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1.
Cell J ; 23(6): 658-664, 2021 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-34939759

RESUMO

Objective: The use of animal or plant exosomes in cancer treatment is promising because of their easy access and low cost. Freshwater crabs are used in traditional Iranian medicine to treat cancer. This study aims to determine the anti-cancer properties of exosomes removed from freshwater crabs on a breast cancer cell line (4T1) compared to bone marrow mesenchymal stem cells (BMSCs). Materials and Methods: In this experimental study, crab haemolymph exosomes were isolated via the precipitation method and characterised by electron microscopy, dynamic light scattering (DLS), and Western blot analysis. The protein concentration and total antioxidant capacity of these exosomes were determined by bicinchoninic acid (BCA) and cupric reducing antioxidant capacity (CUPRAC). The 4T1 cells and BMSCs were treated with exosomes and we assessed the cell survival by the resazurin and MTT assays. The level of nitric oxide (NO) secretion from the 4T1 cells was determined after treatment with the exosomes. Results: Electron microscopy, DLS and Western blot for CD63 confirmed that the isolated exosomes were <100 nm in size and expressed CD63. The total antioxidant capacity in these exosomes was 1.003 µM/ml and the protein concentration was 650 mg/ml. Resazurin and MTT assay results showed a decrease in survival of the 4T1 cells (P≤0.001) after treatment with the exosomes compared to cell growth in the exosome-treated BMSCs. Conclusion: Crab haemolymph contains protein-rich exosomes with antioxidant activities that can have anti-cancer effects on 4T1 cells. These exosomes may be proposed for breast cancer therapeutics.

2.
Cell Tissue Bank ; 2021 Nov 22.
Artigo em Inglês | MEDLINE | ID: mdl-34806123

RESUMO

Numbers of women worldwide face infertility, which will have a significant impact on a couple's life. As a result, assisting with the treatment of these individuals is seen as a critical step. Successful births following uterus and ovary donation have been reported in recent. When immunosuppressive drugs are used in patients who receive donated tissues, there are always problems with the drugs' side effects. In recent years, tissue engineering has mainly been successful in treating infertility using decellularization techniques. Engineered uterus and ovary prevent immunological reactions and do not require immunosuppressive drugs. The most important aspect of using decellularized tissue is its proper function after transplantation. These tissues must be able to produce follicles, secrete hormones and cause pregnancy. This study aimed to investigate research on decellularized tissues and transplanted into the female reproductive system. In this study, just tissues that, after transplantation, have the proper function for fertility were investigated.

3.
Brain Res Bull ; 177: 324-331, 2021 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-34688832

RESUMO

Spinal cord injury (SCI) is a severely debilitating problem leading to substantial decrease in the quality of life. After spinal cord injury, inflammation and oxidative stress plays a key role in initiating the secondary injury cascades leading to progressive tissue degradation and extreme functional deficits. Given that the primary mechanical injuries to spinal cord are rarely repaired, the pharmacological interventions may improve the neurological outcomes caused by secondary injury. Astaxanthin (AST) is considered as a xanthophyll carotenoid with potent antioxidant and anti-inflammatory properties, which has various pharmacological activities. In the present study, we aimed to firstly assess the protective effect of AST, and then to define the AST mechanism of action on a rat model of SCI. Based on the results of von Frey test, AST treatment significantly alleviated the SCI-induced neuropathic pain compared with the control groups (P < 0.05). The expression analysis by western blot shows reduced expression levels of COX-2, TNF-α, IL-1ß, and IL-6 following AST treatment (P < 0.05). The activity of antioxidant enzymes was evaluated using ELISA. Therefore, ELISA experiments showed a significant reduction in the level of oxidative stress in SCI rat following AST treatment (P < 0.05). Furthermore, histopathological evaluations revealed that myelinated white matter and motor neuron number were significantly preserved after treatment with AST (P < 0.05). In conclusion, our study shows that AST could improve SCI through anti-inflammatory and antioxidant effects which leads to decreased tissue damage and mechanical pain after SCI.

4.
J Control Release ; 338: 341-357, 2021 10 10.
Artigo em Inglês | MEDLINE | ID: mdl-34428480

RESUMO

Microneedle arrays have recently received much attention as cancer detection and treatment platforms, because invasive injections and detection of the biopsy are not needed, and drug metabolism by the liver, as well as adverse effects of systemic drug administration, are diminished. Microneedles have been used for diagnosis, vaccination, and in targeted drug delivery of breast cancer. In this review, we summarize the recent progress in diagnosis and targeted drug delivery for breast cancer treatment, using microneedle arrays to deliver active molecules through the skin. The results not only suggest that health and well-being of patients are improved, but also that microneedle arrays can deliver anticancer compounds in a relatively noninvasive manner, based on body weight, breast tumor size, and circulation time of the drug. Moreover, microneedles could allow simultaneous loading of multiple drugs and enable controlled release, thus effectively optimizing or preventing drug-drug interactions. This review is designed to encourage the use of microneedles for diagnosis and treatment of breast cancer, by describing general properties of microneedles, materials used for construction, mechanism of action, and principal benefits. Ongoing challenges and future perspectives for the application of microneedle array systems in breast cancer detection and treatment are highlighted.


