Acta Biomaterialia

Bioprinting of stem cell expansion lattices Publication date: Available online 13 May 2019 Source: Acta Biomaterialia Author(s): Christopher D. Lindsay, Julien G. Roth, Bauer L. LeSavage, Sarah C. Heilshorn Abstract Stem cells have great potential in regenerative medicine, with neural progenitor cells (NPCs) being developed as a therapy for many central nervous system diseases and injuries. However, one limitation to the clinical translation of stem cells is the resource-intensive, two-dimensional culture protocols required for biomanufacturing a clinically-relevant number of cells. This challenge can be overcome in an easy-to-produce and cost-effective 3D platform by bioprinting NPCs in a layered lattice structure. Here we demonstrate that alginate biopolymers are an ideal bioink for expansion lattices and do not require chemical modifications for effective NPC expansion. Alginate bioinks that are lightly crosslinked prior to printing can shield printed NPCs from potential mechanical damage caused by printing. NPCs within alginate expansion lattices remain in a stem-like state while undergoing a 2.5-fold expansion. Importantly, we demonstrate the ability to efficiently remove NPCs from printed lattices for future down-stream use as a cell-based therapy. These results demonstrate that 3D bioprinting of alginate expansion lattices is a viable and economical platform for NPC expansion that could be translated to clinical applications. Graphical abstract

Stimulus-Responsive Polymeric Nanogels as Smart Drug Delivery Systems Publication date: Available online 13 May 2019 Source: Acta Biomaterialia Author(s): Sakineh Hajebi, Navid Rabiee, Mojtaba Bagherzadeh, Sepideh Ahmadi, Mohammad Rabiee, Hossein Roghani-Mamaqani, Mohammadreza Tahriri, Lobat Tayebi, Michael R. Hamblin Abstract Nanogels are three-dimensional nanoscale networks formed by physically or chemically cross-linking polymers. Nanogels have been explored as drug delivery systems due to their advantageous properties, such as biocompatibility, high stability, tunable particle size, drug loading capacity, and possible modification of the surface for active targeting by attaching ligands that recognize cognate receptors on the target cells or tissues. Nanogels can be designed to be stimulus responsive, and react to internal or external stimuli such as pH, temperature, light and redox, thus resulting in the controlled release of loaded drugs. This "smart" targeting ability prevents drug accumulation in non-target tissues and minimizes the side effects of the drug. This review aims to provide an introduction to nanogels, their preparation methods,and to discuss the design of various stimulus-responsive nanogels that are able to provide controlled drug release in response to particular stimuli. Statement of significance Smart and stimulus-responsive drug delivery is a rapidly growing area of biomaterial research. The explosive rise in nanotechnology and nanomedicine, has provided a host of nanoparticles and nanovehicles which may bewilder the uninitiated reader. This review will lay out the evidence that polymeric nanogels have an important role to play in the design of innovative drug delivery vehicles that respond to internal and external stimuli such as temperature, pH, redox, and light. Graphical abstract

Acta Journals Editors Transitioning to Joint Appointments Publication date: Available online 13 May 2019 Source: Acta Biomaterialia Author(s):

Superhydrophobic hierarchical fiber/bead composite membranes for efficient burns treatment Publication date: Available online 13 May 2019 Source: Acta Biomaterialia Author(s): Weichang Li, Qianqian Yu, Hang Yao, Yue Zhu, Paul D. Topham, Kan Yue, Li Ren, Linge Wang Abstract One of the current challenges in burn wound care is the development of multifunctional dressings that can protect the wound from bacteria or organisms and promote skin regeneration and tissue reconstitution. To this end, we report the design and fabrication of a composite electrospun membrane, comprised of electrospun polylactide : poly(vinyl pyrrolidone)/polylactide : poly(ethylene glycol) (PLA:PVP/PLA:PEG) core/shell fibers loaded with bioactive agents, as a functionally integrated wound dressing for efficient burns treatment. Different mass ratios of PLA:PVP in the shell were screened to optimize mechanical, physicochemical, and biological properties, in addition to controlled release profiles of loaded antimicrobial peptides (AMPs) from the fibers for desirable antibacterial activity. Fibroblasts were shown to readily adhere and proliferate when cultured on the membrane, indicating good in vitro cytocompatibility. The introduction of PLA beads by electrospraying on one side of the membrane resulted in biomimetic micro-nanostructures similar to those of lotus leaves. This designer structure rendered the composite membranes with superhydrophobic property to inhibit the adhesion/spreading of exogenous bacteria and other microbes. The administration of the resulting composite fibrous membrane on burnt skin in an infected rat model led to faster healing than a conventional product (sterile silicone membrane) and control detailed herein. These composite fibrous membranes loaded with bioactive drugs provide an integrated strategy for promoting burn wound healing and skin regeneration. Statement of Significance To address an urgent need in complex clinical requirements on developing a new generation of wound dressings with integrated functionalities. This article reports research work on a hierarchical fiber/bead composite membranes design, which combines a lotus-leaf-like superhydrophobic surface with drugs preloaded in the core and shell of fibers for effective burn treatment. This demonstrates a balance between simplified preparation processes and increased multifunctionality of the wound dressings. The creation of hierarchically structured surfaces can be readily achieved by electrospinning, and the composite dressings possessed a considerable mechanical strength, effective wound exudate absorption and permeability, good biocompatibility, broad antibacterial ability and promoting wound healing etc. Thus, our work unveils a promising strategy for the development of functionally integrated wound dressings for burn wound care. Graphical abstract

