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1.
Int J Nanomedicine ; 19: 4007-4019, 2024.
Article in English | MEDLINE | ID: mdl-38715701

ABSTRACT

Introduction: Nanosized outer membrane vesicles (OMVs) from Gram-negative bacteria have attracted increasing interest because of their antitumor activity. However, the antitumor effects of MVs isolated from Gram-positive bacteria have rarely been investigated. Methods: MVs of Staphylococcus aureus USA300 were prepared and their antitumor efficacy was evaluated using tumor-bearing mouse models. A gene knock-in assay was performed to generate luciferase Antares2-MVs for bioluminescent detection. Cell counting kit-8 and lactic dehydrogenase release assays were used to detect the toxicity of the MVs against tumor cells in vitro. Active caspase-1 and gasdermin D (GSDMD) levels were determined using Western blot, and the tumor inhibition ability of MVs was determined in B16F10 cells treated with a caspase-1 inhibitor. Results: The vesicular particles of S. aureus USA300 MVs were 55.23 ± 8.17 nm in diameter, and 5 µg of MVs remarkably inhibited the growth of B16F10 melanoma in C57BL/6 mice and CT26 colon adenocarcinoma in BALB/c mice. The bioluminescent signals correlated well with the concentrations of the engineered Antares2-MVs (R2 = 0.999), and the sensitivity for bioluminescence imaging was 4 × 10-3 µg. Antares2-MVs can directly target tumor tissues in vivo, and 20 µg/mL Antares2-MVs considerably reduced the growth of B16F10 and CT26 tumor cells, but not non-carcinomatous bEnd.3 cells. MV treatment substantially increased the level of active caspase-1, which processes GSDMD to trigger pyroptosis in tumor cells. Blocking caspase-1 activation with VX-765 significantly protected tumor cells from MV killing in vitro and in vivo. Conclusion: S. aureus MVs can kill tumor cells by activating the pyroptosis pathway, and the induction of pyroptosis in tumor cells is a promising strategy for cancer treatment.


Subject(s)
Caspase 1 , Pyroptosis , Staphylococcus aureus , Animals , Female , Mice , Antineoplastic Agents , Bacterial Outer Membrane , Caspase 1/metabolism , Cell Line, Tumor , Colonic Neoplasms , Melanoma, Experimental/metabolism , Mice, Inbred BALB C , Mice, Inbred C57BL , Phosphate-Binding Proteins/metabolism , Staphylococcus aureus/metabolism
2.
Microb Cell Fact ; 23(1): 89, 2024 Mar 25.
Article in English | MEDLINE | ID: mdl-38528536

ABSTRACT

BACKGROUND: Staphylococcus aureus and its single or mixed biofilm infections seriously threaten global public health. Phage therapy, which uses active phage particles or phage-derived endolysins, has emerged as a promising alternative strategy to antibiotic treatment. However, high-efficient phage therapeutic regimens have yet to be established. RESULTS: In this study, we used an enrichment procedure to isolate phages against methicillin-resistant S. aureus (MRSA) XN108. We characterized phage SYL, a new member of the Kayvirus genus, Herelleviridae family. The phage endolysin LysSYL was expressed. LysSYL demonstrated stability under various conditions and exhibited a broader range of efficacy against staphylococcal strains than its parent phage (100% vs. 41.7%). Moreover, dynamic live/dead bacterial observation demonstrated that LysSYL could completely lyse MRSA USA300 within 10 min. Scan and transmission electron microscopy revealed evident bacterial cell perforation and deformation. In addition, LysSYL displayed strong eradication activity against single- and mixed-species biofilms associated with S. aureus. It also had the ability to kill bacterial persisters, and proved highly effective in eliminating persistent S. aureus when combined with vancomycin. Furthermore, LysSYL protected BALB/c mice from lethal S. aureus infections. A single-dose treatment with 50 mg/kg of LysSYL resulted in a dramatic reduction in bacterial loads in the blood, liver, spleen, lungs, and kidneys of a peritonitis mouse model, which resulted in rescuing 100% of mice challenged with 108 colony forming units of S. aureus USA300. CONCLUSIONS: Overall, the data provided in this study highlight the strong therapeutic potential of endolysin LysSYL in combating staphylococcal infections, including mono- and mixed-species biofilms related to S. aureus.


