Plasmonic silver and gold nanoparticles: shape- and structure-modulated plasmonic functionality for point-of-caring sensing, bio-imaging and medical therapy.

Silver and gold nanoparticles have found extensive biomedical applications due to their strong localized surface plasmon resonance (LSPR) and intriguing plasmonic properties. This review article focuses on the correlation among particle geometry, plasmon properties and biomedical applications. It discusses how particle shape and size are tailored via controllable synthetic approaches, and how plasmonic properties are tuned by particle shape and size, which are embodied by nanospheres, nanorods, nanocubes, nanocages, nanostars and core-shell composites. This article summarizes the design strategies for the use of silver and gold nanoparticles in plasmon-enhanced fluorescence, surface-enhanced Raman scattering (SERS), electroluminescence, and photoelectrochemistry. It especially discusses how to use plasmonic nanoparticles to construct optical probes including colorimetric, SERS and plasmonic fluorescence probes (labels/reporters). It also demonstrates the employment of Ag and Au nanoparticles in polymer- and paper-based microfluidic devices for point-of-care testing (POCT). In addition, this article highlights how to utilize plasmonic nanoparticles for in vitro and in vivo bio-imaging based on SERS, fluorescence, photoacoustic and dark-field models. Finally, this article shows perspectives in plasmon-enhanced photothermal and photodynamic therapy.

Visible-light and near-infrared fluorescence and surface-enhanced Raman scattering point-of-care sensing and bio-imaging: a review.

This review article deals with the concepts, principles and applications of visible-light and near-infrared (NIR) fluorescence and surface-enhanced Raman scattering (SERS) in in vitro point-of-care testing (POCT) and in vivo bio-imaging. It has discussed how to utilize the biological transparency windows to improve the penetration depth and signal-to-noise ratio, and how to use surface plasmon resonance (SPR) to amplify fluorescence and SERS signals. This article has highlighted some plasmonic fluorescence and SERS probes. It has also reviewed the design strategies of fluorescent and SERS sensors in the detection of metal ions, small molecules, proteins and nucleic acids. Particularly, it has provided perspectives on the integration of fluorescent and SERS sensors into microfluidic chips as lab-on-chips to realize point-of-care testing. It has also discussed the design of active microfluidic devices and non-paper- or paper-based lateral flow assays for in vitro diagnostics. In addition, this article has discussed the strategies to design in vivo NIR fluorescence and SERS bio-imaging platforms for monitoring physiological processes and disease progression in live cells and tissues. Moreover, it has highlighted the applications of POCT and bio-imaging in testing toxins, heavy metals, illicit drugs, cancers, traumatic brain injuries, and infectious diseases such as COVID-19, influenza, HIV and sepsis.

Macroporous Silk Nanofiber Cryogels with Tunable Properties.

Cryogels are widely used in tissue regeneration due to their porous structures and friendly hydrogel performance. Silk-based cryogels were developed but failed to exhibit desirable tunable properties to adapt various biomedical applications. Here, amorphous short silk nanofibers (SSFs) were introduced to fabricate silk cryogels with versatile cues. Compared to previous silk cryogels, the SSF cryogels prepared under same conditions showed significantly enhanced mechanical properties. The microporous cryogels were achieved under lower silk concentrations, confirming better tunability. Versatile cryogels with the modulus in the range of 0.5-283.7 kPa were developed through adjusting silk concentration and crosslinking conditions, superior to previous silk cryogel systems. Besides better cytocompatibility, the SSF cryogels were endowed with effective mechanical cues to control osteogenetic differentiation behaviors of BMSCs. The mechanical properties could be further regulated finely through the introduction of ß-sheet-rich silk nanofibers (SNFs), which suggested possible optimization of mechanical niches. Bioactive cargo-laden SNFs were introduced to the SSF cryogel systems, bringing biochemical signals without the compromise of mechanical properties. Versatile SNF-based cryogels with different physical and biological cues were developed here to facilitate the applications in various tissue engineering.

Development and Analytical Validation of a Surface-Enhanced Raman Scattering Paper Lateral Flow Immunoassay for Detection of the Ubiquitin C-Terminal Hydrolase-L1 Traumatic Brain Injury Biomarker.

