RESUMO
Tumor-infiltrating lymphocytes (TILs) play a key role in regulating the host immune response and shaping tumor microenvironment. It has been previously shown that T cell infiltration in penile tumors was associated with clinical outcomes. However, few studies have reported the T cell receptor (TCR) repertoire in patients with penile cancer. In the present study, we evaluated the TCR repertoires in tumor and adjacent normal tissues from 22 patients with penile squamous cell carcinoma (PSCC). Analysis of the T cell receptor beta-variable (TRBV) and joining (TRBJ) genes usage and analysis of complementarity determining region 3 (CDR3) length distribution did not show significant differences between tumor and matched normal tissues. Moreover, analysis of the median Jaccard index indicated a limited overlap of TCR repertoire between these groups. Compared with normal tissues, a significantly lower diversity and higher clonality of TCR repertoire was observed in tumor samples, which was associated with clinical characteristics. Further analysis of transcriptional profiles demonstrated that tumor samples with high clonality showed increased expression of genes associated with CD8 + T cells. In addition, we analyzed the TCR repertoire of CD4 + T cells and CD8 + T cells isolated from tumor tissues. We identified that expanded clonotypes were predominantly in the CD8 + T cell compartment, which presented with an exhausted phenotype. Overall, we comprehensively compared TCR repertoire between penile tumor and normal tissues and demonstrated the presence of distinct T cell immune microenvironments in patients with PSCC.
Assuntos
Carcinoma de Células Escamosas , Neoplasias Penianas , Masculino , Humanos , Receptores de Antígenos de Linfócitos T , Neoplasias Penianas/genética , Neoplasias Penianas/metabolismo , Regiões Determinantes de Complementaridade/metabolismo , Receptores de Antígenos de Linfócitos T alfa-beta/metabolismo , Linfócitos T CD8-Positivos , Carcinoma de Células Escamosas/genética , Carcinoma de Células Escamosas/metabolismo , Microambiente TumoralRESUMO
BACKGROUND: The health of elderly individuals is closely linked to their protein intake and the abundance of intestinal microbiota. To investigate the impact of a compound powder made from mulberry leaf and konjac (hereinafter referred to as 'compound powder') on regulating the structure of intestinal microbiota in 15-month-old BALB/c mice that were fed a high-beef-protein diet, 16S rRNA high-throughput sequencing, reverse transcription quantitative polymerase reaction, western blot, and other biochemical methods were used to analyze the differences in intestinal microbiota, protein metabolism-related genes, short-chain fatty acid (SCFA) content, and serum cytokines. RESULTS: The results showed that the compound powder increased the content of SCFAs, reduced the inflammatory reaction of the body, adjusted the abundance of intestinal microbiota (Firmicutes and Bacteroidetes), and increased the ratio of Firmicutes to Bacteroidetes (F/B). Moreover, the compound powder could increase the abundance of Lactobacillus and some non-dominant bacteria that were related to amino acid metabolism and beneficial to human health, such as Eubacterium coprostanoligenes. These beneficial bacteria competitively reduced the abundance of harmful bacteria to protect the intestinal barrier and promote intestinal health, and upregulated the activities of aminopeptidase, proton-coupled oligopeptide transporter 1, and glutamate dehydrogenase at the transcription and translation levels. CONCLUSION: The compound powder could balance the abundance of intestinal microbiota, which may improve the metabolic capacity of old mice on a high-protein diet, and ultimately promoting the well-being of elderly individuals. © 2023 Society of Chemical Industry.
