Platelet Microparticles Research Articles

New perspectives on the role of platelet factors in enhancing wound regeneration

Aim. To analyze the use of biological factors in the stimulation of the wound healing process. In the course of the study, we analysed relevant domestic and foreign literature sources on the given topic.Methods. The literature was reviewed using the key query ‘the role of biological factors in wound healing stimulation’ through the eLIBRARY and PubMed databases.Results. Currently, the range of therapeutic approaches is broad and diverse, incorporating both traditional and experimental methods such as advanced dressings, tissue matrices, growth factors (GFs), cell therapy, and nanotechnology. The wound healing process is regulated by a complex interplay of intercellular, intracellular, and extracellular signalling mechanisms across various phases of healing.Conclusion. The application of platelet-based therapies in different medical fields has shown promising outcomes in certain conditions, such as acute and chronic injuries of bone and cartilage. However, platelet-based preparations have yet to gain widespread clinical use. Several studies have demonstrated the involvement of platelets and related products, such as platelet microparticles (PMPs) and exosomes, in multiple phases of wound healing. The presence of a substantial number of biologically active molecules within platelet granules—exhibiting anti-inflammatory, angiogenic, proliferative, and other properties—renders platelets particularly attractive for use in regenerative medicine, including the stimulation of wound healin

MACE prediction in patients with STEMI and NSTEMI: from classic to novel markers in the estimation of coronary microcirculation disorder

Evaluation of classic and novel markers of coronary microcirculatory disorder (CMD) which have a predictive value on the risk of major cardiovascular event (MACE) in patients with STEMI and NSTEMI. Material and methods. The research included the retrospective analysis of a prospective observational study carried out on 600 patients, of which have been formed equal 4 groups of 80 patients (STEMI+MACE; STEMI-MACE; NSTEMI+MACE; NSTEMI-MACE) in which the collected blood on the 5-th day after angioplasty, the circulating level of 16 markers referring to CMD in the field of estimating endothelial dysfunction and hemostasis was determined: 8 classic markers (hsCRP, IL-6, TNF-α, IL-10, myeloperoxidase, NO , angiopoietin 2, fibrin monomers) and 8 new markers (neopteirn, endocan, syndecan 1, L-Arginine/ADMA, Ang 1-7/Ang II, endothelial microparticles, platelet microparticles, von Willebrand/ADAMTS13). In order to assess the prediction of MACE (cardiac death, re-infarction, stroke) at 1-st year, the relative risk of markers that had significant differences in patients with MACE versus without MACE in both groups (STEMI and NSTEMI) was assessed. The control group consisted of 40 apparently healthy people. Results. The MACE rate in STEMI and NSTEMI patients at 1 year was similar, 29% and 28%, respectively. All markers of endothelial dysfunction explored had significant deviations from control. Of these, however, only 4 markers had significant different values in patients (STEMI and NSTEMI) with MACE vs without MACE: neopterin, endocan, L-Arginine/ADMA and Ang 1-7/Ang II. Remarkably, all 3 markers of hemostasis explored had circulating levels in patients who developed MACE significantly higher than patients without MACE, which significantly exceeded the control marker. The assessment of the relative risk value has allowed the delineation of markers regarding the prediction of MACE such as: von Willebrand/ADAMTS13, platelet microparticles, fibrin monomers, endocan, neopterin, Ang 1-7/ Ang II, L-Arginine/ADAMTS13. Conclusions. 1. Endothelial dysfunction and compromised hemostasis are pathophysiological pillars of CMD which influences the MACE risk, and thus new markers such as von Willebrand/ADAMTS13 ratio, platelet microparticles, endocan, the ratio Ang 1-7/Ang II and L/Arginine/ ADMAs have notable predictive value on MACE.2. Since CMD may be a key mechanism of cardiac ischemia in chronic non-obstructive coronary artery disease (<50%) its evaluation, including through the identified predictors, has a conclusive diagnostic and prognostic relevance.

