Upregulation Of Growth Factors Research Articles

IL-1β–Primed Mesenchymal Stromal Cells Improve Epidermal Substitute Engraftment and Wound Healing via Matrix Metalloproteinases and Transforming Growth Factor-β1

Since the 1980s, deep and extensive skin wounds and burns are treated with autologous split-thickness skin grafts, or cultured epidermal autografts, when donor sites are limited. However, the clinical use of cultured epidermal autografts often remains unsatisfactory because of poor engraftment rates, altered wound healing, and reduced skin functionality. In the past few decades, mesenchymal stromal cells (MSCs) have raised much attention because of their anti-inflammatory, protrophic, and pro-remodeling capacities. More specifically, gingival MSCs have been shown to possess enhanced wound healing properties compared with other tissue sources. Growing evidence also indicates that MSC priming could potentiate therapeutic effects in diverse invitro and invivo models of skin trauma. In this study, we found that IL-1β-primed gingival MSCs promoted cell migration, dermal-epidermal junction formation, and inflammation reduction invitro, as well as improved epidermal substitute engraftment invivo. IL-1β-primed gingival MSCs had different secretory profiles from naive gingival MSCs, characterized by an overexpression of transforming growth factor-β and matrix metalloproteinase (MMP) pathway agonists. Eventually, MMP-1, MMP-9, and transforming growth factor-β1 appeared to be critically involved in IL-1β-primed gingival MSC mechanisms of action.

Electrospun chitosan/PVA/bioglass Nanofibrous membrane with spatially designed structure for accelerating chronic wound healing.

Cutaneous wounds, especially chronic wounds, remain clinical challenges, and this is partially due to the complex healing process composed of four overlapping but distinct stages including hemostasis, inflammation, proliferation and remodeling. Therefore, wound dressings with spatially designed structures which can temporally regulate certain bioactive components to function at specific healing stages might be able to accelerate the healing process. In this study, nanobioglass incorporated chitosan-PVA (polyvinyl alcohol) trilayer nanofibrous membrane (nBG-TFM) was fabricated via sequential electrospinning. This membrane exhibited excellent biocompatibility, antibacterial activity and regeneration promotion effect. Furthermore, spatially designed structure optimized functions of each component and provided more suitable microenvironment as compared with uniform membrane. Rat full-thickness skin defects model and mice diabetic chronic wound model showed that nBG-TFM could achieve significantly accelerated and enhanced healing, in terms of complete re-epithelialization, improved collagen alignment and formation of skin appendages. It was revealed that nBG-TFM functioned through upregulating growth factors including VEGF and TGF-β. Meanwhile inflammatory cytokines such as TNF-α and IL-1β were downregulated. The technology presented in this study shed new light on designing functional wound dressings which can promote healing of chronic wounds.

CGRP and Brain Functioning: Cautions for Migraine Treatment.

Calcitonin gene-related peptide has emerged as a therapeutic target in migraine. Monoclonal antibodies and small molecule receptor antagonists (gepants) directed against CGRP have been approved or are in Phase II or III clinical trials. For monitoring the long-term safety of these drugs, it is helpful to consider the role of CGRP in brain functioning. Qualitative review of the preclinical literature on CGRP in brain physiology and pathophysiology. Within the brain, CGRP is upregulated by stresses such as ischemia, injury, hyperthermia, and seizure, and activates neuroprotective processes. Thus, CGRP buffers intracellular calcium, triggers antiapoptotic signaling, upregulates a number of neurotrophins (including insulin-like growth factor-1/IGF-1, basic fibroblast growth factor/bFGF, and nerve growth factor/NGF), reduces brain edema, and likely increases antioxidant defenses. When released outside the blood-brain barrier, CGRP likely protects the endothelium, upregulates growth factor signaling from the endothelium to the brain parenchyma, strengthens the blood-brain barrier, protects the immune privilege of the brain by inhibiting the movement of neutrophils and monocytes, and facilitates neurogenesis and angiogenesis at stem cell niches. CGRP participates in a wide range of neuroprotective processes. In theory, migraineurs with comorbid brain pathology might be at increased risk from CGRP inhibition. However, the extent of compensating processes is unknown and will determine whether these risks materialize in practice.

