PDGF-BB Research Articles

Mechanisms of cordycepin in the treatment of pulmonary arterial hypertension in rats based on metabonomics and transcriptomics

Pulmonary arterial hypertension (PAH) is a fatal disease featured by high morbidity and mortality. Although Cordycepin is known for its anti-inflammatory, antioxidant and immune-enhancing effects, its role in PAH treatment and the underlying mechanisms remain unclear. The therapeutic effects of Cordycepin on rats with PAH were investigated using a monocrotaline (MCT)-induced rat model. The metabolic effects of Cordycepin were assessed based on the plasma metabolome. The potential mechanisms of Cordycepin in PAH treatment were investigated through transcriptome sequencing and validated in pulmonary artery smooth muscle cells (PASMC). Evaluations included hematoxylin and eosin staining for pulmonary vascular remodeling, CCK-8 assay, EDU, and TUNEL kits for cell viability, proliferation, and apoptosis, respectively, and western blot for protein expression. Cordycepin significantly reduced right ventricular systolic pressure (RVSP) and right ventricular hypertrophy index (RVHI) in PAH rats, and mitigated pulmonary vascular remodeling. Plasma metabolomics showed that Cordycepin could reverse the metabolic disorders in the lungs of MCT-induced PAH rats, particularly impacting linoleic acid and alpha-linolenic acid metabolism pathways. Transcriptomics revealed that the P53 pathway might be the primary pathway involved, and western blot results showed that Cordycepin significantly increased P53 and P21 protein levels in lung tissues. Integrated analysis of transcriptomics and metabolomics suggested that these pathways were mainly enriched in linoleic acid metabolism and alpha-linolenic acid metabolism pathway. In vitro experiments demonstrated that Cordycepin significantly inhibited the PDGFBB (PD)-induced abnormal proliferation and migration of PASMC and promoted PD-induced apoptosis. Meanwhile, Cordycepin enhanced the expression levels of P53 and P21 proteins in PD-insulted PASMC. However, inhibitors of P53 and P21 eliminated these effects of Cordycepin. Cordycepin may activate the P53–P21 pathway to inhibit abnormal proliferation and migration of PASMC and promote apoptosis, offering a potential approach for PAH treatment.

Ethanol Extract of Chromolaena odorata Leaves in Ointment Form: Evaluating Its Efficacy in Healing Excisional Wounds in Male White Rats

The leaves of Chromolaena odorata (C. odorata), which have been traditionally employed by the coastal communities of Aceh for the treatment of small wounds, have significant concentrations of flavonoids and phenolics. The objective of this study was to evaluate the efficacy of an ointment made from ethanol extract of C. odorata leaves in boosting wound healing and increasing the growth and expression of fibroblasts and Platelet Derived Growth Factor (PDGF-BB) in excision wounds of male white rats. On the 15th day, there was a decrease in the size of the wound as follows: The closure rates for Betadine® were measured at 96.07%, whereas the control group had a cure rate of 38.12%. The ointments containing C. odorata extract at concentrations of 4%, 6%, and 8% showed closure rates of 80.93%, 95.38%, and 95.75% respectively. The statistical analysis showed that there was no significant distinction between the 6% and 8% concentrations of the extract and Betadine®. On the seventh day, the number of fibroblast cells per field was 70.60 for Betadine®, 69.20 for the 6% extract, and 48.00 for the control. There were no significant differences between Betadine® and the 6% extract. On day 14, the expression of PDGF-BB was observed. The number of cells per field was 125.20 for Betadine®, 106.40 for the control, and 123.80 for the 6% extract. The results of the extract were similar to those of Betadine®. The results of this study demonstrate that the ointment made from the ethanol extract of C. odorata leaves effectively improves the healing of wounds, stimulates the activity of fibroblasts, and increases the production of PDGF-BB. These effects are comparable to the effectiveness of Betadine® in treating excisional wounds. Keywords: Chromolaena odorata, excision wounds, fibroblasts, PDGF BB expression

Small vessel disease in primary familial brain calcification with novel truncating PDGFB variants.

