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1.
Int J Mol Sci ; 22(2)2021 Jan 08.
Artigo em Inglês | MEDLINE | ID: mdl-33430016

RESUMO

Boron nitride (BN) nanomaterials have been increasingly explored for potential applications in chemistry and biology fields (e.g., biomedical, pharmaceutical, and energy industries) due to their unique physico-chemical properties. However, their safe utilization requires a profound knowledge on their potential toxicological and environmental impact. To date, BN nanoparticles have been considered to have a high biocompatibility degree, but in some cases, contradictory results on their potential toxicity have been reported. Therefore, in the present study, we assessed two commercial 2D BN samples, namely BN-nanopowder (BN-PW) and BN-nanoplatelet (BN-PL), with the objective to identify whether distinct physico-chemical features may have an influence on the biological responses of exposed cellular models. Morphological, structural, and composition analyses showed that the most remarkable difference between both commercial samples was the diameter of their disk-like shape, which was of 200-300 nm for BN-PL and 100-150 nm for BN-PW. Their potential toxicity was investigated using adenocarcinomic human alveolar basal epithelial cells (A549 cells) and the unicellular fungus Saccharomycescerevisiae, as human and environmental eukaryotic models respectively, employing in vitro assays. In both cases, cellular viability assays and reactive oxygen species (ROS) determinations where performed. The impact of the selected nanomaterials in the viability of both unicellular models was very low, with only a slight reduction of S. cerevisiae colony forming units being observed after a long exposure period (24 h) to high concentrations (800 mg/L) of both nanomaterials. Similarly, BN-PW and BN-PL showed a low capacity to induce the formation of reactive oxygen species in the studied conditions. Even at the highest concentration and exposure times, no major cytotoxicity indicators were observed in human cells and yeast. The results obtained in the present study provide novel insights into the safety of 2D BN nanomaterials, indicating no significant differences in the toxicological potential of similar commercial products with a distinct lateral size, which showed to be safe products in the concentrations and exposure conditions tested.


Assuntos
Plaquetas/química , Compostos de Boro/química , Nanoestruturas/química , Estresse Oxidativo/efeitos dos fármacos , Compostos de Boro/efeitos adversos , Humanos , Espécies Reativas de Oxigênio/química
2.
Biochim Biophys Acta Mol Cell Res ; 1868(1): 118886, 2021 01.
Artigo em Inglês | MEDLINE | ID: mdl-33039555

RESUMO

Platelets have been extensively implicated in the progression of cancer and platelet-derived extracellular vesicles (PEVs) are gaining growing attention as potential mediators of the platelet-cancer interplay. PEVs are shed from platelet membrane in response to extracellular stimuli and carry important biological signals for intercellular communication. In this study we demonstrate that PEVs specifically bind to different breast cancer cells and elicit cell-specific functional responses. PEVs were massively internalized by the metastatic cell lines MDA-MB-231 and SKBR3 and the ductal carcinoma cell line BT474, but not by the MCF-7 cell line. In SKBR3 cells, PEVs decreased mitochondrial dehydrogenase activities and altered cell cycle progression without affecting cell viability. Conversely, PEVs potently stimulated migration and invasion of MDA-MB-231, without affecting the distribution in the different phases of the cell cycle. In all the analyzed breast cancer cells, PEVs triggered a sustained increase of intracellular Ca2+, but only in MDA-MB-231 cells, this was associated to the stimulation of selected signaling proteins implicated in migration, including p38MAPK and myosin light chain. Importantly, inhibition of myosin light chain phosphorylation by a Rho kinase inhibitor prevented PEVs-stimulated migration of MDA-MB-231 cells. Our results demonstrate that PEVs are versatile regulators of cancer cell behavior and elicit a variety of different responses depending on the specific breast cancer cell subtype.


Assuntos
Neoplasias da Mama/genética , Movimento Celular/genética , Proliferação de Células/genética , Quinases Associadas a rho/genética , Plaquetas/química , Plaquetas/metabolismo , Neoplasias da Mama/patologia , Comunicação Celular/efeitos dos fármacos , Ciclo Celular/genética , Inibidores Enzimáticos/farmacologia , Vesículas Extracelulares/genética , Feminino , Regulação Neoplásica da Expressão Gênica/efeitos dos fármacos , Humanos , Células MCF-7 , Invasividade Neoplásica/genética , Invasividade Neoplásica/patologia , Fosforilação/efeitos dos fármacos , Proteínas Quinases p38 Ativadas por Mitógeno/genética , Quinases Associadas a rho/antagonistas & inibidores
3.
Methods Mol Biol ; 2158: 295-305, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-32857382

RESUMO

Exosomes are membrane-bound nano-vehicles shed by most eukaryotic cells. Exosomes contain specific proteins and RNAs from parent cells, and they play key signaling roles in cellular development, modulation, and tissue regeneration. Attempts to isolate and modify exosomes to increase their targeting efficiency to specific tissue are still in their infancy. Here, we describe generation of exosomes from biopsy, isolation of exosomes by centrifugal ultrafiltration method, and approaches for manipulation of cardiac homing exosomes by chemical engineering for the treatment of myocardial infarction.