New Lipid Mediators in Retinal Angiogenesis and Retinopathy.

Retinal diseases associated with vascular destabilization and the inappropriate proliferation of retinal endothelial cells have major consequences on the retinal vascular network. In extreme cases, the development of hypoxia, the upregulation of growth factors, and the hyper-proliferation of unstable capillaries can result in bleeding and vision loss. While anti-vascular endothelial growth factor therapy and laser retinal photocoagulation can be used to treat the symptoms of late stage disease, there is currently no treatment available that can prevent disease progression. Cytochrome P450 enzymes metabolize endogenous substrates (polyunsaturated fatty acids) to bioactive fatty acid epoxides that demonstrate biological activity with generally protective/anti-inflammatory and insulin-sensitizing effects. These epoxides are further metabolized by the soluble epoxide hydrolase (sEH) to fatty acid diols, high concentrations of which have vascular destabilizing effects. Recent studies have identified increased sEH expression and activity and the subsequent generation of the docosahexaenoic acid-derived diol; 19,20-dihydroxydocosapentaenoic acid, as playing a major role in the development of diabetic retinopathy. This review summarizes current understanding of the roles of cytochrome P450 enzyme and sEH–derived PUFA mediators in retinal disease.

Upregulation of Fibroblast Growth Factors Caused by Heart and Neural Crest Derivatives Expressed 2 Suppression in Endometriotic Cells: A Possible Therapeutic Target in Endometriosis.

Several features exist that distinguish endometriotic cells from eutopic endometrial cells. Progesterone resistance is one of the main distinguishing features, although how progesterone resistance affects the phenotype of endometriotic cells is not fully elucidated. Heart and neural crest derivatives expressed 2 (HAND2) is a transcriptional factor that plays an important role in maintaining endometrial function in a progesterone-dependent manner. Therefore, we explored whether progesterone-dependent HAND2 is implicated in the progression of endometriosis. HAND2 was less expressed by endometriotic tissues compared to endometrial tissues. Suppression of HAND2 expression induced fibroblast growth factor 1 (FGF1), FGF2, and FGF9 in endometriotic stromal cells and consequently enhanced migration and invasion capacity. AZD4547, a FGF receptor inhibitor, diminished the migration and invasion of endometriotic cells in vitro. In the murine model of endometriosis, AZD4547 showed suppressive effects on the development of endometriotic lesions at a relatively low concentration. In conclusion, we demonstrated that FGF1, FGF2, and FGF9 are downstream effectors of HAND2 in endometriotic cells. Since HAND2-dependent FGFs play roles in enhancing invasive capacity of endometriotic cells, our results suggest that FGF receptor inhibitors, such as AZD4547, can be promising therapeutic targets for endometriosis.

Abstract 3024: Transcriptional and epigenetic regulation of resistance markers in cetuximab sensitive HNSCC cells

