Biofunctionalization Strategies Research Articles

A Surface-Mediated Biomimetic Porous Polyether-Ether-Ketone Scaffold for Regulating Immunity and Promoting Osteogenesis.

The repair of critical-sized bone defects remains a major challenge for clinical orthopedic surgery. Here, we develop a surface biofunctionalized three-dimensional (3D) porous polyether-ether-ketone (PEEK) scaffold that can simultaneously promote osteogenesis and regulate macrophage polarization. The scaffold is created using polydopamine (PDA)-assisted immobilization of silk fibroin (SF) and the electrostatic self-assembly of nanocrystalline hydroxyapatite (nano-HA) on a 3D-printed porous PEEK scaffold. The SF/nano-HA functionalized surface provides a bone-like microenvironment for osteoblastic cells' adhesion, proliferation, mineralization and osteogenic differentiation. Moreover, the biofunctionalized surface can effectively drive macrophages polarization from the pro-inflammatory M1 phenotype to the anti-inflammatory M2 phenotype. Integrin β1-specific cell-matrix binding and the activation of Ca2+ receptor-mediated signaling pathway play critical roles in the regulation of macrophage polarization. Compared with the as-printed scaffold, the SF/nano-HA functionalized porous PEEK scaffold induces minimal inflammatory response, enhanced angiogenesis, and substantial new bone formation, resulting in improved osseointegration in vivo. This study not only develops a promising candidate for bone repair but also demonstrates a facile surface biofunctionalization strategy for orthopedic implants to improve osseointegration by stimulating osteogenesis and regulating immunity.

Lithography-Free Metaplasmonic Sensors Developed by TWD/GLAD Technique

Nowadays, plasmonic sensors are the most widely used and commercialized label-free optical biosensors and have become a widespread tool for studying chemical/biochemical interactions. Label-free, real-time, and direct measurements are the major benefits of the plasmonic sensors, including high-throughput surface bio-functionalization strategies without amplification or pretreatment of the sample [1]. The working principle of plasmonic sensing has been extensively described and reviewed over the decades [2], however, the possibility to develop cheap, effective, and clinically-accepted biosensors has recently become available due to the development of nanotechnology. Briefly, these nanostructures can be localized or can be arranged on 2D arrays of plasmonic metasurfaces, and can be fabricated by single-layer metallic films and a combination of metallic and dielectric films. The most popular plasmonic metals are gold and silver due to their high conductivity and low dielectric losses. Generally, top-down nanofabrication methods based on laser/e-beam lithography have been used, including nanostencil lithography based on shadow-masked nano-pattering and nanoimprint lithography [3]. Although such technologies have a high potential to achieve scalable and cost-effective nanofabrication at the wafer scale, these processes maintain the following main challenge: a master nano-mold/pattern is required to transfer metasurfaces with an associated high cost. Therefore, other technologies are the subject of research that overcomes this limitation and still offer an outstanding quality of metallic films, such as TDW (thermal dewetting) and GLAD (glancing angle deposition) (Fig.1). These techniques do not require master nano-mold/patterns; consequently, they can achieve lithography-free large-scale plasmonic metasurfaces [4]. TDW and GLAD usually generate quasi-ordered plasmonic metasurfaces compared to conventional lithographic methods.In this paper, the experimental results of the optical sensing properties of the sensors developed by the utilization of the combination of these two technologies in a single system are presented. The TWD/GLAD magnetron sputtering system has been designed and manufactured based on the Kurt J. Lesker MAG-Torus magnetrons, supplied with DC/RF power sources and an ECR manipulator that enables deposition at various angles with maximal 20 rpm rotation speed and heating option up to 850oC. The system is controlled by the software that enables deposition of the samples with the same parameters which pave the way for fabrication of the sensors on the industrial scale. The optical-sensing system was built based on an ST-VIS-50 spectrometer (Ocean Optics) and Tungsten Halogen Source (360-2000 nm, 2800 K, Ocean Optics) and optical table from Thorlabs. The obtained reflectance measurement has shown that the utilization of the GLAD technique decreases the reflectance peak in comparison with samples deposited without GLAD (flat samples). At the same time, angles in the range of 82-86oC seem to be preferable for nanostructure fabrication.Additionally, the utilization of the TWD technique, i.e. deposition at lower temperatures in the range of 60-100oC (depending on the substrate) and then annealed at higher temperatures such as 250oC and 300oC in a vacuum and under argon flow in the deposition chamber led to increased normalized response. However, the experiments have shown that the optimal annealing time is between 30-45 min depending on the SPR multi-structure, for example for Ag (6nm), Ti (2nm), and Au (2nm) the 30min at 250oC seems to be the best set. For such compositions, the surface sensitivity (nm/nm) and bulk sensitivity (nm/RIU) are more or less the same - 0.9 and 300, respectively. The obtained results are very promising for developing cheap, rapid, and very effective biosensors for various applications. Therefore, the obtained results seem to be very interesting for the ECS conference audience, for example, the developed Ag/Ti/Au multi-structures can be applied in novel organ-on-a-chip platforms for LADMET (liberations, absorption, distribution, metabolism, excretion, and toxicology) analysis [5].Fig. 1. Schematic diagram representation of lithography free methods for developing quasi-ordered metamaterials from left to right: Thermal dewetting and glancing angle deposition. The insert shows the chiral plasmonic nanospirals fabrication by glanced angle deposition [4].