Paper lateral flow immunoassays combined with surface-enhanced Raman scattering (SERS) technology have gained increasing attention due to their high sensitivity characteristics resulting from the amplified SERS signals of the plasmon-enhanced optical probes. In contrast to conventional colorimetric lateral flow strips, SERS paper lateral flow strips (SERS-PLFSs) are currently not commercially available for widespread use. Analytical validation is the key step for commercialization. In this work, we have developed a PLFS with a hierarchical SERS probe (gold-silver nanoparticle@Raman reporter@silica) for detection of the US Food and Drug Administration (FDA)-approved traumatic brain injury (TBI) protein biomarker, ubiquitin C-terminal hydrolase-L1 (UCH-L1), in blood plasma samples. Analytical validation has been performed on this SERS-PLFS in terms of the limit of detection (LOD), limit of quantification (LOQ), accuracy, precision, selectivity, and stability. The SERS-PLFS exhibits a reportable range of 0.2-100 ng/mL with a LOD of 0.08 ng/mL toward measurement of UCH-L1 in blood plasma. The SERS-PLFS has been applied to clinical TBI samples. The test results were compared with those from enzyme-linked immunosorbent assay (ELISA), demonstrating a strong correlation between the two analytical methods. This study has important implications in the commercialization of SERS-PLFSs for rapid TBI detection in clinical practice.

Blastocyst-Inspired Hydrogels to Maintain Undifferentiation of Mouse Embryonic Stem Cells.

Stem cell fate is determined by specific niches that provide multiple physical, chemical, and biological cues. However, the hierarchy or cascade of impact of these cues remains elusive due to their spatiotemporal complexity. Here, anisotropic silk protein nanofiber-based hydrogels with suitable cell adhesion capacity are developed to mimic the physical microenvironment inside the blastocele. The hydrogels enable mouse embryonic stem cells (mESCs) to maintain stemness in vitro in the absence of both leukemia inhibitory factor (LIF) and mouse embryonic fibroblasts (MEFs), two critical factors in the standard protocol for mESC maintenance. The mESCs on hydrogels can achieve superior pluripotency, genetic stability, developmental capacity, and germline transmission to those cultured with the standard protocol. Such biomaterials establish an improved dynamic niche through stimulating the secretion of autocrine factors and are sufficient to maintain the pluripotency and propagation of ESCs. The mESCs on hydrogels are distinct in their expression profiles and more resemble ESCs in vivo. The physical cues can thus initiate a self-sustaining stemness-maintaining program. In addition to providing a relatively simple and low-cost option for expansion and utility of ESCs in biological research and therapeutic applications, this biomimetic material helps gain more insights into the underpinnings of early mammalian embryogenesis.

Heparin mimics and fibroblast growth factor-2 fabricated nanogold composite in promoting neural differentiation of mouse embryonic stem cells.

The replacement therapy or transplantation using neural cells, which differentiated from stem cells, has emerged as a promising strategy for repairing damaged neural tissues and helping functional recovery in the treatment of neural system diseases. The challenge, however, is how to control embryonic stem cell fate so that neural differentiation can be efficiently directed to enrich a neuron cell population, and meanwhile to maintain their bioactivities. This is a key question and has a very significant impact in regenerative medicine. Here we proposed a new neural-differentiation inductive nanocomposite, containing gold nanoparticles (AuNPs), poly(2-methacrylamido glucopyranose-co-3-sulfopropyl acrylate) (PMS), and basic fibroblast growth factor (FGF2), for the high efficient directional neural-specific differentiation of mouse embryonic stem cells (mESCs). In this AuNP-PMS/FGF2 composite, PMS, playing as the high-active mimic of heparin/heparan sulfate (HS), is covalently anchored to AuNPs and bound with FGF2 on the surface of nanoparticles, forming a HS/FGF2 complex nanomimics to facilitate its binding to FGF receptor (FGFR) and promote high neural-inductive activity of mESCs. The stability, bioactivity and biocompatibility of the composite are investigated in this study. The results showed that the AuNP-PMS/FGF2 composite could maintain a long-term stability at room temperature for at least 8 days, and greatly promote the neural differentiation of mESCs. Compared with the other materials, the AuNP-PMS/FGF2 composite could significantly stimulate the expression of the specific neural differentiation markers (nestin and ß3-tubulin), while obviously down-regulate the mRNA production of pluripotency marker Oct-4 in mESCs. Moreover, the promotion effect of the composite on neuronal maturation marker ß3-tubulin expression achieved maximally at the low concentration of FGF2 (4 ng/mL), which suggested the high efficiency of AuNP-PMS/FGF2 composite in neural differentiation of mESCs. Meanwhile, both mESCs and L929 cells showed desirable growth during the incubation with AuNP-PMS/FGF2 composite. The AuNP-PMS/FGF2 system presents a new way to achieve HS/FGF2 complex nanomimics efficiently for the neural differentiation of mESCs.