Platelet-derived microvesicles in cardiovascular diseases

Extracellular vesicles (EVs) are a family of particles/vesicles present in blood and body fluids, bounded by a lipid dual- layer and non-replicable. They do not contain a functional core and carry a variety of molecules important in mediating cellular communication, thereby modulating crucial cellular processes such as homeostasis, induction/attenuation of inflammation, and support of cellular repair.Their existence, initially suspected in 1946 and confirmed in 1967, led to a sudden increase in the number of scientific publications, given their potential significance in understanding the mechanisms underlying various diseases, such as cancer, cardiovascular, metabolic, neurological diseases and infectious. These researches revealed a role for EVs as promising candidate biomarkers for diagnosis, prognosis, and even therapeutic purposes.The population of platelet-derived vesicles (pEVs) is the largest among other types of circulating EVs.Cell membrane physical characteristics and biological burden define the pivotal role of platelet-derived vesicles in the pathogenesis of cardiovascular (CV) diseases.Platelet-derived EVs are, therefore, key players in mediating inflammatory and coagulation reactions involving endotheliocytes, platelets, smooth muscle cells and inflammatory cells and thus contribute to the development ofatherosclerosis and angiogenesis, dysregulation of the coronary microcirculation in contiguity with endothelial dysfunction. In addition, EVs play an essential role in tissue repair, angiogenesis, and neovascularization through intracellular signaling cascades. In fact, EVs mediate autocrine and paracrine signals that are able to rebuild the homeostatic microenvironment in the heart and vessels.This review aims to provide a brief overview of the biogenesis, characteristics of platelet micro-particles with a special focus on their involvement in cardiovascular disease, but especially on the link between thrombosis, endothelial dysfunction and inflammation.In addition, here we review early experiments, summarize key findings that propelled the field, describe the growth of an organized EV community, and discuss the current state of the field.

Platelet microparticles influence gene expression and modulate biological activities of chronic myeloid leukemia cells (K562).

The current understanding emphasizes the intricate interplay between the Leukemic cell and its environment. Platelet-derived microparticles play a crucial role in facilitating intercellular communication and contribute to the complex landscape of cancer pathology. This study aimed to investigate the influence of platelet-derived microparticles on cell proliferation, apoptosis, and the expression of key genes, including P53, P21, Cyclin D1, Bax, and Bcl-2, within the context of a chronic myeloid leukemia cell line (K562). Platelet-derived microparticles were obtained through centrifugation at various speeds, and their concentration was quantified using the BCA assay. To determine the size and immunophenotypic characteristics of the PMPs, both the DLS technique and flow cytometry were employed. Cell proliferation was assessed using the MTT assay and hemocytometer, and cell cycle analysis was conducted through DNA content evaluation. Real-time PCR was utilized for gene expression analysis of Bax, Bcl-2, Cyclin D1, P53, and P21. Flow cytometry was employed to examine cell apoptosis. The findings revealed that platelet-derived microparticles have the ability to decrease proliferation of the K562 cell line, while not exerting an impact on apoptosis and cell cycle progression. Analysis through real-time PCR indicated an upregulation in the gene expression of P53, P21, and Bcl-2, accompanied by a downregulation in Bax and Cyclin D1. This investigation sheds light on the intricate relationship between chronic myeloid leukemia and its microenvironment, particularly the involvement of platelet-derived microparticles. The study underscores the potential of platelet-derived microparticles to influence cell behavior and gene expression, providing a deeper understanding of their role in CML and its therapeutic implications.

Platelet phosphatidylserine exposure and microparticle production as health bioindicators in marine mammals.

In human medicine, various pathologies, including decompression sickness, thrombocytopenia, and rheumatoid arthritis, have been linked to changes in cellular microparticles (MP) formation, particularly platelet microparticles (PMP). Similar disorders in marine mammals might be attributed to anthropogenic threats or illnesses, potentially impacting blood PMP levels. Thus, detecting platelet phosphatidylserine (PS) exposure and PMP formation could serve as a crucial diagnostic and monitoring approach for these conditions in marine mammals. Our group has developed a methodology to assess real-time PS exposure and PMP formation specifically tailored for marine mammals. This method, pioneered in species such as bottlenose dolphins, beluga whales, walruses, and California sea lions, represents a novel approach with significant implications for both clinical assessment and further research into platelet function in these animals. The adapted methodology for evaluating PS exposure and PMP formation in marine mammals has yielded promising results. By applying this approach, we have observed significant correlations between alterations in PMP levels and specific pathologies or environmental factors. These findings underscore the potential of platelet function assessment as a diagnostic and monitoring tool in marine mammal health. The successful adaptation and application of this methodology in marine mammals highlight its utility for understanding and managing health concerns in these animals.

Improving platelet function following prophylactic platelet transfusion in patients with hematological malignancies.

Platelet transfusion is a standard treatment to prevent bleeding in patients with hematological malignancies. Although transfusions can improve platelet count, their impact on platelet function remains controversial. We conducted flow cytometry to assess platelet function before and after transfusion and performed subgroup analyses to examine differences based on blood type, corrected count increment (CCI), and platelet microparticles. Overall, 50 patients who received prophylactic platelet transfusion were enrolled. CD42b expression increased, whereas CD41 expression decreased after transfusion. Apheresis platelets exhibited the lowest expression of PAC-1 and P-selectin when exposed to agonist stimulations. PAC-1 expression increased under high adenosine diphosphate (ADP) stimulation, while P-selectin expression increased under both high ADP and thrombin receptor-activating peptide stimulation. In the subgroup analysis, patients with a CCI >4500 and those with the same blood types exhibited a more significant increase in PAC-1 and P-selectin expression under agonist stimulation. When comparing apheresis platelets collected on different days, only the percentage of platelet-derived microparticles showed a significantincrease. Prophylactic transfusion improved platelet function. Platelet function significantly improved in patients with a CCI >4500, those with the same blood types as that of apheresis platelets, or those with platelet-derived microparticle levels <4.7%. No significant improvement in platelet function was noted after the transfusion of different blood types with acceptable compatibility or the transfusion of incompatible blood types. Our results suggest that transfusing platelets with the same blood type remains the optimal choice.

