different delivery systems have been developed which are reviewed in this article.

Optimal cell interaction with biomaterial scaffolds is one of the important requirements for the development of successful in vitro tissue-engineered tissues. Fast, efficient and spatially uniform cell adhesion can improve the clinical potential of engineered tissue. Three-dimensional (3-D) solid free form fabrication is one widely used scaffold fabrication technique today. By means of deposition of polymer fibers, scaffolds with various porosity, 3-D architecture and mechanical properties can be prepared. These scaffolds consist mostly of solid round fibers. In this study, it was hypothesized that a corrugated fiber morphology enhances cell adhesion and proliferation and therefore leads to the development of successful in vitro tissue-engineered constructs. Corrugated round fibers were prepared and characterized by extruding poly(ethylene oxide terephthalate)-co-poly(butylene terephthalate) (300PEOT55PBT45) block co-polymer through specially designed silicon wafer inserts. Corrugated round fibers with 6 and 10 grooves on the fiber surface were compared with solid round fibers of various diameters. The culture of mouse pre-myoblast (C2C12) cells on all fibers was studied under static and dynamic conditions by means of scanning electron microscopy, cell staining and DNA quantification. After 7days of culturing under static conditions, the DNA content on the corrugated round fibers was approximately twice as high as that on the solid round fibers. Moreover, under dynamic culture conditions, the cells on the corrugated round fibers seemed to experience lower mechanical forces and therefore adhered better than on the solid round fibers. The results of this study show that the surface architecture of fibers in a tissue engineering scaffold can be used as a tool to improve the performance of the scaffold in terms of cell adhesion and proliferation.

For the treatment of cardiovascular disease, functional arterial blood vessel prostheses with an inner diameter less than 6mm are needed. This article gives an overview of the preparation of such vascular grafts by means of tissue engineering.

The use of nanoparticles in biomedical applications is emerging rapidly. Recent developments have led to numerous studies of noble metal nanoparticles, down to the level of single molecule detection in living cells. The application of noble metal nanoparticles in diagnostics and treatment of early stage carcinomas is the subject of many present studies. Gold nanoparticles are particularly interesting for optical biomedical applications due to their biocompatibility and moreover, their enhanced absorption cross-sections. The latter is a result of surface plasmon resonance, which can be tuned by altering the shape of the nanoparticles enabling usage of the near infrared tissue transparent optical window. This paper presents a brief overview of the variety of shapes, size and surface chemistries of the gold nanoparticles used for cancer detection and treatment, as well as their effects in different tumour models that have recently been investigated, both in vitro and in vivo.

The physiology of vascular endothelial cells is strongly affected by fluid shear stress on their surface. In this study, a microfluidic assay was employed to analyze the alignment of actin filaments in endothelial cells in response to shear stress. When cells were cultured in microfluidic channels and subjected to shear stress, the alignment of filaments in the channel direction was significantly higher than in static cultures. By adding inhibitory drugs, the roles of several signaling proteins in the process of alignment were determined. Thus, it is shown how microfluidic technology can be employed to provide a mechanistic insight into cell physiology.

Vascular endothelial cells form the inner lining of all blood vessels and play a central role in vessel physiology and disease. Endothelial cells are highly responsive to the mechanical stimulus of fluid shear stress that is exerted by blood flowing over their surface. In this study, the immediate micromechanical response of endothelial cells to physiological shear stress was characterized by tracking of ballistically injected, sub-micron, fluorescent particles. It was found that the mean squared displacement (MSD) of the particles decreases by a factor 1.5 within 10 min after the onset of shear stress. This decrease in particle motion is transient, since the MSD returns to control values within 15-30 min after the onset of shear. The immediate micromechanical stiffening is dependent on activation of the vascular endothelial growth factor receptor (VEGFR)-2, because inhibition of the receptor abrogates the micromechanical response. This work shows that the cytoskeleton is actively involved in the acute, functional response of endothelial cells to shear stress.

Acceptance of microfluidic technology in everyday laboratory practice by biologists is still low. One of the reasons for this is that the technology combines poorly with standard cell biological and biochemical analysis tools. Flow cytometry is an example of a conventional analytical tool that is considered to be incompatible with microfluidic technology and its inherent small sample sizes. In this study, it is shown that properly designed microfluidic devices contain cell populations that are large enough to be analyzed by flow cytometry. To illustrate this, the uptake of fluorescent human low‐density lipoprotein (LDL) by human endothelial cells that were cultured in a microfluidic channel was analyzed. It was found that the uptake of LDL by the cells increased linearly over time. Moreover, the uptake decreased when cells were pretreated with fluid shear stress inside the microfluidic devices. This study shows that microfluidic technology can be combined with conventional flow cytometry, while retaining the advantages of working with microfluidics such as low reagent use and dynamic cell culture conditions. This approach of combining microfluidic technology with conventional laboratory tools may contribute to greater acceptance of microfluidic devices in biological research. © 2010 International Society for Advancement of Cytometry

When studying particle uptake in vitro, it is favorable to mimic the in vivo situation as much as possible. In this study we present a microfluidic device to mimic the mechanical stress caused by the flow of blood while studying particle uptake in vitro. Human endothelial cells were treated with liposomes containing fluorescent siRNA. It was found that applying physiologically relevant mechanical stress during transfection diminishes the uptake of liposomes in the cells.

Purpose: The effect of heat treatment in combination with X-irradiation was examined with regard to expression of p53, a tumor suppressor gene product, and Hsp70, a heat-shock protein, in association with the occurrence of programmed cell death (apoptosis).Materials and methods: Three hematopoietic cell lines (HSB2, HL60 and Kasumi-1), which differ in p53 status, were exposed to 42.5 degrees C during one hour and/or X-radiation (total dose 8 Gy). After exposure, both mRNA and protein expression levels of Hsp70 and p53 were investigated by real-time PCR (polymerase chain reaction) and Western blotting. Apoptosis was simultaneously analyzed by observation of cell morphology as well as flowcytometric determination of Annexin V binding to phosphatidylserine and propidium iodide exclusion.Results: Both HL60 and HSB2 cell lines with a low p53 status and a quick response to heat treatment with Hsp70 overexpression are less susceptible to heat-induced apoptosis compared to Kasumi-1 cells with wild-type p53 protein and no Hsp70 response. The combination of first applying X-irradiation followed by heat treatment resulted in the most effective induction of apoptosis due to impairment of the Hsp70 response in all three cell lines.Conclusion: These results indicate that the Hsp70 response and p53 status mediate the susceptibility of hematopoietic cells to undergo heat-induced apoptosis. Therefore, these parameters can be used as markers to predict the effectiveness of hyperthermia in cancer treatment.