Wet-Spun Trojan Horse Cell Constructs for Engineering Muscle

Engineering of 3D regenerative skeletal muscle tissue constructs (skMTCs) using hydrogels containing muscle precursor cells (MPCs) is of potential benefit for repairing Volumetric Muscle Loss (VML) arising from trauma (e.g., road/industrial accident, war injury) or for restoration of functional muscle mass in disease (e.g., Muscular Dystrophy, muscle atrophy). Additive Biofabrication (AdBiofab) technologies make possible fabrication of 3D regenerative skMTCs that can be tailored to specific delivery requirements of VML or functional muscle restoration. Whilst 3D printing is useful for printing constructs of many tissue types, the necessity of a balanced compromise between cell type, required construct size and material/fabrication process cyto-compatibility can make the choice of 3D printing a secondary alternative to other biofabrication methods such as wet-spinning. Alternatively, wet-spinning is more amenable to formation of fibers rather than (small) layered 3D-Printed constructs. This study describes the fabrication of biosynthetic alginate fibers containing MPCs and their use for delivery of dystrophin-expressing cells to dystrophic muscle in the mdx mouse model of Duchenne Muscular Dystrophy (DMD) compared to poly(DL-lactic-co-glycolic acid) copolymer (PLA:PLGA) topically-seeded with myoblasts. In addition, this study introduces a novel method by which to create 3D layered wet-spun alginate skMTCs for bulk mass delivery of MPCs to VML lesions. As such, this work introduces the concept of “Trojan Horse” Fiber MTCs (TH-fMTCs) and 3d Mesh-MTCs (TH-mMTCs) for delivery of regenerative MPCs to diseased and damaged muscle, respectively

Handheld Co-Axial Bioprinting: Application to in situ surgical cartilage repair

Three-dimensional (3D) bioprinting is driving major innovations in the area of cartilage tissue engineering. Extrusion-based 3D bioprinting necessitates a phase change from a liquid bioink to a semi-solid crosslinked network achieved by a photo-initiated free radical polymerization reaction that is known to be cytotoxic. Therefore, the choice of the photocuring conditions has to be carefully addressed to generate a structure stiff enough to withstand the forces phisiologically applied on articular cartilage, while ensuring adequate cell survival for functional chondral repair. We recently developed a handheld 3D printer called Biopen . To progress towards translating this freeform biofabrication tool into clinical practice, we aimed to define the ideal bioprinting conditions that would deliver a scaffold with high cell viability and structural stiffness relevant for chondral repair. To fulfill those criteria, free radical cytotoxicity was confined by a co-axial Core/Shell separation. This system allowed the generation of Core/Shell GelMa/HAMa bioscaffolds with stiffness of 200KPa, achieved after only 10seconds of exposure to 700mW/cm2 of 365nm UV-A, containing \u3e90% viable stem cells that retained proliferative capacity. Overall, the Core/Shell handheld 3D bioprinting strategy enabled rapid generation of high modulus bioscaffolds with high cell viability, with potential for in situ surgical cartilage engineering

In vitro growth and differentiation of primary myoblasts on thiophene based conducting polymers

Polythiophenes are attractive candidate polymers for use in synthetic cell scaffolds as they are amenable to modification of functional groups as a means by which to increase biocompatibility. In the current study we analysed the physical properties and response of primary myoblasts to three thiophene polymers synthesized from either a basic bithiophene monomer or from one of two different thiophene monomers with alkoxy functional groups. In addition, the effect of the dopants pTS- and ClO4 - was investigated. In general, it was found that pTS- doped polymers were significantly smoother and tended to be more hydrophilic than their ClO 4 - doped counterparts, demonstrating that the choice of dopant significantly affects the polythiophene physical properties. These properties had a significant effect on the response of primary myoblasts to the polymer surfaces; LDH activity measured from cells harvested at 24 and 48 h post-seeding revealed significant differences between numbers of cells attaching to the different thiophene polymers, whilst all of the polymers equally supported cell doubling over the 48 h period. Differences in morphology were also observed, with reduced cell spreading observed on polymers with alkoxy groups. In addition, significant differences were seen in the polymers\u27 ability to support myoblast fusion. In general pTS- doped polymers were better able to support fusion than their ClO4 - doped counterparts. These studies demonstrate that modification of thiophene polymers can be used to promote specific cellular response (e.g. proliferation over differentiation) without the use of biological agents. 2013 The Royal Society of Chemistry