Primary familial brain calcification (PFBC) is a neurodegenerative disease characterised by bilateral calcification in the brain, especially in the basal ganglia, leading to neurological and neuropsychiatric manifestations. White matter hyperintensities (WMH) have been described in patients with PFBC and pathogenic variants in the gene for platelet-derived growth factor beta polypeptide (PDGFB), suggesting a manifest cerebrovascular process. We present below the cases of two PFBC families with PDGFB variants and stroke or transient ischaemic attack (TIA) episodes. We examine the possible correlation between PFBC and vascular events as stroke/TIA, and evaluate whether signs for vascular disease in this condition are systemic or limited to the cerebral vessels. Two Swedish families with novel truncating PDGFB variants, p.Gln140* and p.Arg191*, are described clinically and radiologically. Subcutaneous capillary vessels in affected and unaffected family members were examined by light and electron microscopy. All mutation carriers showed WMH and bilateral brain calcifications. The clinical presentations differed, with movement disorder symptoms dominating in family A, and psychiatric symptoms in family B. However, affected members of both families had stroke, TIA, and/or asymptomatic intracerebral ischaemic lesions. Only one of the patients had classical vascular risk factors. Skin microvasculature was normal. Patients with these PDGFB variants develop microvascular changes in the brain, but not the skin. PDGFB-related small vessel disease can manifest radiologically as cerebral haemorrhage or ischaemia, and may explain TIA or stroke in patients without other vascular risk factors.

Tumor-mesothelium HOXA11-PDGF BB/TGF β1-miR-181a-5p-Egr1 feedforward amplifier circuity propels mesothelial fibrosis and peritoneal metastasis of gastric cancer.

A proportion of gastric cancer (GC) patients suffer from peritoneal metastasis (PM) in the late stage of tumor and these patients have a poor prognosis. To provide more care for GC patient with PM, a deeper exploration of the molecular characteristics of GC-PM is needed. Here we performed the in vitro and in vivo study to illustrate the effect of HOXA11 over-expressed GC cells on peritoneal mesothelial cells (HMrSV5), transcriptomics analyses of HMrSV5 cells co-cultured with HOXA11 over-expressed GC cells, counterparts or alone, cytokine array analyses of serum-free culture medium of HOXA11 over-expressed GC cells, we validated our findings through genetic manipulation of HMrSV5 cells and neutralizing antibodies targeting cytokines secreted by HOXA11 over-expressed GC cells in vitro, as well as utilized human peritoneal metastatic lesions to validate expression of potential targets. We identified that HOXA11 over-expressed GC cells strongly propelled mesothelial fibrosis in vivo and in vitro, and HOXA11 regulated paracrine and autocrine of PDGF BB and TGF β1 in GC cells to propel mesothelial fibrosis. Meanwhile, HOXA11 over-expressed GC cells drove PDGF BB and TGF β1 secretion to activate developmental-process related genes in HMrSV5 cells, including Egr1, which processes dependent on miR-181a-5p. Then, Egr1 could mediate peritoneal mesothelial fibrosis. Correspondingly, Egr1 over-expressed HMrSV5 cells supported migration and peritoneal dissemination of GC cells. Together our results suggest that a feedforward amplifier circuity governing GC cells and mesothelial cells in peritoneum contribute to peritoneal metastasis of GC cells.

Diagnostic des sarcomes utérins et tumeurs mésenchymateuses utérines rares à potentiel de malignité. Référentiels du Groupe Sarcome Français et des Tumeurs Rares Gynécologiques

The landscape of uterine sarcomas is becoming more complex with the description of new entities associated with recurrent driver molecular alterations. Uterine sarcomas, in analogy with soft tissue sarcomas, are distinguished into complex genomic and simple genomic sarcomas. Leiomyosarcomas and undifferentiated uterine sarcomas belong to complex genomic sarcomas group. Low-grade and high-grade endometrial stromal sarcomas, other rare tumors associated with fusion transcripts (such as NTRK, PDGFB, ALK, RET ROS1) and SMARCA4-deficient uterine sarcoma are considered simple genomic sarcomas. The most common uterine sarcoma are first leiomyosarcoma and secondly endometrial stromal sarcomas. Three different histological subtypes of leiomyosarcoma (fusiform, myxoid, epithelioid) are identified, myxoid and epithelioid leiomyosarcoma being more aggressive than fusiform leiomyosarcoma. The distinction between low-grade and high-grade endometrial stromal sarcoma is primarily morphological and immunohistochemical and the detection of fusion transcripts can help the diagnosis. Uterine PEComa is a rare tumor, which is distinguished into borderline and malignant, according to a risk assessment algorithm. Embryonal rhabdomyosarcoma of the uterine cervix is more common in children but can also occur in adult women. Embryonal rhabdomyosarcoma of the uterine cervix is almost always DICER1 mutated, unlike that of the vagina which is wild-type DICER1, and adenosarcoma which can be DICER1 mutated but with less frequency. Among the emerging entities, sarcomas associated with fusion transcripts involving the NTRK, ALK, PDGFB genes benefit from targeted therapy. The integration of molecular data with histology and clinical data allows better identification of uterine sarcomas in order to better treat them.

