Levels Of PDGF Research Articles

Pazopanib based oral metronomic therapy for platinum resistant/refractory epithelial ovarian cancer: A phase II, open label, randomized, controlled trial

BackgroundTreatment of patients with platinum resistant/refractory epithelial ovarian cancer (EOC) is an unmet need. We evaluated the role of oral metronomic therapy in this setting. Patients and methodsBetween October 2017 and September 2019 seventy five patients with platinum resistant/refractory EOC were enrolled. Patients received oral etoposide (50 mg, day 1 to 14, cyclophosphamide 50 mg, day 1 to 28, every 4 weeks (Arm A, n = 38). Patients in Arm- B (n = 37) received Pazopanib (400 mg once daily) in addition to etoposide and cyclophosphamide. Quality of life (QoL) was evaluated using the EORTC questionnaire. Serum VEGF and PDGF were estimated at baseline, after 3rd and 6th cycle. The primary endpoint was progression free survival (PFS). Secondary endpoints were overall survival (OS), toxicity and QoL. ResultsPatients characteristics were well matched. Median PFS was higher in arm B, 5.1 months (95% CI 3.13 to10.33) compared to 3.4 months (95% CI 3.0 to 6.53) in arm A, p = 0.045. Median OS has ‘not reached’ in Arm B compared to 11.2 months (95% CI, 5.66 - not reached) in arm A, p = 0.032. Therapy was tolerated well; oral mucositis (p = 0.36) and fatigue (p = 0.08) being more in arm B. QoL assessment revealed modest improvement in ‘symptom scales’ in Arm B. Serum VEGF and PDGF levels decreased with therapy in both arms (Arm A-p < 0.0001, Arm B-p < 0.016). ConclusionAddition of pazopanib to etoposide and cyclophosphamide could be a novel oral combination for metronomic therapy for platinum resistant/refractory EOC. Trial registrationCTRI/2017/10/010219.

A20 Attenuates Lipopolysaccharide-Induced Inflammation Through MAPK/ERK/JNK Pathway in LX-2 Cells

Background: Hepatic stellate cells (HSCs) are liver-specific pericytes that transform into myofibroblasts, which are involved in pathological vascularization in liver fibrosis. We previously suggested that A20 overexpression suppresses lipopolysaccharide (LPS)-induced inflammation in HSC. We aimed to determine the mechanisms of the anti-inflammatory role of A20 in LX-2 cells. Methods: LX-2 cells were transfected with A20-siRNA or control-siRNA and control adenovirus or A20-carrying adenovirus. Quantitative reverse transcription PCR (RT-qPCR) analysis was employed to quantify mRNA levels of α-SMA, col-I, col-III, IL-6, TGF-β, and PDGF in A20-siRNA LX-2 cells stimulated with LPS. Multiple molecular indices of MAPK/ERK/JNK signal pathway were performed by using Western blotting. Results: Relative to control, the fibrosis-related mRNA levels of α-SMA, col-I, and col-III were increased in A20-siRNA LX-2 cells. Meanwhile, A20-siRNA cells significantly increased IL-6, TGF-β, and PDGF mRNA levels. Relative to controls, stimulating A20 overexpressing LX-2 cells with LPS for 5 and 30 minutes significantly reduced the levels of phosphorylated ERK and JNK, respectively. A20 knockdown in LX-2 cells promotes phosphorylated ERK and JNK levels with LPS for 30 minutes. Conclusions: Our data indicate that A20 could be functional in HSCs through the MAPK/ERK/JNK signaling pathway, highlighting a potential novel therapeutic strategy against liver fibrosis.

The Protective Effects of Nebulized Nano-Curcumin Against Bleomycin-Induced Pulmonary Fibrosis in Rats