Assuntos
Neoplasias da Mama , Administração Cutânea , Neoplasias da Mama/tratamento farmacológico , Neoplasias da Mama/metabolismo , Sistemas de Liberação de Medicamentos , Liberação Controlada de Fármacos , Feminino , Humanos , Microinjeções , Agulhas , Pele/metabolismo
5.
Mol Biotechnol ; 63(9): 818-827, 2021 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-34076821

RESUMO

Fabrication method is one of the essential factors which directly affect on the properties of scaffold. Several techniques have been well established to fabricate nanofibrous scaffolds such as electrospinning. However, preparing a three-dimensional (3-D) interconnected macro-pore scaffold essential for transporting the cell metabolites and nutrients is difficult using the electrospinning method. The main aim of this study was developing a highly porous scaffold by poly (L-lactic acid) (PLLA)/chitosan blend using liquid-liquid phase separation (LLPS) technique, a fast and cost-benefit method, in order to use in nerve tissue engineering. In addition, the effect of different polymeric concentrations on morphology, mechanical properties, hydrophilicity, in vitro degradation rate and pH alteration of the scaffolds were evaluated. Moreover, cell attachment, cell viability and cell proliferation of scaffolds as candidates for nerve tissue engineering was investigated. PLLA/chitosan blend not only had desirable structural properties, porosity, hydrophilicity, mechanical properties, degradation rate and pH alteration but also provided a favorable environment for attachment, viability, and proliferation of human neuroblastoma cells, exhibiting significant potential for nerve tissue engineering applications. However, the polymeric concentration in blend fabrication had influence on both characteristics and cell responses. It concluded that PLLA/chitosan nanofibrous 3-D scaffold fabricated by LLPS method as a suitable candidate for nerve tissue engineering.

6.
Cell Tissue Bank ; 2021 Jun 26.
Artigo em Inglês | MEDLINE | ID: mdl-34173897

RESUMO

Since using tissue transplantation has faced limitations all over the world, regenerative medicine has introduced decellularized tissues as natural scaffolds and researchers are trying to improve their efficiency and function. In this study, to increase cell attachment and ultimately cell proliferation on decellularized bovine pericardia, scrophularia striata extract was used. Scrophularia striata is an Iranian traditional medicinal plant. For this aim after decellularization of bovine pericardium and analysis of its morphology, it was incubated in scrophularia striata solution. Next, isolated human adipose-derived mesenchymal stem cells were cultured on the tissue. Finally, MTT assay, nitric oxide assay, and scanning electron microscopy observation were performed. MTT showed an increase in cell survival after treating the tissue with the plant extract after 48 h in a dose dependent manner significantly. The survival of cells in 0.5%, 2.5%, and 5% groups was about 5, 10 and 15 folds higher in comparison to control groups, respectively. Additionally, nitric oxide secretion in 2.5% and 5% samples was three and five folds higher than that in control group, respectively. Moreover, SEM observation indicated an impressive and dose-dependent effect of using Scrophularia striata on tissue biocompatibility. The results of this study showed that using Scrophularia striata increased cell viability and cell attachment on decellularized pericardia which could pave the way for the use of natural extracts of medicinal plants to reduce unwanted effects and make desired changes in decellularized tissues.

7.
Sci Rep ; 11(1): 12397, 2021 06 11.
Artigo em Inglês | MEDLINE | ID: mdl-34117331

RESUMO

Cervical cancer, caused by human papillomavirus (HPV), is the fourth most common type of cancer among women worldwide. While HPV prophylactic vaccines are available, they have no therapeutic effects and do not clear up existing infections. This study aims to design a therapeutic vaccine against cervical cancer using reverse vaccinology. In this study, the E6 and E7 oncoproteins from HPV16 were chosen as the target antigens for epitope prediction. Cytotoxic T lymphocytes (CTL) and helper T lymphocytes (HTL) epitopes were predicted, and the best epitopes were selected based on antigenicity, allergenicity, and toxicity. The final vaccine construct was composed of the selected epitopes, along with the appropriate adjuvant and linkers. The multi-epitope vaccine was evaluated in terms of physicochemical properties, antigenicity, and allergenicity. The tertiary structure of the vaccine construct was predicted. Furthermore, several analyses were also carried out, including molecular docking, molecular dynamics (MD) simulation, and in silico cloning of the vaccine construct. The results showed that the final proposed vaccine could be considered an effective therapeutic vaccine for HPV; however, in vitro and in vivo experiments are required to validate the efficacy of this vaccine candidate.


