LncRNA MEG3 regulates ferroptosis of lens epithelial cells via PTBP1/GPX4 axis to participate in age-related cataract.

Age-related cataract (ARC) is regarded as the principal cause of vision impairment among the aged. The regulatory role of long noncoding RNAs (LncRNAs) in ARC remains unclear. The lncRNA maternally expressed gene 3 (MEG3) has been reported to promote ARC progression, and the underlying mechanism was further investigated in this study. Lens epithelium samples were collected to verify the expression of MEG3. Lens epithelial cells (LECs) were treated with H2O2 to mimic microenvironment of ARC in vitro. Cell viability, reactive oxygen species, and ferroptosis were evaluated during the in viro experiments. In the present work, lncRNA MEG3 was highly expressed in ARC group, compared with normal group. MEG3 was induced, cell viability and glutathione peroxidase 4 (GPX4) level were inhibited, and ferroptosis was promoted in H2O2 treated LECs. LncRNA MEG3 silence reversed the effects of H2O2 on viability and ferroptosis in LECs. Thereafter, lncRNA MEG3 was found to bind to PTBP1 for GPX4 degradation. Silencing of GPX4 reversed the regulation of lncRNA MEG3 inhibition in H2O2-treated LECs. To sum up, lncRNA MEG3 exhibited high expression in ARC. In H2O2-induced LECs, inhibition of lncRNA MEG3 accelerated cell viability and repressed ferroptosis by interaction with PTBP1 for GPX4 messenger RNA decay. Targeting lncRNA MEG3 may be a novel treatment of ARC.

A swellable hydrogel microneedle based on cerium-metal organic frame composite nanozyme for detection of biomarkers.

Skin interstitial fluid (ISF) has been an alternative source in the field of biomarkers analysis. This work developed swellable hydrogel microneedles (MNs) composed of polyvinyl alcohol and sodium alginate by chemical crosslinking (PVA/SA). Here, PVA/SA was firstly used to fabricate hydrogel MNs, achieving a swellable ratio of 150 % and a rapid extraction of 6.4 mg ISF in 15 min. To replace expensive and non-reusable test kits, hydrogel MNs based on composite nanozyme with high oxidase-like activity were successfully developed to recover and detect biomarkers. The nanozyme was composed of MnO2-modified mixed valence cerium-metal organic frame (MCM). MCM was characterized by multiple techniques to further confirm its composition and structure. MCM combined with the reduction reaction of glutathione (GSH) with oxidized substrate to achieve a colorimetric GSH detection, which had a detection limit (LOD, 0.36 µM) of GSH. The hydrogel MNs based on MCM (MCM-MNs) were firstly applied to the rapid detection of GSH in ISF. All in all, this method combines the advantages of nanozyme and hydrogel MNs to achieve a timely and minimally invasive analysis, which provides a new dimension for the in vivo detection of GSH by skin ISF and holds great implications in biomedical and bioanalysis fields.

Swellable colorimetric microneedles for glucose detection based on glucose oxidase-like gold nanoparticles.

BACKGROUND: Regular blood glucose monitoring is very important for diabetic patients. The composition of skin interstitial fluid (ISF) is similar to that of blood, which can be used for daily blood sugar detection and disease care. However, most methods of ISF extraction have complicated steps, may cause skin damage, and can only extract a limited amount of ISF, resulting in low detection efficiency. Therefore, it is very necessary to develop a detection method that can not only extract a large amount of ISF safely, efficiently, and conveniently, but also realize rapid detection of glucose level in ISF. RESULTS: Here, we developed a gold nanoparticle (AuNP)-based swellable colorimetric MN patch with minimally invasive sampling function and real-time ISF glucose analysis ability. The MN patch could quickly absorb a large amount of skin ISF, and 60.2 mg of ISF was extracted within 10 min in vitro. It was divided into two layers: the tip layer was embedded with AuNPs with glucose oxidase (GOx)-like activity, which catalyzed the oxidation of glucose extracted from ISF and produced hydrogen peroxide (H2O2); horseradish peroxidase (HRP) encapsulated in the backing layer catalyzed the oxidation of 3, 3', 5, 5'-tetramethylbenzidine (TMB) by H2O2 to produce oxTMB, which led to a visible color shift in the backing layer. The ISF glucose level was judged by naked eyes and further quantified by color analysis with Image J software. As a result, the colorimetric MN patch successfully identified the normal blood sugar and hyperglycemia state in vivo. SIGNIFICANCE: The colorimetric MN patch combined in-situ colorimetric sensing based on AuNP nanozyme with MN patch, which detected glucose level without blood drawing, increasing patients' compliance and reducing detection steps and time. Compared with the detection methods based on natural nanozymes, our method had better stability and sensitivity to complex environments (extreme pH and high temperature, etc.) in actual detection.

