ACS Applied Bio Materials

Glutathione (GSH) serves many cellular functions, within the range of 1-10 mM in mammalian cells, abnormal levels of which have been related to many pathological processes. Until now, many researches have been devoted to qualitative sensing of intracellular GSH. However, it is still quite difficult to determine the dynamic GSH levels in living cells. To solve this problem, we designed and synthesized a ratiometric fluorescent probe RP-1 for the detection of different GSH levels based on a reversible Michael addition reaction. The Michael addition product RP-GSH was characterized by ESI-MS. The reversibility and kinetics of the reaction of RP-1 with GSH were studied by spectroscopic methods. According to the concentration dependence of the ratio F405/F488 (F405 and F488 are the fluorescence intensities at 505 and 571 nm when RP-1 in the presence of GSH was excited at 405 and 488 nm, respectively), GSH in the range of 0.1-50 mM could be detected by a very low concentration of 2.5 μM RP-1 in the PBS buffer, indicating that the interference of RP-1 to the biological system in living cell imaging would be minimized. Simultaneously, the dissociation constant (Kd) was obtained as 4.89 mM, suggesting its capability to accurately quantify the GSH concentration in the range of 1-10 mM in cells. Moreover, to improve the cell permeability, we converted RP-1 with a carboxylic acid group into RP-ester with an acetoxymethyl ester group. It is noteworthy that the rapid cell uptake rate of the RP-ester made it very promising because the living cells were efficiently stained with high brightness after incubation with RP-ester for only 1 min. Furthermore, the successful ratio imaging (merged image of 405 channel and 488 channel) of different GSH levels by RP-ester in living cells demonstrated the further development and utilization in the future.

Ming Tian

Mian Yang

Yi Liu

Feng-Lei Jiang

Rapid and Reversible Reaction-Based Ratiometric Fluorescent Probe for Imaging of Different Glutathione Levels in Living Cells

Adhesive and stretchable nanofibrous hydrogels have attracted extensive attraction in wound dressings, especially for joint wound treatment. However, adhesive hydrogels tend to display poor stretchable behavior. It is still a significant challenge to integrate excellent adhesiveness and stretchability in a nanofibrous hydrogel. Herein, a highly adhesive, stretchable, and breathable nanofibrous hydrogel was developed via an in situ hybrid cross-linking strategy of electrospun nanofibers comprising dopamine (DA) and gelatin methacryloyl (GelMA). Benefiting from the balance of cohesion and adhesion based on photocross-linking of methacryloyl (MA) groups in GelMA and the chemical/physical reaction between GelMA and DA, the nanofibrous hydrogels exhibited tunable adhesive and mechanical properties through varying MA substitution degrees of GelMA. The optimized GelMA60-DA exhibited 2.0 times larger tensile strength (2.4 MPa) with an elongation of about 200%, 2.3 times greater adhesive strength (9.1 kPa) on porcine skin, and 3.1 times higher water vapor transmission rate (10.9 kg m–2 d–1) compared with gelatin nanofibrous hydrogels. In parallel, the GelMA60-DA nanofibrous hydrogels could facilitate cell growth and accelerate wound healing. This work presented a type of breathable nanofibrous hydrogels with excellent adhesive and stretchable capacities, showing great promise as wound dressings.

Yumin Liu

Qiusheng Wang

Xueting Liu

Pawel Nakielski

Filippo Pierini

Xiaoran Li

Jianyong Yu

Bin Ding

ACS Appl. Bio Mater.

Highly Adhesive, Stretchable and Breathable Gelatin Methacryloyl-based Nanofibrous Hydrogels for Wound Dressings

A synergistic therapy agent (STA) with photothermal, photodynamic, chemodynamic, and starvation therapy (PTT, PDT, CDT, and ST) functions was developed. Hollow, mesoporous, and nearly uniform CeO2 nanoparticles (H-CeO2 NPs) were synthesized using a staged shape templating sol–gel protocol. Chlorin e6 (Ce6) was adsorbed onto H-CeO2 NPs, and a thin polydopamine (PDA) layer was formed on Ce6-adsorbed H-CeO2 NPs. Glucose oxidase (GOx) was bound onto PDA-coated Ce6-adsorbed H-CeO2 NPs to obtain the targeted STA (H-CeO2@Ce6@PDA@GOx NPs). A reversible photothermal conversion behavior with the temperature elevations up to 34 °C was observed by NIR laser irradiation at 808 nm. A cascade enzyme system based on immobilized GOx and intrinsic catalase-like activity of H-CeO2 NPs was rendered on STA for enhancing the effectiveness of PDT by elevation of ROS generation and alleviation of hypoxia in a tumor microenvironment. Glucose-mediated generation of highly toxic hydroxyl radicals (·OH) was evaluated for CDT. The effectiveness of PDT on glioblastoma T98G cells was markedly enhanced by O2 generation started by the decomposition of glucose. A similar increase in cell death was also observed when ST and CDT functions were enhanced by photothermal action. The viability of T98G cells decreased to 10.6% by in vitro synergistic action including ST, CDT, PDT, and PTT without using any antitumor agent.