Background: Idiopathic pulmonary fibrosis (IPF) is a progressive fatal disease affecting the lung, and currently there is no efficient therapy for this condition. Curcumin, as a natural anti-inflammatory and antioxidant agent, could repress the pulmonary fibrosis (PF) caused by Bleomycin (BLM). Objectives: The aim of the research was to evaluate the protective activity of a nano-formulation of curcumin administered by inhalation on BLM-induced PF in rats. Methods: Eighty rats were randomly divided into eight experimental groups. Group one (control) that received saline intratracheally (IT) and subjected to vehicle inhalation. Group two to eight each received a single dose of BLM (5 IU/kg, IT) along with vehicle inhalation, oral prednisolone, oral curcumin, curcumin inhalation, and nano-curcumin inhalation with the doses of 50, 100, and 200 µg/kg, respectively. In the control and other groups, BLM was injected intratracheally on the first day of the experiment. In the treatment groups, curcumin suspension was prepared in distilled water and applied through nebulization for 21 consecutive days after BLM intratracheal administration. Then the rats were euthanized, and inflammatory cytokines (TNF-α, TGF-β, PDGF), hydroxyproline, and IL-10 (as a protective cytokine) were measured. Also, lung histopathological features were examined. Results: The synthesized nano-formulation reduced the overall hydroxyproline content of lungs in BLM-treated rats (P &lt; 0.002). In addition, TNF-α, TGF-β, and PDGF levels significantly increased in the lungs of BLM-instilled rats (P &lt; 0.001). However, the nano-formulation of curcumin (200 µg/kg) significantly decreased the levels of these inflammatory cytokines (P &lt; 0.001) and increased IL-10 level (P = 0.0144) compared with the control group. Conclusions: The nebulization of nano-curcumin is suggested as a novel approach for the treatment of PF induced by BLM in rats. Our findings revealed that the inhalation (as a safe local drug delivery system approach) of the nano-curcumin at a dose of 200 µg/kg (formulated by cyclodextrin) could effectively protect the lung against PF.

Activation of platelet-rich plasma by pulse electric fields: Voltage, pulse width and calcium concentration can be used to control and tune the release of growth factors, serotonin and hemoglobin.

Activated platelet-rich plasma (PRP) has been used in the clinical settings of wound healing and regenerative medicine, with activation typically induced by the addition of bovine thrombin. To eliminate issues with availability, cost and potential side effects associated with bovine thrombin, ex vivo PRP activation using pulse electric fields (PEF) has been proposed and demonstrated. The present study characterizes the effect of PEF voltage and pulse width, in combination with a range of calcium concentrations, on clot formation, growth factor release, and serotonin (5-HT) release from dense granules. The main findings are: 1) increasing calcium concentrations with most PEF conditions leads to increased levels of PDGF and 5-HT release; 2) whether EGF levels increase or decrease with increasing calcium concentration depends on the specific PEF parameters; 3) the pattern of PDGF and EGF levels in supernatants suggest that these molecules are localized differently within platelets; 4) significant levels of PDGF, EGF, and 5-HT can be released without inducing clot formation or hemoglobin release. In conclusion, voltage, pulse width and calcium concentration can be used to control and tune the release of growth factors, serotonin and hemoglobin from PEF-activated PRP. Because growth factor requirements vary for different types of wounds and for wounds at different stages of healing, the unique balance of factors in supernatants of PEF-activated PRP may provide more clinically advantageous than the current standard of bovine thrombin-activated PRP.

Effect of l-carnitine on cardiotoxicity and apoptosis induced by imatinib through PDGF/ PPARγ /MAPK pathways

A tyrosine kinase inhibitor Imatinib (IM) is used in the treatment of different varieties of cancers. The current study was designed to explore the beneficial role of l-carnitine against IM-induced cardiotoxicity in rats. Male albino rats received IM (40 mg/kg, i.p.) either alone or/in combination with l-carnitine (100 mg/kg, i.p.) for 7 days. IM increased serum inflammatory cytokines, concomitant with activation of cardiac MAPK, α-SMA, malondialdehyde (MDA) and nitric oxide(NO), decreased cardiac peroxisome proliferator-activated receptor-γ (PPAR-γ) level, superoxide dismutase (SOD) activity, and glutathione (GSH) content. The expression levels of Bcl-2 and PDGF were significantly decreased, while the expression levels of CTGF and BAX were significantly increased in the IM group. The l-carnitine treatment successfully protected the heart as indicated by the improvement of the biochemical and histopathological parameters. l-carnitine didn't affect the serum concentration of IM and increased intracellular concentration in the combination-treated group as measured by the mass spectrometer. Conclusion: l-carnitine abrogated IM-induced cardiac damage and apoptosis via PDGF/PPARγ/MAPK pathways.

Use of conditioned media (CM) and xeno-free serum substitute on human adipose-derived stem cells (ADSCs) differentiation into urothelial-like cells.