Assuntos
Vacinas Anticâncer/imunologia , Epitopos/imunologia , Vacinas contra Papillomavirus/imunologia , Neoplasias do Colo do Útero/prevenção & controle , Vacinas Anticâncer/química , Biologia Computacional , Epitopos/química , Feminino , Humanos , Imunogenicidade da Vacina , Simulação de Acoplamento Molecular , Proteínas Oncogênicas Virais/química , Proteínas Oncogênicas Virais/imunologia , Proteínas E7 de Papillomavirus/química , Proteínas E7 de Papillomavirus/imunologia , Vacinas contra Papillomavirus/química , Proteínas Repressoras/química , Proteínas Repressoras/imunologia , Linfócitos T Citotóxicos/imunologia , Linfócitos T Auxiliares-Indutores/imunologia , Neoplasias do Colo do Útero/imunologia , Neoplasias do Colo do Útero/virologia
8.
Comput Biol Med ; 133: 104390, 2021 06.
Artigo em Inglês | MEDLINE | ID: mdl-33895459

RESUMO

In December 2019, a new virus called SARS-CoV-2 was reported in China and quickly spread to other parts of the world. The development of SARS-COV-2 vaccines has recently received much attention from numerous researchers. The present study aims to design an effective multi-epitope vaccine against SARS-COV-2 using the reverse vaccinology method. In this regard, structural proteins from SARS-COV-2, including the spike (S), envelope (E), membrane (M), and nucleocapsid (N) proteins, were selected as target antigens for epitope prediction. A total of five helper T lymphocytes (HTL) and five cytotoxic T lymphocytes (CTL) epitopes were selected after screening the predicted epitopes for antigenicity, allergenicity, and toxicity. Subsequently, the selected HTL and CTL epitopes were fused via flexible linkers. Next, the cholera toxin B-subunit (CTxB) as an adjuvant was linked to the N-terminal of the chimeric structure. The proposed vaccine was analyzed for the properties of physicochemical, antigenicity, and allergenicity. The 3D model of the vaccine construct was predicted and docked with the Toll-like receptor 4 (TLR4). The molecular dynamics (MD) simulation was performed to evaluate the stable interactions between the vaccine construct and TLR4. The immune simulation was also conducted to explore the immune responses induced by the vaccine. Finally, in silico cloning of the vaccine construct into the pET-28 (+) vector was conducted. The results obtained from all bioinformatics analysis stages were satisfactory; however, in vitro and in vivo tests are essential to validate these results.


Assuntos
COVID-19 , SARS-CoV-2 , Vacinas contra COVID-19 , China , Epitopos de Linfócito B , Epitopos de Linfócito T , Humanos , Simulação de Acoplamento Molecular , Vacinas de Subunidades
9.
J Tissue Eng Regen Med ; 15(2): 116-128, 2021 02.
Artigo em Inglês | MEDLINE | ID: mdl-33175476

RESUMO

The aim of this study was evaluating the effects of vacuum on microstructure and biocompatibility of bovine decellularized pericardium. So the bovine pericardia were decellularized and then the vacuum was applied for two periods of time; 90 and 180 min. DNA, glucose amino glycan, collagen and elastin content assay, scanning electron microscopy (SEM) examination, hematoxylin and eosin (H&E) and Masson's trichrome stainings performed to evaluate microstructure of tissues. MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) test, subcutaneous implantation, and tensile test were used to assay biocompatibility and mechanical properties of decellularized tissues. The results showed that applying vacuum reduced residual DNA significantly. Vacuum after 180 min reduced more residual DNA. There were no significant differences in the content of glucose amino glycan (GAG), collagen, and elastin between the vacuumed and control groups. SEM examination was revealed that vacuum for 180 min increased pore size and porosity more than 90 min and control groups. H&E and Masson's trichrome stainings revealed extracellular matrix preservation after decellularization in all groups. Cell viability was increased in vacuumed samples significantly after 72 h in vaccumed samples. H&E staining and tensile test after implantation of tissues were showed less inflammation in the vacuum applied tissues and increased durability. The vacuum increased DNA removal, pore size, porosity, and biocompatibility in vitro and in vivo and durability of bovine decellularized pericardium in vivo. Considering the important role of time, more studies should be performed to optimize time, intensity, and method of application of vacuum in decellularization of different tissues as well as bovine pericardium.

