Microarray analysis of tRNA-derived small RNA (tsRNA) in LPS-challenged macrophages treated with metformin.

Metformin, a widely used anti-diabetic drug, has demonstrated its efficacy in addressing various inflammatory conditions. tRNA-derived small RNA (tsRNA), a novel type of small non-coding RNA, exhibits diverse regulatory functions and holds promise as both a diagnostic biomarker and a therapeutic target for various diseases. The purpose of this study is to investigate whether the abundance of tsRNAs changed in LPS versus LPS + metformin-treated cells, utilizing microarray technology. Firstly, we established an in vitro lipopolysaccharide (LPS)-induced inflammation model using RAW264.7 macrophages and assessed the protective effects of metformin against inflammatory damage. Subsequently, we extracted total RNA from both LPS-treated and metformin + LPS-treated cell samples for microarray analysis to identify differentially abundant tsRNAs (DA-tsRNAs). Furthermore, we conducted bioinformatics analysis to predict target genes for validated DA-tsRNAs and explore the biological functions and signaling pathways associated with DA-tsRNAs. Notably, metformin was found to inhibit the inflammatory response in RAW264.7 macrophages. The microarray results revealed a total of 247 DA-tsRNAs, with 58 upregulated and 189 downregulated tsRNAs in the Met + LPS group compared to the LPS group. The tsRNA-mRNA network was visualized, shedding light on potential interactions. The results of bioinformatics analysis suggested that these potential targets of specific tsRNAs were mainly related to inflammation and immunity. Our study provides compelling evidence that metformin exerts anti-inflammatory effects and modulates the abundance of tsRNAs in LPS-treated RAW264.7 macrophages. These findings establish a valuable foundation for using tsRNAs as potential biomarkers for metformin in the treatment of inflammatory conditions.

The Role and Mechanism of Metformin in Inflammatory Diseases.

Metformin is a classical drug used to treat type 2 diabetes. With the development of research on metformin, it has been found that metformin also has several advantages aside from its hypoglycemic effect, such as anti-inflammatory, anti-aging, anti-cancer, improving intestinal flora, and other effects. The prevention of inflammation is critical because chronic inflammation is associated with numerous diseases of considerable public health. Therefore, there has been growing interest in the role of metformin in treating various inflammatory conditions. However, the precise anti-inflammatory mechanisms of metformin were inconsistent in the reported studies. Thus, this review aims to summarize various currently known possible mechanisms of metformin involved in inflammatory diseases and provide references for the clinical application of metformin.

Iodine-Doped 3D Print Ti Alloy for Antibacterial Therapy on Orthopedic Implants.

This study presents a novel approach to mitigating bacterial infections and antibiotic resistance in medical implants through the integration of iodine-doping and 3D printing techniques. Iodine, with its potent antibacterial properties, and titanium alloy (Ti), a popular metal for implants due to its mechanical and biological properties, were combined via electrodeposition on 3D-printed titanium alloy (3D-Ti) implants. Scanning electron microscopy, energy dispersive spectroscopy, and X-ray photoelectron spectroscopy confirmed the successful creation of iodine-doped titanium implants with improved iodine content due to the rough surface of the 3D-printed material. In vitro studies revealed that these implants significantly inhibited bacterial adhesion and biofilm formation and showed favorable release kinetics for iodine ions. Biocompatibility tests demonstrated no cytotoxic effects and good hemocompatibility. The implants demonstrated enhanced antimicrobial efficacy against Escherichia coli (E. coli) and Staphylococcus aureus (S. aureus) bacteria strains. The findings imply that the integration of iodine-doping and 3D printing technologies is a promising strategy for treating postoperative infections associated with medical implants, consequently bettering the prognosis for patients. Future investigations are encouraged to delve into the long-standing impacts and prospective clinical utility of this groundbreaking methodology.

Homogeneous silver nanoparticle loaded polydopamine/polyethyleneimine-coated bacterial cellulose nanofibers for wound dressing.

Utilizing mussel-inspired chemistry is an advanced strategy for surface modification, because dopamine (DA) can form a material-independent adhesive coating and further functionalization can be achieved, including the production of silver nanoparticles (AgNPs). Nevertheless, DA easily aggregates in the nanofiber network structure of bacterial cellulose (BC), which not only blocks the pores in the BC structure but also leads to the formation of large silver particles and the burst release of highly cytotoxic silver ions. Herein, a homogeneous AgNP-loaded polydopamine (PDA)/polyethyleneimine (PEI) coated BC was constructed via a Michael reaction between PDA and PEI. Under the action of PEI, the PDA/PEI coating was uniformly attached to the BC fiber surface with a thickness of approximately 4 nm, and homogeneous AgNPs were produced on the uniform PDA/PEI/BC (PPBC) fiber surface. The sustained release of silver ions was better from AgNPs@PPBC than from AgNPs@PDA/BC. The obtained AgNPs@PPBC exhibited excellent antibacterial activities and cytocompatibility. The results of the in vivo assay indicated that the AgNPs@PPBC dressing could inhibit S. aureus infection and inflammation, promote hair follicle growth, enhance collagen deposition, and accelerate wound healing within 12 days compared with BC. These results illustrate that the homogeneous AgNPs@PPBC dressing has great potential for treating infected wounds.