Abstract Acquired resistance is pervasive in cancers treated with targeted therapies. The understanding of resistance mechanisms and the timing of the driver molecular changes are crucial for alternative interventions while tumors are sensitive. In this study, we focus on cetuximab, the only targeted therapeutic approved to treat head and neck cancers (HNSCC). We and others have shown that signatures of resistance arise in early sensitive states of treatment, including notably EMT and TFAP2 activation. Because TFAP2 regulates both EMT genes and compensatory growth factor receptors and resistance is associated with epigenetic alterations, we hypothesized that epigenetic regulation of TFAP2 is a master regulator of future mechanisms of resistance.To test our central hypothesis, we treated three HNSCC cell lines (SCC1, SCC6 and SCC25) with CTX for 5 days, a time frame that cells are known to be sensitive. We performed RNA-seq and ATAC-seq to understand the global molecular changes induced by CTX. To verify the role of TFAP2A in cell growth and EMT control in response to CTX, we used siRNA for gene silencing and measured cell proliferation and migration ability (scratch assay). In order to identify a potential combination therapy, we also treated the cells with JQ1, a bromodomain inhibitor with a major impact in delaying acquired CTX resistance.RNA-seq analysis shows that CTX sensitive cells up-regulate growth factor receptors and EMT genes. Chromatin changes were also evident in the same cells. Enrichment of accessible chromatin areas for genes from the TFAP2A and EMT pathways suggests epigenetic regulation of CTX resistance associated genes as an early response to therapy. In both, RNA-seq and ATAC-seq analysis, we observed alterations in different sets of genes for each of the cell lines, indicating that resistance signatures vary in the cell models as observed in patients. TFAP2A plays a pivotal role in cell growth. Lack of TFAP2A results in lower proliferation rates in untreated cells and a synergistic effect is observed with CTX, JQ1 and CTX + JQ1 therapies. TFAP2A loss reflected in increased migration ability in SCC1 and SCC25, while in SCC6 we observed less cell motility. However, in all cell lines EMT markers were up-regulated and suggests EMT development, due to down-regulation of TFAP2A, is a cell type and time dependent mechanism with specific cell types requiring more time to translate the transcriptional changes.Overall, we demonstrate that resistance-associated genes in HNSCC sensitive cells are epigenetically regulated. These mechanisms are also cell type and time dependent reflecting observations from patients’ cohorts. We show that TFAP2A regulates growth factor receptors and EMT genes in CTX sensitive cells and could be a potential target for drug development. We also show that the CTX + JQ1 combined therapy is a potential alternative and more effective approach while patients are sensitive to therapy. Citation Format: Luciane T. Kagohara, Fernando Zamuner, Michael Considine, Genevieve Stein-O'Brien, Thomas Sherman, Daria A. Gaykalova, Elana J. Fertig. Transcriptional and epigenetic regulation of resistance markers in cetuximab sensitive HNSCC cells [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 3024.

Electrical stimulation for neuroregeneration in urology: a new therapeutic paradigm.

The present review highlights regenerative electrical stimulation (RES) as potential future treatment options for patients with nerve injuries leading to urological dysfunction, such as urinary incontinence, voiding dysfunction or erectile dysfunction. Additionally, it will highlight the mechanism of nerve injury and regeneration as well as similarities and differences between RES and current electrical stimulation treatments in urology, functional electrical stimulation (FES) and neuromodulation. It has been demonstrated that RES upregulates brain-derived neurotrophic factor (BDNF) and its receptor to facilitate neuroregeneration, facilitating accurate reinnervation of muscles by motoneurons. Further, RES upregulates growth factors in glial cells. Within the past 2 years, RES of the pudendal nerve upregulated BDNF in Onuf's nucleus, the cell bodies of motoneurons that course through the pudendal nerve and accelerated functional recovery in an animal model of stress urinary incontinence. Additionally, electrical stimulation of the vaginal tissue in an animal model of stress urinary incontinence accelerated functional recovery. RES has great potential but future research is needed to expand the potential beneficial effects of RES in the field of urology.

Indications of Intravitreal Injection of Bevacizumab in a sample of Iraqi patients

Background: Anti-vascular endothelial growth factors commonly used intravitreally for treatment of variable retinal disease worldwide, which help to regulate the upregulation of vascular endothelial growth factors due to ischemia or/and other retinal insult.
 Objective: To review all cases of intravitreal bevacizumab given in the retinal unit of the Eye Specialty Private Hospital in Iraq from December 2015 to June 2016, pointing out the common indications.
 Methods: A retrospective study including all cases of intravitreal injections of bevacizumab recorded at the Eye Specialty Private Hospital from December 2015 to June 2016 were retrieved. Age, sex, laterality, diagnosis and the indication for injection were recorded in the data sheet prepared for the study.. The indications for intravitreal injection of bevacizumab are classified into four groups: Group 1: Intravitreal injection for diabetic complications. Group 2: Intravitreal injection for retinal vein occlusion. Group 3: Intravitreal injection for choroidal neovascul-arization Group 4: Intravitreal injection for other retinal disease.
 Results: A total of 306 patients underwent intravitreal injection of bevacizumab were enrolled in the study period. The most common indication was group 1 (intravitreal injection for diabetic complications [213 patients (69.6%)] followed by group 3 (choroidal neovascularization) [45 patients (14.7%)] and then group 2 (intravitreal injection for retinal vein occlusion) [42 patients (13.7%)] and lastly group 4 (intravitreal injection for other retinal disease) represent [6 patients (1.96%)].
 Conclusion The most common indication for intravitreal bevacizumab was diabetic retino-pathy that need more care for early diagnosis and treatment of diabetic retinopathy and good screening programs to avoid complications.

