Biofabrication of functional bone tissue: defining tissue-engineered scaffolds from nature.

Damage to bone leads to pain and loss of movement in the musculoskeletal system. Although bone can regenerate, sometimes it is damaged beyond its innate capacity. Research interest is increasingly turning to tissue engineering (TE) processes to provide a clinical solution for bone defects. Despite the increasing biomimicry of tissue-engineered scaffolds, significant gaps remain in creating the complex bone substitutes, which include the biochemical and physical conditions required to recapitulate bone cells' natural growth, differentiation and maturation. Combining advanced biomaterials with new additive manufacturing technologies allows the development of 3D tissue, capable of forming cell aggregates and organoids based on natural and stimulated cues. Here, we provide an overview of the structure and mechanical properties of natural bone, the role of bone cells, the remodelling process, cytokines and signalling pathways, causes of bone defects and typical treatments and new TE strategies. We highlight processes of selecting biomaterials, cells and growth factors. Finally, we discuss innovative tissue-engineered models that have physiological and anatomical relevance for cancer treatments, injectable stimuli gels, and other therapeutic drug delivery systems. We also review current challenges and prospects of bone TE. Overall, this review serves as guide to understand and develop better tissue-engineered bone designs.

Biomimetic triumvirate nanogel complexes via peptide-polysaccharide-polyphenol self-assembly.

Self-assembled peptide and polysaccharide nanogels are excellent candidates for bioactive delivery vectors. However, there are still significant challenges in the application of nanogels as delivery tools for bioactive elements. This study aims to deliver, and control the release of a hydrophobic bioactive flavonoid hesperidin. Using the self-assembling peptide (SAP) Fmoc-FRGDF, extracellular matrix mimicking nanofibrils were fabricated, which were decorated and bolstered with immunomodulatory polysaccharide strands of fucoidan and infused with hesperidin. The mechanical properties, secondary structure, and microscopic morphologies of the composite hydrogels were characterized using rheometer, FTIR, XRD, and TEM, etc. The encapsulation efficiency (EE) and release behavior of hesperidin were determined. Coassembly of the SAP with fucoidan improved the mechanical properties (from 9.54 Pa of Fmoc-FRGDF hydrogel to 7735 Pa of coassembly hydrogel at 6 mg/mL fucoidan concentration), formed thicker nanofibril bundles at 4 and 6 mg/mL fucoidan concentration, improved the EE of hesperidin from 72.86 % of Fmoc-FRGDF hydrogel to over 90 % of coassembly hydrogels, and showed effectively controlled release of hesperidin in vitro. Intriguingly, the first order kinetic model predicted an enhanced hydrogel retention and release of hesperidin. This study revealed a new approach for bioengineered nanogels that could be used to stabilize and release hydrophobic payloads.

Simple Complexity: Incorporating Bioinspired Delivery Machinery within Self-Assembled Peptide Biogels.

Bioinspired self-assembly is a bottom-up strategy enabling biologically sophisticated nanostructured biogels that can mimic natural tissue. Self-assembling peptides (SAPs), carefully designed, form signal-rich supramolecular nanostructures that intertwine to form a hydrogel material that can be used for a range of cell and tissue engineering scaffolds. Using the tools of nature, they are a versatile framework for the supply and presentation of important biological factors. Recent developments have shown promise for many applications such as therapeutic gene, drug and cell delivery and yet are stable enough for large-scale tissue engineering. This is due to their excellent programmability-features can be incorporated for innate biocompatibility, biodegradability, synthetic feasibility, biological functionality and responsiveness to external stimuli. SAPs can be used independently or combined with other (macro)molecules to recapitulate surprisingly complex biological functions in a simple framework. It is easy to accomplish localized delivery, since they can be injected and can deliver targeted and sustained effects. In this review, we discuss the categories of SAPs, applications for gene and drug delivery, and their inherent design challenges. We highlight selected applications from the literature and make suggestions to advance the field with SAPs as a simple, yet smart delivery platform for emerging BioMedTech applications.

Self-Assembled Peptide Habitats to Model Tumor Metastasis.