Dual Pathway for Promotion of Stem Cell Neural Differentiation Mediated by Gold Nanocomposites.

The neural differentiation of embryonic stem cells (ESCs) is of great value in the treatment of neurodegenerative diseases. On the basis of the two related signaling pathways that direct the neural differentiation of ESCs, we used gold nanoparticles (GNP) as a means of combining chemical and physical cues to trigger the neurogenic differentiation of stem cells. Neural differentiation-related functional units (glyco and sulfonate units on glycosaminoglycans, GAG) were anchored on the GNP surface and were then transferred to the cell membrane surface via GNP-membrane interactions. The functional units were able to activate the GAG-related signaling pathway, in turn promoting differentiation and maturation of stem cells into neuronal lineages. In addition, using the photothermal effect of GNP, the differentiation-inducing factor retinoic acid (RA), could be actively delivered into cells via laser irradiation. The RA-related intracellular signaling pathway was thereby further triggered, resulting in strong promotion of neurogenesis with a 300-fold increase in mature neural marker expression. The gold nanocomposites developed in this work provide the basis for a new strategy directing ESCs differentiation into nerve cells with high efficiency and high purity by acting on two related signaling pathways.

Microfluidic Silk Fibers with Aligned Hierarchical Microstructures.

The hierarchical structure of the ECM provides specific niches for tissues to regulate cell behavior, yet the challenge remains to design biomaterial systems for tissue regeneration to recreate such features in vitro. Here, we achieved this goal through the use of aligned hierarchical structures of native silk fibers, generated through the integration of "bottom-up" and "top-down" strategies to generate regenerated silk fibers with aligned nano- to micro-hierarchical structures. To achieve these designs, we assembled and dispersed silk nanofibers (SNF) in formic acid and spun them into fibers using bioinspired microfluidic chips with a geometry mimicking the native silk gland. The fibers generated using this device exhibited aligned hierarchical structure with fiber mechanical properties superior to fibers derived from more traditional spinning approaches with regenerated silk solutions. Besides the improved mechanical properties, Raman spectroscopic results indicated similarly aligned structures to native fibers and active control of cell proliferation, migration, and aggregate orientation. The results indicate the feasibility of developing bioactive silk fiber materials with hierarchical structures to facilitate utility in a range of cell and tissue regeneration scenarios.

Small addition of Zn2+ in Ca2+@DNA results in elevated gene transfection by aminated PGMA-modified silicon nanowire arrays.

Gene therapy, a promising and effective treatment, has ignited new hope in overcoming difficult-to-cure diseases. The key question in gene therapy is how to efficiently and safely deliver exogenous nucleic acids into the nuclei of target cells. To achieve stable, efficient and safe gene transfer and to ensure efficiency of gene transfer into cell nuclei, a zinc ion-assisted gene delivery nanosystem was proposed in the present study by loading a low concentration of Zn2+ in Ca2+@DNA nanoparticles on ethanolamine-functionalized poly(glycidyl methacrylate) (PGEA)-modified SiNWAs (Zn2+/Ca2+@DNA + SN-PGEA). The results showed that with the help of Zn ions, this composite nanosystem could promote more DNA in the cell nuclei and thus dramatically increased the transfection efficiency by as much as 7-fold. The nanosystem with 0.2 mM Zn2+, 100 mM Ca2+ and PGEA modification on SiNWAs displayed the highest transfection efficiency and good biocompatibility. This new composite nanosystem will have great potential in gene transfection for biomedical research.