Assuntos
Exossomos , Tecnologia Farmacêutica/métodos , Animais , Plaquetas/química , Técnicas de Cultura de Células/métodos , Fracionamento Celular/métodos , Membrana Celular/química , Membrana Celular/metabolismo , Exossomos/ultraestrutura , Humanos , Camundongos , Terapia de Alvo Molecular , Miocárdio/citologia , Peptídeos/análise , Peptídeos/metabolismo , Ratos , Ultracentrifugação , Ondas Ultrassônicas
4.
PLoS One ; 15(8): e0236439, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32813744

RESUMO

Extracellular vesicles (EVs) in human blood are a potential source of biomarkers. To which extent anticoagulation affects their concentration, cellular origin and protein composition is largely unexplored. To study this, blood from 23 healthy subjects was collected in acid citrate dextrose (ACD), citrate or EDTA, or without anticoagulation to obtain serum. EVs were isolated by ultracentrifugation or by size-exclusion chromatography (SEC) for fluorescence-SEC. EVs were analyzed by micro flow cytometry, NTA, TEM, Western blot, and protein mass spectrometry. The plasma EV concentration was unaffected by anticoagulants, but serum contained more platelet EVs. The protein composition of plasma EVs differed between anticoagulants, and between plasma and serum. Comparison to other studies further revealed that the shared EV protein composition resembles the "protein corona" of synthetic nanoparticles incubated in plasma or serum. In conclusion, we have validated a higher concentration of platelet EVs in serum than plasma by contemporary EV methods. Anticoagulation should be carefully described (i) to enable study comparison, (ii) to utilize available sample cohorts, and (iii) when preparing/selecting biobank samples. Further, the similarity of the EV protein corona and that of nanoparticles implicates that EVs carry both intravesicular and extravesicular cargo, which will expand their applicability for biomarker discovery.


Assuntos
Biomarcadores/sangue , Proteínas Sanguíneas/isolamento & purificação , Vesículas Extracelulares/genética , Proteoma/genética , Adulto , Plaquetas/química , Proteínas Sanguíneas/genética , Feminino , Citometria de Fluxo/métodos , Voluntários Saudáveis , Humanos , Masculino , Espectrometria de Massas/métodos , Pessoa de Meia-Idade
5.
Proc Natl Acad Sci U S A ; 117(32): 18969-18976, 2020 08 11.
Artigo em Inglês | MEDLINE | ID: mdl-32719144

RESUMO

Blood platelets are formed by fragmentation of long membrane extensions from bone marrow megakaryocytes in the blood flow. Using lattice-Boltzmann/immersed boundary simulations we propose a biological Rayleigh-Plateau instability as the biophysical mechanism behind this fragmentation process. This instability is akin to the surface tension-induced breakup of a liquid jet but is driven by active cortical processes including actomyosin contractility and microtubule sliding. Our fully three-dimensional simulations highlight the crucial role of actomyosin contractility, which is required to trigger the instability, and illustrate how the wavelength of the instability determines the size of the final platelets. The elasto-hydrodynamic origin of the fragmentation explains the strong acceleration of platelet biogenesis in the presence of an external flow, which we observe in agreement with experiments. Our simulations then allow us to disentangle the influence of specific flow conditions: While a homogeneous flow with uniform velocity leads to the strongest acceleration, a shear flow with a linear velocity gradient can cause fusion events of two developing platelet-sized swellings during fragmentation. A fusion event may lead to the release of larger structures which are observable as preplatelets in experiments. Together, our findings strongly indicate a mainly physical origin of fragmentation and regulation of platelet size in flow-accelerated platelet biogenesis.


Assuntos
Plaquetas/química , Actomiosina/química , Actomiosina/metabolismo , Animais , Biofísica , Velocidade do Fluxo Sanguíneo , Plaquetas/citologia , Hidrodinâmica , Camundongos
6.
Rev Esp Patol ; 53(3): 182-187, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32650969
7.
J Chromatogr A ; 1624: 461206, 2020 Aug 02.
Artigo em Inglês | MEDLINE | ID: mdl-32540064