Subject(s)
Endopeptidases , Methicillin-Resistant Staphylococcus aureus , Staphylococcal Infections , Animals , Mice , Staphylococcus , Staphylococcus aureus , Staphylococcus Phages , Anti-Bacterial Agents/pharmacology , Anti-Bacterial Agents/therapeutic use , Staphylococcal Infections/drug therapy , Staphylococcal Infections/microbiology , Biofilms
3.
Microbiol Res ; 282: 127635, 2024 May.
Article in English | MEDLINE | ID: mdl-38340572

ABSTRACT

Bacteria develop tolerance after transient exposure to antibiotics, and tolerance is a significant driver of resistance. The purpose of this study is to evaluate the mechanisms underlying tolerance formation in vancomycin-intermediate Staphylococcus aureus (VISA) strains. VISA strains were cultured with sub-minimum inhibitory concentrations (sub-MICs) of vancomycin. Enhanced vancomycin tolerance was observed in VISA strains with distinct genetic lineages. Western blot revealed that the VISA protein succinylation (Ksucc) levels decreased with the increase in vancomycin exposure. Importantly, Ksucc modification, vancomycin tolerance, and cell wall synthesis were simultaneously affected after deletion of SacobB, which encodes a desuccinylase in S. aureus. Several Ksucc sites were identified in MurA, and vancomycin MIC levels of murA mutant and Ksucc-simulated (MurA(K69E) and MurA(K191E)) mutants were reduced. The vancomycin MIC levels of K65-MurA(K191E) in particular decreased to 1 mg/L, converting VISA strain K65 to a vancomycin-susceptible S. aureus strain. We further demonstrated that the enzymatic activity of MurA was dependent on Ksucc modification. Our data suggested the influence of vancomycin exposure on bacterial tolerance, and protein Ksucc modification is a novel mechanism in regulating vancomycin tolerance.


Subject(s)
Anti-Bacterial Agents , Staphylococcal Infections , Humans , Anti-Bacterial Agents/pharmacology , Anti-Bacterial Agents/metabolism , Vancomycin/pharmacology , Vancomycin/metabolism , Staphylococcus aureus/genetics , Staphylococcus aureus/metabolism , Vancomycin-Resistant Staphylococcus aureus , Down-Regulation , Microbial Sensitivity Tests , Staphylococcal Infections/microbiology
4.
J Agric Food Chem ; 72(9): 4928-4938, 2024 Mar 06.
Article in English | MEDLINE | ID: mdl-38393975

ABSTRACT

Sliver nanoparticles (AgNPs) have attracted tremendous interest as an alternative to commercially available antibiotics due to their low microbial resistance and broad-spectrum antimicrobial activity. However, AgNPs are highly reactive and unstable and are susceptible to fast oxidation. Synthesizing stable and efficient AgNPs using green chemistry principles remains a major challenge. To address this issue, we establish a facile route to form AgNP-doped zein nanoparticle core-satellite superstructures with ultralow minimum bactericidal concentration (MBC). In brief, polyphenol surface-functionalization of zein nanoparticles was performed, and the epigallocatechin gallate (EGCG) layer on zein nanoparticles served as a reducing-cum-stabilizing agent. We used EGCG-decorated zein nanoparticles (ZE) as a template to direct the nucleation and growth of AgNPs to develop metallized hybrid nanoparticles (ZE-Ag). The highly monodispersed core-satellite nanoparticles (∼150 nm) decorated with ∼4.9 nm AgNPs were synthesized successfully. The spatial restriction of EGCG by zein nanoparticles confined the nucleation and growth of AgNPs only on the surface of the particles, which prevented the formation of entangled clusters of polyphenols and AgNPs and concomitantly inhibited the coalescence and oxidation of AgNPs. Thus, this strategy improved the effective specific surface area of AgNPs, and as a result, ZE-Ag efficiently killed the indicator bacteria, Escherichia coli (E. coli) and Methicillin-resistant Staphylococcus aureus(MRSA) after 20 min of incubation, with MBCs of 2 and 4 µg/mL, respectively. This situation indicated that as-prepared core-satellite nanoparticles possessed potent short-term sterilization capability. Moreover, the simulated wound infection model also confirmed the promising application of ZE-Ag as an efficient antimicrobial composite. This work provides new insights into the synthesis and emerging application of AgNPs in food preservation, packaging, biomedicine, and catalysis.