Impact of surface biofunctionalization strategies on key effector cells response in polyacrylamide hydrogels for bone regeneration

Despite the clinical prevalence of various bone defect repair materials, a full understanding of their influence on bone repair and regeneration remains elusive. This study focuses on poly(acrylamide) (PAAm) hydrogels, popular 2D model substrates, which have regulable mechanical properties within physiological. However, their bio-inert nature requires surface biofunctionalization to enhance cell-material interactions and facilitate the study of bone repair mechanisms. We utilized PAAm hydrogels of varying stiffness (18, 76 and 295 kPa), employed sulfosuccinimidyl-6-(4′-azido-2′-nitropheny-lamino) hexanoate (sulfo-SANPAH) and N-(3-dimethylaminopropyl)-N-ethylcarbodiimide hydrochloride/N-hydroxysuccinimidyl acrylate (EDC/NHS) as crosslinkers, and cultured macrophages, endothelial cells, and bone mesenchymal stem cells on these hydrogels. Our findings indicated that sulfo-SANPAH's crosslinking efficiency surpassed that of EDC/NHS, irrespective of pore size and stiffness. Importantly, we observed that the stiffness and surface biofunctionalization method of hydrogels significantly impacted cell adhesion and proliferation. The collagen-modified hydrogels by EDC/NHS strategy failed to support the normal biological behavior of bone mesenchymal stem cells and hindered endothelial cell spreading. In contrast, these modified hydrogels by the sulfo-SANPAH method showed good cytocompatibility with the three types of cells. This study underscores the critical role of appropriate conjugation strategies for PAAm hydrogels, providing valuable insights for hydrogel surface modification in bone repair and regeneration research.

Near-Infrared Detection of Pathogens Using Single-Walled Carbon Nanotubes

Viral and microbiological infections are a major cause of death that burdens the infrastructure of health systems. To contain the spread of pathogens and assess available treatment options, a number of diagnostic methods have been developed. However, the development of diagnostic tests is typically time consuming and, in the light of current pandemics, the need for adaptable concepts has become more apparent.For sensor applications that require a minimally invasive read out, optical systems offer a rapid detection of biomolecules with a high spatial and temporal resolution. Due to the reduced autofluorescence, absorption and scattering, the near-infrared (NIR, 870 – 2400 nm) tissue transparency window offers an improved signal-to-noise ratio. Biosensors that fluoresce in this region are therefore highly desirable for biological applications and single-walled carbon nanotubes (SWCNTs) have emerged as versatile NIR fluorescent building blocks in this context. Here, we report a biofunctionalization strategy for single walled carbon nanotube (SWCNT) based (bio-)sensors that uses a generic signal transduction mechanism for different recognition elements. It allows the detection of viral compounds in subnanomolar concentrations, which can be used to fingerprint them at 1020 nm in the near infrared. In summary, we present a concept that allows rapid exchange of recognition units and a straightforward method to detect pathogens.