RESUMO

Oxylipins, the oxidation products of polyunsaturated fatty acids, are important signaling molecules in living organisms. Some of them have pro-inflammatory properties, while others act as pro-resolving agents. Oxylipins also play a major role in platelet biology and the progression of thrombo-inflammation. Depending on their structure, they may be pro-thrombotic or anti-thrombotic. For an unbiased biological interpretation, a detailed analysis of a broad spectrum of oxylipins including their stereoisomers is necessary. In our work, we developed for the first time an enantioselective UHPLC-ESI-MS/MS assay which allows quantifying individual oxylipin enantiomers. The assay made use of a sub-2µm particle-based amylose-(3,5-dimethylphenylcarbamate) chiral stationary phase (Chiralpak IA-U) under MS-compatible reversed-phase conditions. It covered 19 enantiomeric pairs of oxylipins and one diasteromeric pair of a lipid mediator: 2 pairs of hydroxyoctadecadienoic acids (HODE), 6 pairs of hydroxyeicosatetraenoic acids (HETE), 5 pairs of hydroxyeicosapentaenoic acids (HEPE), 3 pairs of hydroxydocosahexaenoic acids (HDoHE) and one pair of each: resolvins D1, hydroxyeicosatrienoic acid (HETrE), hydroxyoctadecatrienoic acid (HOTrE) and dihydroxyeicosatetraenoic acid (DiHETE). The new method is fast and showed outstanding peak resolution for most of the isomeric pairs. Excellent method sensitivity (average LOD was equal to 2.7 pg on column) was obtained by using a triple quadrupole instrument as a detector in a targeted, selected reaction monitoring (SRM) mode. The applicability of the method was verified by preliminary validation. It was then applied to analyze oxylipins produced by autoxidation of polyunsaturated fatty acids (PUFA) in air. Multiple oxylipins were found in each of the samples as racemic mixtures and served as reference substances for identification. Finally, the new enantioselective UHPLC method was applied to analyze releasates from platelets in resting state, and following activation with thrombin. The highest abundant oxylipin in the platelet releasate was 12(S)-HETE, but many other oxylipins were found in the thrombin activated samples, usually as single enantiomers (e.g. 12(S)-HEPE, 11(R)-HETE, 9(R)-HODE, 13-(S)-HODE, 14(S)-HDoHE). The latter was detected at about similar concentration in resting platelet releasates as well. 15-HETE showed elevated levels for both R-and S-enantiomers in releasates of thrombin-activated platelets. 12-HETrE was found presumably as both enantiomers, however, retention time inconsistencies indicate that the R-enantiomer is actually a different compound, maybe another constitutional isomer with different double-bond configuration.


Assuntos
Plaquetas/química , Cromatografia Líquida de Alta Pressão/métodos , Oxilipinas/análise , Polissacarídeos/química , Plaquetas/metabolismo , Ácidos Graxos Insaturados/análise , Ácidos Graxos Insaturados/química , Humanos , Limite de Detecção , Oxirredução , Estereoisomerismo , Espectrometria de Massas em Tandem
8.
PLoS One ; 15(4): e0232018, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32352972

RESUMO

INTRODUCTION: In many African countries, laboratory reference values are not established for the local healthy adult population. In Mozambique, reference values are known for young adults (18-24yo) but not yet established for a wider age range. Our study aimed to establish hematological, biochemical and immunological reference values for vaccine trials in Mozambican healthy adults with high-risk for HIV acquisition. METHODS: A longitudinal cohort and site development study in Mozambique between November 2013 and 2014 enrolled 505 participants between 18 to 35 years old. Samples from these healthy participants, were analyzed to determine reference values. All volunteers included in the analysis were clinically healthy and human immunodeficiency virus (HIV), hepatitis B and C virus, and syphilis negative. Median and reference ranges were calculated for the hematological, biochemical and immunological parameters. Ranges were compared with other African countries, the USA and the US National Institute of Health (NIH) Division of AIDS (DAIDS) toxicity tables. RESULTS: A total of 505 participant samples were analyzed. Of these, 419 participants were HIV, hepatitis B and C virus and syphilis negative including 203 (48.5%) females and 216 (51.5%) males, with a mean age of 21 years. In the hematological parameters, we found significant differences between sex for erythrocytes, hemoglobin, hematocrit, MCV, MCH and MCHC as well as white blood cells, neutrophils and platelets: males had higher values than females. There were also significant differences in CD4+T cell values, 803 cells/µL in men versus 926 cells/µL in women. In biochemical parameters, men presented higher values than women for the metabolic, enzymatic and renal parameters: total and direct bilirubin, ALT and creatinine. CONCLUSION: This study has established reference values for healthy adults with high-risk for HIV acquisition in Mozambique. These data are helpful in the context of future clinical research and patient care and treatment for the general adult population in the Mozambique and underline the importance of region-specific clinical reference ranges.