BackgroundCongenital abnormalities, cancers as well as injuries can cause irreversible damage to the urinary tract, which eventually requires tissue reconstruction. Smooth muscle cells, endothelial cells, and urothelial cells are the major cell types required for the reconstruction of lower urinary tract. Adult stem cells represent an accessible source of unlimited repertoire of untransformed cells.AimFetal bovine serum (FBS) is the most vital supplement in the culture media used for cellular proliferation and differentiation. However, due to the increasing interest in manufacturing xeno-free stem cell-based cellular products, optimizing the composition of the culture media and the serum-type used is of paramount importance. In this study, the effects of FBS and pooled human platelet (pHPL) lysate were assessed on the capacity of human adipose-derived stem cells (ADSCs) to differentiate into urothelial-like cells. Also, we aimed to compare the ability of both conditioned media (CM) and unconditioned urothelial cell media (UCM) to induce urothelial differentiation of ADCS in vitro.MethodsADSCs were isolated from human lipoaspirates and characterized by flow cytometry for their ability to express the most common mesenchymal stem cell (MSCs) markers. The differentiation potential was also assessed by differentiating them into osteogenic and adipogenic cell lineages. To evaluate the capacity of ADSCs to differentiate towards the urothelial-like lineage, cells were cultured with either CM or UCM, supplemented with either 5% pHPL, 2.5% pHPL or 10% FBS. After 14 days of induction, cells were utilized for gene expression and immunofluorescence analysis.ResultsADSCs cultured in CM and supplemented with FBS exhibited the highest upregulation levels of the urothelial cell markers; cytokeratin-18 (CK-18), cytokeratin-19 (CK-19), and Uroplakin-2 (UPK-2), with a 6.7, 4.2- and a 2-folds increase in gene expression, respectively. Meanwhile, the use of CM supplemented with either 5% pHPL or 2.5% pHPL, and UCM supplemented with either 5% pHPL or 2.5% pHPL showed low expression levels of CK-18 and CK-19 and no upregulation of UPK-2 level was observed. In contrast, the use of UCM with FBS has increased the levels of CK-18 and CK-19, however to a lesser extent compared to CM. At the cellular level, CK-18 and UPK-2 were only detected in CM/FBS supplemented group. Growth factor analysis revealed an increase in the expression levels of EGF, VEGF and PDGF in all of the differentiated groups.ConclusionEfficient ADSCs urothelial differentiation is dependent on the use of conditioned media. The presence of high concentrations of proliferation-inducing growth factors present in the pHPL reduces the efficiency of ADSCs differentiation towards the urothelial lineage. Additionally, the increase in EGF, VEGF and PDGF during the differentiation implicates them in the mechanism of urothelial cell differentiation.

Evaluation of Platelet Rich Fibrin Obtained Using Different Centrifugation Parameters as a Tool for Regenerative Medicine.

Platelet-rich fibrin (PRF) is a biomaterial widely used in the field of regenerative medicine. The purpose of this work was to analyze the structure and biomolecular characteristics of PRF through nine centrifugation parameters (CP) for its preparation, using a pool of blood samples of five volunteers. The PRF obtained was analyzed by morphological and histological characteristics, as well as electronic and atomic force microscopy and growth factors determinations. A longer time of centrifugation showed taller clots and denser mesh fibrin in comparison with a short time (p < 0.05). The protocols with higher speed of centrifugation showed higher levels of PDGF-BB and VEGF. Higher levels of TGFβ1 were found in protocols with a shorter centrifuge time. The mean platelet count (916.05 ± 23.73 cells x 103 cells x cm3) and its roughness (Ra) (616.5 ± 45.2 nm) did not show significant differences between different CP (p > 0.05). A significant correlation between fibrin density and levels of PDGF (r = 0.57) and VEGF (r = 0.52) was found. Additionally, the size of the clot had a significant correlation (r = -0.47) with TGFβ1 levels. Different centrifugation parameters to obtain PRF have been reported. These results indicate that changes in the conditions to obtain PRF have a significant impact on their fibrin structure, cellular distribution, and biomolecular content, which can be decisive for its choice in the different clinical applications to be used. It is necessary to use a standardized centrifuge and protocol to guarantee high-quality PRF and clinical outcomes with less variability.

Randomized Clinical Trial: Comparing In-Vitro Release of Growth Factors from Combination of Hyaluronic Acid and Advanced Platelet Rich Fibrin (A-PRF) Gel