ADAMTS13, von Willebrand Factor, Platelet Microparticles, Factor VIII, and Impact of Somatic Mutations in the Pathogenesis of Splanchnic Vein Thrombosis Associated with BCR-ABL-Negative Myeloproliferative Neoplasms.

Myeloproliferative neoplasms (MPNs) are often associated with splanchnic vein thrombosis (SVT). Not all the factors involved in the thrombotic tendency are currently known. This study aims to evaluate a possible association between ADAMTS13, von Willebrand factor (VWF), platelet microvesicles (MV), and factor VIII activity (FVIII:C) with thrombotic events in MPN patients. In total, 36 consecutive MPN patients with SVT were enrolled. The MPNs were diagnosed based on clinical characteristics and one or more gene mutations among JAK-2, CALR, and MPL. As controls, 50 randomly selected patients with MPN without thrombosis, 50 patients with deep vein thrombosis without MPNs, and 50 healthy blood donors were evaluated. Complete blood count, ADAMTS13, VWF, MV, and FVIII:C in plasma were measured in all the subjects. The JAK-2 mutation was found in 94% of the patients with SVT, but none were triple-negative for genetic mutations (JAK2 V617F, CALR, MPL, and exon 12). Compared to the normal subjects, in all the MPN patients (with or without SVT), the levels of ADAMTS13 were found to be significantly lower (p < 0.001) and the MV concentrations were significantly higher (p < 0.001). Among the MPN patients, the VWF and FVIII:C levels were significantly higher in the patients with SVT than those without thrombosis (p = 0.007 and p = 0.04, respectively). Splenomegaly was present in 78% of MPN patients with SVT and in 30% of those without SVT (p < 0.001). The ADAMTS13/VWF ratio was reduced in all the patients, but not in the healthy blood donors (p < 0.001). The significant increase in circulating MV, VWF, and FVIII:C in the MPN patients and in the patients with thrombosis supports the role of endothelium damage in promoting thrombotic events. In particular, a significant increase in VWF and FVIII:C levels was found in the MPN patients with SVT.

Platelet Microparticle-Derived MiR-320b Inhibits Hypertension with Atherosclerosis Development by Targeting ETFA.

Hypertension and atherosclerosis often occur simultaneously. This study aimed to explore the role and mechanism of platelet microparticle (PMP) -derived microRNA-320b (miR-320b) in patients with hypertension accompanied by atherosclerosis.We collected samples from 13 controls without hypertension and atherosclerosis and 20 patients who had hypertension accompanied by atherosclerosis. In vitro, platelets were activated by Thrombin receptor-activating peptide to produce PMPs. HUVECs were induced by CoCl2 to mimic a hypoxic environment in vitro. RT-qPCR was employed to detect the expression levels of CD61, miR-320b, and ETFA. The protein expression level of ETFA was evaluated via Western blotting. Furthermore, 3- (4,5-dimethylthiazol-2-yl) -2,5-diphenyltetrazolium bromide, 5-ethynyl-2'-deoxyuridine, and wound healing assays were employed to assess the proliferation and migration of HUVECs. Enzyme-linked immunosorbent assay was used to measure the oxidative stress and inflammation-related factor expression.The expression of miR-320b was reduced in both platelets and PMPs but increased in plasma. MiR-320b promoted CoCl2-induced HUVEC viability, proliferation, and migration. The levels of the oxidative stress factors SOD and GSH as well as the inflammatory factor IL-10 were elevated in the CoCl2 + miR-320b mimics group compared with both the CoCl2 + mimics NC and CoCl2 groups. Conversely, the levels of the oxidative stress factors MDA and ROS as well as the inflammatory factors IL-6, TNF-α, and IL-1β were decreased. These results were regulated by miR-320b targeting ETFA.PMP-derived miR-320b inhibits the development of hypertension accompanied by atherosclerosis by targeting ETFA.

Sorghum phytonutrients and their health benefits: A systematic review from cell to clinical trials.