Nanozymes-Mediated Cascade Reaction System for Tumor-Specific Diagnosis and Targeted Therapy.

Cascade reactions are described as efficient and versatile tools, and organized catalytic cascades can significantly improve the efficiency of chemical interworking between nanozymes. They have attracted great interest in many fields such as chromogenic detection, biosensing, tumor diagnosis, and therapy. However, how to selectively kill tumor cells by enzymatic reactions without harming normal cells, as well as exploring two or more enzyme-engineered nanoreactors for cascading catalytic reactions, remain great challenges in the field of targeted and specific cancer diagnostics and therapy. The latest research advances in nanozyme-catalyzed cascade processes for cancer diagnosis and therapy are described in this article. Here, various sensing strategies are summarized, for tumor-specific diagnostics. Targeting mechanisms for tumor treatment using cascade nanozymes are classified and analyzed, "elements" and "dimensions" of cascade nanozymes, types, designs of structure, and assembly modes of highly active and specific cascade nanozymes, as well as a variety of new strategies of tumor targeting based on the cascade reaction of nanozymes. Finally, the integrated application of the cascade nanozymes systems in tumor-targeted and specific diagnostic therapy is summarized, which will lay the foundation for the design of more rational, efficient, and specific tumor diagnostic and therapeutic modalities in the future.

Applications of cerium-based materials in food monitoring.

With the rapid development of society, food safety to public health has been a topic that cannot be ignored. In recent years, lanthanide-based materials are studied to be potential candidates in the detection of food samples. Cerium (Ce)-based materials (such as Ce ions, CeO2, Ce-metal organic framework (Ce-MOF), etc.) have also attracted more attention in food detection by virtue of colorimetric, fluorescence, sensing, and other methods. This is because the mixed valence of Ce (Ce3+ and Ce4+), the formation of oxygen vacancies, and their optical and electrochemical properties. In this review, Ce-based materials will be introduced and discussed in the field of food detection, including biogenesis, construction, catalytic mechanisms, combination, and applications. In addition, the current challenges and future development trend of these Ce-based materials in food safety detection are also proposed and discussed. Therefore, it is meaningful to explore the Ce-based materials for detection of biomarkers in food samples.

Unignored intracellular journey and biomedical applications of extracellular vesicles.

The intracellular journey of extracellular vesicles (EVs) cannot be ignored in various biological pathological processes. In this review, the biogenesis, biological functions, uptake pathways, intracellular trafficking routes, and biomedical applications of EVs were highlighted. Endosomal escape is a unique mode of EVs release. When vesicles escape from endosomes, they avoid the fate of fusing with lysosomes and being degraded, thus having the opportunity to directly enter the cytoplasm or other organelles. This escape mechanism is crucial for EVs to deliver specific signals or substances. The intracellular trafficking of EVs after endosomal escape is a complex and significant biological process that involves the coordinated work of various cellular structures and molecules. Through the in-depth study of this process, the function and regulatory mechanism of EVs are fully understood, providing new dimensions for future biomedical diagnosis and treatment.

Biomedical applications of supramolecular hydrogels with enhanced mechanical properties.

Supramolecular hydrogels bound by hydrogen bonding, host-guest, hydrophobic, and other non-covalent interactions are among the most attractive biomaterials available. Supramolecular hydrogels have attracted extensive attention due to their inherent dynamic reversibility, self-healing, stimuli-response, excellent biocompatibility, and near-physiological environment. However, the inherent contradiction between non-covalent interactions and mechanical strength makes the practical application of supramolecular hydrogels a great challenge. This review describes the mechanical strength of hydrogels mediated by supramolecular interactions, and focuses on the potential strategies for enhancing the mechanical strength of supramolecular hydrogels and illustrates their applications in related fields, such as flexible electronic sensors, wound dressings, and three-dimensional (3D) scaffolds. Finally, the current problems and future research prospects of supramolecular hydrogels are discussed. This review is expected to provide insights that will motivate more advanced research on supramolecular hydrogels.