Liquid fibrinogen (Advanced platelet-rich fibrin – A-PRF) is an injectable platelet concentrate rich in platelets, leukocytes, and fibrinogen obtained by blood centrifugation. Hyaluronic acid (HA) had been known to induce growth factor release from PRF. This study aimed to analyze and assess the in-vitro release of different growth factors from the combination of HA + A-PRF at different times. The concentration of platelet-derived growth factor-AB (PDGF-AB) and vascular endothelial growth factor (VEGF) released by liquid fibrinogen were examined with ELISA at three-time points (D-0, D-3, D-14). Twenty DFU subjects were involved in this study and randomized into HA + A-PRF group and A-PRF only group. A-PRF was obtained by peripheral blood centrifugation. HA+ A-PRF was prepared by homogenizing PRF and HA with a ratio of 1:0.6. PDGF-AB and VEGF were quantified in HA + A-PRF and the supernatant serum after freezing it in 80 o C while protein profiles were determined by SDS-PAGE. Fibrin gel VEGF level was found to be significantly higher in HA + A-PRF compared PRF alone (on D-3, p=0.010, and D-7, p= 0.043). The PDGF level was found to be significantly higher in fibrin gel on D-3 (p=0.049) and D-7 (p=0.023). Observation of the fibrin gel’s density revealed that the HA + A-PRF group showed a longer melting time compare with A-PRF group (p =0.002). The combination of HA + PRF results in better induction of growth factor release, denser fibrin gel, and better retention of growth factors compared to PRF alone. Funding: This study was funded by Medical Science Doctoral Programme, Universitas Indonesi Ethical Approval: The study has been approved by The Ethics Committee of the Faculty of Medicine Universitas Indonesia ID 0855/UN2.F1/ETIK/2018.

Влияние курса нейромодуляции на профиль нейротрофических факторов у пациентов с хроническими нарушениями сознания

Transcranial magnetic stimulation (TMS) is one of rehabilitation approaches for patients with chronic disorders of consciousness (DOC). The aim of our study was to assess neurotrophic factors and the changes of those after TMS course in patients with chronic DOC. We enrolled 26 patients with chronic DOC of various etiology and 21 heathy volunteers. Blood serum and cerebrospinal fluid (CSF) were collected from all patients before and after the TMS course, the levels of BDNF, NSE, NGF, РDGF, GDNF and NT3 were assessed in the biomaterial. The blood BDNF, NSE, PDGF, GDNF and NT3 in patients with chronic DOC were higher compared to healthy volunteers (p &lt; 0.05). We found no correlations between the type of DOC and neurotrophic factors concentrations in blood and CSF. The CSF level of BDNF in patients after traumatic brain injury (TBI) was higher compared to patients with non-traumatic chronic DOC (p &lt; 0.05). We also found the increase of CSF BDNF after the TMS course in patients after TBI (p &lt; 0.05). No other significant differences between groups and another blood and cerebrospinal fluid biomarker levels were detected. Thus, the serum BDNF, NSE, PDGF, GDNF and NT3 levels in patients with chronic DOC were higher compared to healthy volunteers. The BDNF level in CSF was higher in patients with traumatic DOC, and it also increased after the course of high-frequency TMS in this group. This fact may indicate the long-term neuronal plasticity processes in patients after TBI, as well as more favorable rehabilitation prognosis.

Improved growth factor delivery and cellular activity using concentrated platelet-rich fibrin (C-PRF) when compared with traditional injectable (i-PRF) protocols.

Several studies have recently demonstrated that only marginal improvements in platelet and leukocyte concentrations are achieved following standard injectable platelet-rich fibrin (i-PRF) protocols. Due to these previous findings, a novel harvesting technique was recently developed to collect higher concentrations of platelets/leukocytes specifically from the buffy coat layer (C-PRF) following faster centrifugation protocols. The aim of this study was to investigate the regenerative properties and effects on growth factor release and cellular activity of PRF collected through this novel harvesting technique compared to standard i-PRF protocols. The upper 1-ml layer collected through standard i-PRF protocols at low centrifugation speeds was compared with 1mL of C-PRF collected from the buffy coat layer following high centrifugation protocols (3000×g for 8min on a horizontal centrifuge) to specifically concentrate cells within the platelet/leukocyte-rich buffy coat layer. Thereafter, the expression of seven different growth factors, including PDGF-AA, PDGF-AB, PDGF-BB, TGF-β1, VEGF, IGF-1, and EGF, was characterized for up to 10days. Then, gingival fibroblast biocompatibility was investigated at 24h (live/dead assay); migration was investigated at 24h; proliferation was investigated at 1, 3, and 5days; and the expression of PDGF and TGF-β was investigated at 3days. Collagen 1 immunostaining was also quantified at 14days. At all investigated time periods, a significant increase in growth factor release was observed in C-PRF. In particular, the release of PDGF-AA, TGF-β1, and EGF exhibited the highest increases when compared with that in i-PRF. While both i-PRF and C-PRF exhibited high biocompatibility and induced significantly higher fibroblast migration and proliferation when compared with that of the control tissue culture plastic group, C-PRF showed the greatest potential for cell migration and proliferation. Furthermore, C-PRF induced significantly higher mRNA levels of TGF-β and PDGF levels at 3days and greater collagen 1 staining when compared with induced by i-PRF. In the present study, it was found that C-PRF collected specifically from the buffy coat layer following higher centrifugation protocols exhibited an up to a threefold increase in growth factor release when compared with that exhibited by standard i-PRF. This significantly promoted higher gingival fibroblast migration, proliferation, gene expression, and collagen I synthesis. The findings of the present study demonstrate that a more potent formulation of liquid platelet concentrate than that obtained from the upper plasma layer following a short and slow centrifugation protocol (i-PRF protocol) can be obtained for clinical use by specifically harvesting cells in the platelet- and leukocyte-rich buffy coat layer following an 8-min 3000×g centrifugation protocol (C-PRF protocol).