Cardiac Regeneration by Statin-Polymer Nanoparticle-Loaded Adipose-Derived Stem Cell Therapy in Myocardial Infarction.

Clinical trials with autologous adipose‐derived stem cell (AdSC) therapy for ischemic heart diseases (IHDs) are ongoing. However, little is known about combinational therapeutic effect of AdSCs and statin poly(lactic‐co‐glycolic) acid (PLGA) nanoparticles on the ischemic myocardium. We investigated the hypothesis that statins, which have pleiotropic effects, augment the therapeutic potential of AdSCs and that AdSCs also act as drug delivery tools. Simvastatin‐conjugated nanoparticles (SimNPs) significantly promoted migration activity without changing proliferation activity and upregulated growth factor gene expression in vitro. A small number of intravenously administered SimNP‐loaded AdSCs (10,000 cells per mouse) improved cardiac function following myocardial infarction, inducing endogenous cardiac regeneration in the infarcted myocardium. The de novo regenerated myocardium was thought to be derived from epicardial cells, which were positive for Wilms' tumor protein 1 expression. These findings were attributed to the sustained, local simvastatin release from the recruited SimNP‐loaded AdSCs in the infarcted myocardium rather than to the direct contribution of recruited AdSCs to tissue regeneration. SimNP‐loaded AdSCs may lead to a novel somatic stem cell therapy for IHDs. stem cells translational medicine 2019;8:1055–1067

Hypoxia improves hair inductivity of dermal papilla cells via nuclearNADPHoxidase 4‐mediated reactive oxygen species generation'

Dermal papilla cells (DPCs) play a key role in hair regeneration and morphogenesis. Therefore, tremendous efforts have been made to promote DPC hair inductivity. The aim of this study was to investigate the mitogenic and hair inductive effects of hypoxia on DPCs and examine the underlying mechanism of hypoxia-induced stimulation of DPCs. DPCs' hair inductivity was examined under normoxia (20% O2 ) and hypoxia (2% O2 ). Hypoxia significantly increased the proliferation and delayed senescence of DPCs via Akt phosphorylation and downstream pathways. Hypoxia upregulated growth factor secretion of DPCs through the mitogen-activated protein kinase pathway. Hypoxia-preconditioned DPCs induced the telogen-to-anagen transition in C3 H mice, and also enhanced hair neogenesis in a hair reconstitution assay. Injected green fluorescent protein-labelled DPCs migrated to the outer root sheath of the hair follicle, and hypoxia-preconditioning increased survival and migration of DPCs invivo. Conditioned medium obtained from hypoxia increased the hair length of mouse vibrissa follicles via upregulation of alkaline phosphatase, vascular endothelial growth factor, and glial cell line-derived neurotrophic factor. We examined the mechanism of this hypoxia-induced stimulation, and found that reactive oxygen species (ROS) play a key role. For example, inhibition of ROS generation by N-acetylcysteine or diphenyleneiodonium treatment attenuated DPCs' hypoxia-induced stimulation, but treatment with ROS donors induced mitogenic effects and anagen transition. NADPH oxidase 4 is highly expressed in the DPC nuclear region, and NOX4 knockout by CRISPR-Cas9 attenuated the hypoxia-induced stimulation of DPCs. Our results suggest that DPC culture under hypoxia has great advantages over normoxia, and is a novel solution for producing DPCs for cell therapy. What's already known about this topic? Dermal papilla cells (DPCs) play a key role in hair regeneration and morphogenesis, but they are difficult to isolate and expand for use in cell therapy. Tremendous efforts have been made to increase proliferation of DPCs and promote their hair formation ability. What does this study add? Hypoxia (2% O2 ) culture of DPCs increases proliferation, delays senescence and enhances hair inductivity of DPCs. Reactive oxygen species play a key role in hypoxia-induced stimulation of DPC. What is the translational message? Preconditioning DPCs under hypoxia improves their hair regenerative potential, and is a novel solution for producing DPCs for cell therapy to treat hair loss.