Elucidating Mycosis Fungoides Gene Expression in Skin of Color Patients from Howard University Dermatology

Abstract Mycosis fungoides (MF) is the most common type of cutaneous T cell lymphoma.There are notable differences in MF presentation in Skin of Color (SOC) versus White skin, such as asymptomatic hyperpigmented and hypopigmented lesions resulting from malignant T-cell interaction from keratinocytes and melanocytes. Therefore, this work aims at elucidating MF immunopathogenesis and gene expression in SOC patients and determining whether clinical presentation differences are dependent on the cytokine type produced by the tumor. We used both hypopigmented and hyperpigmented MF biopsy samples from SOC patients with MF from Howard University Dermatology and compared their gene expression to that of biopsy samples from SOC healthy patients. Preliminary data showed an upregulation of genes such as PRDX1, HLA-DRA, CTNNB1, CSTB, and S100A4 in MF samples versus their healthy counterparts. The MF samples also exhibited downregulation of CCL3, CCRL2, PDGFB and HOXD4 genes when compared to their healthy counterparts. Lastly, the pathways ‘cell cycle and apoptosis’, ‘antigen presentation’, and ‘interferon signaling’ were increased in MF and decreased in the healthy samples. Overall, these data will elucidate MF gene expression in SOC patients as well as the immunopathogenesis that results in varying presentations. This may facilitate the development of more concise diagnostic criteria and personalized targeted immunotherapies to better health outcomes within a minority population. Supported by grants from Dermatology Foundation DRSA (SD), the Skin of Color Society Research Grant (JMR), and the American Skin Association Milstein Research Scholar Award for Melanoma/Non-Melanoma Skin Cancer (ASB).

The Effects of Resveratrol Targeting MicroRNA-4325P/PDGF-B to Regulate Tumor Angiogenesis in Osteosarcoma Microenvironment

Objectives Here we sought to determine the specific mechanism by which resveratrol targets microRNA-4325p and PDGFB in an osteosarcoma (OS) cell line and investigate its effect on the OS cell line. Methods As a result of the study related to the effect of resveratrol on the conditioned media of microRNA-4325p mimic transfected MG-63 and HOS (a fibroblast and epithelial-like cell line) cells, we determined that resveratrol decreased survival, and growth, migration, and aortic ring formation of endothelial cells. A combination of miRNA mimics and HOS and MG-63 cell lines were transfected with inhibitors to explore the mechanisms behind angiogenesis formation induced by resveratrol. Results We found that resveratrol improved the activity of microRNA-4325p mimic transfection in OS, thus promoting angiogenesis, as demonstrated by angiogenic parameters and gene expression in OS. Following these results, it is evident that resveratrol can modulate angiogenesis by regulating microRNA-4325p expression by targeting PDGFB genes in OS cells. Conclusion The results demonstrate that resveratrol promotes tumor angiogenesis of OS cells through microRNA-4325p/PDGBB-mediated pathways and provides theoretical and experimental support for the discovery of new bioactive phytochemicals that are effective therapeutic agents in the prevention and treatment of OS diseases.

PDGF inhibits BMP2-induced bone healing

Bone regeneration depends on a pool of bone/cartilage stem/progenitor cells and signaling mechanisms regulating their differentiation. Using in vitro approach, we have shown that PDGF signaling through PDGFRβ inhibits BMP2-induced osteogenesis, and significantly attenuates expression of BMP2 target genes. We evaluated outcomes of treatment with two anabolic agents, PDGF and BMP2 using different bone healing models. Targeted deletion of PDGFRβ in αSMA osteoprogenitors, led to increased callus bone mass, resulting in improved biomechanical properties of fractures. In critical size bone defects BMP2 treatment increased proportion of osteoprogenitors, while the combined treatment of PDGF BB with BMP2 decreased progenitor number at the injury site. BMP2 treatment induced significant bone formation and increased number of osteoblasts, while in contrast combined treatment with PDGF BB decreased osteoblast numbers. This is in vivo study showing that PDGF inhibits BMP2-induced osteogenesis, but inhibiting PDGF signaling early in healing process does not improve BMP2-induced bone healing.

Comprehensive assessment of markers of apoptosis and cell proliferation during progression of atherosclerosis after surgery in patients with peripheral arterial disease.

