Angiogenesis in bone tissue engineering via ceramic scaffolds: A review of concepts and recent advancements.

Due to organ donor shortages, long transplant waitlists, and the complications/limitations associated with auto and allotransplantation, biomaterials and tissue-engineered models are gaining attention as feasible alternatives for replacing and reconstructing damaged organs and tissues. Among various tissue engineering applications, bone tissue engineering has become a promising strategy to replace or repair damaged bone. We aimed to provide an overview of bioactive ceramic scaffolds in bone tissue engineering, focusing on angiogenesis and the effect of different biofunctionalization strategies. Different routes to angiogenesis, including chemical induction through signaling molecules immobilized covalently or non-covalently, in situ secretion of angiogenic growth factors, and the degradation of inorganic scaffolds, are described. Physical induction mechanisms are also discussed, followed by a review of methods for fabricating bioactive ceramic scaffolds via microfabrication methods, such as photolithography and 3D printing. Finally, the strengths and weaknesses of the commonly used methodologies and future directions are discussed.

Modified Poly(ε-caprolactone) with Tunable Degradability and Improved Biofunctionality for Regenerative Medicine.

The use of poly(ε-caprolactone) (PCL) for biomedical applications is well established, particularly for permanent implants, due to its slow degradation rate, suitable mechanical properties, and biocompatibility. However, the slow degradation rate of PCL limits its application for short-term and temporary biomedical applications where bioabsorbability is required. To enhance the properties of PCL and to expand its biomedical applications, we developed an approach to produce PCL membranes with tunable degradation rates, mechanical properties, and biofunctional features. Specifically, we utilized electrospinning to create fibrous PCL membranes, which were then chemically modified using potassium permanganate to alter their degradability while having minimal impact on their fibrous morphology. The effects of the chemical treatments were investigated by treating the samples for different time periods ranging from 6 to 48 h. After the 48 h treatment, the membrane degraded by losing 25% of its mass over 12 weeks in degradation studies, while maintaining its mechanical strength and exhibiting superior biofunctional features. Our results suggest that this approach for developing PCL with tailored properties could have significant potential for a range of biomedical applications.

Bacterial-Nanocellulose-Based Biointerfaces and Biomimetic Constructs for Blood-Contacting Medical Applications.

Understanding the interaction between biomaterials and blood is critical in the design of novel biomaterials for use in biomedical applications. Depending on the application, biomaterials can be designed to promote hemostasis, slow or stop bleeding in an internal or external wound, or prevent thrombosis for use in permanent or temporary medical implants. Bacterial nanocellulose (BNC) is a natural, biocompatible biopolymer that has recently gained interest for its potential use in blood-contacting biomedical applications (e.g., artificial vascular grafts), due to its high porosity, shapeability, and tissue-like properties. To promote hemostasis, BNC has been modified through oxidation or functionalization with various peptides, proteins, polysaccharides, and minerals that interact with the coagulation cascade. For use as an artificial vascular graft or to promote vascularization, BNC has been extensively researched, with studies investigating different modification techniques to enhance endothelialization such as functionalizing with adhesion peptides or extracellular matrix (ECM) proteins as well as tuning the structural properties of BNC such as surface roughness, pore size, and fiber size. While BNC inherently exhibits comparable mechanical characteristics to endogenous blood vessels, these mechanical properties can be enhanced through chemical functionalization or through altering the fabrication method. In this review, we provide a comprehensive overview of the various modification techniques that have been implemented to enhance the suitability of BNC for blood-contacting biomedical applications and different testing techniques that can be applied to evaluate their performance. Initially, we focused on the modification techniques that have been applied to BNC for hemostatic applications. Subsequently, we outline the different methods used for the production of BNC-based artificial vascular grafts and to generate vasculature in tissue engineered constructs. This sequential organization enables a clear and concise discussion of the various modifications of BNC for different blood-contacting biomedical applications and highlights the diverse and versatile nature of BNC as a natural biomaterial.

Super-Repellent and Flexible Lubricant-Infused Bacterial Nanocellulose Membranes with Superior Antithrombotic, Antibacterial, and Fatigue Resistance Properties.