A dual functional Ti-Ga alloy: inhibiting biofilm formation and osteoclastogenesis differentiation via disturbing iron metabolism.

BACKGROUND: Although biomedical implants have been widely used in orthopedic treatments, two major clinical challenges remain to be solved, one is the bacterial infection resulting in biofilm formation, and the other is aseptic loosening during implantation due to over-activated osteoclastogenesis. These factors can cause many clinical issues and even lead to implant failure. Thus, it is necessary to endow implants with antibiofilm and aseptic loosening-prevention properties, to facilitate the integration between implants and bone tissues for successful implantation. To achieve this goal, this study aimed to develop a biocompatible titanium alloy with antibiofilm and anti-aseptic loosening dual function by utilizing gallium (Ga) as a component. METHODS: A series of Ti-Ga alloys were prepared. We examined the Ga content, Ga distribution, hardness, tensile strength, biocompatibility, and anti-biofilm performance in vitro and in vivo. We also explored how Ga3+ ions inhibited the biofilm formation of Staphylococcus aureus (S. aureus) and Escherichia coli (E. coli) and osteoclast differentiation. RESULTS: The alloy exhibited outstanding antibiofilm properties against both S. aureus and E. coli in vitro and decent antibiofilm performance against S. aureus in vivo. The proteomics results demonstrated that Ga3+ ions could disturb the bacterial Fe metabolism of both S. aureus and E. coli, inhibiting bacterial biofilm formation. In addition, Ti-Ga alloys could inhibit receptor activator of nuclear factor-κB ligand (RANKL)-dependent osteoclast differentiation and function by targeting iron metabolism, then suppressing the activation of the NF-κB signaling pathway, thus, showing their potential to prevent aseptic loosening. CONCLUSION: This study provides an advanced Ti-Ga alloy that can be used as a promising orthopedic implant raw material for various clinical scenarios. This work also revealed that iron metabolism is the common target of Ga3+ ions to inhibit biofilm formation and osteoclast differentiation.

Metformin Improves Burn Wound Healing by Modulating Microenvironmental Fibroblasts and Macrophages.

Metformin, a biguanide, exerts different functions through various signaling pathways. In order to investigate the function and mechanism of metformin in burn wounds, we established burn rat models, subcutaneously injected metformin to treat the wounds, and observed the morphologies and the expression of collagen I, collagen III, fibronectin, and pro-inflammatory markers. In vitro experiments were performed to investigate the effects of metformin on the proliferation, migration, and collagen I synthesis of the mouse embryonic fibroblast (NIH 3T3) cell line and on the proliferation, apoptosis, and immune response of the mouse mononuclear macrophage (RAW 264.7) cell line. Finally, we studied the regulatory effects of metformin on a co-culture of RAW 264.7/NIH 3T3 cells. We found that 100 mM of metformin reduced dermal thickness, collagen I deposition, and mRNA expression of IL1ß and CCL2 in rat burn wounds. In vitro experiments revealed that metformin inhibited the proliferation of NIH 3T3 and RAW 264.7 cells. Metformin attenuated NIH 3T3 cell migration via the AMPK/mTOR pathway and attenuated collagen I synthesis through the TGFß1/Smad3 pathway. Metformin inhibited the apoptosis of RAW 264.7 cells induced by 10 µg/mL LPS. Metformin downregulated the mRNA expression of IL1ß and CCL2 in RAW 264.7 cells under 1 µg/mL LPS induction by inhibiting NF-κB p65 phosphorylation. In a RAW 264.7/NIH 3T3 co-culture, metformin attenuated collagen I synthesis in NIH 3T3 cells by inhibiting RAW 264.7 paracrine secretion of TGF-ß1. This provides new evidence related to the development of metformin for potentially improving burn wound healing.

Smart-Temporary-Film-Based Local-Delivery System with Controllable Drug-Release Behavior.

