In vitro vascularized liver tumor model based on a microfluidic inverse opal scaffold for immune cell recruitment investigation.

Liver cancer, characterized as a kind of malignant tumor within the digestive system, poses great health harm, and immune escape stands out as an important reason for its occurrence and development. Chemokines, pivotal in guiding immune cells' migration, is necessary to initiate and deliver an effective anti-tumor immune response. Therefore, understanding the chemotactic environment and identifying chemokines that regulate recruitment of immune cells to the tumor microenvironment (TME) are critical to improve current immunotherapy interventions. Herein, we report a well-defined inverse opal scaffold generated with a microfluidic emulsion template for the construction of a vascularized liver tumor model, offering insights into immune cells' recruitment. Due to the excellent 3D porous morphology of the inverse opal scaffold, human hepatocellular carcinoma cells can aggregate in the pores of the scaffold to form uniform multicellular tumor spheroids. More attractively, the vascularized liver tumor model can be achieved by constructing a 3D co-culture system involving endothelial cells and hepatocellular carcinoma cells. The results demonstrate that the 3D co-cultured tumor cells increase the neutrophil chemokines remarkably and recruit neutrophils to tumor tissues, then promote tumor progression. This approach opens a feasible avenue for realizing a vascularized liver tumor model with a reliable immune microenvironment close to that of a solid tumor of liver cancer.

Sequential Regulation of Local Reactive Oxygen Species by Ir@Cu/Zn-MOF Nanoparticles for Promoting Infected Wound Healing.

Most antimicrobials treat wound infections by an oxidation effect, which is induced by the generation of reactive oxygen species (ROS). However, the potential harm of the prolonged high level of ROS should not be ignored. In this study, we presented a novel cascade-reaction nanoparticle, Ir@Cu/Zn-MOF, to effectively regulate the ROS level throughout the healing progress of the infected wound. The nanoparticles consisted of a copper/zinc-modified metal-organic framework (Cu/Zn-MOF) serving as the external structure and an inner core composed of Ir-PVP NPs, which were achieved through a process known as "bionic mineralization". The released Cu2+ and Zn2+ from the shell structure contributed to the production of ROS, which acted as antimicrobial agents during the initial stage. With the disintegration of the shell, the Ir-PVP NP core was gradually released, exhibiting the property of multiple antioxidant enzyme activities, thereby playing an important role in clearing excessive ROS and alleviating oxidative stress. In a full-layer infected rat wound model, Ir@Cu/Zn-MOF nanoparticles presented exciting performance in promoting wound healing by clearing the bacteria and accelerating neovascularization as well as collagen deposition. This study provided a promising alternative for the repair of infected wounds.

An inflammation-responsive double-layer microneedle patch for recurrent atopic dermatitis therapy.

Seeking a potent therapeutic strategy for alleviating atopic dermatitis (AD) attack and preventing its recurrence is highly desired but remains challenging in clinical practice. Here, we propose an inflammation-responsive double-layer microneedle (IDMN) patch in situ delivering VD3 for recurrent AD therapy. IDMN comprises the backing layer part and the double-layer microneedle part, in which the inner layer is gelatin methacryloyl (GelMA) loaded with VD3 while the outer layer is composed of hyaluronic acid (HA). Introduction of the HA backing layer and outer layer around the GelMA tips can not only provide sufficient mechanical strength to penetrate into hardened AD skin with minimal invasiveness, but also exert a strong moisturizing effect after being rapidly dissolved. The inner layer of GelMA is degraded by the matrix metalloproteinase (MMP) in a dose dependent manner, which is secreted according to the disease progression of AD. The responsive degradation of GelMA tips result in corresponding release of VD3 to treat AD, triggering negative feedback against GelMA degradation. The IDMN administration on AD-bearing mice reveals efficient "curing" performances (including suppress erythema, scaling and lichenification, reduce epidermal thickness, inhibit mast cells infiltration, and down-regulate inflammatory factor secretion), which are basically realized through synergistic effect of the released VD3 and the dissolved HA molecules. Importantly, the residual tips of IDMN with VD3 are retained in the skin after the first AD relief, showing promising "warning" ability to inhibit the recurrence of AD. Hence, the developed IDMN patch is expected to be one of the excellent candidates for AD therapy and other relapsing diseases in clinical fields.

Progress and Perspective of CRISPR-Cas9 Technology in Translational Medicine.