Bacterial nanocellulose (BNC) is a naturally derived hydrogel that has recently paved its way in several biomedical applications. Despite its remarkable tissue-like properties, BNC does not express innate anticoagulant or antimicrobial properties; therefore, appropriate post-modification procedures are required to prevent nonspecific adhesion and enhance the hemocompatibility properties of BNC-based biointerface. Here, we report a new class of flexible, lubricant-infused BNC membranes with superior antithrombotic and antibacterial properties. Using chemical vapor deposition, porous BNC membranes were functionalized with fluorosilane molecules and further impregnated with a fluorocarbon-based lubricant. Compared with unmodified BNC membranes and commercially available poly(tetrafluoroethylene) (PTFE) felts, our developed lubricant-infused BNC samples significantly attenuated plasma and blood clot formation, and prevented bacterial migration, adhesion, and biofilm formation and exhibited superior fat and enzyme repellency properties. Moreover, when subjected to mechanical testing, the lubricant-infused BNC membranes demonstrated a significantly higher tensile strength and greater fatigue resistance when compared with unmodified BNC samples and PTFE felts. Overall, the superior mechanical strength and antithrombotic, antibacterial, and fat/enzyme resistant properties observed in the developed super-repellent BNC-based membranes render their application promising for various biofluid-contacting medical implants and tissue engineering constructs.

Bio-macromolecular design roadmap towards tough bioadhesives.

Emerging sutureless wound-closure techniques have led to paradigm shifts in wound management. State-of-the-art biomaterials offer biocompatible and biodegradable platforms enabling high cohesion (toughness) and adhesion for rapid bleeding control as well as robust attachment of implantable devices. Tough bioadhesion stems from the synergistic contributions of cohesive and adhesive interactions. This Review provides a biomacromolecular design roadmap for the development of tough adhesive surgical sealants. We discuss a library of materials and methods to introduce toughness and adhesion to biomaterials. Intrinsically tough and elastic polymers are leveraged primarily by introducing strong but dynamic inter- and intramolecular interactions either through polymer chain design or using crosslink regulating additives. In addition, many efforts have been made to promote underwater adhesion via covalent/noncovalent bonds, or through micro/macro-interlock mechanisms at the tissue interfaces. The materials settings and functional additives for this purpose and the related characterization methods are reviewed. Measurements and reporting needs for fair comparisons of different materials and their properties are discussed. Finally, future directions and further research opportunities for developing tough bioadhesive surgical sealants are highlighted.

Engineered Hemostatic Biomaterials for Sealing Wounds.

Hemostatic biomaterials show great promise in wound control for the treatment of uncontrolled bleeding associated with damaged tissues, traumatic wounds, and surgical incisions. A surge of interest has been directed at boosting hemostatic properties of bioactive materials via mechanisms triggering the coagulation cascade. A wide variety of biocompatible and biodegradable materials has been applied to the design of hemostatic platforms for rapid blood coagulation. Recent trends in the design of hemostatic agents emphasize chemical conjugation of charged moieties to biomacromolecules, physical incorporation of blood-coagulating agents in biomaterials systems, and superabsorbing materials in either dry (foams) or wet (hydrogel) states. In addition, tough bioadhesives are emerging for efficient and physical sealing of incisions. In this Review, we highlight the biomacromolecular design approaches adopted to develop hemostatic bioactive materials. We discuss the mechanistic pathways of hemostasis along with the current standard experimental procedures for characterization of the hemostasis efficacy. Finally, we discuss the potential for clinical translation of hemostatic technologies, future trends, and research opportunities for the development of next-generation surgical materials with hemostatic properties for wound management.

Enhancing osseointegration and mitigating bacterial biofilms on medical-grade titanium with chitosan-conjugated liquid-infused coatings.