Assuntos
Amorphophallus , Dieta Rica em Proteínas , Microbioma Gastrointestinal , Morus , Bovinos , Camundongos , Humanos , Animais , Idoso , Lactente , Pós , RNA Ribossômico 16S , Bactérias/genética , Folhas de Planta , Dieta HiperlipídicaRESUMO
Ensuring sufficient protein intake, efficient digestion, and optimal absorption are crucial for the elderly. This study aims to investigate the potential of a compound dietary fiber, consisting of mulberry leaf and konjac flour (CMK), to enhance the digestion and absorption of a high-fish-protein diet in elderly mice. Results showed that CMK effectively reduced the number of unique peptide segments, generated short-chain fatty acids (SCFA) in feces, improved the content of glutamic pyruvic transaminase (GPT), glutamic oxaloacetic transaminase (GOT), amino acid, and urea nitrogen in serum, activated the contents of pepsin, trypsin, and erepsin, and enhanced the expression of glutamate dehydrogenase (GDH), peptide transporter 1 (PepT1), and aminopeptidase N (APN). Furthermore, CMK demonstrated its ability to decrease the levels of tumor necrosis factor-α (TNF-α), interleukin-1ß (IL-1ß), interleukin-10 (IL-10), lipopolysaccharide (LPS), and lipopolysaccharide binding protein (LBP), while increase the abundance of beneficial bacteria, such as Lactobacillus and Blautia. In conclusion, CMK proved effective in enhancing the digestion and metabolism of protein in elderly mice through the regulation of gut microbiota structure and intestinal tissue repair.
RESUMO
Lactose intolerance (LI) is a widespread health issue affecting almost 70% of the world population. This study evaluates the potential prebiotic benefits of konjac glucomannan (KGM) and konjac oligogalactomannan (KOGM) in improving LI. Colonic fermentation results indicate that lactase groups of healthy subjects showed lower pH, higher lactic acid content, and lactase activity in fermentation broth compared with LI subjects. Total short-chain fatty acid (SCFAs) content reached 1.71 mmol/L in healthy subjects, whereas it was 1.49 mmol/L in LI subjects. In vivo studies demonstrated that KGM and KOGM intake reduced total cholesterol (T-CHO) and triglyceride (TG) levels in the liver and significantly increased immunoglobulin G (lgG) and immunoglobulin A (lgA) values, while KOGM inclusion led to a significant 23.04% increase in serum free fatty acid (FFA) levels compared to the Blank group (p < 0.05). Furthermore, ileal tissue analysis revealed a marked increase in villus height and intestinal wall thickness (p < 0.05) as well as a decrease in crypt depth (p < 0.05). The composition and proportion of gut microbiota have improved with KGM and KOGM use, notably increasing the abundance of Lactobacillus and Lachnospiraceae_NK4A136_group, respectively (p < 0.05). Compared with the Blank group, Lactobacillus abundance increased by approximately 25.82% in the Drug group, 18.23% in the KGM group, and 8.67% in the KOGM group. These findings suggest that KGM and KOGM can be utilized as prebiotics to alleviate LI symptoms.
RESUMO
The liver is the most important organ for biological transformation in the body and is crucial for maintaining the body's vital activities. Liver injury is a serious pathological condition that is commonly found in many liver diseases. It has a high incidence rate, is difficult to cure, and is prone to recurrence. Liver injury can cause serious harm to the body, ranging from mild to severe fatty liver disease. If the condition continues to worsen, it can lead to liver fibrosis and cirrhosis, ultimately resulting in liver failure or liver cancer, which can seriously endanger human life and health. Therefore, establishing an rodent model that mimics the pathogenesis and severity of clinical liver injury is of great significance for better understanding the pathogenesis of liver injury patients and developing more effective clinical treatment methods. The author of this article summarizes common chemical liver injury models, immune liver injury models, alcoholic liver injury models, drug-induced liver injury models, and systematically elaborates on the modeling methods, mechanisms of action, pathways of action, and advantages or disadvantages of each type of model. The aim of this study is to establish reliable rodent models for researchers to use in exploring anti-liver injury and hepatoprotective drugs. By creating more accurate theoretical frameworks, we hope to provide new insights into the treatment of clinical liver injury diseases.