Upregulation of PDGF Mediates Robust Liver Regeneration after Nanosecond Pulsed Electric Field Ablation by Promoting the HGF/c-Met Pathway.

Nanosecond pulsed electric field (nsPEF) has emerged as a promising tool for hepatocellular carcinoma ablation recently. However, little is known about how nsPEF affects liver regeneration while being applied to eliminate liver lesions. Besides, the impact of nsPEF ablation on liver function should also be taken into consideration in the process. In this paper, we study the impact of nsPEF ablation on liver function by the measurement of serum levels of AST and ALT as well as liver regeneration and relevant molecular mechanisms in vivo. We found that mouse liver function exhibited a temporary injury without weight loss after ablation. In addition, local hepatic nsPEF ablation promoted significant proliferation of hepatocytes of the whole liver with an increase in HGF level. Moreover, the proliferation of hepatocytes was dramatically inhibited by the inhibitor of c-Met. Of interest, the periablational area is characterized by high level of PDGF and a large amount of activated hepatic stellate cells. Furthermore, neutralizing PDGF was able to significantly inhibit liver regeneration, the increased HGF level, and the accumulation of activated HSCs. Our findings demonstrated that nsPEF not only was a safe ablation approach but also could stimulate the regeneration of the whole liver through the activation of the HGF/c-Met pathway by upregulation of PDGF within the periablational zone.

Effects of mesenchymal stem cell and amnion membrane transfer on prevention of pericardial adhesions

Abstract Background To investigate and compare the antiadhesive/antifibrotic effects of mesenchymal stem cells (MSC) and amnion membrane transfer (AMT) in a rat model. Material and methods Three experimental and sham groups were formed using 30 Wistar-Albino rats. AMT and MSC were applied to the related groups. The control group was not treated. After 12 weeks follow-up, intracardiac blood and cardiac-pericardiac tissue samples were taken. The severity of adhesions and fibrosis were scored macroscopically and microscopically with Hematoxylin/Eosin and Masson’s trichrome staining. TNF-α, TGF-β, IL-1, PDGF, FGF, VEGF and Caspase-3 levels were measured with the ELISA method. Results Severe adhesions were observed in the AMT and control groups, but no adhesion was present in the MSC group. Pericardial thickness, increased vascularity, fibrosis, and collagen accumulation were similar between control and AMT groups, but were less in Sham and MSC groups. Between MSC and AMT groups, only Caspase-3 level was different, which is an apoptosis marker. Conclusion The positive effects of MSC on adhesion, which we achieved in our study, suggest that it may prevent adhesion. AMT did not provide a positive effect. The correlation of Caspase-3 with postoperative adhesion/fibrosis should be examined in more detail.

Antiangiogenesis Roles of Exosomes with Fei-Liu-Ping Ointment Treatment are Involved in the Lung Carcinoma with the Lewis Xenograft Mouse Model.

Exosomes display efficient biocompatibility and represent valuable vehicles for drug or effective material delivery in a tumour-therapeutic approach. Following treatment with Fei-Liu-Ping (FLP) ointment, a traditional Chinese herbal formula, which is used for treating lung cancer patients, could inhibit lung carcinoma growth in clinical and animal studies. In the present study, the values of VEGF and PDGF, which were closely related to angiogenesis, were estimated in serum and carcinoma tissue exosomes to unveil the FLP effects on angiogenesis. The common inflammatory factors of IL-6, IL-1β, TNF-α, and TGF-β in serum exosomes were also detected with the Lewis xenograft model. Methods. Male C57BL/6 mice were randomly divided into four groups, namely, normal, model, cyclophosphamide (CTX), and FLP treatment groups. Histological structures were observed and imaged by H&E. CD31 expressions in tumour tissues were detected by immunofluorescence (IF) and western blot (WB). VEGF, PDGF, and PDGFR levels in exosomes, serum, tumour, and lung tissues were detected by enzyme-linked immunosorbent assay (ELISA), immunohistochemistry (IHC), and WB, respectively. IL-6, IL-1β, TNF-α, and TGF-β levels in exosomes were measured by multiplex immunoassay panels. Results. The results showed that FLP had tumour growth inhibition rate (39.31%). CD31 protein expression was obviously decreased in tumour tissues of CTX- and FLP-treated MO mice, compared to that of MO mice (P < 0.05 or P < 0.001). VEGF, PDGF, and PDGFR expression levels with FLP treatment were downregulated in exosomes, serum, tumour, and lung tissues compared to model group (P < 0.05 or P < 0.01). The expressions of IL-6, IL-1β, and TNF-α were downregulated in exosomes compared to the model group (P < 0.05 or P < 0.01). Conclusions. This study suggested that FLP had the ability of inhibiting tumourigenesis in a Lewis lung xenograft mouse model, whose therapeutic mechanisms might relate with the downregulation of angiogenesis factor and tumour inflammatory cytokines levels.