Titanium alloys, in particular, medical-grade Ti-6Al-4 V, are heavily used in orthopaedic applications due to their high moduli, strength, and biocompatibility. Implant infection can result in biofilm formation and failure of prosthesis. The formation of a biofilm on implants protects bacteria from antibiotics and the immune response, resulting in the propagation of the infection and ultimately resulting in device failure. Recently, slippery liquid-infused surfaces (LIS) have been investigated for their stable liquid interface, which provides excellent repellent properties to suppress biofilm formation. One of the current limitations of LIS coatings lies in the indistinctive repellency of bone cells in orthopaedic applications, resulting in poor tissue integration and bone ingrowth with the implant. Here, we report a chitosan impregnated LIS coating that facilitates cell adhesion while preventing biofilm formation. The fabricated coating displayed high contact angles (108.2 ± 5.2°) and low sliding angles (3.56 ± 4.3°). Elemental analysis obtained using X-ray photoelectron spectroscopy (XPS) confirmed the availability of fluorine and nitrogen, indicating the presence of fluorosilane and chitosan in the final coating. Furthermore, our results suggest that chitosan-conjugated LIS increased cell adhesion of osteoblast-like SaOS-2 cells and significantly promoted proliferation (a fourfold increase at 7-day incubation) compared to conventional titanium liquid-infused surfaces. Furthermore, the chitosan conjugated LIS significantly reduced biofilm formation of methicillin-resistant Staphylococcus aureus (MRSA) by up to 50% and 75% when compared to untreated titanium and chitosan-coated titanium, respectively. The engineered coating can be easily modified with other biopolymers or capture molecules to be applied to other biomaterials where tissue integration and biofilm prevention are needed.

Polysiloxane Nanofilaments Infused with Silicone Oil Prevent Bacterial Adhesion and Suppress Thrombosis on Intranasal Splints.

Like all biofluid-contacting medical devices, intranasal splints are highly prone to bacterial adhesion and clot formation. Despite their widespread use and the numerous complications associated with infected splints, limited success has been achieved in advancing their safety and surface biocompatibility, and, to date, no surface-coating strategy has been proposed to simultaneously enhance the antithrombogenicity and bacterial repellency of intranasal splints. Herein, we report an efficient, highly stable lubricant-infused coating for intranasal splints to render their surfaces antithrombogenic and repellent toward bacterial cells. Lubricant-infused intranasal splints were prepared by creating superhydrophobic polysiloxane nanofilament (PSnF) coatings using surface-initiated polymerization of n-propyltrichlorosilane (n-PTCS) and further infiltrating them with a silicone oil lubricant. Compared with commercially available intranasal splints, lubricant-infused, PSnF-coated splints significantly attenuated plasma and blood clot formation and prevented bacterial adhesion and biofilm formation for up to 7 days, the typical duration for which intranasal splints are kept. We further demonstrated that the performance of our engineered biointerface is independent of the underlying substrate and could be used to enhance the hemocompatibility and repellency properties of other medical implants such as medical-grade catheters.

Single and multi-functional coating strategies for enhancing the biocompatibility and tissue integration of blood-contacting medical implants.

Device-associated clot formation and poor tissue integration are ongoing problems with permanent and temporary implantable medical devices. These complications lead to increased rates of mortality and morbidity and impose a burden on healthcare systems. In this review, we outline the current approaches for developing single and multi-functional surface coating techniques that aim to circumvent the limitations associated with existing blood-contacting medical devices. We focus on surface coatings that possess dual hemocompatibility and biofunctionality features and discuss their advantages and shortcomings to providing a biocompatible and biodynamic interface between the medical implant and blood. Lastly, we outline the newly developed surface modification techniques that use lubricant-infused coatings and discuss their unique potential and limitations in mitigating medical device-associated complications.

Lubricant-Infused PET Grafts with Built-In Biofunctional Nanoprobes Attenuate Thrombin Generation and Promote Targeted Binding of Cells.