The development of a simple local drug-delivery system that exhibits the advantages of macro- and microscale carriers with controllable drug-release behavior is still highly desired. Herein, in this work, a smart temporary film was prepared from doxorubicin (DOX)-loaded shape-memory microgels via a simple hot-compression programming method. The temporary film showed a very smooth surface and easy handing, as well as macroscopy mechanical properties, which could disintegrate into the microgels with heating at 45 °C. In this case, the temporary film showed a controllable DOX release behavior when compared with the microgels, which could release the DOX on demand. Consequently, the temporary film exhibited weaker cytotoxicity to normal cells and a much longer antitumor capability, as well as a higher drug-utilization efficiency when compared with microgels. Therefore, the smart temporary film has high potential as a candidate for use as a local drug-delivery system.

Fabrication of gentamicin loaded Col-I/HA multilayers modified titanium coatings for prevention of implant infection.

In this study, gentamicin loaded collagen I/hyaluronic acid multilayers modified titanium coating (TC-AA(C/H)6-G) was fabricated via a layer-by-layer (LBL) covalent immobilization method. The drug releasing properties of collagen I/Hyaluronic acid (Col-I/HA) multilayers and the effect of loaded gentamicin on the antibacterial properties and cytocompatibility of modified TC were investigated. The gentamicin release assay indicated that the Col-I/HA multilayers modified TC exhibited agreeable drug-loading amount (537.22 ± 29.66 µg of gentamicin) and controlled-release performance (240 h of sustained release time). TC-AA(C/H)6-G revealed satisfactory antibacterial activity and inhibited the colonization and biofilm formation of S. aureus. Fortunately, the functions of hMSCs on TC-AA(C/H)6-G did not affected by the loaded gentamicin, and TC-AA(C/H)6-G could improve the adhesion, proliferation and osteogenic differentiation of cells, as well as TC-AA(C/H)6. In vivo animal study indicated that TC-AA(C/H)6-G could effectively control intramedullary cavity infection caused by S. aureus and prevent bone destruction.

Designment of polydopamine/bacterial cellulose incorporating copper (II) sulfate as an antibacterial wound dressing.

Improving antibacterial performance is one of the prerequisites for the clinical application of bacterial cellulose (BC)-based dressings. In this study, a novel copper (Cu) ion loaded BC-based antibacterial wound dressing was prepared via codeposition of polydopamine (PDA) and copper ion. The scanning electron microscope (SEM) results showed that the copper ion/PDA coating was more uniform than the PDA coating, and the 3D nanopore structure of BC was retained in Cu2+@PBC. Cu ions were immobilized by forming a chelate with PDA. The thermal stability and mechanical properties of the Cu2+@PBC dressing decreased with the addition of copper ions. Cu2+@PBC-2 film with a certain amount of copper sulfate used (10 nM) exhibited favorable antibacterial properties against both S. aureus and E. coli without obvious cytotoxicity. The results of the in vivo study also demonstrated that the Cu2+@PBC-2 film can eliminate S. aureus infections and inflammatory response, promote collagen deposition, capillary angiogenesis, hair follicle growth and wound healing. These results suggest that the Cu2+@PBC-2 film has prospective application as a wound dressing.

De novo strategy with engineering a multifunctional bacterial cellulose-based dressing for rapid healing of infected wounds.

The treatment and healing of infected skin lesions is one of the major challenges in surgery. To solve this problem, collagen I (Col-I) and the antibacterial agent hydroxypropyltrimethyl ammonium chloride chitosan (HACC) were composited into the bacterial cellulose (BC) three-dimensional network structure by a novel membrane-liquid interface (MLI) culture, and a Col-I/HACC/BC (CHBC) multifunctional dressing was designed. The water absorption rate and water vapor transmission rate of the obtained CHBC dressing were 35.78 ± 2.45 g/g and 3084 ± 56 g m-2·day-1, respectively. The water retention of the CHBC dressing was significantly improved compared with the BC caused by the introduced Col-I and HACC. In vitro results indicated that the combined advantages of HACC and Col-I confer on CHBC dressings not only have outstanding antibacterial properties against Staphylococcus aureus (S. aureus) compared with BC and CBC, but also exhibit better cytocompatibility than BC and HBC to promote the proliferation and spread of NIH3T3 cells and HUVECs. Most importantly, the results of in vivo animal tests demonstrated that the CHBC dressings fully promoted wound healing for 8 days and exhibited shorter healing times, especially in the case of wound infection. Excellent skin regeneration effects and higher expression levels of collagen during infection were also shown in the CHBC group. We believe that CHBC composites with favorable multifunctionality have potential applications as wound dressings to treat infected wounds.

