Additive biofabrication (3D bioprinting) makes it possible to create scaffolds with precise geometries, control over pore interconnectivity and architectures that are not possible with conventional techniques. Inclusion of cells within the ink to form a "bio-ink" presents the potential to print 3D structures that can be implanted into damaged/diseased tissue to promote highly controlled cell-based regeneration and repair. The properties of an 'ink' are defined by its formulation and critically influence the delivery and integrity of structure formed. Importantly, the ink properties need to conform to biological requirements necessary for the cell system that they are intended to support and it is often challenging to find conditions for printing that facilitate this critical aspect of tissue bioengineering. In this study, alginate (Alg) was selected as the major component of the 'bio-ink' formulations for extrusion printing of cells. The rheological properties of alginate-gelatin (AlgGel) blends were compared with pre-crosslinked alginate and alginate solution to establish their printability whilst maintaining their ability to support optimal cell growth. Pre-crosslinked alginate on its own was liquidlike during printing. However, by controlling the temperature, Alg-Gel formulations had higher viscosity, storage modulus and consistency which facilitated higher print resolution/precision. Compression and indentation testing were used to examine the mechanical properties of alginate compared to Alg-Gel. Both types of gels yielded similar results with modulus increasing with alginate concentration. Decay in mechanical properties over time suggests that Alg-Gel slowly degrades in cell culture media with more than 60% decrease in initial modulus over 7 days. The viability of primary myoblasts delivered as a myoblast/Alg-Gel bio-ink was not affected by the printing process, indicating that the Alg-Gel matrix provides a potential means to print 3D constructs that may find application in myoregenerative applications. control over pore interconnectivity and architectures that are not possible with conventional techniques. Inclusion of cells within the ink to form a "bio-ink" presents the potential to print 3D structures that can be implanted into damaged/diseased tissue to promote highly controlled cell-based regeneration and repair. The properties of an 'ink' are defined by its formulation and critically influence the delivery and integrity of structure formed. Importantly, the ink properties need to conform to biological requirements 10 necessary for the cell system that they are intended to support and it is often challenging to find conditions for printing that facilitate this critical aspect of tissue bioengineering. In this study, alginate (Alg) was selected as the major component of the 'bio-ink' formulations for extrusion printing of cells. The rheological properties of alginate-gelatin (Alg-Gel) blends were compared with pre-crosslinked alginate and alginate solution to establish their printability whil...
We present a new approach which aims to translate freeform biofabrication into the surgical field, while staying true to the practical constraints of the operating theatre. Herein we describe the development of a handheld biofabrication tool, dubbed the 'biopen', which enables the deposition of living cells and biomaterials in a manual, direct-write fashion. A gelatin-methacrylamide/hyaluronic acid-methacrylate (GelMa/HAMa) hydrogel was printed and UV crosslinked during the deposition process to generate surgically sculpted 3D structures. Custom titanium nozzles were fabricated to allow printing of multiple ink formulations in a collinear (side-by-side) geometry. Independently applied extrusion pressure for both chambers allows for geometric control of the printed structure and for the creation of compositional gradients. In vitro experiments demonstrated that human adipose stem cells maintain high viability (>97%) one week after biopen printing in GelMa/HAMa hydrogels. The biopen described in this study paves the way for the use of 3D bioprinting during the surgical process. The ability to directly control the deposition of regenerative scaffolds with or without the presence of live cells during the surgical process presents an exciting advance not only in the fields of cartilage and bone regeneration but also in other fields where tissue regeneration and replacement are critical.
Articular cartilage injuries experienced at an early age can lead to the development of osteoarthritis later in life. In situ three-dimensional (3D) printing is an exciting and innovative biofabrication technology that enables the surgeon to deliver tissue-engineering techniques at the time and location of need. We have created a hand-held 3D printing device (biopen) that allows the simultaneous coaxial extrusion of bioscaffold and cultured cells directly into the cartilage defect in vivo in a single-session surgery. This pilot study assessed the ability of the biopen to repair a full-thickness chondral defect and the early outcomes in cartilage regeneration, and compared these results with other treatments in a large animal model. A standardized critical-sized full-thickness chondral defect was created in the weight-bearing surface of the lateral and medial condyles of both femurs of six sheep. Each defect was treated with one of the following treatments: (i) hand-held in situ 3D printed bioscaffold using the biopen (HH group), (ii) preconstructed bench-based printed bioscaffolds (BB group), (iii) microfractures (MF group) or (iv) untreated (control, C group). At 8 weeks after surgery, macroscopic, microscopic and biomechanical tests were performed. Surgical 3D bioprinting was performed in all animals without any intra- or postoperative complication. The HH biopen allowed early cartilage regeneration. The results of this study show that real-time, in vivo bioprinting with cells and scaffold is a feasible means of delivering a regenerative medicine strategy in a large animal model to regenerate articular cartilage.