Applications of microalga-powered microrobots in targeted drug delivery.

Over the past decade, researchers have proposed a new class of drug delivery systems, bio-hybrid micro-robots, designed with a variety of living cell-driven micro-robots that utilize the unique mobility of natural organisms (bacteria, cells, exosomes, etc.) to transport effective drugs. Microalgae are considered potential drug delivery carriers. Recent studies have shown that microalga-based drug delivery systems exhibit excellent biocompatibility. In addition, microalgae have a large surfactant area, phototaxis, oxygen production, and other characteristics, so they are used as a carrier for the treatment of bacterial infections, cancer, etc. This review summarizes the modification of microalgae including click chemistry and electrostatic adsorption, and can improve the drug loading efficiency through dehydration and hydration strategies. The prepared microalgal drug delivery system can be targeted to different organs by different dosing methods or using external forces. Finally, it summarizes its antibacterial (gastritis, periodontitis, skin wound inflammation, etc.) and antitumor applications.

Novel strategies in melanoma treatment using silver nanoparticles.

Melanoma has remarkably gained extensive attention owing to its high morbidity and mortality. Conventional treatment methods still have some problems and defects. Therefore, more and more novel methods and materials have been continuously developed. Silver nanoparticles (AgNPs) have attracted significant interest in the field of cancer research especially for melanoma treatment because of their excellent properties including antioxidant, antiproliferative, anti-inflammatory, antibacterial, antifungal, and antitumor abilities. In this review, the applications of AgNPs in the prevention, diagnosis, and treatment of cutaneous melanoma are mainly introduced. It also focuses on the therapy strategies of photodynamic therapy (PDT), photothermal therapy (PTT), and chemotherapy for melanoma treatment. Taken together, AgNPs play an increasingly crucial role in cutaneous melanoma treatment, which have promising application in the future.

Functional two-dimensional MXenes as cancer theranostic agents.

Recently, MXenes, as a kind of two-dimensional (2D) layered materials with exceptional performance, have become the research hotspots owing to their unique structural, electronic, and chemical properties. They have potential applications in electrochemical storage, photocatalysis, and biosensors. Furthermore, they have certain characteristics such as large surface area, favorable biocompatibility, and ideal mechanical properties, which can expand their applications in biomedical fields, especially in cancer therapy. To date, several researchers have explored the applications of MXenes in tumor elimination, which exhibited other fantastic properties of those 2D MXenes, such as efficient in vivo photothermal ablation, low phototoxicity, high biocompatibility, etc. In this review, the structures, properties, modifications, and preparation methods are introduced respectively. More importantly, the multifunctional platforms for cancer therapy based on MXenes nanosheets (NSs) are reviewed in detail, including single-modality and combined-modality cancer therapy. Finally, the prospects and challenges of MXenes are prospected and discussed. STATEMENT OF SIGNIFICANCE: In this review, the structures, properties, modifications, and preparation methods of MXenes nanomaterials are introduced, respectively. In addition, the preparation conditions and morphological characterizations of some common MXenes for therapeutic platforms are also summarized. More importantly, the practical applications of MXenes-based nanosheets are reviewed in detail, including drug delivery, biosensing, bioimaging, and multifunctional tumor therapy platforms. Finally, the future prospects and challenges of MXenes are prospected and discussed.

Chitosan-based high-strength supramolecular hydrogels for 3D bioprinting.

The loss of tissues and organs is a major challenge for biomedicine, and the emerging 3D bioprinting technology has brought the dawn for the development of tissue engineering and regenerative medicine. Chitosan-based supramolecular hydrogels, as novel biomaterials, are considered as ideal materials for 3D bioprinting due to their unique dynamic reversibility and fantastic biological properties. Although chitosan-based supramolecular hydrogels have wonderful biological properties, the mechanical properties are still under early exploration. This paper aims to provide some inspirations for researchers to further explore. In this review, common 3D bioprinting techniques and the properties required for bioink for 3D bioprinting are firstly described. Then, several strategies to enhance the mechanical properties of chitosan hydrogels are introduced from the perspectives of both materials and supramolecular binding motifs. Finally, current challenges and future opportunities in this field are discussed. The combination of chitosan-based supramolecular hydrogels and 3D bioprinting will hold promise for developing novel biomedical implants.