Subject(s)
Anti-Infective Agents , Catechin/analogs & derivatives , Metal Nanoparticles , Methicillin-Resistant Staphylococcus aureus , Nanocomposites , Zein , Zein/chemistry , Silver/pharmacology , Silver/chemistry , Escherichia coli , Metal Nanoparticles/chemistry , Anti-Bacterial Agents/pharmacology , Anti-Bacterial Agents/chemistry , Anti-Infective Agents/pharmacology , Polyphenols/pharmacology , Excipients , Microbial Sensitivity Tests
5.
Front Microbiol ; 14: 1254367, 2023.
Article in English | MEDLINE | ID: mdl-37869662

ABSTRACT

Background: Membrane vesicles (MVs) are nanoscale vesicular structures produced by bacteria during their growth in vitro and in vivo. Some bacterial components can be loaded in bacterial MVs, but the roles of the loaded MV molecules are unclear. Methods: MVs of Staphylococcus aureus RN4220 and its derivatives were prepared. Dynamic light scattering analysis was used to evaluate the size distribution, and 4D-label-free liquid chromatography-tandem mass spectrometry analysis was performed to detect protein composition in the MVs. The site-mutation S. aureus RN4220-Δhld and agrA deletion mutant RN4220-ΔagrA were generated via allelic replacement strategies. A hemolysis assay was performed with rabbit red blood cells. CCK-8 and lactate dehydrogenase release assays were used to determine the cytotoxicity of S. aureus MVs against RAW264.7 macrophages. The serum levels of inflammatory factors such as IL-6, IL-1ß, and TNFα in mice treated with S. aureus MVs were detected with an enzyme-linked immunosorbent assay kit. Results: Delta-hemolysin (Hld) was identified as a major loaded factor in S. aureus MVs. Further study showed that Hld could promote the production of staphylococcal MVs with smaller sizes. Loaded Hld affected the diversity of loaded proteins in MVs of S. aureus RN4220. Hld resulted in decreased protein diversity in MVs of S. aureus. Site-mutation (RN4220-Δhld) and agrA deletion (RN4220-ΔagrA) mutants produced MVs (ΔhldMVs and ΔagrAMVs) with a greater number of bacterial proteins than those derived from wild-type RN4220 (wtMVs). Moreover, Hld contributed to the hemolytic activity of wtMVs. Hld-loaded wtMVs were cytotoxic to macrophage RAW264.7 cells and could stimulate the production of inflammatory factor IL-6 in vivo. Conclusion: This study presented that Hld was a major loaded factor in S. aureus MVs, and the loaded Hld played vital roles in the MV-property modification.

6.
mLife ; 2(4): 462-478, 2023 Dec.
Article in English | MEDLINE | ID: mdl-38818266

ABSTRACT

Focal and systemic infections are serious threats to human health. Preclinical models enable the development of new drugs and therapeutic regimens. In vivo, animal bioluminescence (BL) imaging has been used with bacterial reporter strains to evaluate antimicrobial treatment effects. However, high-sensitivity bioluminescent systems are required because of the limited tissue penetration and low brightness of the BL signals of existing approaches. Here, we report that NanoLuc (Nluc) showed better performance than LuxCDABE in bacteria. However, the retention rate of plasmid constructs in bacteria was low. To construct stable Staphylococcus aureus reporter strains, a partner protein enolase (Eno) was identified by screening of S. aureus strain USA300 for fusion expression of Nluc-based luciferases, including Nluc, Teluc, and Antares2. Different substrates, such as hydrofurimazine (HFZ), furimazine (FUR), and diphenylterazine (DTZ), were used to optimize a stable reporter strain/substrate pair for BL imaging. S. aureus USA300/Eno-Antares2/HFZ produced the highest number of photons of orange-red light in vitro and enabled sensitive BL tracking of S. aureus in vivo, with sensitivities of approximately 10 CFU from mouse skin and 750 CFU from mouse kidneys. USA300/Eno-Antares2/HFZ was a powerful combination based on the longitudinal evaluation of the therapeutic efficacy of antibiotics. The optimized S. aureus Eno-Antares2/HFZ pair provides a technological advancement for the in vivo evaluation of antimicrobial treatment.