Metastatic tumours are complex ecosystems; a community of multiple cell types, including cancerous cells, fibroblasts, and immune cells that exist within a supportive and specific microenvironment. The interplay of these cells, together with tissue specific chemical, structural and temporal signals within a three-dimensional (3D) habitat, direct tumour cell behavior, a subtlety that can be easily lost in 2D tissue culture. Here, we investigate a significantly improved tool, consisting of a novel matrix of functionally programmed peptide sequences, self-assembled into a scaffold to enable the growth and the migration of multicellular lung tumour spheroids, as proof-of-concept. This 3D functional model aims to mimic the biological, chemical, and contextual cues of an in vivo tumor more closely than a typically used, unstructured hydrogel, allowing spatial and temporal activity modelling. This approach shows promise as a cancer model, enhancing current understandings of how tumours progress and spread over time within their microenvironment.

Liquid metal polymer composite: Flexible, conductive, biocompatible, and antimicrobial scaffold.

Gallium and its alloys, such as eutectic gallium indium alloy (EGaIn), a form of liquid metal, have recently attracted the attention of researchers due to their low toxicity and electrical and thermal conductivity for biomedical application. However, further research is required to harness EGaIn-composites advantages and address their application as a biomedical scaffold. In this research, EGaIn-polylactic acid/polycaprolactone composites with and without a second conductive filler, MXene, were prepared and characterized. The addition of MXene, into the EGaIn-composite, can improve the composite's electrochemical properties by connecting the liquid metal droplets resulting in electrically conductive continuous pathways within the polymeric matrix. The results showed that the composite with 50% EGaIn and 4% MXene, displayed optimal electrochemical properties and enhanced mechanical and radiopacity properties. Furthermore, the composite showed good biocompatibility, examined through interactions with fibroblast cells, and antibacterial properties against methicillin-resistant Staphylococcus aureus. Therefore, the liquid metal (EGaIn) polymer composite with MXene provides a first proof-of-concept engineering scaffold strategy with low toxicity, functional electrochemical properties, and promising antimicrobial properties.

Replace and repair: Biomimetic bioprinting for effective muscle engineering.

The debilitating effects of muscle damage, either through ischemic injury or volumetric muscle loss (VML), can have significant impacts on patients, and yet there are few effective treatments. This challenge arises when function is degraded due to significant amounts of skeletal muscle loss, beyond the regenerative ability of endogenous repair mechanisms. Currently available surgical interventions for VML are quite invasive and cannot typically restore function adequately. In response to this, many new bioengineering studies implicate 3D bioprinting as a viable option. Bioprinting for VML repair includes three distinct phases: printing and seeding, growth and maturation, and implantation and application. Although this 3D bioprinting technology has existed for several decades, the advent of more advanced and novel printing techniques has brought us closer to clinical applications. Recent studies have overcome previous limitations in diffusion distance with novel microchannel construct architectures and improved myotubule alignment with highly biomimetic nanostructures. These structures may also enhance angiogenic and nervous ingrowth post-implantation, though further research to improve these parameters has been limited. Inclusion of neural cells has also shown to improve myoblast maturation and development of neuromuscular junctions, bringing us one step closer to functional, implantable skeletal muscle constructs. Given the current state of skeletal muscle 3D bioprinting, the most pressing future avenues of research include furthering our understanding of the physical and biochemical mechanisms of myotube development and expanding our control over macroscopic and microscopic construct structures. Further to this, current investigation needs to be expanded from immunocompromised rodent and murine myoblast models to more clinically applicable human cell lines as we move closer to viable therapeutic implementation.

Diamond in the Rough: Toward Improved Materials for the Bone-Implant Interface.

The ability of an orthopedic implant to integrate successfully with the surrounding bone tissue is imperative for optimal patient outcomes. Here, the recent advances and future prospects for diamond-based coatings of conventional osteo-implant materials (primarily titanium) are explored. The ability of these diamond coatings to enhance integration into existing bone, improved implant mechanical properties, facilitate surface chemical functionalization, and provide anti-microbial properties are discussed in context of orthopedic implants. These diamond-based materials may have the additional benefit of providing an osteo-inductive effect, enabling better integration into existing bone via stem cell recruitment and bone regeneration. Current and timely research is highlighted to support the discussion and suggestions in further improving implant integration via an osseoinductive effect from the diamond composite materials.