Assuntos
Doença Hepática Induzida por Substâncias e Drogas , Hepatopatias , Hepatopatia Gordurosa não Alcoólica , Humanos , Fígado/metabolismo , Hepatopatias/patologia , Cirrose Hepática/patologia , Doença Hepática Induzida por Substâncias e Drogas/patologia , Substâncias Protetoras/farmacologia , Hepatopatia Gordurosa não Alcoólica/metabolismoRESUMO
BACKGROUND: Whether Astragalus membranaceus is an effective drug in treatment of ulcerative colitis (UC) and how it exhibit activity effect on UC is unclear. METHODS: TCMSP, GeneCards, String, and DAVID database were used to screening target genes construct PPI network and performed for GO and KEGG pathway enrichment analysis respectively. Molecular docking and animal experiment were performed. The body weight and disease activity index (DAI) of mice were recorded. ELISA kits were used to detect the levels of CAT, SOD, MDA and IL-6, IL-10, TNF-α in the blood of mice. Western blot kits were utilized to measured the expressions of MAPK14, RB1, MAPK1, JUN, ATK1, and IL2 proteins. RESULTS: The active components of Astragalus membranaceus mainly including 7-O-methylisomucronulatol, quercetin, kaempferol, formononetin and isrhamnetin. Astragalus membranaceus may inhibited the expression of TNF-α, IL-6, MDA, and promoted the expression of CAT, SOD, IL-10. The expression levels of MAPK14, RB1, MAPK1, JUN and ATK1 proteins were significantly decreased while IL2 protein increased administrated with Astragalus membranaceus. CONCLUSIONS: Astragalus membranaceus is an effective drug in treatment of UC according to related to above targets that may exhibits the anti-UC effect via its antioxidant pathway and regulating the balance of pro-inflammatory and anti-inflammatory factors.
RESUMO
Acute kidney injury (AKI) stands as a prevalent and economically burdensome condition worldwide, yet its complex molecular mechanisms remain incompletely understood. To address this gap, our study employs a multifaceted approach, combining mass spectrometry and RNA sequencing technologies, to elucidate the intricate molecular landscape underlying nephrotoxin-induced AKI in mice by cisplatin- and LPS-induced. By examining the protein and RNA expression profiles, we aimed to uncover novel insights into the pathogenesis of AKI and identify potential diagnostic and therapeutic targets. Our results demonstrate significant down-regulation of Slc34a1 and Slc34a3, shedding light on their crucial roles in AKI pathology and highlighting their promise as actionable targets for diagnosis and treatment. This comprehensive analysis not only enhances our understanding of AKI pathophysiology but also offers valuable avenues for the development of targeted interventions to mitigate its clinical impact. SIGNIFICANCE: Nephrotoxicity acute kidney injury (AKI) is a common clinical condition whose pathogenesis is the process by which some drugs, chemicals or other factors cause damage to the kidneys, resulting in impaired kidney function. Although it has been proved that different nephrotoxic substances can affect the kidney through different pathways, whether they have a commonality has not been registered. Here, we combined transcriptomics and proteomics to study the molecular mechanism of LPS and cisplatin-induced nephrotoxic acute kidney injury finding that the down-regulation of Slc34a1 and Slc34a3 may be a critical link in nephrotoxic acute kidney injury, which can be used as a marker for its early diagnosis.
Assuntos
Injúria Renal Aguda , Cisplatino , Regulação para Baixo , Proteômica , Transcriptoma , Injúria Renal Aguda/induzido quimicamente , Injúria Renal Aguda/metabolismo , Injúria Renal Aguda/genética , Animais , Camundongos , Proteômica/métodos , Cisplatino/efeitos adversos , Cisplatino/toxicidade , Lipopolissacarídeos/toxicidade , Masculino , Perfilação da Expressão GênicaRESUMO
The aim of the study was to investigated the mechanism of Strychnos nux-vomica L. (Semen Strychni, SS) against papillary carcinoma thyroid (PTC) by combined of network pharmacology and experimental verification. By searching the TCMSP, SEA and SwissTarget Prediction database, the main active ingredients and related targets were obtained. Utilizing Venny 2.1.0 String database and Cytoscape 3.7.2 to screened the intersection target and constructed protein-protein interaction (PPI) network diagram. Using R 4.0.4 software carried out the enrichment analysis of GO and KEGG. HPLC was carried out using LC-20A modular HPLC system to identify the bioactive compound brucine present in SS. Molecular docking was performed using Discovery 2019 software. The inhibition rate was detected by CCK8 method. Western blot was used to detect the expression levels of brucine anti-PTC related pathway proteins. 14 active components were screened out, of which 4 main components showed tight relationship with PTC. SS may play the anti-PTC role by acting on two main pathways (TNF signaling pathway and MAPK signaling pathway) and mediating various biological functions. HPLC analysis revealed that brucine was a suitable marker for standardization of the SS. 4 active components exhibit strong binding energy with core protein. Brucine could significantly reduce the activity of BCPAP cells compared with isobrucine, stigmasterol, (+)-catechin. Brucine may reduce the protein expression levels of IL-6, VEGFA, JUN, TP53, 1L1B, PTGS2, BCL2, CASP3, CASP8, and CASP9 while increase the protein expression levels of BAD, cleaved-CASP3, cleaved-CASP8, and cleaved-CASP9 in BCPAP cells, respectively. The active components of SS against PTC mainly include isobrucine, stigmasterol, (+)-catechin, brucine. Among them, brucine exhibits the strongest anti-PTC activity in BCPAP cells, which may reduce the PTC-related protein expression levels. Therefore, SS may exhibits the anti-PTC activities through multiple targets and pathways.