8.
Mar Drugs ; 20(8)2022 Jul 26.
Article in English | MEDLINE | ID: mdl-35892947

ABSTRACT

The applications of alginate lyase are diverse, but efficient commercial enzymes are still unavailable. In this study, a novel alginate lyase with high activity was obtained from the marine bacteria Vibrio sp. Ni1. The ORF of the algB gene has 1824 bp, encoding 607 amino acids. Homology analysis shows that AlgB belongs to the PL7 family. There are two catalytic domains with the typical region of QIH found in AlgB. The purified recombinant enzyme of AlgB shows highest activity at 35 °C, pH 8.0, and 50 mmol/L Tris-HCl without any metal ions. Only K+ slightly enhances the activity, while Fe2+ and Cu2+ strongly inhibit the activity. The AlgB preferred polyM as substrate. The end products of enzymatic mixture are DP2 and DP3, without any metal ion to assist them. This enzyme has good industrial application prospects.


Subject(s)
Polysaccharide-Lyases , Vibrio , Alginates/metabolism , Bacterial Proteins/metabolism , Cloning, Molecular , Hydrogen-Ion Concentration , Ions , Metals/pharmacology , Polysaccharide-Lyases/metabolism , Substrate Specificity , Vibrio/metabolism
9.
J Oncol ; 2022: 8762717, 2022.
Article in English | MEDLINE | ID: mdl-35190740

ABSTRACT

OBJECTIVE: To analyze the target and potential mechanism of Scutellaria baicalensis (SB) in the treatment of HCC based on bioinformatics, so as to provide suggestions for the diagnosis, treatment, and drug development of hepatocellular carcinoma (HCC). METHODS: The regulated gene targets of SB were screened by gene expression pattern clustering and differential analysis of gene expression data of HepG2 cells treated with SB at 0 h, 1 h, 3 h, 6 h, 12 h, and 24 h. The module genes related to HCC were identified by the weighted gene coexpression network analysis (WGCNA). KEGG and GO enrichment were used to analyze the molecular function and structure of the module, and GSEA was used to evaluate the different functional pathways between normal people and patients with HCC. Then, the module gene was used for univariate Cox proportional hazard analysis and the least absolute shrinkage and selection operator (LASSO) Cox regression analysis to build a prognostic model. The protein-protein interaction network (PPI) was used to analyze the core genes regulated by SB (CGRSB) of the module, and the survival curve revealed the CGRSB impact on patient survival. The CIBERSORT algorithm combined with correlation analysis to explore the relationship between CGRSB and immune infiltration. Finally, the single-cell sequencing technique was used to analyze the distribution of CGRSB at the cellular level. RESULTS: SB could regulate 903 genes, of which 234 were related to the occurrence of HCC. The prognosis model constructed by these genes has a good effect in evaluating the survival of patients. KEGG and GO enrichment analysis showed that the regulation of SB on HCC mainly focused on some cell proliferation, apoptosis, and immune-related functions. GSEA enrichment analysis showed that these functions are related to the occurrence of HCC. A total of 24 CGRSB were obtained after screening, of which 13 were significantly related to survival, and most of them were unfavorable factors for patient survival. The correlation analysis of gene expression showed that most of CGRSB was significantly correlated with T cells, macrophages, and other functions. The results of single-cell analysis showed that the distribution of CGRSB in macrophages was the most. CONCLUSION: SB has high credibility in the treatment of HCC, such as CDK2, AURKB, RRM2, CENPE, ESR1, and PRIM2. These targets can be used as potential biomarkers for clinical diagnosis. The research also shows that the p53 signal pathway, MAPK signal pathway, apoptosis pathway, T cell receptor pathway, and macrophage-mediated tumor immunity play the most important role in the mechanism of SB in treating HCC.