Coiled-Coil-Based Biofunctionalization of 100 nm Gold Nanoparticles with the Trastuzumab Antibody for the Detection of HER2-Positive Cancer Cells.

We compared different biofunctionalization strategies for immobilizing trastuzumab, an IgG targeting the HER2 biomarker, onto 100 nm spherical gold nanoparticles because of the E/K coiled-coil peptide heterodimer. First, Kcoil peptides were grafted onto the gold surface while their Ecoil partners were genetically encoded at the C-terminus of trastuzumab's Fc region, allowing for a strong and specific interaction between the antibodies and the nanoparticles. Gold nanoparticles with no Kcoil peptides on their surface were also produced to immobilize Ecoil-tagged trastuzumab antibodies via the specific adsorption of their negatively charged Ecoil tags on the positively charged gold surface. Finally, the nonspecific adsorption of wild-type trastuzumab on the gold surface was also assessed, with and without Kcoil peptides grafted on it beforehand. We developed a thorough workflow to systematically compare the immobilization strategies regarding the stability of nanoparticles, antibody coverage, and ability to specifically bind to HER2-positive breast cancer cells. All nanoparticles were highly monodisperse and retained their localized surface plasmon resonance properties after biofunctionalization. A significant increase in the amount of immobilized antibodies was observed with the two oriented coil-based strategies compared to nonspecific adsorption. Finally, all biofunctionalization strategies allowed for the detection of HER2-positive breast cancer cells, but among the investigated approaches, we recommend using the E/K coiled-coil-based strategy for gold nanoparticle biofunctionalization because it allows for the qualitative and quantitative detection of HER2-positive cells with a higher contrast compared to HER2-negative cells.

Compact and modular bioprobe: Integrating SpyCatcher/SpyTag recombinant proteins with zwitterionic polymer-coated quantum dots

The development of quantum dot (QD)-based modular bioprobe that has a compact size and enable a facile conjugation of various biofunctional groups is in high demand. To address this, we surface engineered QDs with zwitterion polymer ligands to have an inherent compact size and derivatized them sequentially with the recombinant proteins SpyCatcher/SpyTag (SC/ST) to use their protein ligation system. SC/ST spontaneously form one complex through the isopeptide bond between them. SC-conjugated QDs (QD–SC) were used as base building blocks. Then, ST–biomolecules were added for modular biofunctionalization. We synthesized compact sized (∼15 nm) QD–SC–ST–affibody (antibody-mimicking small protein for tumor detection) conjugates, which showed successful cell-receptor targeting. The target cell-receptor could be easily tuned by changing the type of ST-affibody. We also demonstrated that anti-human-chorionic-gonadotropin mouse IgG1 antibodies can be labeled on the QD surface by mixing QD–SC with the ST–MG1Nb (mouse-IgG1-specific protein). The immunoassay performance of the antibody-labeled QDs was evaluated using a pregnancy test kit, displaying equivalent detection sensitivity to a commercially available kit. This study proposed an innovative strategy for QD biofunctionalization in a modular manner, which can be expanded to a diverse range of colloidal particle derivatization.