Bioavailability of Macro and Micronutrients Across Global Topsoils: Main Drivers and Global Change Impacts

Understanding the chemical composition of our planet\u27s crust was one of the biggest questions of the 20th century. More than 100 years later, we are still far from understanding the global patterns in the bioavailability and spatial coupling of elements in topsoils worldwide, despite their importance for the productivity and functioning of terrestrial ecosystems. Here, we measured the bioavailability and coupling of thirteen macro- and micronutrients and phytotoxic elements in topsoils (3–8 cm) from a range of terrestrial ecosystems across all continents (∼10,000 observations) and in response to global change manipulations (∼5,000 observations). For this, we incubated between 1 and 4 pairs of anionic and cationic exchange membranes per site for a mean period of 53 days. The most bioavailable elements (Ca, Mg, and K) were also amongst the most abundant in the crust. Patterns of bioavailability were biome-dependent and controlled by soil properties such as pH, organic matter content and texture, plant cover, and climate. However, global change simulations resulted in important alterations in the bioavailability of elements. Elements were highly coupled, and coupling was predictable by the atomic properties of elements, particularly mass, mass to charge ratio, and second ionization energy. Deviations from the predictable coupling-atomic mass relationship were attributed to global change and agriculture. Our work illustrates the tight links between the bioavailability and coupling of topsoil elements and environmental context, human activities, and atomic properties of elements, thus deeply enhancing our integrated understanding of the biogeochemical connections that underlie the productivity and functioning of terrestrial ecosystems in a changing world

A common variant near TGFBR3 is associated with primary open angle glaucoma

Primary open angle glaucoma (POAG), a major cause of blindness worldwide, is a complex disease with a significant genetic contribution. We performed Exome Array (Illumina) analysis on 3504 POAG cases and 9746 controls with replication of the most significant findings in 9173 POAG cases and 26 780 controls across 18 collections of Asian, African and European descent. Apart from confirming strong evidence of association at CDKN2B-AS1 (rs2157719 [G], odds ratio [OR] = 0.71, P = 2.81 × 10−33), we observed one SNP showing significant association to POAG (CDC7–TGFBR3 rs1192415, ORG-allele = 1.13, Pmeta = 1.60 × 10−8). This particular SNP has previously been shown to be strongly associated with optic disc area and vertical cup-to-disc ratio, which are regarded as glaucoma-related quantitative traits. Our study now extends this by directly implicating it in POAG disease pathogenesis

Engineering skeletal muscle - from two to three dimensions

Large skeletal muscle defects remain a challenge for pa- tients and surgeons alike. While small injuries resulting in tears and lacerations can be repaired by the body\u27s own regenerative mechanisms, loss of over 20% of muscle will lead to scarring, denervation and loss of function (Turner and Badylak, 2012). This can occur in cases of trauma, tumour, surgery and degenerative disease, which overwhelm the body\u27s repair mechanisms and result in changed anatomy and weakened muscle groups. Loss of limb is another consequence of these disease processes. Restoring mobility and independence is a major recon- structive challenge, for which tissue engineering may of- fer a solution

Can the wet - State conductivity of hydrogels be improved by incorporation of spherical conducting nanoparticles?