Sorghum is key for global food security due to its genetic variability, resilience, and rich phytonutrient content, which are linked to numerous health benefits. A systematic review assessed the health effects of sorghum by analyzing cell (n=22), animal (n=20), and human (n=7) studies across antioxidant, anti-inflammatory, obesity, cancer, cardiovascular, and diabetes outcomes. This review, involving 42 papers and 177 researchers from 12 countries, collected data from sorghum accessions (acc) and significant effects. Studies used 68 identified and 8 unidentified sorghums, 57% red (n=20), brown (n=5), and black (n=17) pericarp colors, and evaluated whole (n=31), brans (n=11), and decorticated grains (n=2). Colored sorghum, richer in phenolic compounds, especially 3-deoxyanthocyanins and tannins, inhibited cancer cell activities, including proliferation, tumor growth, and ROS activity, and promoted cell cycle arrest and apoptosis. Sorghum elevated HO1 and eNOS expression for cardiovascular, health-reduced platelet aggregation, and modulated platelet microparticles. They also suppressed inflammation markers and decreased lipid accumulation. Animal studies indicated sorghum's potential across antioxidant capacity, cancer and inflammation mitigation, and lipid and glucose metabolism. Translating these findings to human scenarios requires caution, especially considering cell studies do not fully represent polyphenol metabolism. Human studies provided mixed results, indicating antioxidant and potential anti-inflammatory benefits and nuanced effects on glucose and lipid metabolism. The main risks of bias highlighted challenges in quantifying phytonutrients, identifying sorghum acc features, and lack of assessors blinding. Nonetheless, sorghum emerges as a promising functional food for countering chronic diseases in Western diets.

Platelet dysfunction reversal with cold-stored vs room temperature–stored platelet transfusions

AbstractPlatelets are stored at room temperature for 5 to 7 days (room temperature–stored platelets [RSPs]). Because of frequent and severe shortages, the US Food and Drug Administration recently approved up to 14-day cold-stored platelets (CSPs) in plasma. However, the posttransfusion function of CSPs is unknown and it is unclear which donors are best suited to provide either RSPs or CSPs. In this study, we sought to evaluate the posttransfusion platelet function and its predictors for platelets stored for the maximum approved storage times (7-day RSPs and 14-day CSPs) in healthy volunteers on acetylsalicylic acid (ASA). We conducted a randomized crossover study in 10 healthy humans. Individuals donated 1 platelet unit, stored at either 22°C or 4°C based on randomization. Before transfusion, participants ingested ASA to inhibit endogenous platelets. Transfusion recipients were tested for platelet function and lipid mediators. Platelet units were tested for lipid mediators only. A second round of transfusion with the alternative product was followed by an identical testing sequence. RSPs reversed platelet inhibition significantly better in αIIbβ3 integrin activation–dependent assays. In contrast, CSPs in recipients led to significantly more thrombin generation, which was independent of platelet microparticles. Lysophosphatidylcholine-O species levels predicted the procoagulant capacity of CSPs. In contrast, polyunsaturated fatty acid concentrations predicted the aggregation response of RSPs. In summary, we provide, to our knowledge, the first efficacy data of extended-stored CSPs in plasma. Our results suggest that identifying ideal RSP and CSP donors is possible, and pave the way for larger studies in the future. This trial is registered at www.ClinicalTrials.gov as #NCT0511102.

Clinical and Serological Associations of Subclinical Atherosclerosis in Spondyloarthropathy

Background: Patients with spondyloarthropathy (SpA) have a higher risk of subclinical atherosclerosis (SCA). We assessed clinical and serological determinants of SCA in Indian SpA patients. Methods: Patients with SpA (fulfilling ASAS 2010 criteria; n = 104) attending our hospital were recruited; mean carotid intima-media thickness (CIMT) was performed by carotid ultrasonography, along with clinical assessment and traditional risk factor evaluation. Microparticles were extracted from plasma and total, as well as endothelial microparticles (EMP), platelet microparticles, T lymphocyte microparticles and B lymphocyte microparticles, were analysed by flow cytometry. Serum samples were analysed for inflammatory cytokines previously implicated in atherosclerosis, namely interleukin 1β (IL-1β), IL-6, IL-17, IL-27, IL-33 and tumour necrosis factor-alpha. Thirty-eight healthy controls were used for comparison. Subgroup analyses compared parameters between SpA with SCA (i.e., with carotid plaque or more than 75th percentile of CIMT for that age and sex in the Indian population) versus those without SCA. Ethical approval and written, informed consent were obtained. Results: Despite significantly younger age, lower body mass index and higher total cholesterol in controls compared to SpA, those with SpA had higher CIMT. Traditional cardiovascular risk factors (older age, higher waist-hip ratio) and novel markers of inflammation (serum IL-1β, IL-6) were associated with SCA. While total microparticles, EMP, T lymphocyte and B lymphocyte microparticles were increased in SpA than in healthy controls, they were not associated with SCA. Conclusions: Traditional risk factors and serum inflammatory cytokines IL-1β and IL-6 are associated with higher SCA in Indian SpA patients.

