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1.
Biomed Mater ; 19(5)2024 Jul 04.
Artículo en Inglés | MEDLINE | ID: mdl-38917828

RESUMEN

The increasing prevalence of bone replacements and complications associated with bone replacement procedures underscores the need for innovative tissue restoration approaches. Existing synthetic grafts cannot fully replicate bone vascularization and mechanical characteristics. This study introduces a novel strategy utilizing pectin, chitosan, and polyvinyl alcohol to create interpenetrating polymeric network (IPN) scaffolds incorporated with extracellular vesicles (EVs) isolated from human mesenchymal stem cells (hMSCs). We assess the osteointegration and osteoconduction abilities of these modelsin vitrousing hMSCs and MG-63 osteosarcoma cells. Additionally, we confirm exosome properties through Transmission Electron Microscopy (TEM), immunoblotting, and Dynamic Light Scattering (DLS).In vivo, chick allantoic membrane assay investigates vascularization characteristics. The study did not includein vivoanimal experiments. Our results demonstrate that the IPN scaffold is highly porous and interconnected, potentially suitable for bone implants. EVs, approximately 100 nm in size, enhance cell survival, proliferation, alkaline phosphatase activity, and the expression of osteogenic genes. EVs-mediated IPN scaffolds demonstrate promise as precise drug carriers, enabling customized treatments for bone-related conditions and regeneration efforts. Therefore, the EVs-mediated IPN scaffolds demonstrate promise as precise carriers for the transport of drugs, allowing for customized treatments for conditions connected to bone and efforts in regeneration.


Asunto(s)
Regeneración Ósea , Proliferación Celular , Vesículas Extracelulares , Células Madre Mesenquimatosas , Osteogénesis , Andamios del Tejido , Humanos , Vesículas Extracelulares/metabolismo , Vesículas Extracelulares/química , Andamios del Tejido/química , Células Madre Mesenquimatosas/citología , Animales , Línea Celular Tumoral , Transducción de Señal , Supervivencia Celular , Ingeniería de Tejidos/métodos , Quitosano/química , Fosfatasa Alcalina/metabolismo , Oseointegración , Polímeros/química , Porosidad
2.
J Immunol ; 2024 Jun 21.
Artículo en Inglés | MEDLINE | ID: mdl-38905108

RESUMEN

Hepatitis E virus (HEV) is a worldwide zoonotic and public health concern. The study of HEV biology is helpful for designing viral vaccines and drugs. Nanobodies have recently been considered appealing materials for viral biological research. In this study, a Bactrian camel was immunized with capsid proteins from different genotypes (1, 3, 4, and avian) of HEV. Then, a phage library (6.3 × 108 individual clones) was constructed using peripheral blood lymphocytes from the immunized camel, and 12 nanobodies against the truncated capsid protein of genotype 3 HEV (g3-p239) were screened. g3-p239-Nb55 can cross-react with different genotypes of HEV and block Kernow-C1/P6 HEV from infecting HepG2/C3A cells. To our knowledge, the epitope recognized by g3-p239-Nb55 was determined to be a novel conformational epitope located on the surface of viral particles and highly conserved among different mammalian HEV isolates. Next, to increase the affinity and half-life of the nanobody, it was displayed on the surface of ferritin, which can self-assemble into a 24-subunit nanocage, namely, fenobody-55. The affinities of fenobody-55 to g3-p239 were ∼20 times greater than those of g3-p239-Nb55. In addition, the half-life of fenobody-55 was nine times greater than that of g3-p239-Nb55. G3-p239-Nb55 and fenobody-55 can block p239 attachment and Kernow-C1/P6 infection of HepG2/C3A cells. Fenobody-55 can completely neutralize HEV infection in rabbits when it is preincubated with nonenveloped HEV particles. Our study reported a case in which a nanobody neutralized HEV infection by preincubation, identified a (to our knowledge) novel and conserved conformational epitope of HEV, and provided new material for researching HEV biology.

4.
J Med Virol ; 96(6): e29691, 2024 Jun.
Artículo en Inglés | MEDLINE | ID: mdl-38783788

RESUMEN

Hepatitis E virus (HEV) is an emerging zoonotic pathogen that is transmitted primarily through the fecal-oral route and can cause acute hepatitis in humans. Since HEV was identified as a zoonotic pathogen, different species of HEV strains have been globally identified from various hosts, leading to an expanding range of hosts. The HEV genome consists of a 5' noncoding region, three open reading frames (ORFs), and a 3' noncoding region. The ORF3 protein is the smallest but has many functions in HEV release and pathogenesis. In this review, we systematically summarize recent progress in understanding the functions of the HEV ORF3 protein in virion release, biogenesis of quasi-enveloped viruses, antigenicity, and host environmental regulation. This review will help us to understand HEV replication and pathogenesis mechanisms better.