Assuntos
Células Sanguíneas/química , Infecções por HIV/prevenção & controle , Testes Hematológicos/normas , Adulto , Plaquetas/química , Estudos de Coortes , Feminino , Infecções por HIV/sangue , Hematócrito/normas , Hemoglobinas/análise , Humanos , Contagem de Leucócitos/normas , Leucócitos/química , Estudos Longitudinais , Masculino , Pessoa de Meia-Idade , Moçambique/epidemiologia , Valores de Referência , Fatores de Risco
9.
PLoS One ; 15(4): e0220163, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32294080

RESUMO

BACKGROUND: Clinical application of mesenchymal stromal cells (MSCs) usually requires an in vitro expansion step to reach clinically relevant numbers. In vitro cell expansion necessitates supplementation of basal mammalian cell culture medium with growth factors. To avoid using supplements containing animal substances, human platelet lysates (hPL) produced from expired and pathogen inactivated platelet concentrates can be used in place of fetal bovine serum. However, globally, most transfusion units are currently not pathogen inactivated. As blood banks are the sole source of platelet concentrates for hPL production, it is important to ensure product safety and standardized production methods. In this proof-of-concept study we assessed the feasibility of producing hPL from expired platelet concentrates with pathogen inactivation applied after platelet lysis by evaluating the retention of growth factors, cytokines, and the ability to support MSC proliferation and tri-lineage differentiation. METHODOLOGY/PRINCIPAL FINDINGS: Bone marrow-derived MSCs (BM-MSCs) were expanded and differentiated using hPL derived from pathogen inactivated platelet lysates (hPL-PIPL), with pathogen inactivation by amotosalen/ultraviolet A treatment applied after lysis of expired platelets. Results were compared to those using hPL produced from conventional expired pathogen inactivated platelet concentrates (hPL-PIPC), with pathogen inactivation applied after blood donation. hPL-PIPL treatment had lower concentrations of soluble growth factors and cytokines than hPL-PIPC treatment. When used as supplementation in cell culture, BM-MSCs proliferated at a reduced rate, but more consistently, in hPL-PIPL than in hPL-PIPC. The ability to support tri-lineage differentiation was comparable between lysates. CONCLUSION/SIGNIFICANCE: These results suggest that functional hPL can be produced from expired and untreated platelet lysates by applying pathogen inactivation after platelet lysis. When carried out post-expiration, pathogen inactivation may provide a valuable solution for further standardizing global hPL production methods, increasing the pool of starting material, and meeting future demand for animal-free supplements in human cell culturing.


Assuntos
Plaquetas/química , Extratos Celulares/farmacologia , Meios de Cultura/farmacologia , Células-Tronco Mesenquimais/efeitos dos fármacos , Plaquetas/efeitos dos fármacos , Plaquetas/efeitos da radiação , Segurança do Sangue/métodos , Técnicas de Cultura de Células/métodos , Diferenciação Celular/efeitos dos fármacos , Extratos Celulares/química , Proliferação de Células/efeitos dos fármacos , Células Cultivadas , Meios de Cultura/química , Furocumarinas/farmacologia , Humanos , Células-Tronco Mesenquimais/citologia
10.
Arterioscler Thromb Vasc Biol ; 40(6): 1432-1440, 2020 06.
Artigo em Inglês | MEDLINE | ID: mdl-32295424

RESUMO

Anucleate platelets, long viewed as merely cell fragments with a limited repertoire of rapid-acting hemostatic functions, are now recognized to have a complex and dynamic transcriptome mirroring that of many nucleated cells. The field of megakaryocyte and platelet transcriptomics has been rapidly growing, particularly with the advent of newer technologies such as next-generation RNA-sequencing. Studies interrogating the megakaryocyte and platelet transcriptome have led to a number of key insights into human health and disease. In this brief focused review, we will discuss some of the recent discoveries made through transcriptome analysis of megakaryocytes and platelets. We will also highlight the utility of integrating ribosome footprint analysis to augment discoveries. Both bulk and single-cell sequencing approaches will be reviewed, along with comparative studies between human and murine platelets under basal healthy settings and during acute systemic inflammatory diseases.


Assuntos
Plaquetas , Perfilação da Expressão Gênica , Megacariócitos , Adulto , Idoso , Envelhecimento , Animais , Plaquetas/química , Plaquetas/metabolismo , Estudos Transversais , Infecções por HIV/sangue , Nível de Saúde , Hemostasia , Sequenciamento de Nucleotídeos em Larga Escala , Humanos , Estudos Longitudinais , Megacariócitos/química , Megacariócitos/metabolismo , Camundongos , Análise de Sequência de RNA , Análise de Célula Única , Especificidade da Espécie
11.
J Biomed Sci ; 27(1): 45, 2020 Mar 23.
Artigo em Inglês | MEDLINE | ID: mdl-32200762