Copper-free click chemistry assisted antibodies for immunodetection of interleukin-10 in saliva

Interleukin-10 (IL-10) is an anti-inflammatory cytokine that is secreted in response to an acute phase inflammation in patients who are suffering from heart failure (HF). The aim of this work was to develop an electrochemical biosensor for determining salivary IL-10 levels. Biofunctionalization strategy was improved through the use of copper-free click chemistry for the developed sensor due to its advantages, leading to high quantitative yields of stable triazoles, rapid reaction, no cytotoxic Cu(I) catalyst requirement, and high specificity of cyclooctynes toward azides. The approach involved in binding of dibenzocyclooctyne acid (DBCO-COOH) to thiol-azide assembled gold microelectrodes, later capturing the monoclonal IL-10 antibody (IL-10 mAb), and ultimately allowing direct detection of IL-10 antigen. Fourier transform infrared spectroscopy (FTIR) and nanoplotter associated with fluorescence microscopy methods have been employed to analyze and prove the biofunctionalization of the gold microelectrodes. Moreover, the electrochemical impedance spectroscopy (EIS) technique was used for detecting IL-10 antigen. The developed immunosensor showed a semi-logarithmic linear range, from 0.1 pg/mL to 5 pg/mL with R2 = 0.9815 and a limit of quantitation (LOQ) of 0.1 pg/mL with relative standard deviation (RSD) of 10.67%. The specificity of the immunosensor was evaluated using an inflammatory cytokine, and none of it generated detectable EIS signals. Finally, the successful analysis of saliva samples from a healthy volunteer without Coronavirus (COVID-19) infection demonstrated the usefulness of the developed immunosensor.

Toward Clinical Transfer of Tumor‐Targeted Ultrasmall Inorganic Nanoparticles

AbstractUltrasmall nanoparticles (USNs) (nanoparticles with hydrodynamic diameter <10 nm) are being widely developed pre‐clinically and started to emerge in clinical trials over the last decade. Most of these USNs display the same features including short retention time in the blood, rapid renal clearance, and relie on passive targeting strategy to reach the tumor. Through this review, the development of AGuIX USNs is focused on because of their clinical usages as passively targeted USN but also because of their possible biofunctionalizations with peptides and monoclonal antibodies which are validated in various pre‐clinical tumor models. As a result, the authors reviewed all the current biofunctionalization strategies that can be employed and confirmed based on a meta‐analysis of the literature that biofunctionalized USNs pharmacokinetic and biodistribution profiles are dictated by the USNs and not the active targeting moiety. Additionally, it is demonstrated that such active targeting strategy improves the tumor targeting efficiency of the AGuIX USN but also increases their tumor retention time in comparison to the passively targeted AGuIX USNs, which may lead to an opportunity to reduce the number of injections/expend the therapeutic benefit of the drug product.

High-throughput biointerfaces for direct, label-free, and multiplexed metaplasmonic biosensing

In recent years, metaplasmonic biosensors have emerged as a novel counterpart of well-established plasmonic biosensors based on thin metallic layers. Metaplasmonic biosensors offer high potential for sensor miniaturization, extreme sensitivity biosensing, and high multiplexing capabilities with detection methods free of coupling optical elements. These capabilities make metaplasmonic biosensors highly attractive for Point-of-Care and handled/portable devices or novel On-Chip devices; as a result, it has increased the number of prototypes and potential applications that emerged during the last years. One of the main challenges to achieving fully operative devices is the achievement of high-throughput biointerfaces for sensitive and selective biodetection in complex media. Despite the superior surface sensitivity achieved by metaplasmonic sensors compared to conventional plasmonic sensors based on metallic thin films, the main limitations to achieving high-throughput and multiplexed biosensing usually are associated with the sensitivity and selectivity of the biointerface and, as a consequence, their application to the direct analysis of real complex samples. This graphical review discusses the potential challenges and capabilities of different biofunctionalization strategies, biorecognition elements, and antifouling strategies to achieve scalable and high-throughput metaplasmonic biosensing for Point-of-Care devices and bioengineering applications like Organs-On-Chip.