Asunto(s)
Virus de la Hepatitis E , Hepatitis E , Proteínas Virales , Virus de la Hepatitis E/genética , Humanos , Proteínas Virales/genética , Proteínas Virales/metabolismo , Hepatitis E/virología , Animales , Replicación Viral , Liberación del Virus , Sistemas de Lectura Abierta , Interacciones Huésped-Patógeno/genética , Genoma Viral
5.
J Virol Methods ; 328: 114954, 2024 Jul.
Artículo en Inglés | MEDLINE | ID: mdl-38763359

RESUMEN

Porcine circovirus type 2 (PCV2) is intensely prevalent in global pig farms. The PCV2 vaccine is an important means of preventing and controlling PCV2. The quality control of PCV2 vaccines is predominantly based on detection techniques such as animal testing and neutralizing antibody titration. Measuring the content of effective proteins in vaccines to measure vaccine efficacy is an excellent alternative to traditional methods, which can greatly accelerate the development speed and testing time of vaccines. In this study, we screened a monoclonal antibody (mAb) that can effectively recognize not only the exogenous expression of PCV2 Cap protein but also PCV2 virus. The double antibody sandwich ELISA (DAS-ELISA) was developed using this mAb that specifically recognize PCV2 Cap. The minimum protein content detected by this method is 3.5 ng/mL. This method can be used for the quality control of PCV2 inactivated vaccine and subunit vaccine, and the detection results are consistent with the results of mice animal experiments. This method has the advantages of simple operation, good sensitivity, high specificity and wide application. It can detect the effective antigen Cap protein content of various types of PCV2 vaccines, which not only shorten the vaccine inspection time but also save costs.


Asunto(s)
Anticuerpos Monoclonales , Anticuerpos Antivirales , Antígenos Virales , Infecciones por Circoviridae , Circovirus , Ensayo de Inmunoadsorción Enzimática , Sensibilidad y Especificidad , Enfermedades de los Porcinos , Vacunas Virales , Circovirus/inmunología , Animales , Ensayo de Inmunoadsorción Enzimática/métodos , Porcinos , Vacunas Virales/inmunología , Anticuerpos Monoclonales/inmunología , Antígenos Virales/inmunología , Antígenos Virales/análisis , Ratones , Anticuerpos Antivirales/sangre , Infecciones por Circoviridae/veterinaria , Infecciones por Circoviridae/diagnóstico , Infecciones por Circoviridae/prevención & control , Enfermedades de los Porcinos/diagnóstico , Enfermedades de los Porcinos/prevención & control , Enfermedades de los Porcinos/virología , Proteínas de la Cápside/inmunología
6.
Colloids Surf B Biointerfaces ; 238: 113925, 2024 Jun.
Artículo en Inglés | MEDLINE | ID: mdl-38657556

RESUMEN

Antibiotic-loaded calcium phosphate cement (CPC) has emerged as a promising biomaterial for drug delivery in orthopedics. However, there are problems such as the burst release of antibiotics, low cumulative release ratio, inappropriate release cycle, inferior mechanical strength, and poor anti-collapse properties. In this research, montmorillonite-gentamicin (MMT-GS) was fabricated by solution intercalation method and served as the drug release pathways in CPC to avoid burst release of GS, achieving promoted cumulative release ratios and a release cycle matched the time of inflammatory response. The results indicated that the highest cumulative release ratio and release concentration of GS in CPC/MMT-GS was 94.1 ± 2.8 % and 1183.05 µg/mL, and the release cycle was up to 504 h. In addition, the hierarchical GS delivery system was divided into three stages, and the kinetics followed the Korsmeyer-Peppas model, the zero-order model, and the diffusion-dissolution model, respectively. Meanwhile, the compressive strength of CPC/MMT-GS was up to 51.33 ± 3.62 MPa. Antibacterial results demonstrated that CPC/MMT-GS exhibited excellent in vitro long-lasting antibacterial properties to E. coli and S. aureus. Furthermore, CPC/MMT-GS promoted osteoblast proliferation and exhibited excellent in vivo histocompatibility. Therefore, CPC/MMT-GS has favorable application prospects in the treatment of bone defects with bacterial infections and inflammatory reactions.