SUN-426 A Case of Pseudo-Apoplexy: An Unexplored Mechanism of Acute Pituitary Failure

A 70-year old male came in with a 2-week history of drowsiness, generalized body weakness, anorexia, occasional nausea and vomiting that started with a sudden onset of headache during his flight from Michigan, USA to the Philippines without any history of trauma. His headache resolved upon arrival in Manila, however, weakness persisted and he had repeated hyponatremia on hospital consults with essentially normal physical and neurologic findings. Hormonal work-up was done and revealed panhyopituitarism. All symptoms resolved within 2 days of initiating hydrocortisone at physiologic dose. Cranial MRI confirmed presence of a pituitary mass (1.7 x 1.7 x 1.5 cm) with pituitary apoplexy for which the patient underwent endoscopic transsphenoidal transpterygoid surgery. However, histopathology report showed a rathke’s cleft cyst without findings of hemorrhage or infection. Evidences of the dynamic nature of rathke’s cleft cysts have all been clinical, circumstantial and scarce due to the difficulty in diagnosis and documentation. However, cases of acute hypopituitarism in the absence of hemorrhage, infarct, infection or trauma, suggests another pathophysiology of disease that has not yet been extensively reported. This case supports a mechanism of either cyst expansion with encroachment on neighboring structures and/or cerebral edema potentially triggered by changes in atmospheric pressure with international commercial flights flying at approximately 30,000 ft. At higher altitudes, lower partial pressure of oxygen causes relative cerebral hypoxia. Consequently, vascular permeability increases due to the upregulation of vascular endothelial growth factors promoting cerebral edema. This has previously been reported with manifestations more commonly known as acute mountain sickness (AMS) and high altitude cerebral edema (HACE) when associated with life-threatening neurologic deterioration. For patients already with intracranial space occupying lesions, development of minimal edema becomes significant. A slight Increase in mass size or volume of cerebrospinal fluid in smaller spaces such as the sella, are more likely to display symptoms. Therefore, caution should be advised for patients with known intracranial masses who are not to undergo surgery and are planning to travel or stay at high altitudes. Furthermore, rathke’s cleft cysts should be considered as a differential diagnosis to apoplexy-like presentations, especially in those patients who have relatively benign or non-persistent symptoms easily resolving with hormone replacement.

Cognitive and Vascular effects of Exercise in patients with Alzheimer's Disease

ObjectTo investigate exercise‐induced effect on cognitive and vascular functions in people with Alzheimer's Disease (AD) and compare it with the standard cognitive therapy.MethodsFifty‐five people with AD (77.6±6.2 years; 23.5±4.8 MMSE) were recruited and randomly assigned to Cognitive Treatment (CT, n=23) or Exercise Treatment (ET, n=22). All subjects performed 72 treatment sessions lasting 90 minutes, 3 times a week. ET included: moderate to high intensity aerobic and strength training. CT included multi‐modal stimuli (visual, verbal, auditive). Before (T0) and after (T1) the 6‐month treatment global cognitive function was measured by means of Mini‐Mental State Examination (MMSE). Vascular function was measured by means of Flow‐Mediated Dilation (FMD, %) and Passive‐Limb movement (PLM, calculating the variation in blood flow: ΔPeak; and Area Under the Curve: AUC) tests. A blood sample was also taken and analyzed for Vascular Endothelial Growth Factor (VEGF)ResultsMMSE did not change after CT nor ET and post‐treatment between‐groups (BG) difference (Diff. of means) were not found. CT group did not exhibit any change in vascular function while ET group significantly increased FMD% (3.64, p=0.003; BG Diff. of means: 4.615, p<0.001), Δpeak (135.88, p=0.004; BG Diff. of means: 160.46, p<0.001) and AUC (38.16, p=0.050; BG Diff. of means: 62.36, p=0.002). VEGF increased significantly in ET group only (8.97, p=0.047; BG Diff. of means: 12.7, p=0.004).ConclusionResults from this study show that both, standard cognitive therapy and exercise help in preserving cognitive function in people with AD. However, only exercise seems to offer the appropriate stimuli to improve also vascular function in this population. Importantly, exercise has a polyhedral and multi‐situ action since the whole organism, brain included, must respond adaptatively when activated during exercise. Most of the exercise‐induced responses include upregulation of growth factors at muscular, vascular and cerebral level as well.Support or Funding InformationThis work was supported by PRIN 2010KL2Y73This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.