RESUMO

BACKGROUND: Human platelets (PLT) and PLT-extracellular vesicles (PEV) released upon thrombin activation express receptors that interact with tumour cells and, thus, can serve as a delivery platform of anti-cancer agents. Drug-loaded nanoparticles coated with PLT membranes were demonstrated to have improved targeting efficiency to tumours, but remain impractical for clinical translation. PLT and PEV targeted drug delivery vehicles should facilitate clinical developments if clinical-grade procedures can be developed. METHODS: PLT from therapeutic-grade PLT concentrate (PC; N > 50) were loaded with doxorubicin (DOX) and stored at - 80 °C (DOX-loaded PLT) with 6% dimethyl sulfoxide (cryopreserved DOX-loaded PLT). Surface markers and function of cryopreserved DOX-loaded PLT was confirmed by Western blot and thromboelastography, respectively. The morphology of fresh and cryopreserved naïve and DOX-loaded PLT was observed by scanning electron microscopy. The content of tissue factor-expressing cancer-derived extracellular vesicles (TF-EV) present in conditioned medium (CM) of breast cancer cells cultures was measured. The drug release by fresh and cryopreserved DOX-loaded PLT triggered by various pH and CM was determined by high performance liquid chromatography. The thrombin activated PEV was analyzed by nanoparticle tracking analysis. The cellular uptake of DOX from PLT was observed by deconvolution microscopy. The cytotoxicities of DOX-loaded PLT, cryopreserved DOX-loaded PLT, DOX and liposomal DOX on breast, lung and colon cancer cells were analyzed by CCK-8 assay. RESULTS: 15~36 × 106 molecules of DOX could be loaded in each PLT within 3 to 9 days after collection. The characterization and bioreactivity of cryopreserved DOX-loaded PLT were preserved, as evidenced by (a) microscopic observations, (b) preservation of important PLT membrane markers CD41, CD61, protease activated receptor-1, (c) functional activity, (d) reactivity to TF-EV, and (e) efficient generation of PEV upon thrombin activation. The transfer of DOX from cryopreserved PLT to cancer cells was achieved within 90 min, and stimulated by TF-EV and low pH. The cryopreserved DOX-loaded PLT formulation was 7~23-times more toxic to three cancer cells than liposomal DOX. CONCLUSIONS: Cryopreserved DOX-loaded PLT can be prepared under clinically compliant conditions preserving the membrane functionality for anti-cancer therapy. These findings open perspectives for translational applications of PLT-based drug delivery systems.


Assuntos
Plaquetas/fisiologia , Criopreservação , Doxorrubicina/química , Vesículas Extracelulares/fisiologia , Células Neoplásicas Circulantes/metabolismo , Plaquetas/química , Humanos
12.
Int J Nanomedicine ; 15: 901-912, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32103945

RESUMO

Background: Aortic valve disease is the most common valvular heart disease leading to valve replacement. The efficacy of pharmacological therapy for aortic valve disease is limited by the high mechanical stress at the aortic valves impairing the binding rate. We aimed to identify nanoparticle coating with entire platelet membranes to fully mimic their inherent multiple adhesive mechanisms and target the sclerotic aortic valve of apolipoprotein E-deficient (ApoE-/-) mice based on their multiple sites binding capacity under high shear stress. Methods: Considering the potent interaction of platelet membrane glycoproteins with components present in sclerotic aortic valves, platelet membrane-coated nanoparticles (PNPs) were synthetized and the binding capacity under high shear stress was evaluated in vitro and in vivo. Results: PNPs demonstrated effectively adhering to von Willebrand factor, collagen and fibrin under shear stresses in vitro. In an aortic valve disease model established in ApoE-/- mice, PNPs exhibited good targeting to sclerotic aortic valves by mimicking platelet multiple adhesive mechanisms. Conclusion: PNPs could provide a promising platform for the molecular diagnosis and targeting treatment of aortic valve disease.


Assuntos
Plaquetas/citologia , Doenças das Valvas Cardíacas/tratamento farmacológico , Nanopartículas/química , Nanopartículas/metabolismo , Animais , Valva Aórtica/efeitos dos fármacos , Valva Aórtica/patologia , Apolipoproteínas E/genética , Plaquetas/química , Membrana Celular/química , Colágeno/metabolismo , Modelos Animais de Doenças , Fibrina/metabolismo , Cardiopatias Congênitas , Doenças das Valvas Cardíacas/patologia , Células Endoteliais da Veia Umbilical Humana , Humanos , Masculino , Camundongos Endogâmicos C57BL , Camundongos Knockout para ApoE , Nanopartículas/uso terapêutico , Copolímero de Ácido Poliláctico e Ácido Poliglicólico/química , Esclerose , Estresse Mecânico , Fator de von Willebrand/metabolismo
13.
Sci Rep ; 10(1): 156, 2020 01 13.
Artigo em Inglês | MEDLINE | ID: mdl-31932650