New surface coatings that enhance hemocompatibility and biofunctionality of synthetic vascular grafts such as expanded poly(tetrafluoroethylene) (ePTFE) and poly(ethylene terephthalate) (PET) are urgently needed. Lubricant-infused surfaces prevent nontargeted adhesion and enhance the biocompatibility of blood-contacting surfaces. However, limited success has been made in incorporating biofunctionality onto these surfaces and generating biofunctional lubricant-infused coatings that both prevent nonspecific adhesion and enhance targeted binding of biomolecules remains a challenge. Here, a new generation of fluorosilanized lubricant-infused PET surfaces with built-in biofunctional nanoprobes is reported. These surfaces are synthesized by starting with a self-assembled monolayer of fluorosilane that is partially etched using plasma modification technique, thereby creating a hydroxyl-terminated fluorosilanized PET surface. Simultaneously, silanized nanoprobes are produced by amino-silanizing anti-CD34 antibody in solution and directly coupling the anti-CD34-aminosilane nanoprobes onto the hydroxyl terminated, fluorosilanized PET surface. The PET surfaces are then lubricated, creating fluorosilanized biofunctional lubricant-infused PET substrates. Compared with unmodified PET surfaces, the designed biofunctional lubricant-infused PET surfaces significantly attenuate thrombin generation and blood clot formation and promote targeted binding of endothelial cells from human whole blood.

Micropatterned biofunctional lubricant-infused surfaces promote selective localized cell adhesion and patterning.

Micropatterned biofunctional surfaces provide a wide range of applications in bioengineering. A key characteristic which is sought in these types of bio-interfaces is prevention of non-specific adhesion for enhanced biofunctionality and targeted binding. Lubricant-infused omniphobic coatings have exhibited superior performance in attenuating non-specific adhesion; however, these coatings completely block the surfaces and do not support targeted adhesion or patterning. In this work, we introduce a novel lubricant-infused surface with biofunctional micropatterned domains integrated within an omniphobic layer. This new class of micropatterned lubricant-infused surfaces simultaneously promotes localized and directed binding of desired targets, as well as repellency of undesired species, especially in human whole blood. Furthermore, this modification method is easily translatable to microfluidic devices offering a wider range of applications and improved performance for immunoassays in whole blood and inhibition of clot formation in microfluidic channels. The biofunctional micropatterned lubricant-infused surfaces were created through a bench-top straight forward process by integrating microcontact printing, chemical vapor deposition (CVD) of self-assembled monolayers (SAMs) of fluorosilanes, and further infusion of the SAMs with a bio-compatible fluorocarbon-based lubricant layer. The developed surfaces, patterned with anti-CD34 antibodies, yield enhanced adhesion and controlled localized binding of target biomolecules (e.g. antibodies) and CD34 positive cells (e.g. HUVECs) inside microfluidic devices, outperforming conventional blocking methods (e.g. bovine serum albumin (BSA) or poly(ethylene glycol) (PEG)) in buffer and human whole blood. These surfaces offer a straightforward and effective way to enhance blocking capabilities while preserving the biofunctionality of a micropatterned system in complex biological environments such as whole blood. We anticipate that these micropatterned biofunctional interfaces will find a wide range of applications in microfluidic devices and biosensors for enhanced and localized targeted binding while preventing non-specific adhesion.

Step-Wise Assessment and Optimization of Sample Handling Recovery Yield for Nanoproteomic Analysis of 1000 Mammalian Cells.

Protein and peptide adhesion is a major factor contributing to sample loss during proteomic sample preparation workflows. Sample loss often has detrimental effects on the quality of proteomic analysis by compromising protein identification and data reproducibility. When starting with a low sample amount, only the most abundant proteins can be identified, which often offers little insights for biological research. Although the general idea about severe sample loss from low amount of starting material is widely presumed in the proteomics field, quantitative assessment on the impact of sample loss has been poorly investigated. In the present study, we have quantitatively assessed sample loss during each step of a conventional in-solution sample preparation workflow using bicinchoninic acid (BCA) and targeted LC/MS/MS protein and peptide assays. According to our assessment, for starting materials of ∼1000 mammalian cells, surface adhesion, along with desalting and speed-vacuum drying steps, all contribute heavily to sample loss, in particular for low-abundance proteins. With this knowledge, we have adapted slippery liquid infused porous surface (SLIPS) treatment, commercial LoBind tubes, and in-line desalting during sample processing. With these improvements, we were able to use a conventional in-solution sample handling method to identify on average 829 proteins with 1000 U2OS osteosarcoma cells (∼100 ng) with 75-min LC/MS/MS runs, an 11-fold increase in protein identification. Our optimized in-solution workflow is straightforward and also much less equipment- and technique-demanding than other advanced sample preparation protocols in the field.