Asunto(s)
Antibacterianos , Bentonita , Cementos para Huesos , Fosfatos de Calcio , Sistemas de Liberación de Medicamentos , Liberación de Fármacos , Escherichia coli , Gentamicinas , Staphylococcus aureus , Bentonita/química , Antibacterianos/farmacología , Antibacterianos/química , Gentamicinas/farmacología , Gentamicinas/química , Gentamicinas/administración & dosificación , Gentamicinas/farmacocinética , Fosfatos de Calcio/química , Cementos para Huesos/química , Cementos para Huesos/farmacología , Animales , Escherichia coli/efectos de los fármacos , Ratones , Staphylococcus aureus/efectos de los fármacos , Pruebas de Sensibilidad Microbiana , Tamaño de la Partícula
7.
Virol Sin ; 39(3): 478-489, 2024 Jun.
Artículo en Inglés | MEDLINE | ID: mdl-38588947

RESUMEN

African swine fever virus (ASFV) poses a significant threat to the global swine industry. Currently, there are no effective vaccines or treatments available to combat ASFV infection in pigs. The primary means of controlling the spread of the disease is through rapid detection and subsequent elimination of infected pig. Recently, a lower virulent ASFV isolate with a deleted EP402R gene (CD2v-deleted) has been reported in China, which further complicates the control of ASFV infection in pig farms. Furthermore, an EP402R-deleted ASFV variant has been developed as a potential live attenuated vaccine candidate strain. Therefore, it is crucial to develop detection methods that can distinguish wild-type and EP402R-deleted ASFV infections. In this study, two recombinant ASFV-p72 and -CD2v proteins were expressed using a prokaryotic system and used to immunize Bactrian camels. Subsequently, eight nanobodies against ASFV-p72 and ten nanobodies against ASFV-CD2v were screened. Following the production of these nanobodies with horse radish peroxidase (HRP) fusion proteins, the ASFV-p72-Nb2-HRP and ASFV-CD2v-Nb22-HRP fusions were selected for the development of two competitive ELISAs (cELISAs) to detect anti-ASFV antibodies. The two cELISAs exhibited high sensitivity, good specificity, repeatability, and stability. The coincidence rate between the two cELISAs and commercial ELISA kits was 98.6% and 97.6%, respectively. Collectively, the two cELISA for detecting antibodies against ASFV demonstrated ease of operation, a low cost, and a simple production process. The two cELISAs could determine whether pigs were infected with wild-type or CD2v-deleted ASFV, and could play an important role in monitoring ASFV infections in pig farms.


Asunto(s)
Virus de la Fiebre Porcina Africana , Fiebre Porcina Africana , Anticuerpos Antivirales , Ensayo de Inmunoadsorción Enzimática , Anticuerpos de Dominio Único , Virus de la Fiebre Porcina Africana/inmunología , Virus de la Fiebre Porcina Africana/genética , Animales , Porcinos , Fiebre Porcina Africana/diagnóstico , Fiebre Porcina Africana/inmunología , Fiebre Porcina Africana/virología , Anticuerpos Antivirales/inmunología , Ensayo de Inmunoadsorción Enzimática/métodos , Anticuerpos de Dominio Único/inmunología , Anticuerpos de Dominio Único/genética , Camelus , Sensibilidad y Especificidad , China , Proteínas de la Cápside/inmunología , Proteínas de la Cápside/genética , Proteínas Virales/inmunología , Proteínas Virales/genética
8.
Med Biol Eng Comput ; 2024 Apr 13.
Artículo en Inglés | MEDLINE | ID: mdl-38609577

RESUMEN

ASTRACT: One of the most common oral diseases affecting millions of people worldwide is periodontitis. Usually, proteins in body fluids are used as biomarkers of diseases. This study focused on hydrogen peroxide, lipopolysaccharide (LPS), and lactic acid as salivary non-protein biomarkers for oxidative stress conditions of periodontitis. Electrochemical analysis of artificial saliva was done using Gamry with increasing hydrogen peroxide, bLPS, and lactic acid concentrations. Electrochemical impedance spectroscopy (EIS) and cyclic voltammetry (CV) were conducted. From EIS data, change in capacitance and CV plot area were calculated for each test condition. Hydrogen peroxide groups had a decrease in CV area and an increase in percentage change in capacitance, lipopolysaccharide groups had a decrease in CV area and a decrease in percentage change in capacitance, and lactic acid groups had an increase of CV area and an increase in percentage change in capacitance with increasing concentrations. These data showed a unique combination of electrochemical properties for the three biomarkers. Scanning electron microscopy (SEM) with energy dispersive spectroscopy (EDS) employed to observe the change in the electrode surface and elemental composition data present on the sensor surface also showed a unique trend of elemental weight percentages. Machine learning models using hydrogen peroxide, LPS, and lactic acid electrochemical data were applied for the prediction of risk levels of periodontitis. The detection of hydrogen peroxide, LPS, and lactic acid by electrochemical biosensors indicates the potential to use these molecules as electrochemical biomarkers and use the data for ML-driven prediction tool for the periodontitis risk levels.