Dose-effect relationship and molecular mechanism by which BMSC-derived exosomes promote peripheral nerve regeneration after crush injury.

BACKGROUND: The development of new treatment strategies to improve peripheral nerve repair after injury, especially those that accelerate axonal nerve regeneration, is very important. The aim of this study is to elucidate the molecular mechanisms of how bone marrow stromal cell (BMSC)-derived exosomes (EXOs) participate in peripheral nerve regeneration and whether the regenerative effect of EXOs is correlated with dose. METHOD: BMSCs were transfected with or without an siRNA targeting Ago2 (SiAgo2). EXOs extracted from the BMSCs were administered to dorsal root ganglion (DRG) neurons in vitro. After 48 h of culture, the neurite length was measured. Moreover, EXOs at four different doses were injected into the gastrocnemius muscles of rats with sciatic nerve crush injury. The sciatic nerve functional index (SFI) and latency of thermal pain (LTP) of the hind leg sciatic nerve were measured before the operation and at 7, 14, 21, and 28 days after the operation. Then, the number and diameter of the regenerated fibers in the injured distal sciatic nerve were quantified. Seven genes associated with nerve regeneration were investigated by qRT-PCR in DRG neurons extracted from rats 7 days after the sciatic nerve crush. RESULTS: We showed that after 48 h of culture, the mean number of neurites and the length of cultured DRG neurons in the SiAgo2-BMSC-EXO and SiAgo2-BMSC groups were smaller than that in the untreated and siRNA control groups. The average number and diameter of regenerated axons, LTP, and SFI in the group with 0.9 × 1010 particles/ml EXOs were better than those in other groups, while the group that received a minimum EXO dose (0.4 × 1010 particles/ml) was not significantly different from the PBS group. The expression of PMP22, VEGFA, NGFr, and S100b in DRGs from the EXO-treated group was significantly higher than that in the PBS control group. No significant difference was observed in the expression of HGF and Akt1 among the groups. CONCLUSIONS: These results showed that BMSC-derived EXOs can promote the regeneration of peripheral nerves and that the mechanism may involve miRNA-mediated regulation of regeneration-related genes, such as VEGFA. Finally, a dose-effect relationship between EXO treatment and nerve regeneration was shown.

The ATP-P2X7 Signaling Pathway Participates in the Regulation of Slit1 Expression in Satellite Glial Cells.

Slit1 is one of the known signaling factors of the slit family and can promote neurite growth by binding to its receptor, Robo2. Upregulation of Slit1 expression in dorsal root ganglia (DRG) after peripheral nerve injury plays an important role in nerve regeneration. Each sensory neuronal soma in the DRG is encapsulated by several surrounding satellite glial cells (SGCs) to form a neural structural unit. However, the temporal and spatial patterns of Slit1 upregulation in SGCs in DRG and its molecular mechanisms are not well understood. This study examined the spatial and temporal patterns of Slit1 expression in DRG after sciatic nerve crush by immunohistochemistry and western blotting. The effect of neuronal damage signaling on the expression of Slit1 in SGCs was studied in vivo by fluorescent gold retrograde tracing and double immunofluorescence staining. The relationship between the expression of Slit1 in SGCs and neuronal somas was also observed by culturing DRG cells and double immunofluorescence labeling. The molecular mechanism of Slit1 was further explored by immunohistochemistry and western blotting after intraperitoneal injection of Bright Blue G (BBG, P2X7R inhibitor). The results showed that after peripheral nerve injury, the expression of Slit1 in the neurons and SGCs of DRG increased. The expression of Slit1 was presented with a time lag in SGCs than in neurons. The expression of Slit1 in SGCs was induced by contact with surrounding neuronal somas. Through injured cell localization, it was found that the expression of Slit1 was stronger in SGCs surrounding injured neurons than in SGCs surrounding non-injured neurons. The expression of vesicular nucleotide transporter (VNUT) in DRG neurons was increased by injury signaling. After the inhibition of P2X7R, the expression of Slit1 in SGCs was downregulated, and the expression of VNUT in DRG neurons was upregulated. These results indicate that the ATP-P2X7R pathway is involved in signal transduction from peripheral nerve injury to SGCs, leading to the upregulation of Slit1 expression.