Adhesive Hemostatic Conducting Injectable Composite Hydrogels with Sustained Drug Release and Photothermal Antibacterial Activity to Promote Full-Thickness Skin Regeneration During Wound Healing.

Developing injectable nanocomposite conductive hydrogel dressings with multifunctions including adhesiveness, antibacterial, and radical scavenging ability and good mechanical property to enhance full-thickness skin wound regeneration is highly desirable in clinical application. Herein, a series of adhesive hemostatic antioxidant conductive photothermal antibacterial hydrogels based on hyaluronic acid-graft-dopamine and reduced graphene oxide (rGO) using a H2 O2 /HPR (horseradish peroxidase) system are prepared for wound dressing. These hydrogels exhibit high swelling, degradability, tunable rheological property, and similar or superior mechanical properties to human skin. The polydopamine endowed antioxidant activity, tissue adhesiveness and hemostatic ability, self-healing ability, conductivity, and NIR irradiation enhanced in vivo antibacterial behavior of the hydrogels are investigated. Moreover, drug release and zone of inhibition tests confirm sustained drug release capacity of the hydrogels. Furthermore, the hydrogel dressings significantly enhance vascularization by upregulating growth factor expression of CD31 and improve the granulation tissue thickness and collagen deposition, all of which promote wound closure and contribute to a better therapeutic effect than the commercial Tegaderm films group in a mouse full-thickness wounds model. In summary, these adhesive hemostatic antioxidative conductive hydrogels with sustained drug release property to promote complete skin regeneration are an excellent wound dressing for full-thickness skin repair.

Exploring the influence of the surface proteins on probiotic effects performed by Lactobacillus pentosus HC-2 using transcriptome analysis in Litopenaeus vannamei midgut

In order to understand the mediation function of surface proteins in probiotic effects executed by Lactobacillus pentosus HC-2 in midgut of Litopenaeus vannamei, the immune and digestion related enzymes and the transcriptome expression were analyzed after shrimp fed with normal HC-2 or with stripped surface proteins HC-2 by lithium chloride (LiCl) treatment. The results showed that the shrimp fed with normal HC-2 produced much higher immune and digestion related enzymes than the control group or LiCl-treated HC-2 group to defense the Vibrio parahaemolyticus E1 infection. We obtained total over 275,099 unigenes from L. vannamei midgut, 981 genes were significant differentially expressed in normal HC-2 group compared with control, 1314 genes were significant differentially expressed in LiCl-treated HC-2 group compared with control, and 1689 genes were significant differentially expressed in LiCl-treated HC-2 group compared with normal HC-2 group. The GO/KEGG enrichment analysis of the significantly different genes demonstrated that L. vannamei fed with normal HC-2 induced immune-related, signal transduction, ion homeostasis, cell-cell adhesion, response stress/stimulus, vascular endothelial growth factor and peritrophin genes up-regulation, which were important genes involved in improving the shrimp intestine immune response, nutrition and growth performance, and bacteria adhesion and colonization, but these genes were suppressed in the midgut of shrimp fed with deprived surface proteins bacteria. Taken together, these results indicated that the surface proteins were essential for HC-2 executing probiotic effects in midgut of shrimp. Our data contribute to improve the current understanding of host - Lactobacillus interaction and the probiotic mechanisms in shrimps.