9.
J Virol ; 98(4): e0164923, 2024 Apr 16.
Artículo en Inglés | MEDLINE | ID: mdl-38548704

RESUMEN

Hepatitis E virus (HEV) is the most common cause of acute viral hepatitis worldwide, responsible for approximately 20 million infections annually. Among the three open reading frames (ORFs) of the HEV genome, the ORF3 protein is involved in virus release. However, the host proteins involved in HEV release need to be clarified. In this study, a host protein, thioredoxin domain-containing protein 5 (TXNDC5), interacted with the non-palmitoylated ORF3 protein by co-immunoprecipitation analysis. We determined that the overexpression or knockdown of TXNDC5 positively regulated HEV release from the host cells. The 17FCL19 mutation of the ORF3 protein lost the ability to interact with TXNDC5. The releasing amounts of HEV with the ORF3 mutation (FCL17-19SSP) were decreased compared with wild-type HEV. The overexpression of TXNDC5 can stabilize and increase ORF3 protein amounts, but not the TXNDC5 mutant with amino acids 1-88 deletion. Meanwhile, we determined that the function of TXNDC5 on the stabilization of ORF3 protein is independent of the Trx-like domains. Knockdown of TXNDC5 could lead to the degradation of ORF3 protein by the endoplasmic reticulum (ER)-associated protein degradation-proteasome system. However, the ORF3 protein cannot be degraded in the knockout-TXNDC5 stable cells, suggesting that it may hijack other proteins for its stabilization. Subsequently, we found that the other members of protein disulfide isomerase (PDI), including PDIA1, PDIA3, PDIA4, and PDIA6, can increase ORF3 protein amounts, and PDIA3 and PDIA6 interact with ORF3 protein. Collectively, our study suggested that HEV ORF3 protein can utilize TXNDC5 for its stability in ER to facilitate viral release. IMPORTANCE: Hepatitis E virus (HEV) infection is the leading cause of acute viral hepatitis worldwide. After the synthesis and modification in the cells, the mature ORF3 protein is essential for HEV release. However, the host protein involved in this process has yet to be determined. Here, we reported a novel host protein, thioredoxin domain-containing protein 5 (TXNDC5), as a chaperone, contributing to HEV release by facilitating ORF3 protein stability in the endoplasmic reticulum through interacting with non-palmitoylated ORF3 protein. However, we also found that in the knockout-TXNDC5 stable cell lines, the HEV ORF3 protein may hijack other proteins for its stabilization. For the first time, our study demonstrated the involvement of TXNDC5 in viral particle release. These findings provide some new insights into the process of the HEV life cycle, the interaction between HEV and host factors, and a new direction for antiviral design.


Asunto(s)
Virus de la Hepatitis E , Hepatitis E , Hepatitis Viral Humana , Humanos , Virus de la Hepatitis E/genética , Factores Inmunológicos , Proteína Disulfuro Isomerasas/genética , Tiorredoxinas/genética , Virión/metabolismo
10.
Thin Solid Films ; 7882024 Jan 15.
Artículo en Inglés | MEDLINE | ID: mdl-38343423