Determination of predictors that can affect development of atherosclerosis progression in the postoperative period is an urgent problem in vascular surgery. Integrated assessment of markers of apoptosis and cell proliferation in atherosclerotic lesions and their progression after surgery in patients with peripheral arterial diseases. The investigation included 30 patients with stage IIB-III peripheral arterial disease. All patients have undergone open surgical interventions on the arteries of the aorto-iliac and femoral-popliteal segments. During these interventions, intraoperative specimens were obtained from the vascular wall with atherosclerotic lesions. The following values were evaluated: VEGF А165, PDGF BB, and sFas. Samples of normal vascular wall were obtained from post-mortem donors and used as a control group. The levels of Bax and p53 were increased (p<0.001) in samples from arterial wall with atherosclerotic plaque, while sFas values were reduced (p<0.001), compared to their levels in control samples. Values of PDGF BB and VEGF A165 were 1.9 and 1.7 times higher in atherosclerotic lesion samples (p=0.001), in comparison with the control group. The levels of p53 and Bax were increased against a background of reduced sFas levels in samples with progression of atherosclerosis compared to their baseline values in samples with atherosclerotic plaque (p<0.05). Initially increased values of the Bax marker against a background of reduced sFas values in vascular wall samples from patients with peripheral arterial disease is associated with risk of atherosclerosis progression in the postoperative period.

The transcriptome analysis of cleft lip/palate-related PTCH1 variants in GMSM-K cells show carcinogenic potential

Cancer progression involves the sonic hedgehog (SHH) pathway, in which the receptor PTCH1 actives the downstream pathways. Dysfunction of PTCH1 can lead to nevoid basal cell carcinoma Syndrome (NBCCs) including neoplastic disease and congenital disorder. To evaluate the relationship between PTCH1 and cancer, we applied the CRISPR/Cas9 system to knock out PTCH1 in oral nontumorous epithelial cells (GMSM-K). Then we screened six PTCH1 variants associated with cleft lip/palate (CL/P), one of the congenital disorders in NBCCs, and generated PTCH1 variant and wild-type recombinant PTCH1−/− GMSM-K cell lines. Transcriptome sequencing was conducted in these cell lines. The results revealed that differentially expressed genes (DEGs) in PTCH1−/− GMSM-K were enriched in extracellular compartments, contributing epithelial diseases by pathway enrichment analysis. RT-PCR confirmed that KRT34, KRT81, KRT86, PDGFB, and WNT10B genes, associated with extracellular compartments were highly expressed in PTCH1−/− . The Kyoto Encyclopedia of Genes and Genomes analysis also suggested that DEGs are closely related to focal adhesion, transcriptional misregulation, and proteoglycans in breast and gastric cancers. Comparative analysis of samples revealed that the CL/P-associated PTCH1 variants A443G and V908G are potentially carcinogenic. These findings provide new insights into the carcinogenic potential of PTCH1 dysfunction.

Влияние применения комбинированной антиоксидантной терапии на результаты эндоваскулярного лечения пациентов с облитерирующим атеросклерозом артерий нижних конечностей

INTRODUCTION: The number of endovascular interventions performed on the arteries of the lower extremities is growing annually in the world, as they provide faster patient recovery and shorten the period of hospital stay. However, the development of restenosis of the reconstruction zone often leads to a need for repeated operations. AIM: To evaluate the effectiveness of combined antioxidant therapy in reducing the risk of restenosis after endovascular interventions. MATERIALS AND METHODS: The study included 84 patients with IIB–III stage peripheral arterial disease (PAD). Group A included 45 patients who underwent endovascular interventions, group B included 39 patients who were prescribed combined antioxidant therapy (Vitamin E, Actovegin®) a week before endovascular intervention and within a week after. On admission to hospital, in the first hours of stay, in 1, 7 days after surgery, the amount of Bax and platelet growth factor (PDGF BB) biomarkers were determined in the blood serum of all the patients. On control visits, the patients underwent duplex scanning of the arteries of the lower extremities to measure the thickness of the neointima and to detect restenosis of the intervention area. RESULTS: When comparing Bax values in the first hours and days after surgery in groups A and B, its growth was noted in patients of the first group (p &lt; 0.01). At the same time, in patients of group A, the increase in PDGF BB was more pronounced (p &lt; 0.01) on the 7th day. In patients of group A with restenosis of the reconstruction zone, the values of Bax marker in the first hours after surgery exceeded those in patients of group B (р &lt; 0.01). On the 7th day, the amount of PDGF BB in these patients of group A was increased relative to group B (р &lt; 0.01). The changes in the number and dynamics of the studied markers in group B patients with restenosis of the reconstruction zone led, firstly, to decrease in the percentage of stenosis in the area of the previous surgical intervention, secondly, to decrease in the severity of the clinical manifestation of this complication, thirdly, to occurrence of restenosis of the reconstruction zone in the later period, and, fourthly, to decrease in the percentage of patients with this complication. CONCLUSION: The use of combined antioxidant therapy targeted at the system of apoptosis, can reduce the thickness of the neointima of the reconstruction zone and reduce the percentage of patients with restenosis after endovascular interventions.