Rational Design of All-Organic Nanoplatform for Highly Efficient MR/NIR-II Imaging-Guided Cancer Phototheranostics.

Organic theranostic nanomedicine has precision multimodel imaging capability and concurrent therapeutics under noninvasive imaging guidance. However, the rational design of desirable multifunctional organic theranostics for cancer remains challenging. Rational engineering of organic semiconducting nanomaterials has revealed great potential for cancer theranostics largely owing to their intrinsic diversified biophotonics, easy fabrication of multimodel imaging platform, and desirable biocompatibility. Herein, a novel all-organic nanotheranostic platform (TPATQ-PNP NPs) is developed by exploiting the self-assembly of a semiconducting small molecule (TPATQ) and a new synthetic high-density nitroxide radical-based amphiphilic polymer (PNP). The nitroxide radicals act as metal-free magnetic resonance imaging agent through shortened longitudinal relaxation times, and the semiconducting molecules enable ultralow background second near-infrared (NIR-II, 1000-1700 nm) fluorescence imaging. The as-prepared TPATQ-PNP NPs can light up whole blood vessels of mice and show precision tumor-locating ability with synergistic (MR/NIR-II) imaging modalities. The semiconducting molecules also undergo highly effective photothermal conversion in the NIR region for cancer photothermal therapy guided by complementary tumor diagnosis. The designed multifunctional organic semiconducting self-assembly provides new insights into the development of a new platform for cancer theranostics.

Engineering a novel antibacterial agent with multifunction: Protocatechuic acid-grafted-quaternized chitosan.

As a mild cationic antibacterial agent, hydroxypropyltrimethyl ammonium chloride chitosan (HACC) could kill gram-positive bacteria and gram-positive drug-resistant bacteria without cytotoxicity. Nevertheless, it was not effective against gram-negative bacteria. Herein, protocatechuic acid (PA) with broad-spectrum antibacterial properties and pharmacological activities was grafted on HACC. PA-g-HACC showed favourable antioxidant capacity and anti-inflammatory properties. Most importantly, the results of antibacterial assay indicated that the antibacterial rates of all PA-g-HACC groups against Staphylococcus aureus (S. aureus) and methicillin-resistant Staphylococcus aureus (MRSA) were above 92 %, and the antibacterial rate of PA-g-HACC against E. coli was increased with the amount of grafted PA. Furthermore, the cytocompatibility of PA-g-HACC was improved by appropriate grafting ratio of PA, while excessive grafted PA can lead to toxicity. We believe that PA-g-HACC in optimum grafting ratio of PA with favorable antibacterial properties, pharmacological activities and cytocompatibility will be potential antibacterial agent for treating infections.

Interpenetrated nano- and submicro-fibrous biomimetic scaffolds towards enhanced mechanical and biological performances.

Developing fibrous scaffolds with hierarchical structures that closely mimic natural extracellular matrix (ECM) is highly desirable. However, fabricating scaffolds with true nanofibers (<100 nm) and submicrofibers (<1 µm) remains a big challenge. In this work, to mimic the fibrillar structure of natural ECM, bacterial cellulose (BC) nanofibers were hybridized with cellulose acetate (CA) submicrofibers for the first time. The interpenetrated nano-submicron fibrous BC/CA scaffold was fabricated using the combined electrospinning and modified in situ biosynthesis method. The BC/CA scaffold has an integrated symmetrical nanostructure in which BC nanofibers (42 nm in diameter) penetrate into the submicrofibrous CA (820 nm in diameter) scaffold. The BC/CA scaffold shows an interconnected porous structure with a high porosity of >90%. Additionally, the combination of CA submicrofibers with BC nanofibers leads to significantly improved mechanical properties over nanofibrous BC and submicrofibrous CA scaffolds and enlarged pores over nanofibrous BC scaffold. In addition, the biological behaviors of prepared BC/CA on MC3T3-E1 cells were investigated. Results suggested that BC/CA scaffold is beneficial for cell migration and proliferation. Moreover, the BC/CA scaffold shows higher alkaline phosphatase (ALP) activity, and calcium depositions. In addition, the hierarchical structures can effectively improve the expression of osteogenic gene (ALP mRNA and Runx2 mRNA) and protein (ALP). We believe that the methodology might provide biomimetic morphological microenvironments for enhanced tissue regeneration.

Optimized Nanointerface Engineering of Micro/Nanostructured Titanium Implants to Enhance Cell-Nanotopography Interactions and Osseointegration.