Bone marrow mesenchymal stem cell-derived exosome uptake and retrograde transport can occur at peripheral nerve endings.

We investigated the occurrence of mesenchymal stem cell (MSC)-derived exosome uptake and retrograde transport at peripheral nerve endings using bone marrow MSCs (bMSCs) transduced with recombinant CD63-green fluorescent protein (GFP) lentiviral plasmid. GFP was used to track the release of bMSC-derived exosomes and the uptake and transport at peripheral nerve terminals, the dorsal root ganglion (DRG), and the spinal cord. In vitro cell culture and injection of a CD63-GFP exosome suspension into the right gastrocnemius muscle of an in vivo rat model were also performed. Fluorescence microscopy of co-cultured CD63-GFP exosomes and SH-SY5Y or BV2 cell lines and primary cultured DRG cells in a separate experiment demonstrated exosome uptake into DRG neurons and glia. Moreover, we observed both retrograde axoplasmic transport and hematogenous transport of exosomes injected into rat models at the DRG and the ipsilateral side of the anterior horn of the spinal cord using fluorescence microscopy, immunohistochemistry, and Western blot analyses. In conclusion, we showed that exosome uptake at peripheral nerve endings and retrograde transport of exosomes to DRG neurons and spinal cord motor neurons in the anterior horn can occur. In addition, our findings propose a novel drug delivery approach for treating neuronal diseases.

The protective effect and underlying mechanism of Hainan papaya water extract against neuronal apoptosis induced by Aß40.

OBJECTIVE: To investigate whether Hainan papayas has protective effects in an Aß40-induced primary neuron injury model and elucidate the underlying molecular mechanism. METHODS: Cultured primary neurons from the dorsal root ganglia (DRG) of Sprague-Dawley (SD) rats were treated with 20 µM Aß40 peptide, 100 µg/L Hainan papaya water extract, peptide plus extract, or culture medium for 24 h. Cell viability was measured by MTT assay, and neuronal apoptosis was evaluated by DAPI staining. ERK signaling pathway-associated molecule activation and changes in Bax expression were analyzed by Western blotting and immunofluorescence. RESULTS: A cell viability rate of (44.11 ± 6.59)% in the Aß40 group was rescued to (79.13 ± 6.64)% by adding different concentrations of the extract. DAPI showed pyknotic nuclei in 39.5% of Aß40-treated cells; the fraction dropped to 17.4% in the 100 µg/L extract group. ERK phosphorylation was observed in the Aß40 group but was ameliorated by pretreatment with 100 µg/L extract. Hainan papaya water extract also prevented Aß40-induced phosphorylation of MEK, RSK1 and CREB associated with ERK signaling and downregulated Bax expression in the neurons. CONCLUSION: The results suggest that Hainan papaya water extract has protective effects on neurons; the mechanism may be related to suppression of ERK signaling activation.

Differential expression of microRNAs in dorsal root ganglia after sciatic nerve injury.

This study investigated the possible involvement of microRNAs in the regulation of genes that participate in peripheral neural regeneration. A microRNA microarray analysis was conducted and 23 microRNAs were identified whose expression was significantly changed in rat dorsal root ganglia after sciatic nerve transection. The expression of one of the downregulated microRNAs, microRNA-214, was validated using quantitative reverse transcriptase-PCR. MicroRNA-214 was predicted to target the 3'-untranslated region of Slit-Robo GTPase-activating protein 3. In situ hybridization verified that microRNA-214 was located in the cytoplasm of dorsal root ganglia primary neurons and was downregulated following sciatic nerve transection. Moreover, a combination of in situ hybridization and immunohistochemistry revealed that microRNA-214 and Slit-Robo GTPase-activating protein 3 were co-localized in dorsal root ganglion primary neurons. Western blot analysis suggested that Slit-Robo GTPase-activating protein 3 was upregulated in dorsal root ganglion neurons after sciatic nerve transection. These data demonstrate that microRNA-214 is located and differentially expressed in dorsal root ganglion primary neurons and may participate in regulating the gene expression of Slit-Robo GTPase-activating protein 3 after sciatic nerve transection.

srGAP3 promotes neurite outgrowth of dorsal root ganglion neurons by inactivating RAC1.