Lab-on-PCB-Based Electrical Immunosensing Platform for Point-of-Care Detection of SARS-CoV-2

The present work shows a multiplexed lab-on-printed circuit board (PCB) platform for label-free immunosensing of SARS-CoV-2 nucleocapsid and spike antigens based on electrical impedance measurements. The sensor consists of an interdigitated electrode of soft gold integrated on a PCB with microwells for sample loading. A mercaptoundecanoic acid–protein-G-based site-specific biofunctionalization strategy is employed to efficiently immobilize dual antibodies on the device surface toward the sensitive and rapid antigen test of SARS-CoV-2. Electrical impedance measurements carried out in a point-of-care setting using the PalmSens Sensit Smart system that could detect nucleocapsid and spike proteins with a detection limit of 40 and 20 pg each. Experiments with nasopharyngeal swab samples from <italic xmlns:mml="http://www.w3.org/1998/Math/MathML" xmlns:xlink="http://www.w3.org/1999/xlink">N</i> = 5 healthy and <italic xmlns:mml="http://www.w3.org/1998/Math/MathML" xmlns:xlink="http://www.w3.org/1999/xlink">N</i> = 14 SARS-CoV-2 positive subjects showed significantly different electrical responses for subjects with high viral load ( <italic xmlns:mml="http://www.w3.org/1998/Math/MathML" xmlns:xlink="http://www.w3.org/1999/xlink">Ct</i> < 25) compared with healthy subjects and control.

Biofunctionalization of Multiplexed Silicon Photonic Biosensors.

Silicon photonic (SiP) sensors offer a promising platform for robust and low-cost decentralized diagnostics due to their high scalability, low limit of detection, and ability to integrate multiple sensors for multiplexed analyte detection. Their CMOS-compatible fabrication enables chip-scale miniaturization, high scalability, and low-cost mass production. Sensitive, specific detection with silicon photonic sensors is afforded through biofunctionalization of the sensor surface; consequently, this functionalization chemistry is inextricably linked to sensor performance. In this review, we first highlight the biofunctionalization needs for SiP biosensors, including sensitivity, specificity, cost, shelf-stability, and replicability and establish a set of performance criteria. We then benchmark biofunctionalization strategies for SiP biosensors against these criteria, organizing the review around three key aspects: bioreceptor selection, immobilization strategies, and patterning techniques. First, we evaluate bioreceptors, including antibodies, aptamers, nucleic acid probes, molecularly imprinted polymers, peptides, glycans, and lectins. We then compare adsorption, bioaffinity, and covalent chemistries for immobilizing bioreceptors on SiP surfaces. Finally, we compare biopatterning techniques for spatially controlling and multiplexing the biofunctionalization of SiP sensors, including microcontact printing, pin- and pipette-based spotting, microfluidic patterning in channels, inkjet printing, and microfluidic probes.

In Situ Hydrogelation of Cellular Monolayers Enables Conformal Biomembrane Functionalization for Xeno-Free Feeder Substrate Engineering.

The intricate functionalities of cellular membranes have inspired strategies for deriving and anchoring cell-surface components onto solid substrates for biological studies, biosensor applications, and tissue engineering. However, introducing conformal and right-side-out cell membrane coverage onto planar substrates requires cumbersome protocols susceptible to significant device-to-device variability. Here, a facile approach for biomembrane functionalization of planar substrates is demonstrated by subjecting confluent cellular monolayer to intracellular hydrogel polymerization. The resulting cell-gel hybrid, herein termed GELL (gelated cell), exhibits extraordinary stability and retains the structural integrity, membrane fluidity, membrane protein mobility, and topology of living cells. In assessing the utility of GELL layers as a tissue engineering feeder substrate for stem cell maintenance, GELL feeder prepared from primary mouse embryonic fibroblasts not only preserves the stemness of murine stem cells but also exhibits advantages over live feeder cells owing to the GELL's inanimate, non-metabolizing nature. The preparation of a xeno-free feeder substrate devoid of non-human components is further shown with HeLa cells, and the resulting HeLa GELL feeder effectively sustains the growth and stemness of both murine and human induced pluripotent stem cells. The study highlights a novel bio-functionalization strategy that introduces new opportunities for tissue engineering and other biomedical applications.

Biofunctionalization of cardiovascular stents to induce endothelialization: Implications for in- stent thrombosis in diabetes.