Assuntos
Catequina , Medicamentos de Ervas Chinesas , Neoplasias da Glândula Tireoide , Humanos , Sêmen , Caspase 3 , Farmacologia em Rede , Simulação de Acoplamento Molecular , Estigmasterol , Câncer Papilífero da Tireoide , Neoplasias da Glândula Tireoide/tratamento farmacológicoRESUMO
Konjac tofu is an irreversible gel formed by removing the acetyl group from konjac glucomannan (KGM) through alkaline heating. This type of food is low in calories, filling, and healthy, making it popular in the market. However, pure konjac tofu has a hard texture and lacks flavor when heated. To improve its taste and appearance, the effects of three varieties of native starch, including corn starch (CS), Canna edulis Ker starch (CKS), and potato starch (PS), on the formation of pasting and rheological properties of the KGM sol were investigated. Konjac tofu samples that incorporated different types and quantities of starch were prepared and analyzed in terms of structure, texture, dehydration, and flavor, with pure konjac tofu serving as a reference. The findings revealed that KGM mixed with a concentration of 4.2% CS, or 0.85% CKS, or 0.85% PS of the total mass produced a gel with the highest viscosity and a steady structure. Texture profile analysis indexes of these combinations were superior to pure KGM, and the konjac-starch tofu had a lamellar network structure. Thus, konjac tofu with the addition of starch has a higher quality texture, lower dehydration, and improved flavor compared to pure KGM gel.
RESUMO
The mixture of proteins, polysaccharides, and other nutrients not only safeguards the nutritional value of the food but also demonstrates superior performance compared to these nutrients when used individually. This study aimed to investigate the effects of konjac glucomannan (KGM) on the properties of whole egg liquid (WEL) and the in vitro fermentation of egg curd (made by the mixture of WEL/KGM). The results revealed that the foaming ability (FA) of the mixture decreased, while the foam stability (FS), emulsifying activity (EA), and emulsion stability (ES) of the mixture increased with increasing concentrations of KGM. The concentration of KGM had a significant effect on the sol-gel transition temperature of WEL. Compared to the fermentation broth of E group (without KGM), KGM decreased the pH from 6.65 to 6.16, free ammonia content from 87.53 µg/g to 72.21 µg/g, and sulfide concentration from 580 µg/L to 470 µg/L in the WEL/KGM mixture (EK group). Moreover, KGM slowed down the gas production of intestinal protein fermentation within 10 h, without affecting the final total gas yield. These findings suggest that adding KGM can be an effective strategy to modify the properties of WEL and improve the intestinal fermentation performance of protein-rich foods.