Graphene oxide containing self-assembling peptide hybrid hydrogels as a potential 3D injectable cell delivery platform for intervertebral disc repair applications Publication date: Available online 12 May 2019 Source: Acta Biomaterialia Author(s): Cosimo Ligorio, Mi Zhou, Jacek K. Wychowaniec, Xinyi Zhu, Cian Bartlam, Aline F. Miller, Aravind Vijayaraghavan, Judith A. Hoyland, Alberto Saiani Abstract Cell-based therapies have shown significant promise in tissue engineering with one key challenge being the delivery and retention of cells. As a result, significant efforts have been made in the past decade to design injectable biomaterials to host and deliver cells at injury sites. Intervertebral disc (IVD) degeneration, a major cause of back pain, is a particularly relevant example where a minimally-invasive cellular therapy could bring significant benefits specifically at the early stages of the disease, when a cell-driven process starts in the gelatinous core of the IVD, the nucleus pulposus (NP). In this present study we explore the use of graphene oxide (GO) as nano-filler for the reinforcement of FEFKFEFK (β-sheet forming self-assembling peptide) hydrogels. Our results confirm the presence of strong interactions between FEFKFEFK and GO flakes with the peptide coating and forming short thin fibrils on the surface of the flakes. These strong interactions were found to affect the bulk properties of hybrid hydrogels. At pH 4 electrostatic interactions between the peptide fibres and the peptide-coated GO flakes are thought to govern the final bulk properties of the hydrogels while at pH 7, after conditioning with cell culture media, electrostatic interactions are removed leaving the hydrophobic interactions to govern hydrogel final properties. The GO-F820 hybrid hydrogel, with mechanical properties similar to the NP, was shown to promote high cell viability and retained cell metabolic activity in 3D over the 7 days of culture and therefore shown to harbour significant potential as an injectable hydrogel scaffold for the in-vivo delivery of NP cells. Statement of significance Short self-assembling peptide hydrogels (SAPHs) have attracted significant interest in recent years as they can mimic the natural extra-cellular matrix, holding significant promise for the ab-initio design of cells' microenvironments. Recently the design of hybrid hydrogels for biomedical applications has been explored through the incorporation of specific nanofillers. In this study we exploited graphene oxide (GO) as nanofiller to design hybrid injectable 3Dscaffolds for the delivery of nucleus pulposus cells (NPCs) for intervertebral disc regeneration. Our work clearly shows the presence of strong interactions between peptide and GO, mimicking the mechanical properties of the NP tissue and promoting high cell viability and metabolic activity. These hybrid hydrogels therefore harbour significant potential as injectable scaffolds for the in-vivo delivery of NPCs. Graphical abstract

Synchrotron tomography of intervertebral disc deformation quantified by digital volume correlation reveals microstructural influence on strain patterns Publication date: Available online 11 May 2019 Source: Acta Biomaterialia Author(s): C.M. Disney, A. Eckersley, J.C. McConnell, H. Geng, A.J. Bodey, J.A Hoyland, P.D. Lee, M.J. Sherratt, B.K. Bay Abstract The intervertebral disc (IVD) has a complex and multiscale extracellular matrix structure which provides unique mechanical properties to withstand physiological loading. Low back pain has been linked to degeneration of the disc but reparative treatments are not currently available. Characterising the disc's 3D microstructure and its response in a physiologically relevant loading environment is required to improve understanding of degeneration and to develop new reparative treatments. In this study, techniques for imaging the native IVD, measuring internal deformation and mapping volumetric strain were applied to an in situ compressed ex vivo rat lumbar spine segment. Synchrotron X-ray micro-tomography (synchrotron CT) was used to resolve IVD structures at microscale resolution. These image data enabled 3D quantification of collagen bundle orientation and measurement of local displacement in the annulus fibrosus between sequential scans using digital volume correlation (DVC). The volumetric strain mapped from synchrotron CT provided a detailed insight into the micromechanics of native IVD tissue. The DVC findings showed that there was no slipping at lamella boundaries, and local strain patterns were of a similar distribution to the previously reported elastic network with some heterogeneous areas and maximum strain direction aligned with bundle orientation, suggesting bundle stretching and sliding. This method has the potential to bridge the gap between measures of macro-mechanical properties and the local 3D micro-mechanical environment experienced by cells. This is the first evaluation of strain at the micro scale level in the intact IVD and provides a quantitative framework for future IVD degeneration mechanics studies and testing of tissue engineered IVD replacements. Statement of Significance Synchrotron in-line phase contrast X-ray tomography provided the first visualisation of native intact intervertebral disc microstructural deformation in 3D. For two annulus fibrosus volumes of interest, collagen bundle orientation was quantified and local displacement mapped as strain. Direct evidence of microstructural influence on strain patterns could be seen such as no slipping at lamellae boundaries and maximum strain direction aligned with collagen bundle orientation. Although disc elastic structures were not directly observed, the strain patterns had a similar distribution to the previously reported elastic network. This study presents technical advances and is a basis for future X-ray microscopy, structural quantification and digital volume correlation strain analysis of soft tissue. Graphical abstract