Translational perspective on bone-derived cytokines in inter-organ communications

Translational perspective on bone-derived cytokines in inter-organ communications

Unilateral Primary Breast Dermatofibrosarcoma Protuberans with Negative COL1A1-PDGFB Fusion on Fluorescence In Situ Hybridization

Abstract Introduction/Objective Dermatofibrosarcoma protuberans (DFSP) is a superficial and locally aggressive cutaneous soft tissue sarcoma. In most patients, DFSP arises from the trunk and proximal extremities with breast being an uncommon location. Although they rarely metastasize (&amp;lt;5%), DFSP variants tend to recur locally. Most molecularly characterized cases have been defined by the translocation t(17;22)(q21.3;q13.1), which yields a fusion of the COL1A1 and PDGFB genes. In routine practice, dual fusion or break-apart fluorescence in situ hybridization (FISH) for the COL1A1-PDGFB fusion gene is the most widely used screening method for DFSP. However, in ~4% of typical DFSP cases, routine FISH screening is negative. In some of the DFSP cases negative for COL1A1-PDGFB fusion gene by FISH, RNA sequencing has identified two novel gene fusions, COL6A3-PDGFD and EMILIN2-PDGFD, with the former having a predilection for the female breast. Methods/Case Report We present a 32-year-old woman who underwent a biopsy for a 3.4 cm echogenic firm mass in her right breast. Histology showed a cytologically-uniform spindle cell neoplasm with fibroblastic morphology. The neoplastic cells displayed a storiform growth pattern and honeycomb-like infiltration of the adipose tissue. Immunohistochemically, the neoplastic cells expressed CD34 and vimentin and were negative for AE1/AE3, p63, S100, CD31, SMA, β-catenin, and STAT6. PDGFB break-apart FISH was negative. Based on the histologic and immunohistochemical findings, a diagnosis of DFSP was rendered. FISH for PDFGD rearrangements or RNA sequencing targeting PDGFD was not performed at this time. Results (if a Case Study enter NA) NA. Conclusion Awareness is warranted for the rare variant of DSFP with a predilection for female breast, which is characterized by a novel gene fusion not detectable by routine FISH used for DFSP screening.

Transcriptome changes in stages of non-alcoholic fatty liver disease.

BACKGROUNDNon-alcoholic fatty liver disease (NAFLD) is the most common chronic liver disease in the United States and globally. The currently understood model of pathogenesis consists of a ‘multiple hit’ hypothesis in which environmental and genetic factors contribute to hepatic inflammation and injury. AIMTo examine the genetic expression of NAFLD and non-alcoholic steatohepatitis (NASH) tissue samples to identify common pathways that contribute to NAFLD and NASH pathogenesis.METHODSWe employed the Search Tag Analyze Resource for Gene Expression Omnibus platform to search the The National Center for Biotechnology Information Gene Expression Omnibus to elucidate NAFLD and NASH pathology. For NAFLD, we conducted meta-analysis of data from 58 NAFLD liver biopsies and 60 healthy liver biopsies; for NASH, we analyzed 187 NASH liver biopsies and 154 healthy liver biopsies. RESULTSOur results from the NAFLD analysis reinforce the role of altered metabolism, inflammation, and cell survival in pathogenesis and support recently described contributors to disease activity, such as altered androgen and long non-coding RNA activity. The top upstream regulator was found to be sterol regulatory element binding transcription factor 1 (SREBF1), a transcription factor involved in lipid homeostasis. Downstream of SREBF1, we observed upregulation in CXCL10, HMGCR, HMGCS1, fatty acid binding protein 5, paternally expressed imprinted gene 10, and downregulation of sex hormone-binding globulin and insulin-like growth factor 1. These molecular changes reflect low-grade inflammation secondary to accumulation of fatty acids in the liver. Our results from the NASH analysis emphasized the role of cholesterol in pathogenesis. Top canonical pathways, disease networks, and disease functions were related to cholesterol synthesis, lipid metabolism, adipogenesis, and metabolic disease. Top upstream regulators included pro-inflammatory cytokines tumor necrosis factor and IL1B, PDGF BB, and beta-estradiol. Inhibition of beta-estradiol was shown to be related to derangement of several cellular downstream processes including metabolism, extracellular matrix deposition, and tumor suppression. Lastly, we found riciribine (an AKT inhibitor) and ZSTK-474 (a PI3K inhibitor) as potential drugs that targeted the differential gene expression in our dataset. CONCLUSIONIn this study we describe several molecular processes that may correlate with NAFLD disease and progression. We also identified ricirbine and ZSTK-474 as potential therapy.