Procoagulant Phospholipid Activity and MPV Values in Acute Ischemic Stroke

Ischemic stroke can be caused by atherothrombosis or embolism. Atherothrombosis occurs due to the rupture of an atherosclerotic plaque causing platelet activation. There are several markers of platelet activation, including platelet microparticles. The levels of platelet microparticles can be measured by examining procoagulant phospholipid (PPL) activity. It is not yet known exactly what activates thrombopoiesis, which can be assessed by an increase in Mean Platelet Volume (MPV). This study aimed to know whether there is an increase in PPL activity and MPV values in acute ischemic stroke, whether platelet activation is influenced by differences in ischemic stroke subtypes, and whether there is a relationship between PPL activity and MPV values in acute ischemic stroke. The study design was cross-sectional and involved 60 subjects. PPL activity in acute ischemic stroke (65.14±13.35 seconds) tends to be higher (shorter clotting time) than in healthy individuals (68.59±8.56 seconds), however, this difference was not statistically and clinically significant. The MPV value in acute ischemic stroke (9.83±0.72 fL) compared to healthy individuals (9.65±0.86 fL) however this difference was not statistically significant. PPL activity in the SAO subtype (61.66±1.31 seconds) tends to be higher than LAA (68.62±14.57 seconds), however, this difference was also not statistically significant. There was a weak correlation between PPL activity (seconds) and MPV value (fL) in acute ischemic stroke (r =0.34, p-value=0.03).

Gelatin In Situ Zymography to Study Gelatinase Activity in Colon Cancer Cells Treated with Platelet Microparticles (PMPs).

The platelet-derived microparticles (PMPs) have been connected with tumor progression and metastatic dissemination. PMPs infiltrate solid tumors and transfer platelet-derived cargo to cancer cells. The functional roles of PMPs in cancer progression are still poorly understood. PMPs, incorporated by colorectal cancer (CRC) cells, were shown to upregulate the expression and activity of matrix metalloproteases (MMPs). To investigate the impact of PMPs on the invasive potential of CRC, we established the protocol of dequenched gelatin(DG), fluorescein conjugate assay. The procedure confirms the activity of two gelatinases, namely, MMP2 and MMP9, that digest denatured collagen (gelatin). This "step-by-step" protocol, with notes and comments implemented to human CRC lines with different phenotypes and migratory potentials, should be sufficient to obtain representative and elegant results.

Differential Expression of the hTERT Gene in Umbilical Cord-Derived Mesenchymal Stem Cells Cocultured with B Cell Precursor Leukemia Cell Microparticles or CD41+/CD61+ Platelet Microparticles.

Several investigations are being done to increase the short lifetime of mesenchymal stem cells (MSCs). One of the crucial genes involved in the immortalization of MSCs, hTERT (human telomerase reverse transcriptase), is activated in most publications using viral-based techniques. In this work, we investigated the use of platelet-derived (PMPs) and B cell precursor leukemia-derived microparticles as a nonviral method to trigger and compare the expression of the hTERT gene in MSCs. MSCs were extracted from the umbilical cord for the current investigation and identified using a flow cytometry approach and an inverted microscope. The Nalm-6 cell line and platelet concentrate were used to isolate microparticles (MPs). MSCs and MPs were cocultured for 14days at 25-, 50-, and 100μg/ml concentrations. qRT-PCR was used to research the expression of the hTERT gene. SPSS 26.0's t test was used to compare the outcomes. After coculture with platelet MPs, MSCs had higher levels of hTERT gene expression than the control group. In contrast, this gene's expression was concurrently decreased in MSCs exposed to MPs generated from Nalm-6. We demonstrated that following 14-day treatment, PMP significantly boosted the hTERT gene expression in MSCs, while the Nalm-6 MPs lowered the gene expression. However, additional studies are necessary due to the stability of hTERT gene expression and the immortalization of MSCs following exposure.

Phosphodiesterase 3A Acts As a Tumor Activator and Its Inhibitor Anagrelide Combined with Idarubicin Represents a Potential Therapeutic Approach for PDE3A High Expressed Acute Myeloid Leukemia through Pyroptosis By Triggering Cleavage of Gasdermin E