Multifunctional Sutures with Temperature Sensing and Infection Control.

The next-generation sutures should provide in situ monitoring of wound condition such as temperature while reducing surgical site infection during wound closure. In this study, functionalized nanodiamond (FND) and reduced graphene oxide (rGO) into biodegradable polycaprolactone (PCL) are incorporated to develop a new multifunctional suture with such capabilities. Incorporation of FND and rGO into PCL enhances its tensile strength by about 43% and toughness by 35%. The sutures show temperature sensing capability in the range of 25-40 °C based on the shift in zero-splitting frequency of the nitrogen-vacancy (NV- ) centers in FND via optically detected magnetic resonance, paving the way for potential detection of infection or excessive inflammation in healing wounds. The suture surface readily coats with antibiotics to reduce bacterial infection risk to the wounds. The new suture thus is promising in monitoring and supporting wound closure.

Osteoblast Cell Response on Polycrystalline Diamond-Coated Additively Manufactured Scaffolds.

Additive manufacturing of metals using selective laser melting can create customized parts with various degrees of complexity and geometry for medical implants. However, challenges remain in accepting orthopedic implants due to the bio-inert surface of metal scaffolds, resulting in a lack of osseointegration. Here, we show that polycrystalline diamond (PCD) coatings on selective laser melted titanium (SLM-Ti) scaffolds can improve the cell-to-material interaction of osteoblasts. The results show that by controlling the uniformity of the diamond coatings, we can mediate the biological response of osteoblasts, such as cell adhesion, proliferation, and spreading. The osteoblasts show favorable cell adhesion and spreading on non-planar PCD-coated scaffolds compared to the un-coated SLM-Ti scaffold. This study plays an important role in understanding the key physicochemical behavior of bone cell growth on customized orthopedic implant materials.

Polypropylene-nanodiamond composite for hernia mesh.

Commercial hernia mesh is commonly made from polypropylene (PP), due to its inertness, biocompatibility, physical properties, ease of processing and versatility for conversion into flexible shape. However, reportedly hernia mesh prepared from PP experienced issues such as diminished long-term strength, foreign body rejection, lack of biocompatibility and high adhesion to the abdomen wall. Infiltration of the mesh by soft tissue (called remodeling) results in an integration of mesh into the body, leading to a rapid reduction in mesh mechanical properties and potential infection. Here, this study addresses these issues through the incorporation of nanodiamond (ND) into PP filament and coating on the surface of plasma-treated PP-ND mesh. The results show that the dynamic modulus of the PP-ND mesh increased significantly, without compromising its flexibility. Coating PP-ND mesh with hydroxylated ND led to a reduction in nonspecific protein adsorption onto the surface of nanocomposite, which is an important characteristic for hernia mesh to prevent foreign body reaction, attachment of mesh to the abdominal wall and nearby organs. In-vitro study with mammalian cells shows that coated PP-ND mesh with functionalized ND exhibits a significant increase in the number of adhered cells with more elongated morphology in comparison with other PP meshes, due to the better hydrophilicity. Therefore, the ND coated nanocomposite mesh can be a promising candidate for hernia repair in the future; however, more investigation is required.

3D-Printed Diamond-Titanium Composite: A Hybrid Material for Implant Engineering.

Diamond-based implant materials make up an emerging research area where the materials could be prepared to promote cellular functions, decrease bacteria attachment, and be suitable for potential in situ imaging. Up until now, diamond implants have been fabricated using coating technologies or embedding diamond nanoparticles in polymer matrices. Here we demonstrated a method of manufacturing diamond implants using laser cladding technology to 3D print a composite of diamond and fused titanium material. Using this method, we could prepare composite scaffolds of up to 50% diamond, which has never been achieved before. We next investigated the interfacial properties of these scaffolds for potential applications in implants. The addition of diamond to the biomaterial results in a 30% decrease in the water contact angle, making the scaffolds more hydrophilic and improving cellular adhesion and proliferation.

Hybrid diamond/ carbon fiber microelectrodes enable multimodal electrical/chemical neural interfacing.