New predictors of atherosclerosis progression after surgical interventions on the arteries of the lower extremities

BACKGROUNDː Thanks to experiments on animals, the influence of biomarkers of apoptosis, proliferation and endothelial dysfunction on the pathogenesis of atherosclerosis was proved. Aim: to identify the effect of these biomarkers on the progression of atherosclerosis in patients with peripheral arterial disease after surgery. METHODSː Patients were divided into two groups: A - 40 patients who have undergone endovascular interventions, B -38 patients, who have undergone hybrid surgery on the arteries of the lower extremities. Biomarkers Bax, sFas, PDGF BB, and nitric oxide (NO) were studied before intervention, on the 1st, 7th, 30th day after the operation. RESULTSː When comparing initial values of indicators and their values in the dynamics, the difference between group A and B was not found (p &gt; 0.05). Nine patients of group В developed progression of atherosclerosis after 17 months. Roc analysis showed that initial sFas ≤ 0,92 ng / ml, initial Вах ≥ 25,3 ng / ml, NO values ≤128.4 mmol / ml, sFas ≤ 0.645 ng / ml, Вах ≥ 33,8 ng / ml on the first postoperative day, and PDGF BB amount on the 7th day ≥ 28 ng / ml is associated with progression of atherosclerosis. CONCLUSIONSː The dynamics of biomarkers of apoptosis, proliferation and endothelium dysfunction in patients after endovascular and hybrid interventions did not differ significantly. The activation of two pathways markers of apoptosis against the background of reduced synthesis of NO leads to an enhanced proliferative response and is associated with progression of atherosclerosis after surgery.

Markers of the Development of the Reconstruction Area Restenosis After Endovascular Interventions in Lower Limbs Arteries

Restenosis of the reconstruction zone is one of the main postoperative complications of vascular reconstructions, which develops in 18–40% of cases and leads to an increase in the risk of limb loss to 20–25%. The search for new markers for predicting the development of this complication is an urgent problem today.Aim of study. To assess the dynamics of markers of apoptosis and cell proliferation at different times after endovascular intervention and their role in the development of restenosis of the reconstruction zone in patients with peripheral arterial disease (PAD).Material and methods. The clinical study included 30 patients with PAD stage III disease. After further examination, the patients underwent endovascular intervention on the arteries of the femoral-popliteal segment. If restenosis developed, the patients were re-operated. In patients before surgery, within the first hour, and then on days 1, 7, 14, 21, 30 after the operation, venous blood was taken to determine the markers Bax, sFas, PDGF BB using enzyme immunoassay.Results. After endovascular intervention, the first wave of apoptosis was triggered with an increase in the amount of proapoptotic protein Bax (p=0,00003) from 1 to 24 hours, followed by a decrease by day 7 (p=0.0008) compared to the amount on day 1. The PDGF BB level increased from day 1 after surgery (p=0.03) with maximum values on day 7 (p=0.0002) compared to the level on day 1. Then the second wave was triggered with a peak decrease in the level of the apoptosis inhibitor sFas on day 21 (p=0.002). After 9-12 months, restenosis of the intervention zone with a return of limb ischemia developed in 10 patients. During the first hour (p=0.004) in patients with restenosis, the level of Bax protein was significantly increased, with an increase in the level of PDGF BB by day 7 (p=0.011), and sFas by day 21 (p=0.0001), PDGF BB by the end of 1 month (p=0.004) compared to values in patients without this complication.Conclusion. 1. Endovascular intervention causes two waves of apoptosis in the postoperative period. The first wave is associated with an increase in Bax protein in the first hours, followed by an increase in PDGF BB on day 7. The second wave of apoptosis is due to a decrease in the inhibitor of apoptosis — sFas for 21 days against the background of the shift of the PDGF BB to the initial level. 2. An increase in Bax protein within the first hours after surgery in the course of PDGF BB growth on day 7 with an increased amount of sFas on day 21 leads and PDGF BB by the end of 1 month leads to the development of restenosis of the intervention area.