OBJECTIVE: To explore effect of srGAP3 promotes neurite outgrowth of dorsal root ganglion neurons. METHODS: In this study, expression of Slit1 was observed predominantly in the glia, while expression of Robo2 and srGAP3 was detected in sensory neurons of postnatal rat cultured dorsal root ganglion (DRG). Furthermore, upregulation of srGAP3 following sciatic nerve transection was detected by immunohistochemistry and Western blotting. RESULTS: It was observed that inhibition of neurite outgrowth in cultured adult DRG neurons following treatment with anti-srGAP3 or anti-Robo2 was more effectively (1.5-fold higher) than that following treatment with an anti-BDNF positive control antibody. It demonstrated that srGAP3 interacted with Robo2 and Slit1 protein to decrease Rac1-GTP activity in cultured adult rat DRG neurons and the opposite effect on Rac1-GTP activity was detected by co-immunoprecipitation and immunoblotting analyses following treatment with anti-Robo2 or anti-srGAP3. These data demonstrated a role for srGAP3 in neurite outgrowth of DRG sensory neurons. CONCLUSIONS: Our observations suggest that srGAP3 promotes neurite outgrowth and filopodial growth cones by interacting with Robo2 to inactivate Rac1 in mammalian DRG neurons.

Slit-Robo GTPase-activating proteins are differentially expressed in murine dorsal root ganglia: modulation by peripheral nerve injury.

The Slit-Robo GTPase-activating proteins (srGAPs) play an important role in neurite outgrowth and axon guidance; however, little is known about its role in nerve regeneration after injury. Here, we studied the expression of srGAPs in mouse dorsal root ganglia (DRG) following sciatic nerve transection (SNT) using morphometric and immunohistochemical techniques. Reverse transcriptase polymerase chain reaction and Western blot analysis indicated that srGAP1 and srGAP3, but not srGAP2, were expressed in normal adult DRG. Following unilateral SNT, elevated mRNA and protein levels of srGAP1 and srGAP3 were detected in the ipsilateral relative to contralateral L(3-4) DRGs from day 3 to day 14. Immunohistochemical results showed that srGAP1 and srGAP3 were largely expressed in subpopulations of DRG neurons in naïve DRGs. However, after SNT, srGAP3 in neurons was significantly increased in the ipsilateral relative to contralateral DRGs, which peaked at day 7 to day 14. Interestingly, DRG neurons with strong srGAP3 labeling also coexpressed Robo2 after peripheral nerve injury. These results suggest that srGAPs are differentially expressed in murine DRG and srGAP3 are the predominant form. Moreover, srGAP3 may participate in Slit-Robo signaling in response to peripheral nerve injury or the course of nerve regeneration.

MicroRNAs 144, 145, and 214 are down-regulated in primary neurons responding to sciatic nerve transection.

MicroRNAs (miRNAs) play an important role in the development, differentiation, proliferation, survival, and oncogenesis of cells and organisms including nervous system. However, the role of miRNAs in primary neurons of dorsal root ganglion (DRG) after injury was not clear. In this study, a miRNA microarray analysis was performed, and a total of 21 miRNAs were found to be down-regulated following unilateral sciatic nerve transection. The miR-144, miR-145, and miR-214 were further validated using quantitative reverse transcriptase PCR (qRT-PCR). Moreover, in situ hybridization (ISH) experiments using locked nucleic acid (LNA)-modified DNA oligonucleotide probes verified that miR-144, miR-145, and miR-214 were expressed in primary neurons of DRG and down-regulated following sciatic nerve transection. Predictions of potential miRNA targets involved were identified by performing a bioinformatics analysis. These predictions were tested using miRNA luciferase reporter vectors, with Robo2 and srGAP2 evaluated as the potential targets of miR-145 and miR-214, respectively. The role of miR-145 in cultured primary neurons was also investigated, and the result found that miR-145 miR-145 inhibited neurite growth and down-regulated Robo2 expression. Finding from this study suggested that miRNAs of DRG can mediated the course of regeneration including through Slit-Robo-srGAP signaling pathway after injury.