Tissue Response and Biodistribution of Injectable Cellulose Nanocrystal Composite Hydrogels.

Interest in cellulose nanocrystal (CNC)-based hydrogels for drug delivery, tissue engineering, and other biomedical applications has rapidly expanded despite the minimal in vivo research reported to date. Herein, we assess both in vitro protein adsorption and cell adhesion as well as in vivo subcutaneous tissue responses and CNC biodistribution of injectable CNC-poly(oligoethylene glycol methacrylate) (POEGMA) hydrogels. Hydrogels with different PEG side chain lengths, CNC loadings, and with or without in situ magnetic alignment of the CNCs are compared. CNC loading has a minimal impact on protein adsorption but significantly increases cell adhesion. In vivo, both CNC-only and CNC-POEGMA injections largely stay at their subcutaneous injection site over one month, with minimal bioaccumulation of CNCs in any typical clearance organ. CNC-POEGMA hydrogels exhibit mild acute and chronic inflammatory responses, although significant fibroblast penetration was observed with the magnetically aligned hydrogels. Collectively, these results suggest that CNC-POEGMA hydrogels offer promise in practical biomedical applications.

Biofunctional Lubricant-Infused Vascular Grafts Functionalized with Silanized Bio-Inks Suppress Thrombin Generation and Promote Endothelialization.

The ongoing problem with the thrombogenicity and poor tissue integration of synthetic vascular grafts demands the design of new surfaces that simultaneously suppress thrombosis and promote endothelialization. Lubricant-infused surfaces have shown outstanding results in preventing clot formation; however, their innate ability to completely block the surface, averts targeted binding of desired biomolecules. We report a new class of expanded polytetrafluoroethylene (ePTFE) vascular grafts that prevent blood coagulation and concurrently promote endothelial cell adhesion. This is made possible by direct silanization of anti-CD34 antibody with the coupling agent and subsequent conjugation of the silanized antibody to the ePTFE surface. In contrast to the conventional methods, we eliminated the need to chemically modify the ePTFE substrate for attaching the capturing ligand, and as a result preserved the innate surface properties of the ePTFE substrate. This is crucial for infiltrating the fluorine-based ePTFE substrate with a biocompatible perfluorocarbon-based lubricant and ultimately creating a functional and stable lubricant-infused layer. Compared to commercially available ePTFE vascular grafts and the ones coated using conventional methods, our developed ePTFE grafts significantly attenuate thrombin generation and blood clot formation and specifically capture endothelial cells from human whole blood while preventing nonspecific adhesion of undesirable proteins and cells. The developed technology can be applied to other biomarkers and biomaterials and can be tailored toward different biomedical applications where biofunctionality and targeted binding are of importance.

Lubricant-Infused Surfaces with Built-In Functional Biomolecules Exhibit Simultaneous Repellency and Tunable Cell Adhesion.

Lubricant-infused omniphobic surfaces have exhibited outstanding effectiveness in inhibiting nonspecific adhesion and attenuating superimposed clot formation compared with other coated surfaces. However, such surfaces blindly thwart adhesion, which is troublesome for applications that rely on targeted adhesion. Here we introduce a new class of lubricant-infused surfaces that offer tunable bioactivity together with omniphobic properties by integrating biofunctional domains into the lubricant-infused layer. These novel surfaces promote targeted binding of desired species while simultaneously preventing nonspecific adhesion. To develop these surfaces, mixed self-assembled monolayers (SAMs) of aminosilanes and fluorosilanes were generated. Aminosilanes were utilized as coupling molecules for immobilizing capture ligands, and nonspecific adhesion of cells and proteins was prevented by infiltrating the fluorosilane molecules with a thin layer of a biocompatible fluorocarbon-based lubricant, thus generating biofunctional lubricant-infused surfaces. This method yields surfaces that (a) exhibit highly tunable binding of anti-CD34 and anti-CD144 antibodies and adhesion of endothelial cells, while repelling nonspecific adhesion of undesirable proteins and cells not only in buffer but also in human plasma or human whole blood, and (b) attenuate blood clot formation. Therefore, this straightforward and simple method creates biofunctional, nonsticky surfaces that can be used to optimize the performance of devices such as biomedical implants, extracorporeal circuits, and biosensors.