Advanced platelet-rich fibrin promotes the paracrine function and proliferation of adipose-derived stem cells and contributes to micro-autologous fat transplantation by modulating HIF-1α and VEGF

BackgroundThe micro-autologous fat transplantation (MAFT) technique has demonstrated its feasibility in multiple medical fields, such as facial rejuvenation. Advanced platelet-rich fibrin (APRF), an autologous platelet concentrated on a fibrin membrane without added external factors, has shown significant potential for tissue restoration. However, the role of APRF in the modulation of MAFT remains unclear. Here, we aimed to explore the effect of APRF on MAFT.MethodsAdipose-derived stem cells (ASCs) were isolated from human gastric subcutaneous fat and treated with APRF. ELISA assays measured cytokines. The proliferation of ASCs was analyzed by CCK-8 assays. The levels of hypoxia-inducible factor-1α (HIF-1α), heat shock protein 70 (HSP70), insulin like growth factor 2 (IGF-2), interleukin-6 (IL-6), interleukin-8 (IL-8), and vascular endothelial growth factor (VEGF) were measured by ELISA assays, quantitative reverse transcription-PCR (qRT-PCR), and Western blot analysis. The effect of APRF/HIF-1α/VEGF on MAFT in vivo was analyzed in Balb/c nude mice. The BALB/c mice were subcutaneously co-transplanted with fat, APRF, and control shRNA, HIF-1α shRNA, or VEGF shRNA into the dorsal area. The serum and protein levels of the above cytokines were analyzed by ELISA assays and Western blot analysis. Lipid accumulation was measured by Oil Red O staining. The expression of CD34 was assessed by immunohistochemical staining.ResultsAPRF continuously secreted multiple cytokines, including epidermal growth factor (EGF), FGF-2, insulin like growth factor 1 (IGF-1), interleukin-1beta (IL-1β), interleukin-4 (IL-4), platelet-derived growth factor alpha polypeptide b (PDGF-AB), platelet-derived growth factor beta polypeptide b (PDGF-BB), transforming growth factor-beta (TGF-β), and VEGF. APRF was able to promote the proliferation of ASCs. APRF dose-dependently activated the expression of HIF-1α, HSP70, IGF-2, IL-6, IL-8, and VEGF in ASCs. APRF regulated the paracrine function of ASCs by modulating HIF-1α and VEGF. APRF increased the survival of MAFT by modulating HIF-1α and VEGF in vivo.ConclusionsAPRF promotes the paracrine function and proliferation of ASCs and contributes to MAFT by modulating HIF-1α and VEGF. Our findings provide new insights into the mechanism by which APRF regulates MAFT.

The effect of the geometry of the proximal anastomosis, markers of apoptosis and cell proliferation on the long-term patency following reconstructive interventions on the femoropopliteal arterial segment

Aim. To study changes in the topography of the orifice of the deep femoral artery (DFA), markers of proliferation, and apoptosis in patients after open interventions on the femoropopliteal arterial segment.&#x0D; Methods. The study included 35 patients with atherosclerotic peripheral arterial disease (PAD), femoral-popliteal occlusion, stage IIBIII of the disease according to the classification of A.V. PokrovskyFontaine, who underwent open surgery. The average age of the patients was 694.6 years. These patients included 26 men. Patients were divided into two groups: group A included 18 patients who underwent femoral-popliteal prosthetics (distal End-To-End bypass anastomoses), group B included 17 patients with femoral-popliteal bypass surgery (distal End-To-Side bypass anastomoses). The groups were comparable in terms of age and disease severity (p 0.05). Determination of serum platelet-derived growth factor BB (PDGF BB) and soluble form Fas (sFas) levels was carried out immediately before the intervention, on the 1st, 7th days, and 1 month after the operation. Duplex scanning (DS) was performed on day 7, after 1 and 18 months. Statistica 10.0 software was used for statistical data processing. The significance of differences between unrelated samples was assessed using the Student's t-test. The correlations between variables were analyzed by using Pearson's method.&#x0D; Results. On 1st day, there was a decrease in soluble Fas in patients of group A compared with group B (0.41 ng/ml vs 0.78 ng/ml, p=0.01). On the 7th day, the levels of serum platelet-derived growth factor BB were increased in patients of group A compared with group B (35.2 ng/ml vs 23.2 ng/ml, p=0.00001). After 1 month, the level of serum platelet-derived growth factor BB in patients of group A remained elevated compared with those in patients of group B (22.8 ng/ml vs 14.4 ng/ml, p=0.0003).&#x0D; Conclusion. Femoropopliteal prosthetics leads to a change in branching angle of the deep femoral artery up to 7080%, accompanied by changing dynamics of apoptotic markers and cell proliferation, leading to an increase in the thickness of neointimal hyperplasia and the progression of atherosclerosis.