10.
Biochem Biophys Res Commun ; 582: 125-130, 2021 12 10.
Article in English | MEDLINE | ID: mdl-34710827

ABSTRACT

CYP76AH1 is the key enzyme in the biosynthesis pathway of tanshinones in Salvia miltiorrhiza, which are famous natural products with activities against various heart diseases and others. CYP76AH1 is a membrane-associated typical plant class II cytochrome P450 enzyme and its catalytic mechanism has not to be clearly elucidated. Structural determination of eukaryotic P450 enzymes is extremely challenging. Recently, we solved the crystal structures of CYP76AH1 and CYP76AH1 in complex with its natural substrate miltiradiene. The structure of CYP76AH1 complexed with miltiradiene is the first plant cytochrome P450 structure in complex with natural substrate. The studies revealed a unique array pattern of amino acid residues, which may play an important role in orienting and stabilizing the substrate for catalysis. This work would provide structural insights into CYP76AH1 and related P450s and the basis to efficiently improve tanshinone production by synthetic biology techniques.


Subject(s)
Abietanes/biosynthesis , Cytochrome P-450 Enzyme System/chemistry , Diterpenes/chemistry , Plant Proteins/chemistry , Salvia miltiorrhiza/chemistry , Abietanes/genetics , Binding Sites , Cloning, Molecular , Crystallography, X-Ray , Cytochrome P-450 Enzyme System/genetics , Cytochrome P-450 Enzyme System/metabolism , Diterpenes/metabolism , Escherichia coli/genetics , Escherichia coli/metabolism , Gene Expression , Genetic Vectors/chemistry , Genetic Vectors/metabolism , Models, Molecular , Plant Proteins/genetics , Plant Proteins/metabolism , Protein Binding , Protein Conformation, alpha-Helical , Protein Conformation, beta-Strand , Protein Interaction Domains and Motifs , Recombinant Proteins/chemistry , Recombinant Proteins/genetics , Recombinant Proteins/metabolism , Salvia miltiorrhiza/enzymology , Secondary Metabolism/genetics , Substrate Specificity
11.
Exp Ther Med ; 22(3): 1001, 2021 Sep.
Article in English | MEDLINE | ID: mdl-34345283

ABSTRACT

Cyclooxygenase-2 (COX-2) is a common factor in inflammation, and its specific regulatory mechanism has not been fully elucidated. The present study aimed to investigate COX-2 mRNA and protein expression levels in synovium tissues and synovial fluid from patients with knee osteoarthritis (KOA), and determine the molecular mechanism by which microRNA (miRNA/miR)-758 regulates KOA via COX-2. A total of 37 patients with KOA and 29 patients with acute knee trauma (control group) were enrolled in the present study. Reverse transcription-quantitative PCR analysis was performed to detect miR-758-3p and COX-2 mRNA expression, while western blotting and ELISA were performed to detect COX-2 protein expression in synovium and synovial fluid, respectively. The dual-luciferase reporter assay was performed to verify the interaction between miR-758-3p and the 3'-untraslated region (UTR) of COX-2 mRNA. Synovial cells were transfected with agomiR-758-3p, and the MTT assay was performed to assess cell proliferation. The results demonstrated that COX-2 expression was higher in patients with KOA than those with acute knee trauma. Conversely, miR-758-3p expression was lower in patients with KOA than those with acute knee trauma. Notably, miR-758-3p interacted with the 3'-UTR of COX-2 mRNA to regulate its expression. Overexpression of miR-758-3p inhibited the expression and release of COX-2, as well as the proliferation of human KOA synovial cells. Taken together, these results suggest that COX-2 expression is upregulated in synovium tissues and synovial fluid from patients with KOA, which is associated with downregulated miR-758-3p expression. In addition, miR-758-3p affects the proliferation of synovial cells and the expression of relevant proteins in these cells, thus promoting the occurrence and development of KOA.