Acute myeloid leukemia (AML) is a clinically aggressive and heterogeneous hematologic neoplasm with a poor overall outcome. Platelets can interact with malignant cells including leukemic blasts and play a crucial role in cancer progression, metastasis and angiogenesis, budding of platelet microparticles can transfer a plethora of bioactive molecules to cancer cells upon internalization and in AML this can induce chemoresistance. Anti-platelet therapy using Phosphodiesterase 3A (PDE3A) inhibitors, such as anagrelide (ANA), holds promise in tumor elimination. Additionally, upregulation of PDE3A in solid tumors is associated with a poor prognosis in patients and activates inflammatory pathways, impacting cancer cell stemness. However, the role of PDE3A in AML remains unclear. This study aims to investigate the correlation between PDE3A expression and AML prognosis, as well as to characterize the feasibility and mechanism of ANA as a potential anti-leukemia therapy and its synergistic effect to chemotherapeutic drugs. To explore the correlation between PDE3A expression and AML prognosis, we initially analyzed its expression in AML patients and healthy donors from the Children's Hospital of Soochow University. The results demonstrated significantly higher levels of PDE3A in bone marrow mononuclear cells (BMMNCs) from AML patients, both at the protein and mRNA levels, compared to BMMNCs from healthy donors. Subsequently, we investigated PDE3A expression and survival data of 152 AML patients from our center, revealing that patients with high PDE3A expression exhibited significantly lower overall survival (OS), relapse-free survival (RFS), and event-free survival (EFS) compared to those with low PDE3A expression. These findings were further corroborated by data from other databases (R2 and GEPIA), confirming the significant role of PDE3A in AML. Among AML cell lines, HEL and MOLM-16 cells exhibited notably higher PDE3A expression at both the protein and mRNA levels, which were chosen for further study. ANA treatment specifically inhibited the proliferation of PDE3A-high-expressing AML cells but not low expression cells in vitro, which was further confirmed in the HEL-mediated xenograft leukemia model. Notably, the RNA sequencing data revealed ANA treatment significantly downregulated several chemotherapy-resistant genes, including PAK1, HMGCS1, p-SRC, DUSP16 and EGR1, indicating ANA has the potential to synergize with chemotherapeutic drugs for AML therapy. Therefore, we combine ANA and chemotherapy drugs to investigate the synergetic inhibitory effect. IDA with ANA simultaneously exerted the strongest synergistic effect among the six anti-leukemia drugs depend on the judgement of CI value and had the synergistic effect in HEL, MOLM-16 and primary cells from PDE3A high expressed AML patients. The GSEA analysis of RNA sequencing data showed that the IDA-ANA combination treatment led to a significant enrichment of the inflammatory response. Moreover, at the protein level, pyroptosis-related proteins including NLRP3, Cleaved-CASP3, Cleaved-CASP8 and GSDME-N were notably upregulated in IDA-ANA combination group compared to IDA or ANA group. Finally, we established an AML xenograft mouse model using the HEL cell line and administered different treatments: vehicle (control), IDA (0.5 mg/kg), ANA (5 mg/kg), or the indicated combination of IDA and ANA. The results demonstrated that the IDA-ANA combination treatment significantly extended the survival time of the mice and reduced the leukemia burden when compared to the individual treatments with either IDA or ANA. In second HEL xenograft mice study, separate cohorts of vehicle and drug treated mice were sacrificed and leukemic livers as well as spleens were harvested and weighed, the IDA-ANA combination treated mice exhibited smaller and lighter livers and spleens than the other groups and controls. HE results explained the histological decrease of HEL cells infiltration of IDA-ANA combination treatment in the livers, spleens and bone marrow compared with the treatment of IDA or ANA. Congruently, the analysis of IHC confirmed the same result. In summary, high expression of PDE3A related to poor prognosis in AML and PDE3A inhibitor ANA combined with IDA exerted anti-leukemia effect through pyroptosis on PDE3A high expressed AML. Our findings represent a potential therapeutic regimen for AML.

Buyang Huanwu Decoction alleviates blood stasis, platelet activation, and inflammation and regulates the HMGB1/NF-κB pathway in rats with pulmonary fibrosis