Assuntos
Mananas , Polissacarídeos , Fermentação , Mananas/química , IntestinosRESUMO
An effective and comprehensive quality evaluation method for Liuwei Dihuang pills (LDP) was established by the simultaneous determination of 8 active components in LDP by the quantitative analysis of multicomponents by single marker (QAMS) method and high-performance liquid chromatography (HPLC) fingerprint combined with chemometrics. These 8 active components were determined by QAMS and the external standard method (ESM), and the quantitative results of the two methods were compared to validate the accuracy and feasibility of the QAMS method. 8 active components showed good linear relationships within their ranges, whose average recoveries were 99.7â¼102.3%. No significant difference was found (P > 0.05) in the quantitative results determined by QAMS and ESM. Furthermore, the fingerprint of LDP was also established, with 11 common peaks identified, and the similarity of the fingerprints of 21 batches of LDP was greater than 0.95. The 21 batches of LDP were basically divided into 3 groups by hierarchical cluster analysis (HCA) and principal component analysis (PCA), and 3 differential markers were screened out by orthogonal partial least squares discriminant analysis (OPLS-DA). The established QAMS method is accurate, economical, fast, and convenient and can simultaneously determine the content of 8 active components in LDP. HPLC fingerprint combined with chemometric analysis more comprehensively evaluated the quality consistency of different batches of LDP and analyzed the markers that cause quality differences between batches. It can provide a scientific basis and reference of quality consistency evaluation for the manufacturers and drug regulatory departments of the preparation.
RESUMO
Acute liver injury (ALI) leads to abnormal liver function and damage to liver cells. Syringin (syr) and costunolide (cos) are the major extracts from Dolomiaea souliei (Franch.) C.Shih (D. souliei), showing diverse biological functions in various biological processes. We explored the underlying hepatoprotective effects of syr+cos against LPS-induced ALI. Cell viability and proliferation were assessed using an MTT assay and immunofluorescence staining. Flow cytometry analysis was used to detect cell cycle distribution and apoptosis. ELISA was utilized to measure liver function and antioxidant stress indexes. qRT-PCR and western blotting was performed to determine mRNA and protein levels respectively. Using shRNA approach to Rac1 analyzed transcriptional targets. The results showed that syr+cos promoted L-02 cell proliferation, inhibiting the cell apoptosis and blocking cell cycle in G1 and G2/M phase. Syr+cos decreased the production of ALT, AST, LDH, MDA and ROS while increased SOD and CAT activities. Pretreated with syr+cos may decrease expressions of caspase-3,7,9, NF-κB, TNF-α proteins, Cyclin B, CDK1 and p-IκB proteins while p-IκB increased. Silencing of Rac-1 may protect the liver by increasing AKT, S473, T308 and reducing p-AKT proteins. Syr+cos exhibits anti-ALI activity via Rac1/AKT/NF-κB signaling pathway which might act as an effective candidate drug for the treatment of ALI.
Assuntos
NF-kappa B , Proteínas Proto-Oncogênicas c-akt , NF-kappa B/metabolismo , Proteínas Proto-Oncogênicas c-akt/metabolismo , Lipopolissacarídeos/farmacologia , Transdução de Sinais , Fígado/metabolismoRESUMO
Objective: To explore the potential molecular mechanism of Pueraria Lobata Radix (RP) and Salviae Miltiorrhizae Radix (RS) in the treatment of type 2 diabetes mellitus (T2DM) based on network pharmacology and molecular docking. Methods: The chemical constituents and core targets of RP and RS were searched by Traditional Chinese Medicine System Pharmacology (TCMSP); target genes related to T2DM were obtained through GeneCards database, component target network diagram was constructed, intersection genes of active compounds and T2DM were synthesized, protein-protein interaction (PPI) relationship was obtained, and core targets were screened by using Cytoscape 3.7.2. Gene Ontology (GO) biological process and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway were analyzed utilizing R studio 4.0.4 according to David database. Based on molecular docking, the screened active components of RP and RS were verified by molecular docking with the core target using Discovery Studio 2019. Results: There were totally 92 components and 29 corresponding targets in the component target network of RP and RS drug pair, of which 6 were the core targets of RP and RS in the treatment of T2DM. Molecular docking results showed that the active compounds of puerarin, formononetin, tanshinone iia, and luteolin had better binding activity with AKT1, VEGFA, NOS3, PPARG, MMP9, and VCAM1, respectively. Among them, puerarin showed significant effects in activating NOS3 pathway and luteolin exhibited significant effects in activating MMP9 pathway, respectively. The main biological processes mainly including xenobiotic stimulus, response to peptide, gland development, response to radiation, cellular response to chemical stress, response to oxygen levels, and the main signal pathways include response to xenobiotic stimulus, cellular response to chemical stress, response to peptide, gland development, and response to oxygen levels. Conclusion: Network pharmacology is an effective tool to explain the action mechanism of Traditional Chinese Medicine (TCM) from the overall perspective. RP and RS pair could alleviate T2DM via the molecular mechanism predicted by the network pharmacology, which provided new ideas and further research on the molecular mechanism of T2DM.