Stent thrombosis remains one of the main causes that lead to vascular stent failure in patients undergoing percutaneous coronary intervention (PCI). Type 2 diabetes mellitus is accompanied by endothelial dysfunction and platelet hyperactivity and is associated with suboptimal outcomes following PCI, and an increase in the incidence of late stent thrombosis. Evidence suggests that late stent thrombosis is caused by the delayed and impaired endothelialization of the lumen of the stent. The endothelium has a key role in modulating inflammation and thrombosis and maintaining homeostasis, thus restoring a functional endothelial cell layer is an important target for the prevention of stent thrombosis. Modifications using specific molecules to induce endothelial cell adhesion, proliferation and function can improve stents endothelialization and prevent thrombosis. Blood endothelial progenitor cells (EPCs) represent a potential cell source for the in situ-endothelialization of vascular conduits and stents. We aim in this review to summarize the main biofunctionalization strategies to induce the in-situ endothelialization of coronary artery stents using circulating endothelial stem cells.

Surface Bio-Functionalization of Anti-Bacterial Titanium Implants: A Review

Titanium (Ti) and titanium alloy have been widely used in orthopedics. However, the successful application of titanium implants is mainly limited due to implant-associated infections. The implant surface contributes to osseointegration, but also has the risk of accelerating the growth of bacterial colonies, and the implant surfaces infected with bacteria easily form biofilms that are resistant to antibiotics. Biofilm-related implant infections are a disastrous complication of trauma orthopedic surgery and occur when an implant is colonized by bacteria. Surface bio-functionalization has been extensively studied to better realize the inhibition of bacterial proliferation to further optimize the mechanical functions of implants. Recently, the surface bio-functionalization of titanium implants has been presented to improve osseointegration. However, there are still numerous clinical and non-clinical challenges. In this review, these aspects were highlighted to develop surface bio-functionalization strategies for enhancing the clinical application of titanium implants to eliminate implant-associated infections.

Silk Fibroin/Hydroxyapatite Coating Improved Osseointegration of Porous Titanium Implants under Diabetic Conditions via Activation of the PI3K/Akt Signaling Pathway.

The application of three-dimensional printed porous titanium implants (TIs) is compromised in patients suffering from diabetes mellitus (DM), which disturbs the normal process of implant osseointegration, resulting in fixation failure. It was possibly because of reactive oxygen species (ROS) overproduction at the bone-implant interface. A silk fibroin-based hydroxyapatite (SF/HA) hybrid material emerged as a novel biological material for accelerating new bone formation. We proposed that the SF/HA hybrid coated titanium implant (SHT) could mitigate DM-mediated impaired osseointegration, which had never been reported previously. To test this assumption and further elucidate the mechanisms, primary rabbit osteoblasts were seeded on TIs or SHTs and cultured with normal serum, diabetic serum (DS), DS + N-acetyl-L-cysteine (NAC) (a potent ROS inhibitor), and DS + LY294002 (a specific PI3K/Akt inhibitor) for osteoblast behavior examinations. An animal study was performed on diabetic rabbits implanted with the two kinds of implants for osseointegration tests. DM-mediated ROS overproduction caused osteoblastic biological dysfunctions and apoptotic injury, associated with suppression of PI3K/Akt signaling in osteoblasts cultured on a TI substrate. Of note, the SHT substrate significantly suppressed ROS overproduction under diabetic conditions, improved osteoblast functional recovery including ameliorative osteoblast adhesion and morphology, improved cellular proliferation and differentiation, and abrogated apoptosis, which exhibited the same effect as NAC administration on the TI. The in vitro results were further corroborated in vivo by enhanced osteogenesis and osseointegration of SHTs in diabetic rabbits. Moreover, the aforesaid promotive effects afforded by the SF/HA coating were totally abolished with administration of LY294002 for blocking PI3K/Akt signaling. The above results collectively demonstrated that the SF/HA hybrid coating significantly ameliorated DM-mediated impaired osseointegration of the TI via reactivation of the ROS-mediated PI3K/Akt signaling pathway. The hybrid coating elicited a novel surface biofunctionalization strategy to attain favorable clinical performance of TI in diabetics.