Self-Cleaning Ceramic Tiles Produced via Stable Coating of TiO2 Nanoparticles.

The high photocatalytic power of TiO2 nanoparticles has drawn great attention in environmental and medical applications. Coating surfaces with these particles enables us to benefit from self-cleaning properties and decomposition of pollutants. In this paper, two strategies have been introduced to coat ceramic tiles with TiO2 nanoparticles, and the self-cleaning effect of the surfaces on degradation of an organic dye under ultraviolent (UV) exposure is investigated. In the first approach, a simple one-step heat treatment method is introduced for coating, and different parameters of the heat treatment process are examined. In the second method, TiO2 nanoparticles are first aminosilanized using (3-Aminopropyl)triethoxysilane (APTES) treatment followed by their covalently attachment onto CO2 plasma treated ceramic tiles via N-(3-Dimethylaminopropyl)-N′-ethylcarbodiimide hydrochloride (EDC) and N-Hydroxysuccinimide (NHS) chemistry. We monitor TiO2 nanoparticle sizes throughout the coating process using dynamic light scattering (DLS) and characterize developed surfaces using X-ray photoelectron spectroscopy (XPS). Moreover, hydrophilicity of the coated surfaces is quantified using a contact angle measurement. It is shown that applying a one-step heat treatment process with the optimum temperature of 200 °C for 5 h results in successful coating of nanoparticles and rapid degradation of dye in a short time. In the second strategy, the APTES treatment creates a stable covalent coating, while the photocatalytic capability of the particles is preserved. The results show that coated ceramic tiles are capable of fully degrading the added dyes under UV exposure in less than 24 h.

Injectable and Degradable Poly(Oligoethylene glycol methacrylate) Hydrogels with Tunable Charge Densities as Adhesive Peptide-Free Cell Scaffolds.

Injectable, dual-responsive, and degradable poly(oligo ethylene glycol methacrylate) (POEGMA) hydrogels are demonstrated to offer potential for cell delivery. Charged groups were incorporated into hydrazide and aldehyde-functionalized thermoresponsive POEGMA gel precursor polymers via the copolymerization of N,N'-dimethylaminoethyl methacrylate (DMAEMA) or acrylic acid (AA) to create dual-temperature/pH-responsive in situ gelling hydrogels that can be injected via narrow gauge needles. The incorporation of charge significantly broadens the swelling, degradation, and rheological profiles achievable with injectable POEGMA hydrogels without significantly increasing nonspecific protein adsorption or chronic inflammatory responses following in vivo subcutaneous injection. However, significantly different cell responses are observed upon charge incorporation, with charged gels significantly improving 3T3 mouse fibroblast cell adhesion in 2D and successfully delivering viable and proliferating ARPE-19 human retinal epithelial cells via an "all-synthetic" matrix that does not require the incorporation of cell-adhesive peptides.

An omniphobic lubricant-infused coating produced by chemical vapor deposition of hydrophobic organosilanes attenuates clotting on catheter surfaces.

ABSTARCT: Catheter associated thrombosis is an ongoing problem. Omniphobic coatings based on tethering biocompatible liquid lubricants on self-assembled monolayers of hydrophobic organosilanes attenuate clotting on surfaces. Herein we report an efficient, non-invasive and robust process for coating catheters with an antithrombotic, omniphobic lubricant-infused coating produced using chemical vapor deposition (CVD) of hydrophobic fluorine-based organosilanes. Compared with uncoated catheters, CVD coated catheters significantly attenuated thrombosis via the contact pathway of coagulation. When compared with the commonly used technique of liquid phase deposition (LPD) of fluorine-based organosilanes, the CVD method was more efficient and reproducible, resulted in less disruption of the outer polymeric layer of the catheters and produced greater antithrombotic activity. Therefore, omniphobic coating of catheters using the CVD method is a simple, straightforward and non-invasive procedure. This method has the potential to not only prevent catheter thrombosis, but also to prevent thrombosis on other blood-contacting medical devices.