Assuntos
Diabetes Mellitus Tipo 2 , Medicamentos de Ervas Chinesas , Pueraria , Humanos , Luteolina , Metaloproteinase 9 da Matriz , Medicina Tradicional Chinesa , Simulação de Acoplamento Molecular , Farmacologia em Rede , Xenobióticos , Salvia/químicaRESUMO
Gut microbiota diversity is a result of co-evolution between microorganisms and their hosts. However, there are few studies on the evolution of the gut microbiota of wild boars and domestic pigs. Therefore, this study aimed to analyze the composition and function of the gut microbiota of wild boars and domestic pigs using 16S rRNA gene V3-V4 region sequencing, 16S rRNA gene full-length sequencing, and metagenomic sequencing. This study showed that after a long evolution, as compared to wild boars, the domestic pigs exhibited significantly increased relative abundances of Lactobacillus, Lactobacillus reuteri, Lactobacillus johnsonii, Lactobacillus sp.DJF_WC5, and Lactobacillus; s_uncultured bacterium, while the relative abundances of Bifidobacterium and Methanococcaceae decreased significantly. In addition, the relative abundances of "carbohydrate metabolism", "starch and sucrose metabolism", "valine, leucine, and isoleucine biosynthesis", "lysine biosynthesis", and starch-degrading CAZymes were significantly increased in the domestic pigs, while the relative abundances of "environmental adaptation", "immune system", "fatty acid degradation and synthesis", and cellulose-hemicellulose-degrading CAZymes were significantly increased in the wild boars. Finally, the diversity of ARGs and the "antimicrobial resistance genes" in domestic pigs also increased significantly. This study illustrates that the gut microbiota composition and function of wild boars and domestic pigs changed during the long evolution process. These findings provide a basic research theory for the evolution of gut microbiota and the treatment of health and disease.
RESUMO
OBJECTIVE: To detect the recombinant intermediates of hepatitis B virus (HBV) between genotype B and C in vitro. METHODS: Vector Plenti6/V5-D-topo-X was genetically modified by genotype B and C to transfect HepG2 cells. Then the HepG2 cells were amplified and sequence of the nucleic acid after the transinfection was tested and compared with RDP3Beta40 software package and bootscanning procedure in SimPlot program package. RESULTS: Three recombinant intermediates of HBV between genotype B and C were identified in vitro. Genotype C in the precore region plus the core gene spanning nucleotide positions from 1740-1838 to 2443-2485 contributed to the recombination with genotype B. Isolate R1 recombinant intermediate had 2 break points at nt2170-2172 and nt2188-2189. Nucleic acid changed from CAC to TGT and from GA to AC, respectively. Isolate R2 recombinant intermediate had a break point at nt1740-1 838, and 3 bases changed in different nucleic acid sites: from A to T at nt1740, from C to T at nt1753, and from G to A at nt1838, respectively. Isolate R3 recombinant intermediate had a break point at nt2443-2483, and 4 bases changed in different nucleic acid sites: from C to T at nt2443, from A to G at nt2452, from T to C at nt2480, and from C to T at nt2483, respectively. CONCLUSION: The recombinant intermediates of HBV between genotype B and C have been detected in vitro and the changes have been identified in the precore region plus the core gene in genotype B and C.