Modular Polymers as a Platform for Cell Surface Engineering: Promoting Neural Differentiation and Enhancing the Immune Response.

Regulating cell behavior and cell fate are of great significance for basic biological research and cell therapy. Carbohydrates, as the key biomacromolecules, play a crucial role in regulating cell behavior. Herein, "modular" glycopolymers were synthesized by reversible addition-fragmentation chain transfer polymerization. These glycopolymers contain sugar units (glucose), anchoring units (cholesterol), "guest" units (adamantane) for host-guest interaction, and fluorescent labeling units (fluorescein). It was demonstrated that these glycopolymers can insert into cell membranes with high efficiency and their residence time on the membranes can be regulated by controlling their cholesterol content. Furthermore, the behavior of the engineered cells can be controlled by modifying with different functional ß-cyclodextrins (CD-X) via host-guest interactions with the adamantane units. Host-guest interactions with the modular polymers were demonstrated using CD-RBITC (X = a rhodamine B isothiocyanate). The glycopolymers were modified with CD-S (X = seven sulfonate groups) and CD-M (X = seven mannose groups) and were then attached, respectively, to the surfaces of mouse embryonic stem cells for the promotion of neural differentiation and to the surfaces of cancer cells for the enhancement of the immune response. The combination of multiple anchors and host-guest interactions provides a widely applicable cell membrane modification platform for a variety of applications.

Structure-Chemical Modification Relationships with Silk Materials.

Chemical modifications used with silk materials can be challenging due to heterogeneous reactions, in part due to the assembly state of the protein chains. Here, we assess factors that determine the efficiency of chemical modifications with silk materials. Unlike other natural macromolecules, silk presents changeable self-assembled or aggregation states in aqueous solution, which affect the chemical reactions based on reactive group distribution or accessibility. To confirm this hypothesis, silk nanofibers in various conformation and aggregation states in solution were exposed to the same reaction conditions. Amorphous silk nanofibers provided improved control for consistent chemical modification outcomes, while silk nanofibers with control of structure could be utilized to generate bifunctional materials through multiple chemical modifications. The results of the chemical modifications demonstrated that control of the conformational transitions of silk nanofibers provided a feasible strategy for developing multifunctional silk materials with improved chemical outcomes.

Microfluidic channels with renewable and switchable biological functionalities based on host-guest interactions.

Poly(dimethylsiloxane) (PDMS)-based microfluidic systems are gaining increasing attention due to their ease of fabrication, optical transparency and mechanical properties. However, the inherent hydrophobicity and chemical inertness of PDMS hinder its wider application in microfluidic systems. There is thus a strong need for methods for surface modification of PDMS-based microfluidic channels. In this work, oligo(ethylene glycol)methacrylate (OEGMA) and adamantane-containing OEGMA (OEGMA-Ada) were graft copolymerized on PDMS microchannel surfaces using a simple photochemical process to give PDMS-POA. OEGMA was chosen for its resistance to non-specific protein adsorption, and OEGMA-Ada was chosen for its subsequent attachment of mannose with bacteria binding affinity or biotin with avidin binding affinity. ß-CD decorated with biotin (CD-B) and/or mannose (CD-M) was attached to the PDMS-POA microchannels via host-guest interactions between the adamantane and ß-CD moieties. The data obtained suggest that the functions of the PDMS-POA/CD-B and PDMS-POA/CD-M microchannels with respect to biotin binding and bacterial adhesion were renewable. In addition, the biofunction of the PDMS-POA microchannels could be switched by treatment with SDS to release the CD component followed by treatment with a different ß-CD derivative. Different from previous surface modification strategies for PDMS-based microfluidic channels, the combination of visible light-induced grafting and host-guest chemistry provides modified PDMS microchannels with renewable and switchable biofunctions for the detection and measurement of specific proteins and bacteria.