Convergent Arrangement of sgRNA and Cas9 in CRISPRsome for Transcellular Trafficking

The emergence of clustered regularly interspaced short palindromic repeats (CRISPR)-based genome editing systems has led to the appreciation of CRISPR as a versatile and powerful gene-editing tool in the molecular biology laboratory and a potentially potent therapeutic for the treatment of a variety of intractable diseases. Nonetheless, one of the most onerous obstacles that restricts the wide availability of CRISPR technology from wide practical applications is the lack of appropriate delivery vehicles for the simultaneous transportation of multicomponent CRISPR systems to the intracellular targets. Herein, we attempted to create an intriguing nanometer-scaled vesicle platform (CRISPRsome) that, with the aid of a block copolymer of poly(ethylene glycol)-polylysine, simultaneously combined the fundamental components of the CRISPR system into the CRISPRsome, particularly the ribonucleic sgRNA and proteinic Cas9 endonuclease precisely located at the intermediate membrane and the internal reservoir regions of the nanometer-scaled vesicles, respectively. Furthermore, biofunctionalization strategies, including using redox-responsive disulfide linkages between lysine units to cross-link the well-defined CRISPRsome structures and surface functionalization with the cyclic Arg-Gly-Asp ligand to stimulate receptor-mediated endocytosis, were proposed in the well-defined CRISPRsome. The subsequent results validated its appreciable gene knockout efficacies toward cancerous cells with the green fluorescent protein (GFP) reporter gene, thereby highlighting the prosperous potential of CRISPRsome to facilitate the use of CRISPR technology in a variety of practical applications.

Improved cell adhesion to activated vapor silanization-biofunctionalized Ti-6Al-4V surfaces with ECM-derived oligopeptides

Titanium implants are widely used in traumatology and various orthopedic fields. Titanium and other metallic-based implants have limited structural and functional integration into the body, which translates into progressive prosthesis instability and the need for new surgical interventions that have enormous social and economic impacts. To enhance the biocompatibility of titanium implants, numerous biofunctionalization strategies have been developed. However, the problem persists, as more than 70% of implant failures are due to aseptic loosening. In this study we addressed the problem of improving the physiological engraftability and acceptability of titanium-based implants by applying a robust and versatile functionalization method based on the covalent immobilization of extracellular matrix (ECM)-derived oligopeptides on Ti-6Al-4V surfaces treated by activated vapor silanization (AVS). The feasibility of this technique was evaluated with two oligopeptides of different structures and compositions. These oligopeptides were immobilized on Ti-6Al-4V substrates by a combination of AVS and N-(3-dimethylaminopropyl)-N′-ethylcarbodiimide hydrochloride/N-hydroxysuccinimide (EDC/NHS) crosslinking chemistry. The immobilization was shown to be stable and resistant to chemical denaturing upon sodium dodecyl sulfate treatment. On Ti-6Al-4V surfaces both peptides increased the attachment, spreading, rearrangement and directional growth of mesenchymal stem and progenitor cells (MSC) with chondro- and osteo-regenerative capacities. We also found that this biofunctionalization method (AVS-EDC/NHS) increased the attachment capacity of an immortalized cell line of neural origin with poor adhesive properties, highlighting the versatility and robustness of this method in terms of potential oligopeptides that may be used, and cell lineages whose anchorage to the biomaterial may be enhanced. Collectively, this novel functionalization strategy can accelerate the development of advanced peptide-functionalized metallic surfaces, which, in combination with host or exogenously implanted stem cells, have the potential to positively affect the osteoregenerative and osteointegrative abilities of metallic-based prostheses.

Designing Cardiovascular Implants Taking in View the Endothelial Basement Membrane.