RESUMO

Platelet microvesicles (pMVs) are submicron-sized heterogeneous vesicles released upon activation and contain several membrane receptors and proteins (CD41, CD61, CD62, CXCR4, PAR-1, etc.). We have revealed their ability to adhere to the triblock copolymer pluronic-F127 (PF127) and form a platelet microvesicular nanocloud which has the potential to enhance the transvascular migration of hematopoietic stem cells across the sinusoidal endothelium to the bone marrow. Besides, the pMVs nanoclouds bestow survival benefits when present on the cells used for infusion, particularly with PF127-stabilized with chitosan-alginate (PF127-CA HSCs). The vesicles were found to be firmly associated with PF127 in the nanocloud, which was detected by confocal laser scanning microscopy. The abrogation of CXCR4/SDF-1 axis regulating the transmigration of the cells by antagonist AMD3100 revealed that the enriched CXCR4 receptors on pMVs robustize the transmigration of the infused cells. The homing of the cells led to effective engraftment and faster regeneration of the critical blood lineages, which elicited 100% survival of the mice receiving lethal doses of radiation. The Human Long-Term Culture Initiating Cells (LTC-ICs), Severe Combined Immunodeficient (SCID) - Repopulating Cells (SRCs) and Colony Forming Cells (CFCs) responsible for the regeneration, but present in extremely low numbers in the infused cell dose, have enabled the cells to reach the bone marrow in high numbers. This potential of the PF127 to sequester the pMVs and its application to achieve over 10-fold delivery of HSCs across the trans-endothelial checkpoint has so far not been reported. Thus, this mechanistic innovation is a potential post-exposure life-saving regimen capable of circumventing the irreparable damage to the bone marrow caused by lethal doses of radiation.


Assuntos
Plaquetas/química , Células da Medula Óssea/citologia , Micropartículas Derivadas de Células/química , Transplante de Células-Tronco Hematopoéticas , Células-Tronco Hematopoéticas/citologia , Nanocompostos/administração & dosagem , Polietilenos/química , Polipropilenos/química , Animais , Apoptose , Células da Medula Óssea/metabolismo , Células da Medula Óssea/efeitos da radiação , Movimento Celular , Proliferação de Células , Células Cultivadas , Feminino , Humanos , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Endogâmicos NOD , Camundongos Nus , Camundongos SCID , Nanocompostos/química
14.
Sci Rep ; 10(1): 1032, 2020 01 23.
Artigo em Inglês | MEDLINE | ID: mdl-31974417

RESUMO

We analyzed the potential antibacterial effects of two different PdB against methicillin-resistant S. aureus and P. aeruginosa. The third-degree burn wound healing effects of PdB was also studied. Blood samples were obtained from 10 healthy volunteers and biological assays of the PdB were performed and the antimicrobial activity against MRSA and P. aeruginosa was determined using disk diffusion (DD), broth microdilution (BMD), and time-kill assay methods. 48 Wistar albino rats were burned and infected with MRSA. Two groups were injected PdB, the control groups were treated with plasma and received no treatment respectively. In the next step, the rats were euthanized and skin biopsies were collected and histopathologic changes were examined. The results of DD and BMD showed that both PdB performed very well on MRSA, whereas P. aeruginosa was only inhibited by F-PdB and was less susceptible than MRSA to PdBs. The time-kill assay also showed that F-PdB has an antibacterial effect at 4 hours for two strains. Histopathological studies showed that the treated groups had less inflammatory cells and necrotic tissues. Our data suggest that PdB may possess a clinical utility as a novel topical antimicrobial and wound healing agent for infected burn wounds.


Assuntos
Antibacterianos/uso terapêutico , Plaquetas/química , Extratos Celulares/uso terapêutico , Cicatrização/efeitos dos fármacos , Infecção dos Ferimentos/tratamento farmacológico , Animais , Materiais Biocompatíveis/uso terapêutico , Queimaduras/tratamento farmacológico , Queimaduras/microbiologia , Testes de Sensibilidade a Antimicrobianos por Disco-Difusão , Humanos , Masculino , Staphylococcus aureus Resistente à Meticilina/efeitos dos fármacos , Infecções por Pseudomonas/tratamento farmacológico , Pseudomonas aeruginosa/efeitos dos fármacos , Ratos , Ratos Wistar , Infecções Estafilocócicas/tratamento farmacológico
15.
Int J Mol Sci ; 21(3)2020 Jan 26.
Artigo em Inglês | MEDLINE | ID: mdl-31991927