Assuntos
Vírus da Hepatite B/classificação , Vírus da Hepatite B/genética , Recombinação Genética , Transfecção , Sequência de Bases , Vetores Genéticos , Genótipo , Células Hep G2 , Antígenos do Núcleo do Vírus da Hepatite B/genética , Humanos , Dados de Sequência Molecular , Mutagênese Insercional , Vírus Reordenados/classificação , Vírus Reordenados/genética , Análise de Sequência de DNARESUMO
Osteoporosis is the most common cause of fractures in middle-aged and elderly people. Fracture repair can be difficult due to the decreased bone volume in osteoporosis patients and implants are often required. In this study, a slow-release system for microstructured titanium (Micro-Ti) was designed to promote osteogenesis and inhibit osteoclastogenesis. Firstly, Micro-Ti was prepared on titanium surfaces by dual acid etching. Micro-Ti was covered with naringin (NA), chitosan (CHI) and gelatin (GEL) multilayers through layer by layer technique, which is denoted as LBL (NA) coated-Ti. Osteoblasts (ME3T3-E1) and macrophages (RAW 264.7) were cultured on untreated and treated titanium surfaces in vitro. Osteoblasts grown on LBL (NA) coated-Ti showed higher alkaline phosphatase (ALP) and mineralization, consistent with qRT-PCR analysis of osteoblast genes including runt-related transcription factor 2 (Runx2), ALP, collagen I (Col I), osteocalcin (OCN), osteopontin (OPN), and osteoprotegerin (OPG). In contrast, acid tartarate-resistant phosphatase activity and the expression of osteoclastic differentiation related genes comprising of cathepsin K (CTSK), nuclear factor of activated T cells (NFAT), tartrate resistant acid phosphatase (TRAP) and V-ATPase (VATP) in osteoclasts were significantly reduced on LBL (NA) coated-Ti surfaces compared with other groups. These results indicate that microstructured titanium functionalized by naringin inserted multilayers enhanced the differentiation of osteoblasts and inhibited osteoclast formation. The proposed approach in this research provides a novel way to modify titanium-based implants for fracture repair in osteoporosis patients.
Assuntos
Flavanonas , Osteogênese , Idoso , Diferenciação Celular , Flavanonas/farmacologia , Humanos , Pessoa de Meia-Idade , Osteoblastos , Propriedades de Superfície , TitânioRESUMO
Controlling macrophage response to biomaterials is critical for the reduction of inflammation after implantation. Here we designed a sustained release system from TiO2 nanotubes (TNTs) to improve osteogenesis on titanium implants with anti-inflammatory properties. TNTs (around 70 nm diameter) were first fabricated on titanium surfaces by anodization, directly filled with the anti-inflammatory drug, dexamethasone (DEX) and then covered by chitosan (CHI) multilayer films. Primary osteoblast and macrophage (RAW 264.7) cells were cultured on untreated and treated titanium surfaces in vitro. Osteoblasts grown on CHI-coated Dex-filled TNTs surfaces displayed higher alkaline phosphatase (ALP) and mineralization, which was consistent with qRT-PCR analysis of osteoblastic genes including collagen type I (Col I), osteocalcin (OCN), osteopontin (OPN) and runt related transcription factor 2 (Runx2). In contrast, protein levels of nitric oxide (NO) and proinflammatory cytokines (TNF-α and IL-1ß) from macrophages on Dex-filled TNTs, CHI-coated TNTs and CHI-coated Dex-filled TNTs were significantly lower, especially on CHI-coated Dex-filled TNTs surfaces compared to levels on titanium and TNTs. These results indicate that CHI-coated Dex-filled TNTs enhanced osteoblast differentiation and decreased the inflammatory response of macrophages. The approach presented here provides new insight into the modification of TNTs for the development of titanium-based implants.
Assuntos
Quitosana , Nanotubos , Animais , Anti-Inflamatórios/farmacologia , Diferenciação Celular , Quitosana/farmacologia , Preparações de Ação Retardada/farmacologia , Dexametasona/farmacologia , Camundongos , Osteoblastos , Propriedades de Superfície , Titânio/farmacologiaRESUMO
Inflammatory response is an essential part of optimal tissue-implant integration and the regeneration process. Due to their highly plastic properties, macrophages display phenotypic changes during inflammatory signaling. Investigating these changes on implant surfaces is essential for evaluating implant stability and longevity. In order to control macrophage polarization, IL-4 was conjugated to titanium dioxide nanotubes (TNTs) through polydopamine, and successful fabrication was checked by scanning electron microscopy (SEM), atomic force microscopy (AFM), X-ray photoelectron spectroscopy (XPS) and contact angle, respectively. In vitro experiments including immunofluorescence staining, cell proliferation, the expression of genes associated with pro-inflammatory M1 phenotype (tumor necrosis factor-alpha (TNF-α), Interleukin-18 (IL-18)) and cytokines related to the anti-inflammatory M2 phenotype (IL-4 and IL-10), and the production of nitric oxide (NO) and cytokines TNF-α, IL-10 were detected. Macrophage response showed that IL-4 functionalized TNTs favored macrophage polarization towards an anti-inflammatory M2-phenotype. This study provides a new strategy for use in medical devices and the development of advanced nano-biomaterials in immunotherapy applications.