12.
Materials (Basel) ; 14(12)2021 Jun 17.
Article in English | MEDLINE | ID: mdl-34204347

ABSTRACT

Excessive bleeding induces a high risk of death and is a leading cause of deaths that result from traffic accidents and military conflict. In this paper, we developed a novel porous chitosan-CaCO3 (CS-CaCO3) composite material and investigated its hemostatic properties and wound healing performance. The CS-CaCO3 composites material was prepared via a wet-granulation method. Granulation increases the infiltrating ability of the CS-CaCO3 composites material. The improved water absorption ability was enhanced to 460% for the CS-CaCO3 composites material compared to the CaCO3 or chitosan with only one single component. The coagulation studies in vivo illustrated that the blood clotting time was greatly reduced from 31 s for CaCO3 to 16 s for the CS-CaCO3 composite material. According to the results of the wound healing experiments in rats, it was found that the CS-CaCO3 composite material can promote wound healing. The CS-CaCO3 composite material could accelerate wound healing to a rate of 9 days, compared with 12 days for the CaCO3. The hemostatic activity, biocompatibility, and low cost of CS-CaCO3 composite material make it a potential agent for effective hemostatic and wound healing materials.

13.
Chem Sci ; 12(11): 3885-3889, 2021 Feb 17.
Article in English | MEDLINE | ID: mdl-34163657

ABSTRACT

Chemo-enzymatic cascade processes are invaluable due to their ability to rapidly construct high-value products from available feedstock chemicals in a one-pot relay manner. However, they have proven to be challenging because of the mutual inactivation of both catalysts. A conceptually novel strategy based on Pickering interfacial catalysis (PIC) is proposed here to address this challenge. This study aimed to construct a protein-stabilized Pickering system for biphasic cascade catalysis, enabled by phosphorylated zein nanoparticles (ZCPOPs) immobilized in gold nanoparticles (Au NCs). Ultra-small Au NCs, 1-2 nm in diameter, were integrated into ZCPOPs at room temperature. Then, the as-synthesized ZCPOPs-Au NCs were used to stabilize the oil-in-water (o/w) Pickering emulsion. Besides their excellent catalytic activity and recycling ability in a variety of oil phases, ZCPOPs-Au NCs possess unpredictable catalytic activity and exhibit mimicking properties of horseradish peroxidase. Particularly, the cascade reaction is well achieved using a metal catalyst and a biocatalyst at the oil-water interface. The result showed that such a combination of chemo- and biocatalysis improved the catalytic yield more than two times compared with that of sole metal catalysis. This study opened a new avenue to design nanomaterials using the combination of chemo- and biocatalysis in a Pickering emulsion system for multistep syntheses.

14.
Food Chem ; 360: 130152, 2021 Oct 30.
Article in English | MEDLINE | ID: mdl-34034052

ABSTRACT

Protein-based Pickering emulsions have received considerable attention as nutraceutical vehicles. However, the oral bioavailability of nutraceuticals encapsulated in Pickering emulsions was not well established. In this work, a simulated gastrointestinal tract/Caco-2 cell culture model was applied to investigate the oral bioavailability of quercetin encapsulated in zein-based Pickering emulsions with quercetin in zein particles as the control. Pickering emulsions with shell (ZCP-QE) and core quercetin (ZCPE-Q) were constructed, and quercetin bioaccessibility, cell uptake and secretion, and the overall bioavailability were evaluated and compared. The overall oral bioavailability of quercetin was increased from 2.71% (bulk oil) to 38.18% (ZCPs-Q) and 18.97% (ZCPE-Q), particularly reached 41.22% for ZCP-QE. This work took new insights into the contributions of bioaccessibility and absorption (cell uptake plus secretion) to the overall oral bioavailability of quercetin. A schematic representation is proposed to relate the types of colloidal nanostructures in the digesta to the uptake, cell absorption, and overall oral bioavailability of quercetin. This study provided an attractive basis for identifying effective strategies to improve the oral bioavailability of hydrophobic nutraceuticals.


Subject(s)
Emulsions/chemistry , Quercetin/metabolism , Zein/chemistry , Biological Availability , Caco-2 Cells , Cell Survival/drug effects , Digestion , Humans , Microscopy, Confocal , Particle Size , Quercetin/chemistry , Quercetin/pharmacology
15.
Front Robot AI ; 8: 599650, 2021.
Article in English | MEDLINE | ID: mdl-33898528