Why PRP works only on certain patients with tennis elbow? Is PDGFB gene a key for PRP therapy effectiveness? A prospective cohort study

BackgroundThere is variability in individual response to platelet-rich plasma (PRP) therapy in tennis elbow treatment. Genetic variation, especially within genes encoding growth factors may influence the observed inter-individual differences. The purpose of this study was to identify polymorphic variants of the platelet-derived growth factor beta polypeptide gene (PDGFB) that determine an improved individual response to PRP therapy in tennis elbow patients.MethodsThis prospective cohort study was designed in accordance with STROBE and MIBO guidelines. A cohort of 107 patients (132 elbows, 25 bilateral) was studied, including 65 females (77 elbows) and 42 males (55 elbows), aged 24–64 years (median 46.00 ± 5.50), with lateral elbow tendinopathy treated with autologous PRP injection. The effectiveness of PRP therapy was recorded in all subjects at 2, 4, 8, 12, 24 and 52 weeks after PRP injection using the Visual Analog Scale (VAS), quick version of Disabilities of the Arm, Shoulder and Hand score (QDASH) and Patient-Rated Tennis Elbow Evaluation (PRTEE). In order to determine the PDGFB variants with the best response to PRP therapy, patient reported outcome measures were compared between individual genotypes within studied polymorphic variants (rs2285099, rs2285097, rs2247128, rs5757572, rs1800817 and rs7289325). The influence of single nucleotide polymorphisms on blood and PRP parameters, including the concentration of PDGF-AB and PDGF-BB proteins was also analyzed.ResultsOur analysis identified genetic variants of the PDGFB gene that lead to a better response to PRP therapy. The TT (rs2285099) and CC (rs2285097) homozygotes had higher concentration of platelets in whole blood than carriers of other genotypes (p = 0.018) and showed significantly (p < 0.05) lower values of VAS (weeks 2–12), QDASH and PRTEE (weeks 2–24). The rs2285099 and rs2285097 variants formed strong haplotype block (r2 = 98, D’=100). The AA homozygotes (rs2247128) had significantly lower values of VAS (weeks 4–52), QDASH and PRTEE (weeks 8, 12).ConclusionsPDGFB gene’s polymorphisms increase the effectiveness of PRP therapy in tennis elbow treatment. Genotyping two polymorphisms of the PDGFB gene, namely rs2285099 (or rs2285097) and rs2247128 may be a helpful diagnostic tool while assessing patients for PRP therapy and modifying the therapy to improve its effectiveness.

The level of markers of apoptosis and cell proliferation in the area of restenosis after lower extremity arterial reconstruction

Aim. To assess the number of markers of apoptosis and cell proliferation, as well as their relationships in the area of restenosis of arterial reconstructions.&#x0D; Methods. The study included 14 patients with a diagnosis of arteriosclerosis obliterans of the lower extremities. Post-thrombotic occlusion of femoropopliteal bypass. All patients were males with stage III disease according to the Fontaine classification modified by A.V. Pokrovsky. The average age of the patients was 653.4 years. The mean disease duration was 92.5 months after the initial intervention. Intraoperative material distal anastomosis of femoropopliteal bypass was taken from patients during arterial reconstructions. As a control, we used arterial wall samples obtained at organ procurement from postmortem donors without arteriosclerosis obliterans of the lower extremities. The number of samples is 8. The site of their collection is the popliteal artery. After sampling, they were crushed, and a homogenate was prepared, followed by the determination of the amount of p53, PDGF BB, Bcl2, and Bax proteins using the enzyme immunoassay. Statistical analysis was performed using the Statistica 10.0 software. Group differences were assessed by using the MannWhitney test. Correlation coefficients were determined using the Spearman test. Data are presented as medians and interquartile ranges.&#x0D; Results. In tissue samples of restenosis, the amount of p53 protein was 0.07 units/mg and was significantly reduced compared with the control samples 0.14 units/mg (р=0,015). The amount of platelet-derived growth factor PDGF BB was 0.17 ng/mg (р=0.05), Bcl2 1.61 ng/mg (р=0.008), Bax 6.0 ng/mg (р=0.25) in the restenosis area and was increased in comparison with the control samples (0.04 ng/mg, 0.9 ng/mg, 4.4 ng/mg, respectively). A relationship between p53 and platelet-derived growth factor BB (r=0.724, p=0.002), platelet-derived growth factor BB and Bcl2 (r=0.672, p=0.003) was revealed in samples from restenosis tissue obtained during arterial reconstructions.&#x0D; Conclusion. The decreased apoptosis, expressed in a low level of p53 protein, with an increased Bax/Bcl-2 ratio is associated with an increase in the proliferative response of vascular wall cells in the area of restenosis of arterial reconstruction.