RESUMEN

The present study aimed to investigate the feasibility of using acoustic emission (AE) as a detection method for identifying failure mechanisms at the modular junction interface in total hip replacements (THRs) subjected to fretting corrosion. The experimental setup involved simulating fretting corrosion using a Ti6Al4V disc representing the femoral neck and a ZrO2 pin representing the femoral head. Mechanical testing provided insights into the wear and frictional behavior occurring at the modular junction interface. The results revealed that for all three potential conditions, a fretting condition of partial slip was observed. These findings highlight the importance of understanding the mechanical interactions and their influence on the overall performance and longevity of THRs. Electrochemical analysis shed light on the corrosion behavior under different potentiostatic conditions. High potentials in the anodic condition led to increased corrosion and ion transfer due to the breakdown of the passive oxide layer. Conversely, the cathodic potential condition exhibited a regrowth of the passive oxide layer, protecting the Ti6Al4V surface from further corrosion. The mid-range corrosion potential condition showed a dynamic equilibrium between corrosion and passivation processes. These electrochemical insights enhance our understanding of the mechanisms involved in fretting corrosion. The AE data proved to be promising in detecting and monitoring the onset and progression of failure mechanisms. The AE signals exhibited distinctive patterns that correlated with the severity of fretting corrosion. Notably, the hit driven data results, derived from AE signals, demonstrated the ability to differentiate between different levels of fretting conditions. This suggests that AE can serve as a valuable diagnostic tool for early detection and continuous monitoring of implant failure in THRs.

11.
J Virol ; 98(2): e0165023, 2024 Feb 20.
Artículo en Inglés | MEDLINE | ID: mdl-38271227

RESUMEN

Vaccination is the most effective method to protect humans and animals from diseases. Anti-idiotype vaccines are safer due to their absence of pathogens. However, the commercial production of traditional anti-idiotype vaccines using monoclonal and polyclonal antibodies (mAb and pAb) is complex and has a high failure rate. The present study designed a novel, simple, low-cost strategy for developing anti-idiotype vaccines with nanobody technology. We used porcine circovirus type 2 (PCV2) as a viral model, which can result in serious economic loss in the pig industry. The neutralizing mAb-1E7 (Ab1) against PCV2 capsid protein (PCV2-Cap) was immunized in the camel. And 12 nanobodies against mAb-1E7 were screened. Among them, Nb61 (Ab2) targeted the idiotype epitope of mAb-1E7 and blocked mAb-1E7's binding to PCV2-Cap. Additionally, a high-dose Nb61 vaccination can also protect mice and pigs from PCV2 infection. Epitope mapping showed that mAb-1E7 recognized the 75NINDFL80 of PCV2-Cap and 101NYNDFLG107 of Nb61. Subsequently, the mAb-3G4 (Ab3) against Nb61 was produced and can neutralize PCV2 infection in the PK-15 cells. Structure analysis showed that the amino acids of mAb-1E7 and mAb-3G4 respective binding to PCV2-Cap and Nb61 were also similar on the amino acids sequences and spatial conformation. Collectively, our study first provided a strategy for producing nanobody-based anti-idiotype vaccines and identified that anti-idiotype nanobodies could mimic the antigen on amino acids and structures. Importantly, as more and more neutralization mAbs against different pathogens are prepared, anti-idiotype nanobody vaccines can be easily produced against the disease with our strategy, especially for dangerous pathogens.IMPORTANCEAnti-idiotype vaccines utilize idiotype-anti-idiotype network theory, eliminating the need for external antigens as vaccine candidates. Especially for dangerous pathogens, they were safer because they did not contact the live pathogenic microorganisms. However, developing anti-idiotype vaccines with traditional monoclonal and polyclonal antibodies is complex and has a high failure rate. We present a novel, universal, simple, low-cost strategy for producing anti-idiotype vaccines with nanobody technology. Using a neutralization antibody against PCV2-Cap, a nanobody (Ab2) was successfully produced and could mimic the neutralizing epitope of PCV2-Cap. The nanobody can induce protective immune responses against PCV2 infection in mice and pigs. It highlighted that the anti-idiotype vaccine using nanobody has a very good application in the future, especially for dangerous pathogens.


Asunto(s)
Infecciones por Circoviridae , Circovirus , Anticuerpos de Dominio Único , Vacunas Virales , Animales , Humanos , Ratones , Proteínas de la Cápside , Infecciones por Circoviridae/prevención & control , Infecciones por Circoviridae/veterinaria , Epítopos , Porcinos , Vacunas Virales/química , Vacunas Virales/inmunología
12.
J Virol ; 98(1): e0131923, 2024 Jan 23.
Artículo en Inglés | MEDLINE | ID: mdl-38084961