There has been an emerging interest in stretchable power sources compatible with flexible/wearable electronics. Such power sources must be able to withstand large mechanical strains and still maintain function. Here we report a highly stretchable H3PO4-poly(vinyl alcohol) (PVA) polymer electrolyte obtained by optimizing the polymer molecular weight and its weight ratio to H3PO4 in terms of conductivity and mechanical properties. The electrolyte demonstrates a high conductivity of 3.4 x 10-3 S cm-1, and a high fracture strain at 410% elongation. It is mechanically robust with a tensile strength of 2 MPa and a Young's modulus of 1 MPa, and displays a small plastic deformation (5%) after 1000 stretching cycles at 100% strain. A stretchable supercapacitor device has been developed based on buckled polypyrrole electrodes and the polymer electrolyte. The device shows only a small capacitance loss of 5.6% at 30% strain, and can retain 81% of the initial capacitance after 1000 cycles of such stretching. ABSTRACT: There has been an emerging interest in stretchable power sources compatible with flexible/wearable electronics. Such power sources must be able to withstand large mechanical strains and still maintain function. Here we report a highly stretchable H 3 PO 4 -poly(vinyl alcohol) (PVA) polymer electrolyte obtained by optimizing the polymer molecular weight and its weight ratio to H 3 PO 4 in terms of conductivity and mechanical properties. The electrolyte demonstrates a high conductivity of 3.4×10 -3 S cm -1 , and a high fracture strain at 410% elongation. It is mechanically robust with a tensile strength of 2 MPa and a Young's modulus of 1 MPa, and displays a small plastic deformation (5%) after 1000 stretching cycles at 100% strain. A stretchable supercapacitor device has been developed based on buckled polypyrrole electrodes and the polymer electrolyte. The device shows only a small capacitance loss of 5.6% at 30%strain, and can retain 81% of the initial capacitance after 1000 cycles of such stretching.2
Cellulose nanofibrils (CNFs) in the form of hydrogels stand out as a platform biomaterial in bioink formulation for 3D printing because of their low cytotoxicity and structural similarity to extracellular matrices. In the present study, 3D scaffolds were successfully printed with low-concentration inks formulated by 1 w/v % 2,2,6,6-tetramethylpiperidine-1-oxyl radical (TEMPO)-oxidized CNF with less than 1 w/v % gelatin methacrylate (GelMA). Quartz crystal microbalance with dissipation monitoring (QCM-D) measurements showed strong interaction between the two biopolymers. The UV cross-linking ability of GelMA (≤1 w/v %) was enhanced in the presence of TEMPO-oxidized CNFs. Multiple factors including strong physical interaction between CNF and GelMA, in situ cross-linking of CNF by Ca 2+ , and UV cross-linking of GelMA enabled successful 3D printing of low-concentration inks of CNF/GelMA into scaffolds possessing good structural stability. The mechanical strength of the scaffolds was tuned in the range of 2.5 to 5 kPa. The cell culture with 3T3 fibroblasts revealed noncytotoxic and biocompatible features for the formulated inks and printed scaffolds. More importantly, the incorporated GelMA in the CNF hydrogel promoted the proliferation of fibroblasts. The developed low-concentration CNF/GelMA formulations with a facile yet effective approach to fabricate scaffolds showed great potential in 3D printing for wound healing application.
Frequency domain fluorescence lifetime imaging microscopy (FLIM) has been used in combination with laser scanning confocal microscopy to study the cellular uptake behavior of the antitumor drug doxorubicin (DOX) and micellar-encapsulated DOX (PLyAd-DOX). The endocytosis uptake process of PLyAd-DOX was monitored over 72 hours using confocal microscopy, with a maximum fluorescence recorded at incubation periods around 24 hours. The micellar structure was not found to release the encapsulated DOX during the time course of imaging. FLIM revealed single lifetime distributions of PLyAd-DOX during accumulation in the cytoplasm. The free DOX in contrast was observed both in the cytoplasm and the nuclear domain of the cell, showing bimodal lifetime distributions. There was a marked dependence of the measured free-DOX lifetime on concentration within the cell, in contrast to reference experiments in aqueous solution, where no such dependence was found. The results suggest the formation of macromolecular structures inside the living cells. © 2008 Elsevier Inc. All rights reserved.
In this review, we provide insight into protein interactions with organic conducting polymers, a class of “intelligent” and dynamic materials that offer unique strategies for controlling protein interactions as a prelude to developing a wide range of bioapplications. Following a general introduction on the importance of protein interactions, this review initially focuses on the areas of bioseparation and biosensor applications. These applications amount to an extensive body of work; however, we provide only a brief overview for the purpose of introducing palpable examples of translating the ability to control organic conducting polymer–protein interactions into practical and useful applications. Because organic conducting polymers are breaking new ground for implantable electrodes and tissue regeneration/engineering, we duly turn to the importance of protein interactions and role organic conducting polymers can play in advancing these applications. Lastly, for those not familiar with organic conducting polymers, we take a step back and examine the unique properties underlying their innate ability to control protein interactions, particularly the use of external electrical control to reversibly switch the physical surface properties. Several characterization techniques identified as being critical to our understanding at the macroscopic down to the single molecule level are also highlighted.
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