The success of orthopedic implants requires rapid and complete osseointegration which relies on an implant surface with optimal features. To enhance cellular function in response to the implant surface, micro- and nanoscale topography have been suggested as essential. The aim of this study was to identify an optimized Ti nanostructure and to introduce it onto a titanium plasma-sprayed titanium implant (denoted NTPS-Ti) to confer enhanced immunomodulatory properties for optimal osseointegration. To this end, three types of titania nanostructures, namely, nanowires, nanonests, and nanoflakes, were achieved on hydrothermally prepared Ti substrates. The nanowire surface modulated protein conformation and directed integrin binding and specificity in such a way as to augment the osteogenic differentiation of bone marrow-derived mesenchymal stem cells (BMSCs) and induce a desirable osteoimmune response of RAW264.7 macrophages. In a coculture system, BMSCs on the optimized micro/nanosurface exerted enhanced effects on nonactivated or lipopolysaccharide-stimulated macrophages, causing them to adopt a less inflammatory macrophage profile. The enhanced immunomodulatory properties of BMSCs grown on NTPS-Ti depended on a ROCK-medicated cyclooxygenase-2 (COX2) pathway to increase prostaglandin E2 (PGE2) production, as evidenced by decreased production of PGE2 and concurrent inhibition of immunomodulatory properties after treatment with ROCK or COX2 inhibitors. In vivo evaluation showed that the NTPS-Ti implant resulted in enhanced osseointegration compared with the TPS-Ti and Ti implants. The results obtained in our study may provide a prospective approach for enhancing osseointegration and supporting the application of micro/nanostructured Ti implants.

Studies on bacterial cellulose/poly(vinyl alcohol) hydrogel composites as tissue-engineered corneal stroma.

Corneal transplantation is currently the major solution in the treatment of severe corneal diseases. However, it is restricted due to the limited number of corneal donors. A tissue-engineered cornea is a potential substitute which could help overcome this limitation. This research envisages the development of a novel tissue-engineered corneal stroma consisting of bacterial cellulose (BC)/poly(vinyl alcohol) (PVA) hydrogel composites for reconstructing the cornea. It was found that the properties of BC/PVA were better suited for use as a corneal stroma material than the BC hydrogel. The human corneal stromal cells (hCSCs) were used to evaluate the cytotoxicity of the materials, wherein BC/PVA displayed excellent biocompatibility with these cells. Furthermore, in the in vivo studies, the BC/PVA was transplanted intrastromally in rabbits. After four weeks, the cornea remained almost transparent, and without obvious inflammation, sensitization or neovascularization, as confirmed by the clinical and histological examinations. Our results demonstrate that BC/PVA was well-tolerated in the rabbit cornea, and may be a potential substitute for corneal stroma.

Improved antibacterial properties of collagen I/hyaluronic acid/quaternized chitosan multilayer modified titanium coatings with both contact-killing and release-killing functions.

Implant infection is one of the most severe complications after orthopedic surgery. The construction of an antibacterial coating on orthopedic implants with release-killing or contact-killing is one of the most efficient strategies to prevent implant-related infections. Here we reported a hydroxypropyltrimethyl ammonium chloride chitosan (HACC) based multilayer modified plasma-sprayed porous titanium coating generated via the layer-by-layer covalent-immobilized method. We demonstrated that the multilayer coating inhibited the colonization and biofilm formation of several bacterial strains, including Staphylococcus aureus (ATCC 25923), methicillin-resistant Staphylococcus aureus (MSRA, ATCC 43300) and clinical isolates of methicillin-resistant Staphylococcus epidermidis (MRSE 287), in vitro. HACC in the multilayer was released slowly with the degradation of the coating under the action of collagenase, further killing the planktonic bacteria, while the remaining HACC could kill the colonized bacteria. In a rat model of femur implants, the HACC-based multilayer-modified TCs effectively controlled the infection caused by MRSA and prevented bone destruction. Therefore, the HACC-based multilayer modified TCs with multiple antimicrobial properties could be a new potential ideal surface modification strategy to prevent implant associated infections.

An in vivo study on the effect of coating stability on osteointegration performance of collagen/hyaluronic acid multilayer modified titanium implants.

Aseptic loosening of implant is one of the main causes of Ti-based implant failure. In our previous work, a novel stable collagen/hyaluronic acid (Col/HA) multilayer modified titanium coatings (TCs) was developed by layer-by-layer (LBL) covalent immobilization technique, which showed enhanced biological properties compared with TCs that were physically absorbed with Col/HA multilayer in vitro. In this study, a rabbit model with femur condyle defect was employed to compare the osteointegration performance of them. Results indicated that Col/HA multilayer with favourable stability could better facilitate osteogenesis around implants and bone-implant contact. The Col/HA multilayer covalent-immobilized TC may reduce aseptic loosening of implant.