Translational medicine aims to improve human health by exploring potential treatment methods developed during basic scientific research and applying them to the treatment of patients in clinical settings. The advanced perceptions of gene functions have remarkably revolutionized clinical treatment strategies for target agents. However, the progress in gene editing therapy has been hindered due to the severe off-target effects and limited editing sites. Fortunately, the development in the clustered regularly interspaced short palindromic repeats associated protein 9 (CRISPR-Cas9) system has renewed hope for gene therapy field. The CRISPR-Cas9 system can fulfill various simple or complex purposes, including gene knockout, knock-in, activation, interference, base editing, and sequence detection. Accordingly, the CRISPR-Cas9 system is adaptable to translational medicine, which calls for the alteration of genomic sequences. This review aims to present the latest CRISPR-Cas9 technology achievements and prospect to translational medicine advances. The principle and characterization of the CRISPR-Cas9 system are firstly introduced. The authors then focus on recent pre-clinical and clinical research directions, including the construction of disease models, disease-related gene screening and regulation, and disease treatment and diagnosis for multiple refractory diseases. Finally, some clinical challenges including off-target effects, in vivo vectors, and ethical problems, and future perspective are also discussed.

Aligned electrospun fiber film loaded with multi-enzyme mimetic iridium nanozymes for wound healing.

A film with elaborate microstructures that offers biomimetic properties and multi functionalities is highly desired in wound healing. Here, we develop an aligned hydrogel fiber film integrated with multi-active constituents to promote wound healing. Such fiber films are designed and constructed by photo-crosslinking the methacrylate gelatin (GelMA) doped with silver nanoparticles (Ag NPs) and iridium nanoparticles coated with polyvinylpyrrolidone (PVP-Ir NPs) in the precursor solution using electrospinning. The nature of GelMA hydrogel and the aligned arrangement of nanofibers endow the film with high-water content, self-degradability, improved bionic characteristics, oriented cell growth, and improved cell proliferation and migration. Moreover, the encapsulated nanozymes and Ag NPs offer the fiber film with superior reactive oxygen species (ROS) scavenging and antibacterial capability. The infected wound model shows that the multi-active hydrogel fiber film can reduce inflammation by killing bacteria and decomposing ROS, which accelerates the growth of new blood vessels and granulation tissue. Benefitting from these features, the versatile aligned GelMA fiber film demonstrates the clinically translational potential for wound healing.

Erratum to "Responsive Inverse Opal Scaffolds with Biomimetic Enrichment Capability for Cell Culture".

[This corrects the article DOI: 10.34133/2019/9783793.].

Droplet microfluidics-based biomedical microcarriers.

Droplet microfluidic technology provides a new platform for controllable generation of microdroplets and droplet-derived materials. In particular, because of the ability in high-throughput production and accurate control of the size, structure, and function of these materials, droplet microfluidics presents unique advantages in the preparation of functional microcarriers, i.e., microsized liquid containers or solid particles that serve as substrates of biomolecules or cells. These microcarriers could be extensively applied in the areas of cell culture, tissue engineering, and drug delivery. In this review, we focus on the fabrication of microcarriers from droplet microfluidics, and discuss their applications in the biomedical field. We start with the basic principle of droplet microfluidics, including droplet generation regimes and its control methods. We then introduce the fabrication of biomedical microcarriers based on single, double, and multiple emulsion droplets, and emphasize the various applications of microcarriers in biomedical field, especially in 3D cell culture, drug development and biomedical detection. Finally, we conclude this review by discussing the limitations and challenges of droplet microfluidics in preparing microcarriers. STATEMENT OF SIGNIFICANCE: Because of its precise control and high throughput, droplet microfluidics has been employed to generate functional microcarriers, which have been widely used in the areas of drug development, tissue engineering, and regenerative medicine. This review is significant because it emphasizes recent progress in research on droplet microfluidics in the preparation and application of biomedical microcarriers. In addition, this review suggests research directions for the future development of biomedical microcarriers based on droplet microfluidics by presenting existing shortcomings and challenges.

Porous MOF Microneedle Array Patch with Photothermal Responsive Nitric Oxide Delivery for Wound Healing.