ABSTRACT

We introduce a soft robot actuator composed of a pre-stressed elastomer film embedded with shape memory alloy (SMA) and a liquid metal (LM) curvature sensor. SMA-based actuators are commonly used as electrically-powered limbs to enable walking, crawling, and swimming of soft robots. However, they are susceptible to overheating and long-term degradation if they are electrically stimulated before they have time to mechanically recover from their previous activation cycle. Here, we address this by embedding the soft actuator with a capacitive LM sensor capable of measuring bending curvature. The soft sensor is thin and elastic and can track curvature changes without significantly altering the natural mechanical properties of the soft actuator. We show that the sensor can be incorporated into a closed-loop "bang-bang" controller to ensure that the actuator fully relaxes to its natural curvature before the next activation cycle. In this way, the activation frequency of the actuator can be dynamically adapted for continuous, cyclic actuation. Moreover, in the special case of slower, low power actuation, we can use the embedded curvature sensor as feedback for achieving partial actuation and limiting the amount of curvature change.

16.
J Biol Eng ; 15(1): 5, 2021 Feb 08.
Article in English | MEDLINE | ID: mdl-33557881

ABSTRACT

BACKGROUND: Synthetic biomaterials assist in modulating the vascular response in an injured bone by serving as delivery vehicles of pro-angiogenic molecules to the site of injury or by serving as mimetic platforms which offer support to cell growth and proliferation. METHODS: This study applied natural phospholipid modified protein technologies together with low temperature three-dimensional printing technology to develop a new model of three-dimensional artificial bone scaffold for potential use in repairing body injuries. The focus was to create a porous structure (PS) scaffold of two components, Bone Morphogenetic Protein-2 and Human Beta Defensin-3 (BMP2 and hBD3), which can synchronously realize directional bone induction, angiogenesis and postoperative antibacterial effects. BMP2 induces osteogenesis, whereas hBD3 is antibacterial. RESULTS: Our data showed that in the BMP2-hBD3-PS or hBD3-PS scaffolds, BMP2 had a slow-release rate of about 40% in 30 days, ensuring that BMP2 could penetrate into stem cells for osteogenic differentiation for a long time. The scaffolds promoted cell growth when in combination with BMP2, thus showing its importance in promoting cell growth. Alkaline Phosphatase (ALP) staining showed that the ALP content of BMP2-hBD3-PS and BMP2-PS had a significant increase in samples that contained BMP2, thus showing that these scaffolds promoted osteogenic differentiation. In all the constructs that had hBD3, they displayed antibacterial properties with hBD3, having a slow release of about 35% in 30 days, thus ensuring they provided protection. CONCLUSION: Based on this study, the 3D printed BMP2 scaffolds show a great potential for the development of biodegradable bone implants. LEVEL OF EVIDENCE: Level II, experimental comparative design.

17.
Mol Pharm ; 18(1): 377-385, 2021 01 04.
Article in English | MEDLINE | ID: mdl-33295773

ABSTRACT

RNAi therapy has been developed and explored for treating retinal conditions since last decades. The progression of retinal diseases including the age-related macular degeneration and glaucoma is associated with the malfunction of specific retinal cells. Therefore, to deliver therapeutic RNAi to selective retinal tissues with desired gene downregulation is crucial for the treatment of retinal diseases via RNAi therapy. Lipid-based nanoparticles are potent delivery vectors for RNAi therapeutics to achieve high gene silencing efficiency. The surface charge has been demonstrated to affect the intraocular behaviors and retinal distribution of intravitreally administered lipid nanoparticles (LNPs), which could subsequently affect the gene knockdown efficiency in specific retinal layers. Here, we evaluated three charged LNPs for their ability to deliver siRNA and facilitate gene downregulation both in vitro and in vivo. LNPs with different surface charges ranging from neutral to positive (5-34 mV) were successfully formulated. All types of charged LNPs managed gene knockdown in both mammalian cell line and primary neurons. At 48 h post intravitreal injection, neutral LNPs (6.2 mV) and mildly positive LNPs (15.9 mV) mediated limited retinal gene suppression (<10%) and the more positive LNPs (31.2 mV) led to ∼25% gene suppression in the retinal ganglion cell (RGC) layer. No gene silencing in the retinal pigmented epithelium layer was facilitated by any LNPs independent of the charges. In summary, this study has shown that positive LNPs with an optimized charge managed specific gene downregulation in the RGC layer. These RNAi carriers hold potential for the treatment of RGC-associated retinal diseases.