RESUMEN

Porcine reproductive and respiratory syndrome (PRRS) is a serious global pig industry disease. Understanding the mechanism of viral replication and developing efficient antiviral strategies are necessary for combating with PRRS virus (PRRSV) infection. Recently, nanobody is considered to be a promising antiviral drug, especially for respiratory viruses. The present study evaluated two nanobodies against PRRSV nucleocapsid (N) protein (PRRSV-N-Nb1 and -Nb2) for their anti-PRRSV activity in vitro and in vivo. The results showed that intracellularly expressed PRRSV-N-Nb1 significantly inhibited PRRSV-2 replication in MARC-145 cells (approximately 100%). Then, the PRRSV-N-Nb1 fused with porcine IgG Fc (Nb1-pFc) as a delivering tag was produced and used to determine its effect on PRRSV-2 replication in porcine alveolar macrophages (PAMs) and pigs. The inhibition rate of Nb1-pFc against PRRSV-2 in PAMs could reach >90%, and it can also inhibit viral replication in vivo. Epitope mapping showed that the motif Serine 105 (S105) in PRRSV-2 N protein was the key amino acid binding to PRRSV-N-Nb1, which is also pivotal for the self-interaction of N protein via binding to Arginine 97. Moreover, viral particles were not successfully rescued when the S105 motif was mutated to Alanine (S105A). Attachment, entry, genome replication, release, docking model analysis, and blocking enzyme-linked immunosorbent assay (ELISA) indicated that the binding of PRRSV-N-Nb1 to N protein could block its self-binding, which prevents the viral replication of PRRSV. PRRSV-N-Nb1 may be a promising drug to counter PRRSV-2 infection. We also provided some new insights into the molecular basis of PRRSV N protein self-binding and assembly of viral particles.IMPORTANCEPorcine reproductive and respiratory syndrome virus (PRRSV) causes serious economic losses to the swine industry worldwide, and there are no highly effective strategies for prevention. Nanobodies are considered a promising novel approach for treating diseases because of their ease of production and low costing. Here, we showed that PRRSV-N-Nb1 against PRRSV-N protein significantly inhibited PRRSV-2 replication in vitro and in vivo. Furthermore, we demonstrated that the motif Serine 105 (S105) in PRRSV-N protein was the key amino acid to interact with PRRSV-N-Nb1 and bond to its motif R97, which is important for the self-binding of N protein. The PRRSV-N-Nb1 could block the self-interaction of N protein following viral assembly. These findings not only provide insights into the molecular basis of PRRSV N protein self-binding as a key factor for viral replication for the first time but also highlight a novel target for the development of anti-PRRSV replication drugs.


Asunto(s)
Proteínas de la Nucleocápside , Síndrome Respiratorio y de la Reproducción Porcina , Virus del Síndrome Respiratorio y Reproductivo Porcino , Anticuerpos de Dominio Único , Animales , Aminoácidos , Línea Celular , Proteínas de la Nucleocápside/metabolismo , Síndrome Respiratorio y de la Reproducción Porcina/tratamiento farmacológico , Virus del Síndrome Respiratorio y Reproductivo Porcino/metabolismo , Serina , Anticuerpos de Dominio Único/farmacología , Porcinos , Proteínas no Estructurales Virales/metabolismo , Replicación Viral/efectos de los fármacos
13.
Adv Appl Ceram ; 122(3-4): 236-249, 2023.
Artículo en Inglés | MEDLINE | ID: mdl-38108047

RESUMEN

Carbide-derived carbon (CDC) was previously proposed as a surface modification method for hip implant applications since it showed excellent tribocorrosion performance under open-circuit potential (OCP) conditions. Nonetheless, a systematic evaluation of CDC's tribocorrosion properties was still missing. Therefore, our objective is to test CDC's tribocorrosion performance under various electrochemical conditions and to identify the synergism between wear and corrosion. Based on the findings, the variations in OCP for CDC (0.626 mV) is smaller than Ti6Al4V (1.91 mV), and CDC showed lower induced current than T6Al4V for all potentials, suggesting CDC is more stable than Ti6Al4V under tribocorrosive conditions. Eventually, the weight loss of Ti6Al4V (50.662±5.19 µg) was found to be significantly higher than that of CDC (4.965±5.19 µg), which agrees with the electrochemical results. In summary, CDC showed better tribocorrosion performance than Ti6Al4V and was determined as an Antagonism regime.