Nonlinear elasticity of the lung extracellular microenvironment is regulated by macroscale tissue strain Publication date: Available online 11 May 2019 Source: Acta Biomaterialia Author(s): Ignasi Jorba, Gabriel Beltrán, Bryan Falcones, Béla Suki, Ramon Farré, José Manuel García-Aznar, Daniel Navajas Abstract The extracellular matrix (ECM) of the lung provides physical support and key mechanical signals to pulmonary cells. Although lung ECM is continuously subjected to different stretch levels, detailed mechanics of the ECM at the scale of the cell is poorly understood. Here, we developed a new polydimethylsiloxane (PDMS) chip to probe nonlinear mechanics of tissue samples with atomic force microscopy (AFM). Using this chip, we performed AFM measurements in decellularized rat lung slices at controlled stretch levels. The AFM revealed highly nonlinear ECM elasticity with the microscale stiffness increasing with tissue strain. To correlate micro- and macroscale ECM mechanics, we also assessed macromechanics of decellularized rat lung strips under uniaxial tensile testing. The lung strips exhibited exponential macromechanical behavior but with stiffness values one order of magnitude lower than at the microscale. To interpret the relationship between micro- and macromechanical properties, we carried out a finite element (FE) analysis which revealed that the stiffness of the alveolar cell microenvironment is regulated by the global strain of the lung scaffold. The FE modeling also indicates that the scale dependence of stiffness is mainly due to the porous architecture of the lung parenchyma. We conclude that changes in tissue strain during breathing result in marked changes in the ECM stiffness sensed by alveolar cells providing tissue-specific mechanical signals to the cells. Statement of significance The micromechanical properties of the extracellular matrix (ECM) are a major determinant of cell behavior. The ECM is exposed to mechanical stretching in the lung and other organs during physiological function. Therefore, a thorough knowledge of the nonlinear micromechanical properties of the ECM at the length scale that cells probe is required to advance our understanding of cell-matrix interplay. We designed a novel PDMS chip to perform atomic force microscopy measurements of ECM micromechanics on decellularized rat lung slices at different macroscopic strain levels. For the first time, our results reveal that the microscale stiffness of lung ECM markedly increases with macroscopic tissue strain. Therefore, changes in tissue strain during breathing result in variations in ECM stiffness providing tissue-specific mechanical signals to lung cells. Graphical abstract

Quantum Dots in Biomedical Applications Publication date: Available online 11 May 2019 Source: Acta Biomaterialia Author(s): Angela M. Wagner, Jennifer M. Knipe, Gorka Orive, Nicholas A. Peppas Abstract Semiconducting nanoparticles, more commonly known as quantum dots, possess unique size and shape dependent optoelectronic properties. In recent years, these unique properties have attracted much attention in the biomedical field to enable real-time tissue imaging (bioimaging), diagnostics, single molecule probes, and drug delivery, among many other areas. The optical properties of quantum dots can be tuned by size and composition, and their high brightness, resistance to photobleaching, multiplexing capacity, and high surface-to-volume ratio make them excellent candidates for intracellular tracking, diagnostics, in vivo imaging, and therapeutic delivery. We discuss recent advances and challenges in the molecular design of quantum dots are discussed, along with applications of quantum dots as drug delivery vehicles, theranostic agents, single molecule probes, and real-time in vivo deep tissue imaging agents. We present a detailed discussion of the biodistribution and toxicity of quantum dots, and highlight recent advances to improve long-term stability in biological buffers, increase quantum yield following bioconjugation, and improve clearance from the body. Last, we present an outlook on future challenges and strategies to further advance translation to clinical application. Statement of Significance Semiconducting nanoparticles, commonly known as quantum dots, possess unique size and shape dependent electrical and optical properties. In recent years, they have attracted much attention in biomedical imaging to enable diagnostics, single molecule probes, and real-time imaging of tumors. This review discusses recent advances and challenges in the design of quantum dots, and highlights how these strategies can further advance translation to clinical applications. Graphical abstract