Ethnopharmacological relevanceQi deficiency and blood stasis are identified to be pathological factors of pulmonary fibrosis (PF) in traditional Chinese medicine (TCM) theory. Buyang Huanwu Decoction (BYHWD) is a traditional Chinese prescription ameliorating Qi deficiency and blood stasis. Aim of the studyThe objective of this study was to investigate the anti-fibrosis effect of BYHWD and the potential molecular mechanism in rats. Materials and methodsBleomycin was used to construct PF rat models. 27 PF rats were randomly divided into three groups based on treatments: model group (saline solution, n = 9), low-dose BYHWD group (3.5 g/kg, n = 9), and high-dose BYHWD group (14.0 g/kg, n = 9). Moreover, 9 normal rats were used as the blank group. The blood viscosity, coagulation indexes (APTT, TT, PT, and FIB), platelet-related parameters (PLT, PDW, MPV, PCT, and PLCR), platelet microparticles (PMPs), and inflammatory factors (IL-2, IL-10, IL-1β, IL-6, IL-8, IL-17, IFN-γ, TNF-α, PAC-1, HMGB1, NF-κB, and TF) were determined. The lung tissue samples of rats were observed after hematoxylin-eosin (HE) staining. The full component analysis of the BYHWD extract was performed using the ultra-high performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) method. The signaling pathway included into the study was selected on the basis of bioinformatics analysis and the results of the phytochemical analysis. The expression levels of genes and proteins involved in the selected signaling pathway were detected. ResultsCompared to the blank group, the whole blood viscosity, PLR, PDW, MPV, PCT, PLCR, PMPs, and the levels of IL-1β, IL-6, IL-8, IL-17, TNF-α, PAC-1, HMGB1, NF-κB, and TF were increased, while the levels of IL-2 and IL-10 were decreased in the model group. Both low-dose BYHWD and high-dose BYHWD reversed these PF-induced effects in spite of the fact that low-dose BYHWD had no significant effect on the level of NF-κB. In addition, BYHWD ameliorated PF-induced inflammation in the rat lung tissue. The phytochemical analysis of the BYHWD extract combined with the bioinformatics analysis suggested that the therapeutical effect of BYHWD on PF was related to the HMGB1/NF-κB pathway, which consisted of NF-κB, IKBKB, ICAM1, VCAM1, HMGB1, and TLR4. Both RT-qPCR and western blot analyses showed that PF induced increases in the expression levels of NF-κB, ICAM1, VCAM1, HMGB1, and TLR4, but a decrease in the expression level of IKBKB. Moreover, both low-dose BYHWD and high-dose BYHWD exerted the opposite effects, and recovered the expression levels of NF-κB, ICAM1, VCAM1, HMGB1, TLR4, and IKBKB, despite the fact that low-dose BYHWD had no effects on the mRNA expression levels of NF-κB or TLR4. ConclusionsIn summary, BYHWD alleviated PF-induced blood stasis, platelet activation, and inflammation in the rats. Our study suggested BYHWD had a therapeutic effect on PF and was a good alternative for the complementary therapy of PF, and the potential molecular mechanism was modulation of HMGB1/NF-κB signaling pathway, and it needs further study.

The study of platelet microparticles and P-selectin expression in patients with the peripheral arterial diseases

BACKGROUND: High platelet reactivity leads to the progression of atherosclerosis and its complications. Activated platelets adhere to the site of endothelium damage on the vessel wall and initiate the formation of an arterial thrombus, followed by acute ischemia of the organ. Biochemical and cellular marker such as platelet microvesicles and P-selectin can be analyzed using flow cytometry, which is based on a specific antigen-antibody interaction. Patients with peripheral arterial disease are at significantly greater risk of cardiovascular and cerebrovascular complications than individuals of the same age and sex. The use of antiplatelet agents is an important part of pathogenetic therapy and prevention of cardiovascular diseases and their complications.&#x0D; AIM: This study was to evaluate the level of platelet microparticles and P-selectin expression in patients with peripheral arterial disease receiving antiplatelet therapy.&#x0D; MATERIALS AND METHODS: The study included 49 people, which included three study groups: patients with obliterating disease of the arteries of the lower extremities (n = 14) on the background of long-term use (more than 14 days) of double (75 mg clopidogrel and 100 mg Acetylsalicylic acid) antiplatelet therapy, patients with COVID-19 (n = 15), who made up the positive control group in the determination of microparticles of platelet origin, and healthy volunteers (n = 20) without signs of acute respiratory disease, without a history of cardiovascular and thromboembolic episodes, not taking antiplatelet drugs. The functional activity of platelets was assessed by two methods: using light aggregometry and analysis of P-selectin expression on platelet surface by flow cytometry. The number of platelets microparticles in blood plasma was also determined using flow cytometry.&#x0D; RESULTS: A significant decrease in platelet aggregation was found in patients with peripheral arterial disease taking antiplatelet agents, compared with controls by used light aggregometry. Similar changes were obtained when analyzing the expression of P-selectin on platelets. A higher percentage of platelets microparticles with the CD9+CD41+ phenotype was found in patients with severe inflammation compared with peripheral arterial disease patients treated with antiplatelet agents and compared with the healthy controls.&#x0D; CONCLUSIONS: Thus, our study reflects the consistency of the results of three different laboratory tests in assessing the platelets reactivity in patients with the peripheral arterial diseases taking antiplatelet drugs.