Sauromatum guttatum extract promotes wound healing and tissue regeneration in a burn mouse model via up-regulation of growth factors

Contexts: Sauromatum guttatum (Wall.) Schott (Araceae) has been traditionally used for the treatment of wounds.Objectives: This study evaluates the healing and tissue regeneration potential of S. guttatum extract in burn wounds.Materials and methods: S. guttatum extract was analysed using various chemical tests, thin layer chromatography (TLC) and high-performance liquid chromatography (HPLC). Moreover, the extract was tested against burn associated bacteria and minimum inhibitory concentration (MIC) was also calculated. Wound healing and tissue regeneration potential was assessed using a thermally induced burn BALBc mouse model. S. guttatum extract (2% w/w) prepared in petroleum jelly, vehicle and positive control [silver sulfadiazine (SD)] groups was applied three times a day. The treatment was continued for 15 d and wound closure was measured and photographed on day 5, 10 and 15. The burnt tissues excised from wounds were subjected to histological and comparative gene expression analysis.Results: The results of the chemical tests indicated the presence of alkaloids, saponins, phenols, phytosterols, tannins, and flavonoids, while TLC and HPLC analysis indicated the presence of various compounds. The extract showed excellent activity against the tested pathogens. The lowest MIC (125 µg/mL) was observed against Staphylococcus aureus. A considerable decrease in wound area (72%) was observed in extract-treated group. Histological examination of extract-treated group showed good signs of wound healing with complete re-epithelialization and better tissue regeneration. Comparative gene expression analysis revealed the up-regulation of wound healing related PDGF, EGF and FGF genes.Conclusions: S. guttatum extract may be used to isolate bioactive constituents for the treatment of burn wounds.

Diabetic Macular Edema: Current Understanding, Pharmacologic Treatment Options, and Developing Therapies.

Diabetic retinopathy and diabetic macular edema comprise a major source of visual disability throughout the developed world. The etiology and pathogenesis of macular edema is intricate and multifactorial, in which the hyperglycemic state in diabetes induces a microangiopathy. Through several inflammatory and vasogenic mediators, including vascular endothelial growth factor (VEGF) upregulation and inflammatory cytokines and chemokines, pathologic changes are induced in the vascular endothelium triggering breakdown of the blood retinal barrier, causing extravasation of fluid into the extracellular space and manifesting clinically as macular edema, resulting in visual loss. The advent of medications targeting the VEGF pathway has led to great clinical improvements compared with the previous standard of care of laser therapy alone, as shown in studies such as RISE, RIDE, VIVID, VISTA, and DRCR. However, analyses have shown that many patients have inadequate response or are nonresponders to anti-VEGF therapy, demonstrating the need for additional therapies to more comprehensively treat this disease. Although corticosteroid treatments and implants have demonstrated some efficacy in adjunctive and supplemental treatment, the need to more adequately treat macular edema remains. Our knowledge of diabetic macular edema continues to grow, leading to new currently available and emerging pharmacotherapies to further enhance our treatment and restore vision in those affected by diabetic macular edema. This review will discuss the pathogenesis of diabetic macular edema and the pharmacologic therapies available for its treatment, including anti-VEGF, steroids, and newer therapies still in development, such as angiopoietin antagonists, Tie2 agonists, kallikrein inhibitors, interleukin inhibitors, and others.

A murine model of acute lung injury identifies growth factors to promote tissue repair and their biomarkers.