Development of biocompatible and fully bioabsorbable PLA/Mg films for tissue regeneration applications Publication date: Available online 11 May 2019 Source: Acta Biomaterialia Author(s): A. Ferrández-Montero, M. Lieblich, J.L. González-Carrasco, R. Benavente, V. Lorenzo, R. Detsch, A.R. Boccaccini, B. Ferrari Abstract During recent years, Mg reinforced polylactic acid (PLA) composites have emerged as potential biocompatible and bioabsorbable materials for biomedical applications. It has been shown that Mg particles added to a matrix based on a biodegradable polymer can address the lack of bioactivity and the low mechanical properties of the polymers and, furthermore, it can counteract the detrimental effects associated to the high degradation rate of Mg, as alcalinization and elevated H2 release. Additionally, the polymer can protect the Mg particles, by tailoring their degradation rate. Former processing of these composites performed by extrusion, compression and injection molding employed Mg contents up to 10 wt.%. Higher amounts of Mg resulted in heterogeneous materials and thermally degraded matrices, with the corresponding higher degradation rate. In the present work, Mg reinforced PLA films with Mg content as high as 50 wt.% were obtained without compromising the thermal stability of the polymer. Firstly, a successful dispersion of Mg microparticles was achieved by a breakthrough in processing introducing a colloidal step where organic additives were added to modify the Mg particle surface and promote a chemically stable suspension. The resulting colloidal suspension was then used as feedstock to obtain composite films by tape casting. The films show advantageous in vitro behaviour in terms of degradation, hydrogen release and oxygen permeability. In addition, the viability with fibroblastic cells (MEF) opens a window of opportunity for these composite films as bioabsorbable material for tissue engineering and wound dressing applications. Statement of Significance Magnesium materials have extraordinary biodegradable properties and bioactive behavior due to release of Mg2+ ions, which offer a promising opportunity for their applicability as biomaterials for tissue regeneration. However, Mg is one of the most reactive metals with a high degradation rate. In contact with water produces H2, associated with a risk of failure of the implant. One alternative to minimize this drawback is the use of Mg particles surrounded by a biodegradable biocompatible polymer such as polylactic acid (PLA) to obtain PLA/Mg composites. In this work we processed Mg reinforced PLA in the shape of films that would be suitable for tissue regeneration. In vitro behavior of PLA/Mg films demonstrated that Mg2+ ions increase the fibroblastic cells growth. Graphical abstract

Osteogenic and pH Stimuli-responsive Self-healing Coating on Biomedical Mg-1Ca Alloy Publication date: Available online 11 May 2019 Source: Acta Biomaterialia Author(s): Pan Xiong, Zhaojun Jia, Wenhao Zhou, Jianglong Yan, Pei Wang, Wei Yuan, Yangyang Li, Yan Cheng, Zhenpeng Guan, Yufeng Zheng Abstract Various coatings have been used to slow down the corrosion rate of biomedical magnesium alloys. However, these coatings usually act only as passive barriers. It is much more desirable to endow such coatings with active, biocorrosion-responsive self-repairing capacity. In the present work, a self-healing coating system (denoted as "silk-PA") was constructed in the form of a sandwich architecture of fluoride precoating (bottom), silk-phytic acid (PA) coating (middle), and silk fibroin coating (top). Here, PA was loaded in the middle coating as a corrosion inhibitor by harnessing its strong chelating ability toward dissolving Mg2+ and Ca2+ ions. The self-healing property was evaluated by scratch and SVET tests, and the corrosion resistance was evaluated by in vitro immersion and electrochemical measurements. The results showed that the silk-PA manifested intriguing self-healing capacity with pH responsiveness, hence profiting the corrosion resistance of the Mg-1Ca alloy. The biocompatibility and osteogenic activity of the coating system were further evaluated using MC3T3-E1 cells, and it demonstrated favorable responses in multiple cellular behaviors, i.e., adherence, spreading, proliferation, and differentiation. These findings open new opportunities in the study of self-healing coatings for protection against corrosion in biomedical Mg alloys. Statement of significance In the present study, a self-healing coating system with pH stimuli-responsiveness as well as osteogenic activity was fabricated on Mg-1Ca alloy by integrating a silk fibroin barrier coating, a silk fibrin/phytic acid composite coating, and a fluoride pre-coating. This coating system demonstrated interesting self-healing ability in comparison to traditional surface modification layers. Furthermore, self-healing ability enhanced corrosion resistance of biomedical magnesium alloys, while effective compositions of the coating system endowed the substrate with osteogenic activity. This work gives some new insights into smart surface modification for biomedical Mg alloys. Graphical abstract