T cell-mimicking platelet-drug conjugates

•Platelets act as “Trojan horse” killers to target and inhibit tumor metastasis •Platelet-drug conjugates mimic the natural cytotoxic mechanisms of T cells •Engineered platelets explore the potential of immune-independent cell therapies Cancer metastasis is the leading cause of cancer-related deaths. The indispensable role of platelets in tumor metastasis inspires the targeted delivery strategy by using platelet-drug conjugates (PDCs). Here, we armed platelets with granzyme B and perforin nanocomplexes to suppress cancer metastasis by leveraging the natural cytotoxic mechanism of T cells. Upon systemic administration, PDCs naturally capture circulating tumor cells (CTCs) and actively target nascent metastatic lesions, thus enhancing the targeted delivery efficiency. Subsequently, PDCs activated in situ by tumor cells can produce platelet microparticles (PMPs) that can actively target tumor cells by the expression of adhesion molecules such as P-selectin and thrombospondin-1 (TSP-1) and by secreting cytotoxic protein complexes mimicking cytotoxic lymphocytes. We demonstrated that the PDCs significantly suppressed lung metastasis in two tumor models, B16F10 melanomas and 4T1 breast carcinomas. Cancer metastasis is the leading cause of cancer-related deaths. The indispensable role of platelets in tumor metastasis inspires the targeted delivery strategy by using platelet-drug conjugates (PDCs). Here, we armed platelets with granzyme B and perforin nanocomplexes to suppress cancer metastasis by leveraging the natural cytotoxic mechanism of T cells. Upon systemic administration, PDCs naturally capture circulating tumor cells (CTCs) and actively target nascent metastatic lesions, thus enhancing the targeted delivery efficiency. Subsequently, PDCs activated in situ by tumor cells can produce platelet microparticles (PMPs) that can actively target tumor cells by the expression of adhesion molecules such as P-selectin and thrombospondin-1 (TSP-1) and by secreting cytotoxic protein complexes mimicking cytotoxic lymphocytes. We demonstrated that the PDCs significantly suppressed lung metastasis in two tumor models, B16F10 melanomas and 4T1 breast carcinomas.

Shear-Mediated Platelet Microparticles Demonstrate Phenotypic Heterogeneity as to Morphology, Receptor Distribution, and Hemostatic Function.

Implantable Cardiovascular Therapeutic Devices (CTD), while lifesaving, impart supraphysiologic shear stress to platelets, resulting in thrombotic and bleeding coagulopathy. We previously demonstrated that shear-mediated platelet dysfunction is associated with downregulation of platelet GPIb-IX-V and αIIbβ3 receptors via generation of Platelet-Derived MicroParticles (PDMPs). Here, we test the hypothesis that sheared PDMPs manifest phenotypical heterogeneity of morphology and receptor surface expression and modulate platelet hemostatic function. Human gel-filtered platelets were exposed to continuous shear stress. Alterations of platelet morphology were visualized using transmission electron microscopy. Surface expression of platelet receptors and PDMP generation were quantified by flow cytometry. Thrombin generation was quantified spectrophotometrically, and platelet aggregation was measured by optical aggregometry. Shear stress promotes notable alterations in platelet morphology and ejection of distinctive types of PDMPs. Shear-mediated microvesiculation is associated with the remodeling of platelet receptors, with PDMPs expressing significantly higher levels of adhesion receptors (αIIbβ3, GPIX, PECAM-1, P-selectin, and PSGL-1) and agonist receptors (P2Y12 and PAR1). Sheared PDMPs promote thrombin generation and inhibit platelet aggregation induced by collagen and ADP. Sheared PDMPs demonstrate phenotypic heterogeneity as to morphology and defined patterns of surface receptors and impose a bidirectional effect on platelet hemostatic function. PDMP heterogeneity suggests that a range of mechanisms are operative in the microvesiculation process, contributing to CTD coagulopathy and posing opportunities for therapeutic manipulation.

Quantifying Stent Wall Shear Stress for Assessing Stent and Flow Diverter Performance for Treating Intracranial Aneurysms

AbstractAnalysis of flow diverting intracranial aneurysm repair devices has traditionally focused on reducing intrasaccular blood flow velocity and aneurysm wall shear stress (WSS) as the primary metrics for improved perceived device performance. However, the interpretation of this data has been debated, particularly with regard to the specific biological benefits of high or low aneurysm WSS. Therefore, this research proposes an additional parameter of WSS at the stent struts that could provide valuable insight regarding the device's potential to promote occlusion at the aneurysm neck by indicating locations of increased platelet activation and microparticle shedding. Fluid flow effects were evaluated for two flow diverters (Pipeline™ and FRED™) and three stents (Enterprise™, Atlas™, and LVIS™) using computational fluid dynamics (CFD) models developed from two patient-derived CTA datasets with aneurysms in the middle cerebral artery (MCA) and basilar artery (BA), respectively. The device WSS data provides an additional metric for evaluating the ability of the device to constrain the blood flow within the main vessel, as well as indicating potential locations of the initiation of aneurysm occlusion. It is hypothesized that high device WSS close to the aneurysm neck creates a higher likelihood of thrombus formation and aneurysm occlusion due to platelet activation and microparticle shedding, while high device WSS proximal or distal on the device would indicate a higher likelihood of undesirable daughter vessel occlusion. Conversely, low-to-moderate device WSS throughout the device length could be interpreted as a reduced likelihood of complete occlusion of the aneurysm over time, resulting in lesser device performance.