Effect of Resistance Training and Berberine Chloride Supplementation on IGF-1, PDGF, and VEGF Gene Expression in Hippocampal Tissue of Diazinon-Poisoned Rats

Background: One of the most common agricultural pesticides is organophosphorus pesticides. Diazinon is one of the most widely used organophosphorus pesticides used in the fight against pesticides in the country. Much research has recently been done on the impact of exercise on brain function and several biological mechanisms have been proposed for the effects of exercise and physical activity on brain function. It is said that resistance training increases neuronal plasticity due to an increase in insulin-like growth factor-1 (IGF-1). Research has shown that IGF-1 can cross the blood-brain barrier and play an important role as a neurotrophic factor, especially in the hippocampus, and activate various important central mechanisms related to cognitive processes. Exercise also affects the expression of angiogenic genes such as PDGF and VEGF. Methods: We randomly divided 40 male Wistar rats weighing 250 ± 50 g into eight groups of five and treated them with diazinon, berberine chloride, and resistance training. The groups were treated for five weeks. At the end of the fifth week, the levels of IGF-1, PDGF, and VEGF gene expression in hippocampal tissue homogenates were measured using Quantitative real-time PCR. Data were analyzed using ANOVA and Tukey’s post hoc test. Results: The results showed that resistance training and berberine chloride significantly increased the levels of IGF-1, PDGF, and VEGF gene expression in poisoned rats. Conclusions: Based on the results of this study, resistance training and berberine supplementation have significant effects on reducing the harmful effects of diazinon toxin (P < 0.05) and increasing IGF-1, PDGF, and VEGF gene expression in poisoned rats.

The Effect of Platelet Rich Fibrin (PRF) and Bone Graft (Carbonate Apatite) Applications on Platelet Derived Growth Factor-BB Post Flap Surgery in Patients with Chronic Periodontitis

Chronic periodontitis characterized by a periodontal pocket over 5 mm, alveolar bone resorption accompanied by tooth mobility, requires periodontal surgical treatment. Conventional surgical treatment can not achieve the regeneration of periodontal tissue and thus requires regenerative procedures, such as with the addition of PRF. PDGF-BB is a growth factor that plays an important role in wound healing. Developments in additional bioactive ingredients in surgical procedures are used to regulate inflammation and increase the speed of the healing process. The purpose of this study was to determine the effect of PRF and bone graft (carbonate apatite) on PDGF-BB levels post-surgical flap in patients with chronic periodontitis. The experimental study with open label and consecutive sampling was conducted on 10 subjects of chronic periodontitis who received surgical flap treatment with bone graft and PRF; and 10 subjects without PRF. GCF samples were collected immediately before surgery and 7 days post flap surgery for examination of PDGF-BB levels by ELISA method. The data were calculated statistically by Mann-Whitney test (p &lt;0.05). The results showed that the mean difference of PDGF-BB level in PRF group was 9.44 pg/ml (-1.20 – 37.79 pg/ml) with p value (0.013) and Non-PRF group was 7.97 pg/ml (-6.62 - 44.42 pg/ml) with p value (0.059). The p value in the PRF group &gt; 0.05 means a significant increase in PDGF-BB levels on the 7th day post flap surgery. Examination of PDGF levels, especially PDGF-BB, should be done in each healing phase of the wound from the inflammatory phase to the maturation and remodeling phase, for example on the 3rd, 7th, 14th and 28th days when PDGF levels approach zero and the healing process it's almost over, referring to the results of Alzahrani's research (2018) and also Matsuoka and Grotendorst (1989) which showed higher levels of PDGF-BB and VEGF in GCF on days 1, 3, 7 and 14 after surgery with minimally invasive surgery (MIS) technique. The conclusions of this study showed that the application of Platelet Rich Fibrin (PRF) and bone graft had an effect on the level of Platelet Derived Growth Factor (PDGF-BB) post-surgical flap in patients with chronic periodontitis, and elevated levels of PDGF-BB post-surgical flap with PRF application higher than surgical flap without the PRF application. Increased levels of PGDF-BB in the group applied to the PRF were higher than those that were not applied PRF, although statistically significant differences were not found, there was a tendency for the healing process of the PRF group to be better than the Non PRF group.