Çağıl Zeynep Süngü Akdogan

Esin Akbay Çetin

Mehmet Ali Onur

Selis Önel

Ali Tuncel

In Vitro Synergistic Photodynamic, Photothermal, Chemodynamic, and Starvation Therapy Performance of Chlorin e6 Immobilized, Polydopamine-Coated Hollow, Porous Ceria-Based, Hypoxia-Tolerant Nanozymes Carrying a Cascade System

氯in e6固定化、聚多巴胺包覆的中空多孔氧化铈基、耐缺氧纳米酶携带级联系统的体外协同光动力、光热、化学动力和饥饿治疗性能

Microalgae show great promise for producing valuable molecules like biofuels, but their large-scale production faces challenges, with harvesting being particularly expensive due to their low concentration in water, necessitating extensive treatment. While methods such as centrifugation and filtration have been proposed, their efficiency and cost-effectiveness are limited. Flotation, involving air-bubbles lifting microalgae to the surface, offers a viable alternative, yet the repulsive interaction between bubbles and cells can hinder its effectiveness. Previous research from our group proposed using an amphiphilic chitosan derivative, polyoctyl chitosan (PO-chitosan), to functionalize bubbles used in dissolved air flotation (DAF). Molecular-scale studies performed using atomic force microscopy (AFM) revealed that PO-chitosan's efficiency correlates with cell surface properties, particularly hydrophobic ones, raising the question of whether this molecule can in fact be used more generally to harvest different microalgae. Evaluating this, we used a different strain of Chlorella vulgaris and first characterized its surface properties using AFM. Results showed that cells were hydrophilic but could still interact with PO-chitosan on bubble surfaces through a different mechanism based on specific interactions. Although force levels were low, flotation resulted in 84% separation, which could be explained by the presence of AOM (algal organic matter) that also interacts with functionalized bubbles, enhancing the overall separation. Finally, flocculation was also shown to be efficient and pH-independent, demonstrating the potential of PO-chitosan for harvesting microalgae with different cell surface properties and thus for further sustainable large-scale applications.

Irem Demir-Yilmaz

Michaela Pappa

Sanjaya Lama

Pascal Guiraud

Dries Vandamme

Cécile Formosa-Dague

The Biophysical Properties of Microalgal Cell Surfaces Govern Their Interactions with an Amphiphilic Chitosan Derivative Used for Flocculation and Flotation

微藻细胞表面的生物物理特性决定了它们与用于絮凝和浮选的两亲性壳聚糖衍生物的相互作用

Superior flexibility and toughness can be achieved in bioactive hydrogels by the use of a double polymer network with complementary properties. Inspired by this design principle, we here combine polyacrylic acid (PAA) and sodium alginate (SA) to obtain a dual-reinforced double interpenetrating network (d-DIPN) hydrogel. The dual reinforcement involves ionic cross-linking and introduction of SiO2 nanoparticles, which leads to extraordinary improvements in strength and toughness. Compared with the standard PAA hydrogel that offers an elongation of 240% and a breakage stress of 0.03 MPa, the prepared SA(Ca2+)–PAA–SiO2 hydrogel shows an elongation above 1000% and a breakage stress of 1.62 MPa. Moreover, the combination of strong covalent cross-links and weak reversible interactions provides the d-DIPN hydrogel with swelling resistance and self-healing behavior, adhesive abilities, and shape memory performance. Furthermore, we show that the biocompatibility and bone cell proliferation ability of the hydrogels can be improved through a mineralization process despite an observed reduction in breakage strain and stress. Taken as a whole, our work paves the way for the design of strong and tough hydrogels, with potential applications within biomedicine and particularly tissue engineering.