10.
J Biomed Mater Res A ; 109(7): 1275-1285, 2021 07.
Artigo em Inglês | MEDLINE | ID: mdl-33058428

RESUMO

The two-dimensional (2D) models of breast cancer still exhibit a limited success. Whereas, three-dimensional (3D) models provide more similar conditions to the tumor for growth of cancer cells. In this regard, a 3D in vivo model of breast cancer using 4 T1 cells and chitosan-based thermosensitive hydrogel were designed. Chitosan/ß-glycerol phosphate hydrogel (Ch/ß-GP) was prepared with a final ratio of 2% and 10%. The hydrogel properties were examined by Fourier transformed infrared spectroscopy, MTT assay, pH, scanning electron microscopy, and biodegradability assay. 3D model of breast cancer was induced by injection of 1 × 106 4 T1 cells in 100 µl hydrogel and 2D model by injection of 1 × 106 4 T1 cells in 100 µl phosphate-buffered saline (PBS) subcutaneously. After 3 weeks, induced tumors were evaluated by size and weight determination, ultrasound, hematoxylin- and eosin and Masson's trichrome staining and evaluating of cancer stem cells with CD44 and CD24 markers. The results showed that hydrogel with physiological pH had no cytotoxicity. In 3D model, tumor size and weight increased significantly (p ≤ .001) in comparison with 2D model. Histological and ultrasound analysis showed that 3D tumor model was more similar to breast cancer. Expression of CD44 and CD24 markers in the 3D model was more than 2D model (p ≤ .001). This 3D in vivo model of breast cancer mimicked native tumor and showed malignant tissue properties. Therefore, the use of such models can be effective in various cancer studies, especially in the field of cancer stem cells.

11.
Polim Med ; 50(1): 41-51, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-33150750

RESUMO

BACKGROUND: Skin, the first barrier to pathogens, loses its integrity and function after an injury. The presence of an antibacterial dressing at the wound site may prevent bacterial invasion and also improve the healing process. OBJECTIVES: The current study aimed to fabricate a biomimetic membrane with antibacterial properties for healing chronic wounds. MATERIAL AND METHODS: The membranes, fabricated through electrospinning, are comprised of poly(ethylene oxide) (PEO) and zinc oxide nanoparticles (ZnO-NPs) as the main biomaterial and antibacterial agent, respectively. Antibacterial activity, cell attachment and viability were tested to evaluate the biological properties of the membranes. The optimal cell compatible concentration of ZnO-NPs was determined for further studies. In vitro characterization of the membranes was performed to confirm their suitable properties for wound healing. RESULTS: The antibacterial PEO/ZnO-NP membrane containing 2% of nanoparticles showed no cell toxicity, and human fibroblast cells were able to adhere and proliferate on the scaffold. The in vitro results from the tensile test, wettability, porosity, and protein adsorption revealed appropriate properties of the membrane as a scaffold for skin tissue engineering. CONCLUSIONS: Synthetic polymers have been widely used for tissue engineering applications. The proper characteristics of PEO nanofibers, including a high ratio of surface/volume, moderate hydrophilicity and good mechanical properties, make this polymer interesting for skin regeneration. The results demonstrate the potential of the antibacterial PEO/ZnO-NP membrane to be used as an engineered scaffold to improve the wound healing process.


Assuntos
Quitosana , Nanofibras , Polietilenoglicóis , Tecidos Suporte , Óxido de Zinco , Antibacterianos/uso terapêutico , Células Cultivadas , Etilenos , Fibroblastos/citologia , Humanos , Cicatrização
12.
Int J Biol Macromol ; 164: 871-883, 2020 Dec 01.
Artigo em Inglês | MEDLINE | ID: mdl-32682041

RESUMO

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) caused COVID-19 disease in China. So far, no vaccine has licensed to protect against infection with COVID-19, therefore an effective COVID-19 vaccine needed. The aim of this study was to predict antigenic peptides of SARS-CoV-2 for designing the COVID-19 vaccine using immunoinformatic analysis. In this study, T and B-cell epitopes of S protein were predicted and screened based on the antigenicity, toxicity, allergenicity, and cross-reactivity with human proteomes. The epitopes were joined by the appropriate linker. LT-IIc as an adjuvant was attached to the end of the structure. The secondary and 3D structure of the vaccine was predicted. The refinement process was performed to improve the quality of the 3D model structure; the validation process is performed using the Ramachandran plot and ProSA z-score. The proposed vaccine's binding affinity to the HLA-A11:01 and HLA-DRB1_01:01 molecule was evaluated by molecular docking. Using molecular dynamics, the stability of vaccine-HLA complexes was also evaluated. Finally, in silico gene cloning was performed in the pET30a (+) vector. The findings suggest that the current vaccine may be a promising vaccine to prevent SARS-CoV-2 infection.