RESUMO

Wound repair is a dynamic process during which crucial signaling pathways are regulated by growth factors and cytokines released by several kinds of cells directly involved in the healing process. However, the limited applications and heterogeneous clinical results of single growth factors in wound healing encouraged the use of a mixture of bioactive molecules such as platelet derivatives for best results in wound repair. An interesting platelet derivative, obtained from blood samples, is platelet lysate (PL), which has shown potential clinical application. PL is obtained from freezing and thawing of platelet-enriched blood samples. Intracellular calcium (Ca2+) signals play a central role in the control of endothelial cell survival, proliferation, motility, and differentiation. We investigated the role of Ca2+ signaling in the PL-driven endothelial healing process. In our experiments, the functional significance of Ca2+ signaling machinery was highlighted performing the scratch wound assay in presence of different inhibitors or specific RNAi. We also pointed out that the PL-induced generation of intracellular ROS (reactive oxygen species) via NOX4 (NADPH oxidase 4) is necessary for the activation of TRPM2 and the resulting Ca2+ entry from the extracellular space. This is the first report of the mechanism of wound repair in an endothelial cell model boosted by the PL-induced regulation of [Ca2+]i.


Assuntos
Plaquetas/química , Sinalização do Cálcio , Células Endoteliais/metabolismo , Endotélio Vascular/metabolismo , Animais , Diferenciação Celular , Linhagem Celular Transformada , Movimento Celular , Proliferação de Células , Sobrevivência Celular , Camundongos
16.
Colloids Surf B Biointerfaces ; 188: 110769, 2020 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-31918157

RESUMO

Polysulfone (PSf) membrane is widely employed in blood purification fields, but the blood compatibility of PSf membrane is not adequate. To improve the hemocompatibility of PSf membrane, 4-(chloromethyl)benzoic acid (CMBA) and sulfonated hydroxypropyl chitosan (SHPCS) were grafted onto PSf membrane surface. In our strategy, CMBA was firstly grafted on the PSf membrane surface through the Friedel-Crafts alkylation reaction, and the product was named BAPSf membrane. Then, SHPCS was grafted onto the BAPSf membrane surface by esterification, and the product was named SHPCS-BAPSf membrane. The effects of temperature and reaction time on the productivity of BAPSf and the grafting density of carboxyl and the effects of reaction time on the grafting density of SHPCS grafted onto the BAPSf membrane surface were studied. The SHPCS-BAPSf membranes are investigated by ATR-FTIR, XPS, contact angle measurements and evaluated by blood compatibility in vitro. The results reveal that the hydrophilicity of SHPCS-BAPSf membranes were grealy improved and the evaluation of protein adsorption, hemolysis test, platelet adhesion plasma recalcification time(PRT), activated partial thromboplastin time(APTT), prothrombin time(PT) and thrombin time(TT) confirmed that the SHPCS-BAPSf membranes have remarkable blood compatibility.


Assuntos
Ácido Benzoico/química , Materiais Biocompatíveis/química , Quitosana/química , Polímeros/química , Sulfonas/química , Adsorção , Animais , Testes de Coagulação Sanguínea , Plaquetas/química , Bovinos , Humanos , Interações Hidrofóbicas e Hidrofílicas , Teste de Materiais , Tamanho da Partícula , Adesividade Plaquetária , Soroalbumina Bovina/química , Propriedades de Superfície
17.
J Immunoassay Immunochem ; 41(2): 184-194, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-31842683

RESUMO

Lymphocyte-related parameters in Alcohol Use Disorder (AUD) have recently been investigated. However, knowledge of platelet to lymphocyte ratio (PLR) in AUD is limited. In this study, we compared complete blood count values of 31 AUD male patients and 31 healthy male controls. There was no significant difference between the two groups in terms of PLR (p = .123). When the age was controlled, there was a negative correlation between the duration of alcohol use and PLR (r = -0.567; p = .005). The significance of the parameters in the AUD group was found to be related to the duration of alcohol use.


Assuntos
Alcoolismo/sangue , Alcoolismo/diagnóstico , Plaquetas , Linfócitos , Adulto , Biomarcadores/análise , Plaquetas/química , Estudos de Coortes , Humanos , Contagem de Linfócitos , Linfócitos/química , Masculino , Contagem de Plaquetas , Estudos Retrospectivos
18.
Mol Biol Rep ; 47(1): 521-531, 2020 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-31721019

RESUMO

Diabetic complications are associated with the glycation and formation of advanced glycation end products (AGEs) which leads to structural modifications of biomolecules further affecting cells. Carbonyl compounds such as methylglyoxal and glyceraldehyde-3-phosphate are highly reactive and form an elevated amount of AGEs as compared to glucose and fructose. The investigation of glycation modifications by different compounds may be important to assess the specific pattern of biomolecular and cellular modifications and compare their glycation potential. The present work aims to comprehensively and comparatively examine the effect of glycating agents (glucose, fructose, ribose, methylglyoxal, and glyceraldehyde) on plasma, erythrocytes, platelets, and blood DNA. Glycation of plasma, cells, and DNA was initiated by incubating them with glycating agents for 24-48 h at 37 °C. Negative control samples (without glycating agents) were maintained simultaneously. After treatment, plasma and DNA samples were dialyzed and cell lysate was prepared. Markers of glycation (fructosamine), structural modifications (free amino, ß-amyloid, absorption spectra), antioxidant indices (catalase activity, glutathione) and erythrocyte hemolysis were estimated. In the presence of glycating agents, there was a significant increase in the formation of fructosamine, structural modification markers and depletion in antioxidant indices. Overall results suggest that among all glycating agents; methylglyoxal and glyceraldehyde have more potency of glycation induced structural modifications in plasma and vascular cells. This indicates the specific glycation modifications in plasma and vascular cells by various glycating agents may be investigated further for controlling diabetic pathological changes.