Wei Fan

Lars R. Jensen

Yibing Dong

Abigail J. Deloria

Bengang Xing

Donghong Yu

Morten M. Smedskjaer

Highly Stretchable, Swelling-Resistant, Self-Healed, and Biocompatible Dual-Reinforced Double Polymer Network Hydrogels

高延展性、抗溶胀、自愈合且生物相容的双增强双重聚合物网络水凝胶

Bacterial infections pose an increasingly worrisome threat to the health of humankind, with antibiotic resistance contributing significantly to this burden. With current conventional antibiotics perpetuating the problem, and a paucity in developing antibiotics, drug delivery systems incorporating nanotechnology appear promising. As such, a dual enzyme-responsive multifunctional nanostructured lipid carrier (NLC) incorporating farnesol (FAN) and triglycerol monostearate (TGMS), was conceptualized for the codelivery of vancomycin (VCM) and antimicrobial peptide (AMP) to enhance the antibacterial activity of VCM. In silico studies and Microscale Thermophoresis demonstrated the strong binding relationships between the NLC constituents and two enzymes that exist in higher concentrations during host infection, namely lipase and a matrix metalloproteinase (MMP). The formulated nanosystem, VCM-AMP-TF-NLCs, had a particle size, polydispersity index, zeta potential, and entrapment efficiency of 149.00 ± 2.97 nm, 0.07 ± 0.01, −5.51 ± 1.21 mV, and 86.20% ± 1.47%, respectively. The NLCs, which showed stability, and biocompatibility, also demonstrated lipase- and MMP-responsiveness. The in vitro antibacterial studies revealed 2-fold and 8-fold reductions in the minimum inhibitory concentration for the NLCs compared to bare VCM, against methicillin-resistant Staphylococcal aureus (MRSA) and Escherichia coli, respectively. Furthermore, in vivo studies revealed that tissues treated with the VCM-AMP-TF-NLCs displayed significantly reduced bacterial burdens (up to 8.73-fold) and less histopathological cellular injury, edema, and necrosis compared to the tissues treated with bare VCM alone. The results support the superiority of the VCM-AMP-TF-NLCs as a multifunctional dual enzyme-responsive NLC compared to bare VCM.

Kerisha Chetty

Xylia Q. Peters

Calvin A. Omolo

Eman A. Ismail

Mohammed A. Gafar

Eman Elhassan

Sania Z. F. Kassam

Jasoda Govender

Sbongumusa Dlamini

Thirumala Govender

Multifunctional Dual Enzyme-Responsive Nanostructured Lipid Carriers for Targeting and Enhancing the Treatment of Bacterial Infections

多功能双酶响应纳米结构脂质载体用于靶向增强细菌感染治疗

Chemo/photothermal combination therapy is a promising and practical approach for cancer treatment which calls for certain nanovehicles to achieve the spatiotemporal co-occurrence of photothermal conversion and drug delivery. Herein, we developed a montmorillonite-based Pickering emulsion equipped with a near-infrared photothermal agent (indocyanine green, ICG) and anticarcinogen (paclitaxel, PTX). With both montmorillonite and ICG functioning as interfacial stabilizers, the Pickering emulsion showed good stability and nanoscale droplet size, which were favored for cellular applications. Due to the vast oil–water interface, where the majority of amphiphilic ICG was prone to distribute, the Pickering nanoemulsion could achieve a higher local concentration of ICG than the aqueous solution, therefore leading to a higher local photothermal performance under near-infrared irradiation. The Pickering nanoemulsion exhibited fast cell penetration, which promoted the photothermal therapeutic effect of ICG. Moreover, the inner phase of the Pickering nanoemulsion also facilitated the loading of PTX, further improving its killing efficacy against cancer cells under near-infrared irradiation, because the photothermal conversion of the Pickering nanoemulsion could not only cause heat damage by itself but also promote the loaded PTX to diffuse out and induce cell death. Therefore, this clay-based Pickering nanoemulsion as a nanovehicle could realize the synergy of chemo- and photothermal therapy.

Yonghui Zan

Jiaren Liu

Ziwei Zhao

Yi Wei

Ning Yang

Hean Zhang

Xiaoyu Wang

Yuetong Kang

A Montmorillonite-Based Pickering Nanoemulsion for the Integration of Photothermal Therapy and NIR-Responsive Drug Delivery