Implantable medical devices are now in regular use to treat or ameliorate medical conditions, including movement disorders, chronic pain, cardiac arrhythmias, and hearing or vision loss. Aside from offering alternatives to pharmaceuticals, one major advantage of device therapy is the potential to monitor treatment efficacy, disease progression, and perhaps begin to uncover elusive mechanisms of diseases pathology. In an ideal system, neural stimulation, neural recording, and electrochemical sensing would be conducted by the same electrode in the same anatomical region. Carbon fiber (CF) microelectrodes are the appropriate size to achieve this goal and have shown excellent performance, in vivo. Their electrochemical properties, however, are not suitable for neural stimulation and electrochemical sensing. Here, we present a method to deposit high surface area conducting diamond on CF microelectrodes. This unique hybrid microelectrode is capable of recording single-neuron action potentials, delivering effective electrical stimulation pulses, and exhibits excellent electrochemical dopamine detection. Such electrodes are needed for the next generation of miniaturized, closed-loop implants that can self-tune therapies by monitoring both electrophysiological and biochemical biomarkers.

Engineering the Interface: Nanodiamond Coating on 3D-Printed Titanium Promotes Mammalian Cell Growth and Inhibits Staphylococcus aureus Colonization.

Additively manufactured selective laser melted titanium (SLM-Ti) opens the possibility of tailored medical implants for patients. Despite orthopedic implant advancements, significant problems remain with regard to suboptimal osseointegration at the interface between the implant and the surrounding tissue. Here, we show that applying a nanodiamond (ND) coating onto SLM-Ti scaffolds provides an improved surface for mammalian cell growth while inhibiting colonization of Staphylococcus aureus bacteria. Owing to the simplicity of our methodology, the approach is suitable for coating SLM-Ti geometries. The ND coating achieved 32 and 29% increases in cell density of human dermal fibroblasts and osteoblasts, respectively, after 3 days of incubation compared with the uncoated SLM-Ti substratum. This increase in cell density complements an 88% reduction in S. aureus detected on the ND-coated SLM-Ti substrata. This study paves a way to create facile antifouling SLM-Ti structures for biomedical implants.

Nanodiamond/poly-ε-caprolactone nanofibrous scaffold for wound management.

We describe preparation, characterization and cytocompatibility of nanodiamond (ND) dispersed in poly (ε-caprolactone) (PCL) based nanofibrous scaffold. The results show that this unique scaffold potentially provides essential properties for wound healing by enhancing proliferation of epithelial cells, in addition to restricting the microbial activities. Electrospinning technique was used to fabricate and develop PCL-NDs nanocomposite scaffold. The developed nanocomposites were characterized for morphology, thermal, surface and biological properties. The incorporation of ND into the PCL matrix resulted in better moisture management and higher thermal stability. Transmission electron microscopy images and attenuated total reflectance-Fourier transform infrared (ATR-FTIR) spectroscopy showed existence of ND particles on the surface of the nanofibers. The aggregation of ND particles increased with the increase in their concentration in nanofiber. The developed scaffolds showed no cytotoxicity and, due to improved hydrophilicity, better cellular activities with Chinese hamster ovarian (CHO) cells, 43%, 38% and 22% more cell proliferation for PCL-5% ND for 1, 3- and 7-days incubations in compare with PCL. Furthermore, Staphylococcus aureus (S. aureus) showed significantly less affinity to the scaffold surface with the increase in ND concentration, ~56% less for PCL-5% ND in compare with PCL, indicating that such ND dispersed nanofibrous scaffold maybe asuitable choice for complex wound management.

Polycrystalline Diamond Coating of Additively Manufactured Titanium for Biomedical Applications.

Additive manufacturing using selective laser melted titanium (SLM-Ti) is used to create bespoke items across many diverse fields such as medicine, defense, and aerospace. Despite great progress in orthopedic implant applications, such as for "just in time" implants, significant challenges remain with regards to material osseointegration and the susceptibility to bacterial colonization on the implant. Here, we show that polycrystalline diamond coatings on these titanium samples can enhance biological scaffold interaction improving medical implant applicability. The highly conformable coating exhibited excellent bonding to the substrate. Relative to uncoated SLM-Ti, the diamond coated samples showed enhanced mammalian cell growth, enriched apatite deposition, and reduced microbial S. aureus activity. These results open new opportunities for novel coatings on SLM-Ti devices in general and especially show promise for improved biomedical implants.