Assuntos
Interleucina-4 , Nanotubos , Macrófagos , Propriedades de Superfície , TitânioRESUMO
Chlorin e6 (Ce6) is a second generation photosensitizer for photodynamic therapy (PDT). However, free Ce6 still has some defects leading to reduced clinical efficacy, such as easy agglomeration in a physiological environment and poor accumulation in tumor tissue. In order to solve these problems, a hyaluronic acid (HA) modified zeolitic imidazolate framework-8 (ZIF-8) based Ce6 (ZIF-8@Ce6-HA) therapeutic agent is constructed for PDT by one-pot encapsulation and self-assembly. ZIF-8@Ce6-HA exhibits acceptable encapsulation efficiency, effective cell uptake and good biocompatibility. Moreover, the results of in vitro anticancer experiments demonstrated that the ZIF-8@Ce6-HA group exhibited greater cytotoxicity after irradiation than the free Ce6 group, which caused about 88.4% of HepG2 cells to die since ROS is produced by PDT. Additionally, the data of inductively coupled plasma mass spectrometry indicated that modification of HA increased the blood circulation time and reduced the systemic toxicity of ZIF-8@Ce6. In summary, this work created an interesting Ce6 therapeutic agent for PDT and provided the data for HA regarding the improvement in biocompatibility and biological half-life of metal organic frameworks.
Assuntos
Antineoplásicos/farmacologia , Materiais Biocompatíveis/farmacologia , Estruturas Metalorgânicas/farmacologia , Fotoquimioterapia , Fármacos Fotossensibilizantes/farmacologia , Polissacarídeos/química , Porfirinas/farmacologia , Animais , Antineoplásicos/síntese química , Antineoplásicos/química , Materiais Biocompatíveis/síntese química , Materiais Biocompatíveis/química , Sobrevivência Celular/efeitos dos fármacos , Clorofilídeos , Sistemas de Liberação de Medicamentos , Feminino , Células Hep G2 , Humanos , Neoplasias Hepáticas Experimentais/tratamento farmacológico , Neoplasias Hepáticas Experimentais/patologia , Estruturas Metalorgânicas/síntese química , Estruturas Metalorgânicas/química , Camundongos , Camundongos Endogâmicos BALB C , Tamanho da Partícula , Fármacos Fotossensibilizantes/química , Porfirinas/química , Propriedades de Superfície , Distribuição TecidualRESUMO
Thermosensitive in situ gel is a novel drug delivery system which can form gel in situ after injection of the polymer solution into the body and releases the drug in a controlled manner, thus provides a promising strategy for localized drug delivery. The aim of the present work is to investigate the characteristics including gelation temperature, sol-gel transition temperature (T(s-g)), gel strength, stable viscosity, erosion and drug release behavior of the thermosensitive in situ gel which are composed of different concentrations of poloxamer Pluronic F127 and F68. The gelation temperature was determined by tube-reverse method. Rheological measurements were carried out to evaluate T(s-g), stable viscosity and gel strength. Erosion of the gels and release of dexamethasone sodium phosphate (DSP) from the gels were investigated by membrane-free method and HPLC. Increased F127 concentration in gel decreased the gelation temperature, T(s-g) as well as erosion of the gel and drug release rate, while viscosity and gel strength rose accordingly. However, increased F68 in gel could lead to the opposite result. The poloxamer solution below T(s-g) is Newtonian fluid with comparatively low viscosity, but shows the characteristics of the pseudoplastic fluid when temperature rises near to T(s-g). Drug release was controlled by the erosion of the gel matrix, and both of them followed the zero-order kinetics. An optimized formation containing 22.5% F127 and 2.5% F68 showed more desirable characteristics which meet the clinical requirements and is of potential in future clinical therapy.