Patches with the capacity of controllable delivering active molecules toward the wound bed to promote wound healing are expectant all along. Herein, a novel porous metal-organic framework (MOF) microneedle (MN) patch enabling photothermal-responsive nitric oxide (NO) delivery for promoting diabetic wound healing is presented. As the NO-loadable copper-benzene-1,3,5-tricarboxylate (HKUST-1) MOF is encapsulated with graphene oxide (GO), the resultant NO@HKUST-1@GO microparticles (NHGs) are imparted with the feature of near-infrared ray (NIR) photothermal response, which facilitate the controlled release of NO molecules. When these NHGs are embedded in a porous PEGDA-MN, the porous structure, larger specific surface area, and sufficient mechanical strength of the integrated MN could promote a more accurate and deeper delivery of NO molecules into the wound site. By applying the resultant NHG-MN to the wound of a type I diabetic rat model, the authors demonstrate that it is capable of accelerating vascularization, tissue regeneration, and collagen deposition, indicating its bright prospect applied in wound healing and other therapeutic scenarios.

Bio-inspired wettability patterns for biomedical applications.

Benefiting from the remarkable wettability heterogeneity, bio-inspired wettability patterns present a progressive and versatile platform for manipulating and patterning liquids, which provides an emerging strategy for operating liquid samples with crucial values in biomedical applications. In this review, we present a general summary of bio-inspired wettability patterns. After a compendious introduction of natural wettability phenomena and their underlying mechanisms, we summarize the general design principles and fabrication methods for preparing artificial wettability materials. Next, we shift to patterned surface wettability with an emphasis on the fabrication approaches. Then, we discuss in detail the various practical applications of wettability patterns in the biomedical field, including cell culture, drug screening and biosensors. Critical thinking about the current challenges and future outlook is also provided. We believe that this review would propel the prosperous development of bio-inspired wettability patterns to flourish in the field of biomedical engineering.

Aptamer-decorated porous microneedles arrays for extraction and detection of skin interstitial fluid biomarkers.

The detection of biomarkers in body fluids plays a great role in the diagnosis, treatment, and prognosis of diseases. Here, we present novel aptamer-decorated porous microneedles (MNs) arrays to realize the extraction and detection of biomarkers in skin interstitial fluid (ISF) in situ. The porous MNs arrays are fabricated by replicating the negative molds comprising glass microspheres with a UV-curable ethoxylated trimethylolpropane triacrylate (ETPTA). As the MNs arrays combine the superiorities of porous structure and aptamers, their specific surface area increased significantly to 6.694 m2/g, thus vast of stable aptamer probes with a concentration of 0.9459 µM could be immobilized. In addition, the MNs arrays could extract skin ISF into their porous structure on the basis of the capillarity principle, and subsequently capture and detect skin ISF biomarkers without sample post-process. Taking advantage of these features, we further demonstrated a highly sensitive and rapid detection of ISF endotoxin in the concentration ranges of 0.0342 EU/mL to 8.2082 EU/mL from rats model injected with endotoxin via tail vein by using such aptamer-decorated porous MNs arrays, with the limit of detection (LOD) of 0.0064 EU/mL. These results indicated that the aptamer-decorated porous MNs arrays possess great potential for non-invasive extraction and detection of biomarkers in clinical applications.

Zn-MOF Encapsulated Antibacterial and Degradable Microneedles Array for Promoting Wound Healing.

An infected skin wound caused by external injury remains a serious challenge in clinical practice. Wound dressings with the properties of antibacterial activity and potent regeneration capacity are highly desirable for wound healing. In this paper, a degradable, ductile, and wound-friendly Zn-MOF encapsulated methacrylated hyaluronic acid (MeHA) microneedles (MNs) array is fabricated through the molding method for promoting wound healing. Due to the damage capability against the bacteria capsule and oxidative stress of the zinc ion released from the Zn-MOF, such MNs array presents excellent antibacterial activity, as well as considerable biocompatibility. Besides, the degradable MNs array composed of photo-crosslinked MeHA possesses the superior capabilities to continuously and steadily release the loaded active ingredients and avoid secondary damage to the wound. Moreover, the low molecular weight hyaluronic acid (HA) generated by hydrolysis of MeHA is also conducive to tissue regeneration. Benefiting from these features, it has been demonstrated that the Zn-MOF encapsulated degradable MNs array can dramatically accelerate epithelial regeneration and neovascularization. These results indicate that the combination of MOFs and degradable MNs array is of great value for promoting wound healing.

Chinese herb microneedle patch for wound healing.