Type II alveolar epithelial cells (AEC2s) play a crucial role in the regeneration of type I AECs after acute lung injury. The mechanisms underlying the regeneration of AEC2s are not fully understood. To address this issue, here, we investigated a murine model of acute lung injury using mice expressing human Diphtheria Toxin Receptor (DTR) under the control of Lysozyme M promoter (LysM-DTR). DT injection induced the depletion of AEC2s, alveolar macrophages, and bone marrow (BM)-derived myeloid cells in LysM-DTR mice, and the mice died within 6days after DT injection. Apoptotic AEC2s and bronchiolar epithelial cells appeared at 24hr, whereas Ki67-positive proliferating cells appeared in the alveoli and bronchioles in the lung of LysM-DTR mice at 72-96hr after DT injection. Transfer of wild-type BM cells into LysM-DTR mice accelerated the regeneration of AEC2s along with the up-regulation of several growth factors. Moreover, several metabolites were significantly decreased in the sera of LysM-DTR mice compared with WT mice after DT injection, suggesting that these metabolites might be biomarkers to predict AEC2s injury. Together, LysM-DTR mice might be useful to identify growth factors to promote lung repair and the metabolites to predict the severity of lung injury.

Adipose-Derived Stem Cell Coculturing Stimulates Integrin-Mediated Extracellular Matrix Adhesion of Melanocytes by Upregulating Growth Factors.

Coculture with adipose-derived stem cells (ADSCs) can stimulate proliferation and migration of melanocytes. To enhance outcomes of skin disorders caused by melanocyte loss or death, mixed transplantation with ADSCs has been suggested. However, role of cocultured ADSCs in proliferation and migration of melanocytes remains unclear. This study determined the effect of ADSCs on production of growth factors and expression levels of intergrins in primary culture of adult human melanocytes with or without ADSCs and in nude mice grafted with such melanocytes. Higher amounts of growth factors for melanocytes, such as bFGF and SCF were produced and released from ADSCs by coculturing with melanocytes. Relative levels of integrins β1, α5, and α6 as well as adhesion to fibronectin and laminin were increased in melanocytes cocultured with ADSCs. Such increases were inhibited by neutralization of bFGF or SCF. Relative levels of bFGF, SCF and integrins were increased in nude mice skin after grafting with melanocyte+ADSC cocultures. Collectively, these results indicate that ADSCs can stimulate proliferation and migration of melanocytes by increasing expression of integrins in melanocytes through upregulation of production/release of melanocyte growth factors such as bFGF and SCF.

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Proteomic profiling of senescent human diploid fibroblasts treated with gamma-tocotrienol

BackgroundReplicative senescence of human diploid fibroblasts (HDFs) has been used as a model to study mechanisms of cellular aging. Gamma-tocotrienol (γT3) is one of the members of vitamin E family which has been shown to increase proliferation of senescent HDFs. However, the modulation of protein expressions by γT3 in senescent HDFs remains to be elucidated. Therefore, this study aimed to determine the differentially expressed proteins (DEPs) in young and senescent HDFs; and in vehicle- and γT3-treated senescent HDFs using label-free quantitative proteomics.MethodsWhole proteins were extracted and digested in-gel with trypsin. Peptides were detected by Orbitrap liquid chromatography mass spectrometry. Mass spectra were identified and quantitated by MaxQuant software. The data were further filtered and analyzed statistically using Perseus software to identify DEPs. Functional annotations of DEPs were performed using Panther Classification System.ResultsA total of 1217 proteins were identified in young and senescent cells, while 1218 proteins in vehicle- and γT3-treated senescent cells. 11 DEPs were found in young and senescent cells which included downregulation of platelet-derived growth factor (PDGF) receptor beta and upregulation of tubulin beta-2A chain protein expressions in senescent cells. 51 DEPs were identified in vehicle- and γT3-treated senescent cells which included upregulation of 70 kDa heat shock protein, triosephosphate isomerase and malate dehydrogenase protein expressions in γT3-treated senescent cells.ConclusionsPDGF signaling and cytoskeletal structure may be dysregulated in senescent HDFs. The pro-proliferative effect of γT3 on senescent HDFs may be mediated through the stimulation of cellular response to stress and carbohydrate metabolism. The expressions and roles of these proteins in relation to cellular senescence are worth further investigations. Data are available via ProteomeXchange with identifier PXD009933.