Evaluation of biological integration and inflammatory response to blood vessels produced by tissue engineering: experimental model in rabbit

The cardiovascular disease is the main cause of mortality in the western population and the Peripheral Arterial Disease (PAD) evolves, in large proportion, to the amputation of the affected limb. This study aimed to synthesize blood vessels using scaffolds of rabbit’s Inferior Vena Cava (IVC) and test its interaction with the receiver tissue and test inflammatory responses. Methods: The IVC were obtained from 8 rabbits to decellularization or in natura veins. The descellularized veins (DV) were obtained through protocols of decellularization established previously, using sodium dodecyl sulfate 1% (SDS) and mechanical agitation for 2 hours.12 animals were used to the experiment in vivo (3 animals in each group), being the product implanted in the interescapular dorsal area of each animal. The established groups are: Group 1- in natura allogeneic vein; Group 2- DV-SDS no cells added; Group 3- DV- SDS with 1x105 adipose tissue allogeneic Mesenchymal Stem Cells (MSC) added; Group 4- DV-SDS + autologous MSC. The (MSC) of the autologous receptors were collected 21 days before the implant and expanded in vitro. The explants were analyzed by histomorphological/immunohistochemistry and the peripheral blood was collected in the pre operatory in 1d, 7d, 14d, 30d and 60 day post operatory to dose the inflammatory and anti-inflammatory interleukins. Results: IL-10 and PDGF levels were significantly higher in group 4, which also showed neovascularization and large endothelial reconstruction. We may conclude the existence of an inflammatory response to the use of allogeneic grafts, which is lower when associated with autologous MSC.

Role of Cytokines in Ranibizumab Therapy for Macular Edema in Patients with Central Retinal Vein Occlusion.

Purpose: To investigate aqueous humor levels of 11 factors/cytokines in patients with central retinal vein occlusion (CRVO) and macular edema (ME) receiving anti-vascular endothelial growth factor (VEGF) therapy, as well as correlations between changes of functional or morphological parameters and aqueous cytokine levels. Methods: In 32 CRVO patients scheduled to receive 2 consecutive doses of intravitreal ranibizumab, aqueous samples were obtained at the time of injecting each dose. Aqueous levels of VEGF, soluble VEGF receptor (sVEGFR)-1, sVEGFR-2, platelet-derived growth factor-AA (PDGF-AA), placental growth factor (PlGF), interleukin-6, and monocyte chemotactic protein-1 (MCP-1) were measured using a suspension array. Results: Aqueous humor levels of VEGF, sVEGFR-1, PDGF, PlGF, interleukin-6, and MCP-1 were all significantly lower at 1 month after the initial dose of intravitreal ranibizumab compared with baseline. A significant negative correlation was noted between the change of ME and the changes of aqueous humor VEGF or interleukin-6 levels after intravitreal ranibizumab. The change of VEGF also showed a significant negative correlation with improvement of visual acuity. Conclusions: In patients with CRVO, the changes of visual acuity and ME after intravitreal ranibizumab were associated with inhibition of intraocular VEGF production. VEGF could be a useful marker for the response of ME to treatment.

Ampelopsin E Reduces the Invasiveness of the Triple Negative Breast Cancer Cell Line, MDA-MB-231.

Breast cancer is the most common and the second leading cause of cancer-related deaths in women. It has two distinctive hallmarks: rapid abnormal growth and the ability to invade and metastasize. During metastasis, cancer cells are thought to form actin-rich protrusions, called invadopodia, which degrade the extracellular matrix. Current breast cancer treatments, particularly chemotherapy, comes with adverse effects like immunosuppression, resistance development and secondary tumour formation. Hence, naturally-occurring molecules claimed to be less toxic are being studied as new drug candidates. Ampelopsin E, a natural oligostilbene extracted from Dryobalanops species, has exhibited various pharmacological properties, including anticancer and anti-inflammatory activities. However, there is yet no scientific evidence of the effects of ampelopsin E towards metastasis. Scratch assay, transwell migration and invasion assays, invadopodia and gelatin degradation assays, and ELISA were used to determine the effects of ampelopsin E towards the invasiveness of MDA-MB-231 cells. Strikingly in this study, ampelopsin E was able to halt migration, transmigration and invasion in MDA-MB-231 cells by reducing formation of invadopodia and its degradation capability through significant reduction (p < 0.05) in expression levels of PDGF, MMP2, MMP9 and MMP14. In conclusion, ampelopsin E reduced the invasiveness of MDA-MB-231 cells and was proven to be a potential alternative in treating TNBC.