Traditional Chinese medicine and Chinese herbs have a demonstrated value for disease therapy and sub-health improvement. Attempts in this area tend to develop new forms to make their applications more convenient and wider. Here, we propose a novel Chinese herb microneedle (CHMN) patch by integrating the herbal extracts, Premna microphylla and Centella asiatica, with microstructure of microneedle for wound healing. Such path is composed of sap extracted from the herbal leaves via traditional kneading method and solidified by plant ash derived from the brine induced process of tofu in a well-designed mold. Because the leaves of the Premna microphylla are rich in pectin and various amino acids, the CHMN could be imparted with medicinal efficacy of heat clearing, detoxicating, detumescence and hemostatic. Besides, with the excellent pharmaceutical activity of Asiatic acid extracted from Centella asiatica, the CHMN is potential in promoting relevant growth factor genes expression in fibroblasts and showing excellent performance in anti-oxidant, anti-inflammatory and anti-bacterial activity. Taking advantages of these pure herbal compositions, we have demonstrated that the derived CHMN was with dramatical achievement in anti-bacteria, inhibiting inflammatory, collagen deposition, angiogenesis and tissue reconstruction during the wound closure. These results indicate that the integration of traditional Chinese herbs with progressive technologies will facilitate the development and promotion of traditional Chinese medicine in modern society.

Multifunctional Composite Inverse Opal Film with Multiactives for Wound Healing.

A film with an elaborate microstructure and multifunctions is urgently needed in wound healing. Here, we present a multiactive encapsulated inverse opal film with a monitorable delivery system for chronic wound healing. The inverse opal film is prepared by using poly(lactic-co-glycolic acid) to negatively replicate a colloidal crystal template, which presents a high specific surface area and interconnected nanopores. It could be imparted with a potent antibacterial effect and promote angiogenesis by loading the vascular endothelial growth factor into the nanopores and encapsulating by chitosan. In addition, it is demonstrated that the structure color change of the film could intuitively reflect the drug release progress from the nanopores, which made the film a real-time drug monitoring system. In the affected wound model, the properties of the multifunctional film in promoting wound healing are certified by the faster healing speed, more granulation tissue, less inflammation, and even a distribution of new blood vessels and collagen. These results indicate that the resultant multifunctional film has a practical application value in clinical wound care.

Metformin loaded porous particles with bio-microenvironment responsiveness for promoting tumor immunotherapy.

PD1/PD-L1 antibody blockade-based immunotherapy has been widely recognized in the field of cancer treatment; however, only a small number of cancer patients have been shown to respond well due to the PD1/PD-L1 antibody hydrolysis induced substandard immunotherapeutic efficacy and the low immunogenicity and immunosuppressive tumor microenvironment of the patients. Here, we present a novel tumor microenvironment (TME) responsive particle delivery system with a metformin-loaded chitosan (CS) inverse opal core and a manganese dioxide (MnO2) shell (denoted as CS-metformin@MnO2 particles) for inhibiting the PD-1/PD-L1 signaling pathway and promoting tumor immunotherapy. Benefiting from the interconnected porous structure of the inverse opal, metformin can be easily extensively loaded into the CS particles. With the coating of the TME responsive MnO2 shells, the particle delivery system was imparted with an intelligent "trigger" to prevent premature leaking of the drug until it reaches the tumor tissue. We have demonstrated that CS-metformin@MnO2 particles were able to promote the apoptosis of tumor cells through immunotherapeutic means both in vivo and in vitro. Specifically, the viability of tumor cells in the drug carrier-treated group was nearly 20% less than in the untreated group. In addition, the CS particles could serve as scaffolds for the regeneration of normal tissues and promote post-surgical wound healing due to their biocompatibility and antibacterial ability. These results make CS-metformin@MnO2 particles an excellent delivery system in tumor immunotherapy and post-surgical wound healing applications.

Distance-based quantification of miRNA-21 by the coffee-ring effect using paper devices.

Enabled by the coffee-ring effect, a paper-based signal transduce method is employed for catalytic hairpin assembly (CHA) amplification and hybridization chain reaction (HCR) to achieve miRNA quantification. Once the target miRNAs appeared, it was circularly used by CHA to initiate HCR amplification to produce a large number of G-quadruplex, which is combined with hemin to form a hemin/G-quadruplex DNAzyme. The DNAzyme catalyzes a colorimetric reaction to produce colored nanoparticles, which were converted to the end edge of the paper by evaporation-driven flow, forming a visible colored band. Higher concentration of miRNA led to more colored nanoparticles and thus a longer colored band that can simply be measured by a ruler. The results of determination of miRNA in samples demonstrate that the relative standard deviation of the proposed approach is 5.2%, highly sensitive and repeatable, with a working range 1.0 to 1000 pM and a LOD of 0.2 pM. The paper-based analytical device as a novel platform offers a new signal transduce pathway toward the detection of low-abundance biomarkers for diagnosis.Graphical abstract Schematic representation of the principle for quantification of miRNA on paper based on the coffee-ring effect.