Subject(s)
Lipids/chemistry , Nanoparticles/administration & dosage , Nanoparticles/chemistry , RNA, Small Interfering/administration & dosage , RNA, Small Interfering/chemistry , Retinal Ganglion Cells/drug effects , Animals , Gene Silencing/drug effects , HEK293 Cells , Humans , Male , Mice , Mice, Inbred C57BL , RNA Interference/drug effects , RNAi Therapeutics/methods
18.
Polymers (Basel) ; 13(1)2020 Dec 23.
Article in English | MEDLINE | ID: mdl-33374509

ABSTRACT

It is important to emphasize that the adjustment of an organic-inorganic interfacial chemical environment plays an important role during the separation performance of composite materials. In this paper, a series of hybrid membranes were prepared by blending polyvinyl alcohol (PVA) solution and sulfonated nano-TiO2 (SNT) suspension. The effects of different interfacial chemical surroundings on ions transfer were explored by regulating the dosage content of SNT. The as-prepared membranes exhibited high thermal and mechanical stability, with initial decomposition temperatures of 220-253 °C, tensile strengths of 31.5-53.4 MPa, and elongations at break of 74.5-146.0%. The membranes possessed moderate water uptake (WR) values of 90.9-101.7% and acceptable alkali resistances (swelling degrees were 187.2-206.5% and weight losses were 10.0-20.8%). The as-prepared membranes were used for the alkali recovery of a NaOH/Na2WO4 system via the diffusion dialysis process successfully. The results showed that the dialysis coefficients of OH- (UOH) were in a range of 0.013-0.022 m/h, and separate factors (S) were in an acceptable range of 22-33. Sulfonic groups in the interfacial regions and -OH in the PVA main chains were both deemed to play corporate roles during the transport of Na+ and OH-.

19.
Polymers (Basel) ; 12(10)2020 Oct 14.
Article in English | MEDLINE | ID: mdl-33066612

ABSTRACT

Carboxyl-type boronic acid copolymers (CBACs) were synthesized by a radical polymerization method and used for the preparation of polyvinyl alcohol (PVA)-based composite membranes via a solution mixture method. The as-prepared composite membranes exhibited a water uptake (WR) of 122.6-150.0%, an ion exchange capacity (IEC) of 0.0147-0.0518 mmol g-1, and excellent mechanical (elongation at break (Eb) of 103.8-148.4%, tensile strength (TS) of 38.7-58.6 MPa) and thermal stability. The alkali resistances of the as-prepared membranes were tested by immersing the samples into 2 mol L-1 NaOH solutions at 25 °C for 60 h, and the results were encouraging: the mass loss and swelling degree of the as-prepared membranes were in the ranges of 1.9-5.9% and 222.6-241.9%, respectively. The separation performances of the as-prepared membranes were evaluated by the diffusion dialysis (DD) process with an NaOH/Na2WO4 mixture at room temperature. The results demonstrated that the dialysis coefficients of hydroxide (UOH) were in the range of 0.0147-0.0347 m h-1, and the separation factors (S) were in the range of 29.5-62.6. The introduced carboxyl groups from CBACs and the -OH groups from PVA were both deemed to play significant roles in the promotion of ion transport: the -COO- groups formed negatively charged transport channels for Na+ by electrostatic attraction, and the -OH groups promoted the transport of OH- via hydrogen bonding.

20.
Nat Commun ; 11(1): 2233, 2020 05 06.
Article in English | MEDLINE | ID: mdl-32376823

ABSTRACT

Soft robots are primarily composed of soft materials that can allow for mechanically robust maneuvers that are not typically possible with conventional rigid robotic systems. However, owing to the current limitations in simulation, design and control of soft robots often involve a painstaking trial. With the ultimate goal of a computational framework for soft robotic engineering, here we introduce a numerical simulation tool for limbed soft robots that draws inspiration from discrete differential geometry based simulation of slender structures. The simulation incorporates an implicit treatment of the elasticity of the limbs, inelastic collision between a soft body and rigid surface, and unilateral contact and Coulombic friction with an uneven surface. The computational efficiency of the numerical method enables it to run faster than real-time on a desktop processor. Our experiments and simulations show quantitative agreement and indicate the potential role of predictive simulations for soft robot design.

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