Assuntos
Betacoronavirus/imunologia , Infecções por Coronavirus/prevenção & controle , Epitopos/química , Pandemias/prevenção & controle , Pneumonia Viral/prevenção & controle , Vacinas Virais/imunologia , Sequência de Aminoácidos , COVID-19 , Vacinas contra COVID-19 , Infecções por Coronavirus/imunologia , Infecções por Coronavirus/virologia , Epitopos/imunologia , Epitopos de Linfócito B/imunologia , Epitopos de Linfócito T/química , Humanos , Simulação de Acoplamento Molecular , Simulação de Dinâmica Molecular , Pneumonia Viral/imunologia , Pneumonia Viral/virologia , Conformação Proteica , SARS-CoV-2 , Glicoproteína da Espícula de Coronavírus/química , Glicoproteína da Espícula de Coronavírus/imunologia , Vacinas Virais/química
13.
Heliyon ; 5(8): e02253, 2019 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-31517085

RESUMO

Aims: The aim of this study was to present a new method for removing Sodium dodecyl sulfate (SDS) detergent from decellularized bovine pericardium using vacuum. Materials and Methods: The cows' pericardia were collected and decellularized. The samples were incubated with SDS1% for 48 h at 40 °C. To perform vacuum washing (VW: negative pressure was used to wash and remove detergents), every decellularized tissue was cut in 75mm diameter and fixed via a stainless-steel ring with 60mm diameter in the center of filtration Buchner Funnel which was connected to glass filtration flask The system was connected to a vacuum pump by a hose, and a negative pressure of -100 mmHg was applied for 15 min. Then, the samples were shaken and washed at 40-rpm in 100 ml of distilled water for 45 min. This process was repeated for samples of each group (6 times for sample VW6h, 12 times for sample VW12h, and 24 times for sample VW24h). At the end of every cycle, the effluent was collected to take a sample for SDS measurement. The normal washing (NW) group containing distilled water (NWd) and PBS (Phosphate buffered saline) (NWp) were used to wash and remove detergents. SDS measurements, MTT Assay, histological and tensile test, to compare two methods were used. Results: The highest SDS in the effluent was in groups VW12h and VW24h (P ≤ 0.001) and the lowest residual SDS in scaffold was in two groups of VW12h and VW24h (P ≤ 0.001). MTT assay showed that cell survival in the VW12h and VW24h groups was higher than other groups and there' was no significant difference between cell survival in the VW12h and VW24h groups. Histological study showed destruction of tissue in the VW24h group. The results of the tensile test were shown that the native group had the highest module and the lowest amount was the VW24h sample which was reported with P ≤ 0.001 significance for all groups. Conclusion: VW12h can be used as an effective method for SDS removal from decellularized pericardium which morphologically demonstrated a good structure in ECM.

14.
J Mech Behav Biomed Mater ; 62: 282-290, 2016 09.
Artigo em Inglês | MEDLINE | ID: mdl-27232830

RESUMO

Biocompatible Zinc-hydroxyapatite-titania and Zinc-hydroxyapatite nanocomposite coatings have been prepared by electrodeposition on NiTi shape memory alloy. Structures of coatings were characterized using X-ray diffraction (XRD). It was found that addition of TiO2 particles cause to reduction of crystallite size of coating. Scanning Electronic Microscope (SEM) observation showed that the Zn-HA/TiO2 coating consists of plate-like regions which can express that this plate-like structure can facilitate bone growth. X-ray photoelectron microscope (XPS) was performed to investigation of chemical state of composite coating and showed that Zinc matrix was bonded to oxygen. high-resolution transmission electron microscope (HRTEM) result illustrated the crystalline structure of nanocomposite coating. Mechanical behavior of coating was evaluated using microhardness and ball on disk wear test. The TiO2 incorporated composite coatings exhibited the better hardness and anti-wear performance than the Zn-HA coatings. Polarization measurements have been used to evaluate the electrochemical coatings performance. The Zn-HA/TiO2 composite coatings showed the highest corrosion resistance compared with Zn-HA and bare NiTi.


Assuntos
Materiais Revestidos Biocompatíveis/química , Durapatita/química , Nanocompostos/química , Titânio/química , Zinco/química , Corrosão , Propriedades de Superfície , Difração de Raios X
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