Wearable capillary microfluidics for continuous perspiration sensing.

Perspiration contains valuable information indicating physiological health. For most wearable perspiration sensors, the sensing element contacts with skin directly. Yet lack of precise fluidic manipulation unit limits accurate and continuous analysis considering perspiration aggregation, evaporation loss and electrolyte reabsorption by sweat glands. The potential skin irritation caused by the chemicals in the sensor is also a safety concern. In this work, we report a wearable microfluidic device with fluidic manipulation unit based on capillary force to address these issues. Inspired by wicking materials for wiping perspiration in our daily life, herein, we use biocompatible threads to collect perspiration by capillary absorption. Then the collected perspiration was spontaneously delivered to a hydrophilic microfluidic channel, forming a continuous flow. Electrodes were embedded in the microfluidic channel for continuous electrochemical analysis and to avoid the direct skin contact. On-body tests demonstrated that continuous perspiration collection, transportation and analysis of Na+ as a proof-of-concept analyte can be achieved using the pump-free epidermal microfluidic device.

Antibacterial and angiogenic chitosan microneedle array patch for promoting wound healing.

A patch with the capability of avoiding wound infection and promoting tissue remolding is of great value for wound healing. In this paper, we develop a biomass chitosan microneedle array (CSMNA) patch integrated with smart responsive drug delivery for promoting wound healing. Chitosan possesses many outstanding features such as the natural antibacterial property and has been widely utilized for wound healing. Besides, the microstructure of microneedles enables the effective delivery of loaded drugs into the target area and avoids the excessive adhesion between the skin and the patch. Also, vascular endothelial growth factor (VEGF) is encapsulated in the micropores of CSMNA by temperature sensitive hydrogel. Therefore, the smart release of the drugs can be controllably realized via the temperature rising induced by the inflammation response at the site of wounds. It is demonstrated that the biomass CSMNA patch can promote inflammatory inhibition, collagen deposition, angiogenesis, and tissue regeneration during the wound closure. Thus, this versatile CSMNA patch is potentially valuable for wound healing in clinical applications.

Smart Microneedles for Therapy and Diagnosis.

Microneedles represent a cutting-edge and idea-inspiring technology in biomedical engineering, which have attracted increasing attention of scientific researchers and medical staffs. Over the past decades, numerous great achievements have been made. The fabrication process of microneedles has been simplified and becomes more precise, easy-to-operate, and reusable. Besides, microneedles with various features have been developed and the microneedle materials have greatly expanded. In recent years, efforts have been focused on generating smart microneedles by endowing them with intriguing functions such as adhesion ability, responsiveness, and controllable drug release. Such improvements enable the microneedles to take an important step in practical applications including household drug delivery devices, wearable biosensors, biomedical assays, cell culture, and microfluidic chip analysis. In this review, the fabrication strategies, distinctive properties, and typical applications of the smart microneedles are discussed. Recent accomplishments, remaining challenges, and future prospects are also presented.

Correction: A bio-inspired photonic nitrocellulose array for ultrasensitive assays of single nucleic acids.

Correction for 'A bio-inspired photonic nitrocellulose array for ultrasensitive assays of single nucleic acids' by Junjie Chi, et al., Analyst, 2018, 143, 4559-4565.

Bio-inspired photonic crystals for naked eye quantification of nucleic acids.

Herein, a chip imitating the desert beetle shell was presented for naked eye nucleic acid quantification. The hydrophobic photonic crystal substrate treated by ultraviolet local irradiation could effectively disperse the sample into hundreds of droplets for digital loop-mediated isothermal amplification (dLAMP). Pyrophosphate (PPI), a by-product of the LAMP reaction, combined with magnesium ions to form a poorly soluble precipitate. It could be fixed on a silica substrate due to complexation, resulting in the disappearance of the structural color of the photonic crystals. The number of points without structural color contains the information of the copy number of nucleic acids in the sample. This chip could achieve the naked eye quantitative detection of Salmonella DNA without fluorescence or other chromogenic reagents. Thus, the chip designed in this study can help the development of digital nucleic acid detection under limited resource settings (LRS) and can be suitable for POCT (point of care test) standards.