基于蒙脱石的Pickering纳米乳液用于光热治疗与近红外响应药物递送的整合

Skin wounds are extremely frequent injuries related to many etiologies. They are a burden on healthcare systems worldwide. Skin dressings are the most popular therapy, and collagen is the most commonly used biomaterial, although new sources of collagen have been studied, especially spongin-like from marine sponges (SPG), as a promising source due to a similar composition to vertebrates and the ability to function as a cell-matrix adhesion framework. Despite evidence showing the positive effects of SPG for tissue healing, the effects of skin dressings manufactured are still limited. In this context, this study aimed at investigating the effects of collagen skin dressings in an experimental model of skin wounds in rats. For this purpose, SEM, FTIR, cell viability, morphological and morphometric aspects, collagen deposition, and immunostaining of TGF-β and FGF were evaluated. The results demonstrated micro- and macropores on the rough surface, peak characteristics of collagen, and no cytotoxicity for the skin dressing. Also, the control group (CG) after 5 and 10 days exhibited an intense inflammatory process and the presence of granulation tissue, while the treated group (TG) exhibited re-epithelialization after 10 days. The evaluation of granulation tissue and neoepithelial length had an intragroup statistical difference (p = 0.0216) and no intergroup difference. Birefringence demonstrated an organized mesh arranged in a network pattern, presenting type I and type III collagen fibers in all groups. Moreover, in the morphometric evaluation, there were no statistical differences in intergroups or time points for the different types of collagen evaluated. In conclusion, these findings may indicate that the dressing has not exacerbated the inflammatory process and may allow faster healing. However, further studies using a critical wound healing injury model should be used, associated with longer experimental periods of evaluation, to further investigate the effects of these promising therapeutic approaches throughout the skin repair process.

Amanda de Souza

Cintia C. S. Martignago

Lívia Assis

Fernanda Vieira Botelho Delpupo

Marcelo Assis

Karolyne S. J. Sousa

Lais Caroline Souza e Silva

Laura O. Líbero

Flavia de Oliveira

Ana Claudia Muniz Renno

Casting Skin Dressing Containing Extractions of the Organic Part of Marine Sponges for Wound Healing

含海洋海绵有机提取物的伤口愈合用敷料

Stem cell-based therapies are emerging as significant approaches in tissue engineering and regenerative medicine, applicable to both fundamental scientific research and clinical practice. Despite remarkable results in clinical studies, challenges such as poor standardization of graft tissues, limited sources, and reduced functionality have hindered the effectiveness of these therapies. In this review, we summarize the engineering approaches involved in fabricating stem cell assisted patches and the substantial strategies for designing stem cell-laden engineered patches (SCP) to complement the existing stem cell-based therapies. We then outline the potential applications of SCP in advancing tissue regeneration and regenerative medicine. By combining living stem cells with engineered patches, SCP can enhance the functions of both components, particularly for tissue engineering applications. Finally, we addressed current challenges, such as ethical considerations, high costs, and regulatory hurdles and proposed future research directions to overcome these barriers.

Seyeong Heo

Minhyeok Noh

Yeonseo Kim

Sunho Park

Stem Cell-Laden Engineered Patch: Advances and Applications in Tissue Regeneration

干细胞负载的工程化补片：组织再生的进展与应用

Proteins are important biological macromolecules that perform a wide variety of functions in the cell and human body, and can serve as important biomarkers for early diagnosis and prognosis of human diseases as well as monitoring the effectiveness of disease treatment. Hence, sensitive and accurate detection of proteins in human biospecimens is imperative. However, at present, there is no ideal method available for the detection of proteins in clinical samples, many of which are present at ultralow (less than 1 pM) concentrations and in complicated matrices. Herein, we report an ultrasensitive and selective DNA-assisted CRISPR-Cas12a enhanced fluorescent assay (DACEA) for protein detection with detection limits reaching as low as attomolar concentrations. The high assay sensitivity was accomplished through the combined DNA barcode amplification (by using dual-functionalized AuNPs) and CRISPR analysis, while the high selectivity and high resistance to the matrix effects of our method were accomplished via the formation of protein-antibody sandwich structure and the specific recognition of Cas12a (under the guidance of crRNA) toward the designed target ssDNA. Given its ability to accurately and sensitively detect trace amounts of proteins in complicated matrices, the DACEA protein assay platform pioneered in this work has a potential application in routine protein biomarker testing.

Sathishkumar Munusamy

Haiyan Zheng

Rana Jahani

Shuo Zhou

Jun Chen

Juanhua Kong

Xiyun Guan

DNA-Assisted CRISPR-Cas12a Enhanced Fluorescent Assay for Protein Detection in Complicated Matrices

DNA辅助CRISPR-Cas12a增强型荧光测定法用于复杂基质中的蛋白质检测

Abbreviation	ACS Appl. Bio Mater.
Journal Impact	4.42
Quartiles(Global)	NANOSCIENCE & NANOTECHNOLOGY(Q2)

ISSN	2576-6422
h-index	55

Publication Information	Publisher: American Chemical Society，Publishing cycle: ，Journal Type: journal，Open Access Journals: No
Basic data	Year of publication: 2018，Proportion of original research papers: 91.64%，Self Citation Rate:2.20%， Gold OA Rate: 7.29%

ACS Applied Bio Materials

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