14.
Res Sq ; 2023 Oct 05.
Artículo en Inglés | MEDLINE | ID: mdl-37886457

RESUMEN

Fretting-corrosion is one of the failure processes in many applications, including biomedical implants. For example, the modern design of hip implants with multiple components offers better flexibility and inventory storage. However, it will trigger the fretting at the implant interfaces with a small displacement amplitude (< 5 µm) and usually in a partial slip region. Although many studies have been reported on the fretting, they have high displacement amplitude and are in the gross slip region. It is imperative to have an apparatus to overcome such limitations, specifically for hip implant applications. Therefore, this study describes the development of a fretting-corrosion apparatus with low micro-motion (≤ 5 µm) that can simultaneously monitor the corrosion process. Initial experiments with Ti6Al4V-Ti6Al4V in 0.9% saline, Ti6Al4V-Ti6Al4V in bovine calf serum (BCS), and ZrO2-Ti6Al4V in BCS were conducted to validate the system. As a result, the fretting regime of all groups remained partially slip region throughout the 3600 cycles, and the possible failure mechanisms are proposed in this manuscript.

15.
Ann Biomed Eng ; 51(12): 2749-2761, 2023 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-37530907

RESUMEN

Titanium-based implants utilized in total joint arthroplasties could restore primary musculoskeletal function to patients suffering from osteoarthritis and other conditions. Implants are susceptible to failure stemming from aseptic loosening and infection at the joint site, eventually requiring revision surgery. We hypothesized that there might be a feedback loop by which metal degradation particles and ions released from the implant decrease cell viability and increase immune response, thereby creating biochemical conditions that increase the corrosion rate and release more metal ions. This study focused on the synergistic process through cell viability assays and electrochemical tests. From the results, inflammatory conditions from ion release resulting in cell death would further increase the corrosion rate at the metal implant site. The synergistic interaction in the implant surroundings in which infectious conditions produce Ti ions that contribute to more infection, creating a potential cycle of accelerating corrosion.


Asunto(s)
Metales , Titanio , Humanos , Prótesis e Implantes , Iones , Corrosión
16.
Virology ; 587: 109849, 2023 10.
Artículo en Inglés | MEDLINE | ID: mdl-37515945

RESUMEN

Myosin heavy chain 9 (MYH9) has been identified as a crucial factor in gammaherpesvirus infection. Murine gammaherpesvirus 68 (MHV-68) was used as an appropriate viral model for investigating gammaherpesviruses in vivo and developing antiviral treatments. However, the roles of MYH9 in MHV-68 infection have not been documented. In the study, the relationship between the expression of MYH9 and MHV-68 infection and MYH9 as the antiviral target were analyzed. The results revealed that MYH9 was enriched on the cell surface and co-localized with MHV-68 upon viral infection. Knocking down MYH9 with siRNA or using the specific inhibitor of MYH9 activity, Blebbistatin, resulted in the decreasing of MHV-68 infection. Furthermore, polyclonal antibodies against MYH9 reduced infection by approximately 74% at a dose of 100 µg/ml. The study determined that MYH9 contributes to MHV-68 infection by interacting with viral glycoprotein 150 (gp150) in the BHK-21 cell membrane. The specific region of MYH9, amino acids 1811-1960 (C-150), was identified as the key domain involved in the interaction with MHV-68 gp150 and was found to inhibit MHV-68 infection. Moreover, C-150 was also shown to decrease HSV-1 infection in Vero cells by approximately 73%. Both C-150 and Blebbistatin were found to inhibit MHV-68 replication and reduce histopathological lesions in vivo in C57BL/6J mice. Taken together, these findings suggested that MYH9 is crucial for MHV-68 infection through its interaction with viral gp150 and that C-150 may be a promising antiviral target for inhibiting MHV-68 infection in vitro and in vivo.


Asunto(s)
Gammaherpesvirinae , Infecciones por Herpesviridae , Rhadinovirus , Animales , Ratones , Aminoácidos , Antivirales/metabolismo , Chlorocebus aethiops , Gammaherpesvirinae/genética , Ratones Endogámicos C57BL , Cadenas Pesadas de Miosina/genética , Cadenas Pesadas de Miosina/metabolismo , Rhadinovirus/genética , Células Vero , Proteínas Virales/genética , Proteínas Virales/metabolismo
17.
Mater Today Bio ; 18: 100500, 2023 Feb.
Artículo en Inglés | MEDLINE | ID: mdl-36471893