Insufficient endothelialization of cardiovascular grafts is a major hurdle in vascular surgery and regenerative medicine, bearing a risk for early graft thrombosis. Neither of the numerous strategies pursued to solve these problems were conclusive. Endothelialization is regulated by the endothelial basement membrane (EBM), a highly specialized part of the vascular extracellular matrix. Thus, a detailed understanding of the structure–function interrelations of the EBM components is fundamental for designing biomimetic materials aiming to mimic EBM functions. In this review, a detailed description of the structure and functions of the EBM are provided, including the luminal and abluminal interactions with adjacent cell types, such as vascular smooth muscle cells. Moreover, in vivo as well as in vitro strategies to build or renew EBM are summarized and critically discussed. The spectrum of methods includes vessel decellularization and implant biofunctionalization strategies as well as tissue engineering-based approaches and bioprinting. Finally, the limitations of these methods are highlighted, and future directions are suggested to help improve future design strategies for EBM-inspired materials in the cardiovascular field.

Immunosensing Based on Optical Fiber Technology: Recent Advances.

The evolution of optical fiber technology has revolutionized a variety of fields, from optical transmission to environmental monitoring and biomedicine, given their unique properties and versatility. For biosensing purposes, the light guided in the fiber core is exposed to the surrounding media where the analytes of interest are detected by different techniques, according to the optical fiber configuration and biofunctionalization strategy employed. These configurations differ in manufacturing complexity, cost and overall performance. The biofunctionalization strategies can be carried out directly on bare fibers or on coated fibers. The former relies on interactions between the evanescent wave (EW) of the fiber and the analyte of interest, whereas the latter can comprise plasmonic methods such as surface plasmon resonance (SPR) and localized SPR (LSPR), both originating from the interaction between light and metal surface electrons. This review presents the basics of optical fiber immunosensors for a broad audience as well as the more recent research trends on the topic. Several optical fiber configurations used for biosensing applications are highlighted, namely uncladded, U-shape, D-shape, tapered, end-face reflected, fiber gratings and special optical fibers, alongside practical application examples. Furthermore, EW, SPR, LSPR and biofunctionalization strategies, as well as the most recent advances and applications of immunosensors, are also covered. Finally, the main challenges and an outlook over the future direction of the field is presented.

State of the Art and Perspectives on the Biofunctionalization of Fluorescent Metal Nanoclusters and Carbon Quantum Dots for Targeted Imaging and Drug Delivery.

The interface of nanobio science and cancer nanomedicine is one of the most important current frontiers in research, being full of opportunities and challenges. Ultrasmall fluorescent metal nanoclusters (MNCs) and carbon quantum dots (CQDs) have emerged as promising fluorescent nanomaterials due to their unique physicochemical and optical properties, facile surface functionalization, good photostability, biocompatibility, and aqueous dispersity. These characteristics make them advantageous over conventional fluorophores such as organic dye molecules and semiconductor quantum dots (QDs) for the detection, diagnosis, and treatment of various diseases including cancer. Recently, researchers have focused on the biofunctionalization strategy of the MNCs and CQDs which can tailor their physicochemical and biological properties and, in turn, can empower these biofunctionalized nanoprobes for diverse applications including imaging, drug delivery, theranostics, and other biomedical applications. In this invited feature article, we first discuss some fundamental structural and physicochemical characteristics of the fluorescent biocompatible quantum-sized nanomaterials which have some outstanding features for the development of multiplexed imaging probes, delivery vehicles, and cancer nanomedicine. We then demonstrate the diverse surface engineering of these fluorescent nanomaterials with reactive target specific functional groups which can help to construct multifunctional nanoprobes with improved targeting capabilities having minimal toxicity. The promising future of the biofunctionalized fluorescent quantum-sized nanomaterials in the field of bioanalytical and biomedical research is elaborately demonstrated, showing selected recent works with relevant applications. This invited feature article finally ends with a short discussion of the current challenges and future prospects of the development of these bioconjugated/biofunctionalized nanomaterials to provide insight into this burgeoning field of MNC- and CQD-based diagnostics and therapeutic applications.