In nerve and muscle regeneration applications, the incorporation of conducting elements into biocompatible materials has gained interest over the last few years, as it has been shown that electrical stimulation of some regenerating cells has a positive effect on their development. A variety of different materials, ranging from graphene to conducting polymers, have been incorporated into hydrogels and increased conductivities have been reported. However, the majority of conductivity measurements are performed in a dry state, even though material blends are designed for applications in a wet state, in vivo environment. The focus of this work is to use polypyrrole nanoparticles to increase the wet-state conductivity of alginate to produce a conducting, easily processable, cell-supporting composite material. Characterization and purification of the conducting polymer nanoparticle dispersions, as well as electrochemical measurements, have been performed to assess conductivity of the nanoparticles and hydrogel composites in the wet state, in order to determine whether filling an ionically conducting hydrogel with electrically conductive nanoparticles will enhance the conductivity. It was determined that the introduction of spherical nanoparticles into alginate gel does not increase, but rather slightly reduces conductivity of the hydrogel in the wet state

Development of self-assembling peptide inspired bioinks for neural tissue engineering applications

© 2019 Omnipress - All rights reserved. Statement of Purpose: The ability to fabricate artificial tissue constructs offers significant promise to regenerative medicine and tissue engineering. Advances in additive manufacturing have enabled the bioprinting of spatially defined cell-laden constructs capable of providing spatiotemporal presentation of biological and physical cues to cells in a multicomponent structure. 1 Despite significant advance in bioprinting techniques, a key challenge remains in developing soft biomaterials that can adequately recapitulate the complexities of native extracellular matrix (ECM) whilst facilitating bioprinting. Self-Assembling Peptides (SAPs) are a unique class of hydrogels which spontaneously immobilize surrounding solvents through formation of biomimetic structures, resulting in hydrogel networks highly reminiscent of native ECM. Through strategic engineering of SAP sequences, ECM inspired bioactive motifs may be introduced. IKVAV, a laminin motif, can be used to generate Fmoc-DIKVAV (a fibril forming SAP) shown to support the survival and migration of transplanted cortical neural progenitor cells in vivo. 2 Despite Fmoc-DIKVAV’s excellent biocompatibility, the inherent lack of covalent bonds between fibrils results in poor printability and extensive swelling in vitro. For this reason, there is a key need to modify this material to enhance printability without compromising bioactivity

Cell compatible encapsulation of filaments into 3D hydrogels

Tissue engineering scaffolds for nerve regeneration, or artificial nerve conduits, are particularly challenging due to the high level of complexity the structure of the nerve presents. The list of requirements for artificial nerve conduits is long and includes the ability to physically guide nerve growth using physical and chemical cues as well as electrical stimulation. Combining these characteristics into a conduit, while maintaining biocompatibility and biodegradability, has not been satisfactorily achieved by currently employed fabrication techniques. Here we present a method combining pultrusion and wet-spinning techniques facilitating incorporation of pre-formed filaments into ionically crosslinkable hydrogels. This new biofabrication technique allows the incorporation of conducting or drug-laden filaments, controlled guidance channels and living cells into hydrogels, creating new improved conduit designs

Lubricin on Platinum Electrodes: A Low-Impedance Protein-Resistant Surface Towards Biomedical Implantation

Biofouling on surfaces compromises the function of biomedical devices whose function involves contact with biological fluids. In the context of electrochemical devices, proteins are attracted to the surface via coaction of various forces (hydrogen bond, hydrophobic effect, and other polar interactions) and protein interaction with the surface can significantly alter the surface chemistry. In response to this issue, we have developed an efficient anti-biofouling surface that employs a glycoprotein, lubricin (LUB), and which generates low impedance layers compatible with electrochemical applications. Herein, we investigate how different LUB densities on platinum (Pt) electrodes affect the surface electrochemistry and its ability to prevent nonspecific adsorption of protein to the surface. Surfaces with higher densities of LUB were more resistant to protein adsorption. The LUB modified Pt electrodes were challenged in artificial perilymph (AP) media under passive and electrically stimulated conditions over 7-day periods throughout which the LUB layer retained its anti-biofouling and surface coating stability