RESUMEN

Polymethyl methacrylate (PMMA) bone cement (PBC) is commonly used in orthopaedic surgery. However, polymerization volumetric shrinkage, exothermic injury, and low bioactivity prevent PBC from being an ideal material. The developed expandable P(MMA-AA-St) well overcomes the volumetric shrinkage of PBC. However, its biomechanical properties are unsatisfactory. Herein, graphene oxide (GO), a hydrophilic material with favourable biomechanics and osteogenic capability, was added to P(MMA-AA-St) to optimize its biomechanics and bioactivity. The GO-modified self-expandable P(MMA-AA-St)-GO nanocomposite (PGBCs) exhibited outstanding compressive strength (>70 â€‹MPa), water absorption, and volume expansion, as well as a longer handling time and a reduced setting temperature. The cytocompatibility of PGBCs was superior to that of PBC, as demonstrated by CCK-8 assay, live-dead cell staining, and flow cytometry. In addition, better osteoblast attachment was observed, which could be attributed to the effects of GO. The improved level of osteogenic gene and protein expression further illustrated the improved cell-material interactions between osteoblasts and PGBCs. The results of an in vivo study performed by filling bone defects in the femoral condyles of rabbits with PGBCs demonstrated promising intraoperative handling properties and convenient implantation. Blood testing and histological staining demonstrated satisfactory in vivo biosafety. Furthermore, bone morphological and microarchitecture analyses using bone tissue staining and micro-CT scanning revealed better bone-PGBCs contact and osteogenic capability. The results of this study indicate that GO modification improved the physiochemical properties, cytocompatibility, and osteogenic capability of P(MMA-AA-St) and overcame the drawbacks of PBC, allowing its material derivatives to serve as effective implantable biomaterials.

18.
Artículo en Inglés | MEDLINE | ID: mdl-38525435

RESUMEN

Tribocorrosion is an integration of two areas-tribology and corrosion. It can be defined as the material degradation caused by the combined effect of corrosion and tribological process at the material interfaces. Significant development has occurred in the field of tribocorrosion over the past years. This development is due to its applications in various fields, such as aerospace, marine, biomedical, and space. Focusing on biomedical applications, tribocorrosion finds its applications in the implants used in cardiovascular, spine, orthopedics, trauma, and dental areas. It was reported that around 7.2 million Americans are living with joint implants. Implant surgery is a traumatic and expensive procedure. Tribocorrosion can affect the lifespan of the implants, thus leading to implant failure and a potential cause of revision surgery. Hence, it is essential to understand how tribocorrosion works, its interaction with the implants, and what procedures can be implemented to protect materials from tribocorrosion. This paper discusses how tribocorrosion research has evolved over the past 11 years (2010-2021). This is a comprehensive overview of tribocorrosion research in biomedical applications.

19.
J Biol Chem ; 298(12): 102709, 2022 12.
Artículo en Inglés | MEDLINE | ID: mdl-36402446

RESUMEN

Circulation of influenza A virus (IAV), especially within poultry and pigs, continues to threaten public health. A simple and universal detecting method is important for monitoring IAV infection in different species. Recently, nanobodies, which show advantages of easy gene editing and low cost of production, are a promising novel diagnostic tool for the monitoring and control of global IAVs. In the present study, five nanobodies against the nucleoprotein of H9N2 IAV were screened from the immunized Bactrian camel by phage display and modified with horseradish peroxidase (HRP) tags. Out of which, we determined that H9N2-NP-Nb5-HRP can crossreact with different subtypes of IAVs, and this reaction is also blocked by positive sera for antibodies against different IAV subtypes. Epitope mapping showed that the nanobody-HRP fusion recognized a conserved conformational epitope in all subtypes of IAVs. Subsequently, we developed a nanobody-based competitive ELISA (cELISA) for detecting anti-IAV antibodies in different species. The optimized amount of coating antigen and dilutions of the fusion and testing sera were 100 ng/well, 1:4000, and 1:10, respectively. The time for operating the cELISA was approximately 35 min. The cELISA showed high sensitivity, specificity, reproducibility, and stability. In addition, we found that the cELISA and hemagglutination inhibition test showed a consistency of 100% and 87.91% for clinical and challenged chicken sera, respectively. Furthermore, the agreement rates were 90.4% and 85.7% between the cELISA and commercial IEDXX ELISA kit. Collectively, our developed nanobody-HRP fusion-based cELISA is an ideal method for monitoring IAV infection in different species.


Asunto(s)
Subtipo H9N2 del Virus de la Influenza A , Gripe Humana , Anticuerpos de Dominio Único , Animales , Humanos , Anticuerpos Antivirales , Ensayo de Inmunoadsorción Enzimática/métodos , Subtipo H9N2 del Virus de la Influenza A/aislamiento & purificación , Gripe Humana/diagnóstico , Reproducibilidad de los Resultados , Porcinos , Aves de Corral
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