Inflammatory cytokines and growth factors were not associated with psychosis liability or childhood trauma.

Psychosis is a multifactorial condition arising from an interaction between genetic liability and exposure to environmental risk factors, in particular childhood trauma. Furthermore, accumulating evidence supports a role for the immune system in the aetiology of psychosis. Increased peripheral levels of pro-inflammatory cytokines and reduced neurotrophic factors are found in patients with psychosis. Childhood trauma is highly prevalent in psychosis patients and is also associated with increased pro-inflammatory cytokines and reduced neurotrophic factors. Recent studies suggest the increase in pro-inflammatory cytokines and decrease in neurotrophic factors seen in psychosis may be attributable to the effects of child maltreatment. The aim of this study was to improve understanding of the relation between childhood trauma, inflammation and psychosis. We examined separate and interaction effects of psychosis liability and childhood trauma on serum levels of BDNF, CCL-2, CRP, IFN-γ, IGFBP2, IL-6, PDGF, SCF and TNF-α in 40 patients with recent onset psychosis, 13 patients at Ultra-High Risk (UHR) for psychosis, 31 unaffected siblings of psychosis patients and 41 healthy controls. Childhood trauma was assessed retrospectively with the Childhood Trauma Questionnaire (CTQ). No statistically significant effects of psychosis liability or childhood trauma on concentrations of cytokines or growth factors in peripheral blood were found, nor were there any statistically significant interaction effects of psychosis liability with childhood trauma on serum levels of cytokines and growth factors.

SUN-134 CYTOSKELETON-ASSOCIATED PROTEIN 4 (CKAP4), A NEW PLAYER IN THE REGULATION OF MESANGIAL CELL PROLIFERATION

Cytoskeleton-associated protein 4 (CKAP4) has recently attracted attention within cancer research, where its overexpression has been associated with increased lung, pancreatic and kidney cancer cell proliferation and a worsened prognosis. However, the potential role of CKAP4 in mesangioproliferative diseases such as IgA nephropathy (IgAN) and diabetic nephropathy (DN) has not previously been studied. The main growth factor known to induce proliferation in mesangial cells is PDGF, and the mesangial cells are the only glomerular cells known to express PDGF receptor beta (PDGFRβ). PDGF is normally secreted by the glomerular endothelial cells as well as the mesangial cells themselves resulting in a paracrine and autocrine regulation of mesangial cell proliferation. Increased PDGF levels in the glomeruli leads to mesangial proliferation as seen in DN and IgAN. Our hypothesis is that CKAP4 is involved in the regulation of mesangial proliferation. Expression of CKAP4 in glomeruli was investigated using immunofluorescence and immunoEM on human renal sections. CKAP4 was silenced using shRNA in primary human mesangial cells. Westernblot and masspectrometry was used to study protein expression and BrdU proliferation assay to determine proliferation. In order to understand protein interaction co-immunoprecipitation was used. Finally, two different mouse models of DN was used to investigate CKAP4 expression in disease. Immunohistochemistry as well as immunoEM showed expression of CKAP4 in glomerular mesangial cells and in the cell bodies of podocytes. Silencing CKAP4 resulted in an abated mitogenic response to the growth factor PDGF-BB, and further experiments revealed a significantly decreased expression of the PDGF receptor, both on RNA and protein level. Preliminary results from co-immunoprecipitation experiments indicate that PDGFRβ and CKAP4 interact. Analysis of the total protein content in the silenced cells using mass spectrometry showed only small or no changes in expression of other growth factor receptors such as EGFR and TGFbRs. In two different mouse models of DN CKAP4 was found to be significantly upregulated in later stages of disease with established mesangial expansion and other key features of the disease. We here report for the first time that CKAP4 is expressed in mesangial cells and that the expression of CKAP4 influences the cellular proliferative response to PDGF-BB and the expression of PDGFRβ. CKAP4 has previously been reported to interact with the EGF receptor on the cell surface of hepatocellular carcinomas and thereby influencing the growth of these cells. The EGF receptor and the PDGF receptor has several similarities and are tyrosine kinase receptors. We suggest that the interaction between the PDGF receptor and CKAP4 resembles the one of CKAP4 and EGFR. CKAP4 could be a potential mediator and target of mesangial proliferation in glomerular kidney disease.