Assuntos
Plaquetas , Eritrócitos , Glicosilação/efeitos dos fármacos , Monossacarídeos/farmacologia , Antioxidantes/análise , Plaquetas/química , Plaquetas/efeitos dos fármacos , Plaquetas/metabolismo , DNA/química , DNA/efeitos dos fármacos , Eritrócitos/química , Eritrócitos/efeitos dos fármacos , Eritrócitos/metabolismo , Frutosamina/análise , Hemólise/efeitos dos fármacos , Humanos , Plasma/química , Plasma/efeitos dos fármacos , Aldeído Pirúvico/farmacologia
19.
Biochem Biophys Res Commun ; 521(4): 821-826, 2020 01 22.
Artigo em Inglês | MEDLINE | ID: mdl-31706576

RESUMO

Lanternfish, a family Myctophidae, use ventro-lateral body photophores for camouflage of the ventral silhouette, a strategy called counterillumination. While other deep-sea fishes possess pigmented filters and silver reflectors to match sunlight filtering down through the depths, myctophids developed a blue-green reflector for this purpose. In this study, we showed in a lanternfish Diaphus watasei that the reflector comprised monolayered iridophores containing multilayered guanine crystals which enable high reflection with light interference colouration. Platelets shape in body photophores is an unique near-regular hexagonal, probably to allow the homogeneity of reflection angle of the luminescence from photocytes. Focus point of the parabola-like reflector is positioned on the photocytes that ensures the light produced from the photocytes is redirected to the ventral direction. In vitro luminescence reaction using purified luciferase and the substrate coelenterazine showed the light emission at λmax 454 nm, while reflection spectra of the iridophores exhibit peaks at longer wavelength, which accomplish to alter the luminescence emitted from photocytes to longer wavelength to fit the mesopelagic light environment. Taken together, we revealed multiple mechanistic elaborations in myctophid body photophores to achieve effective control of biochemical luminescence for counterillumination.


Assuntos
Peixes/fisiologia , Animais , Mimetismo Biológico/fisiologia , Plaquetas/química , Plaquetas/fisiologia , Peixes/anatomia & histologia , Guanina/química , Imidazóis/metabolismo , Luciferases/metabolismo , Luminescência , Pirazinas/metabolismo , Espectroscopia de Infravermelho com Transformada de Fourier , Difração de Raios X
20.
Clin Chim Acta ; 501: 48-52, 2020 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-31809747

RESUMO

OBJECTIVE: This study was designed to retrospectively analyze the value of the hematological parameter platelet to lymphocyte ratio (PLR) and hemoglobin to platelet ratio (HPR) in patients with colon cancer. METHODS: The hematological parameters and clinical data of 354 cases patients with colon cancer, 108 cases patients with benign colon tumors and 123 healthy controls were collected from our hospital electronic medical records. RESULTS: Compared with the colon benign tumor group and the healthy control group, the colon cancer group had an increased PLR value and a decreased HPR value. The correlation between the clinicopathological features and the laboratory parameters of colon cancer patients was analyzed, and the results showed that both PLR and HPR were associated with tumor invasion and tumor size. Compared with PLR (AUC = 0.725, 95%CI: 0.682-0.765), HPR (AUC = 0.752, 95%CI: 0.710-0.790) or carcinoembryonic antigen (CEA) (AUC = 0.710, 95%CI: 0.666-0.751) used alone, the combination with PLR and CEA (AUC = 0.790, 95%CI: 0.750-0.826) or with HPR and CEA (AUC = 0.814, 95%CI: 0.775-0.848) can improve specificity and produce greater AUC in differentiating colon cancer from benign colon cancer. CONCLUSION: Combined application of PLR, HPR, and CEA may improve the diagnostic efficacy of distinguishing between colon cancer and benign colon tumors.


Assuntos
Análise Química do Sangue , Plaquetas/química , Neoplasias Colorretais/diagnóstico , Hemoglobinas/análise , Linfócitos/patologia , Neoplasias Colorretais/sangue , Feminino , Humanos , Contagem de Linfócitos , Masculino , Pessoa de Meia-Idade , Contagem de Plaquetas , Estudos Retrospectivos
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