Human Neonatal Fibroblasts Research Articles

Spontaneous Efficient Differentiation of Human Pluripotent Stem Cells (hPSC) Upon Co-culture of hPSCs with Human Neonatal Foreskin Fibroblasts in 3D.

Pluripotent stem cells (PSCs) form well-formed embryoid bodies (EBs) in 3D culture. These EBs are formed in culture media lacking leukemia inhibitory factor (LIF) or basic fibroblast growth factor (bFGF) in mouse and human PSCs, respectively. EBs are excellent technical tools for understanding developmental biology and inducing controlled differentiation in succeeding experimental steps. Technically speaking, EBs are spontaneously differentiated PSCs in 3D and exhibit all three lineages in a time-point/sequential manner. For example, ectoderm will form first, followed by mesoderm and endoderm. We have attempted to co-culture human neonatal foreskin-derived fibroblast cells in our laboratory with the PSCs first in 2D conditions followed by the induction of EBs (PSC+fibroblasts co-cultured) in low attachment dishes. We also performed spontaneous differentiation of such EBs (co-cultured with fibroblasts). We checked the presence of markers of various lineages, namely, ectoderm, mesoderm, and endoderm in days 6, 10, and 12day EBs. We have also compared the fibroblast co-cultured EBs, along with control EBs (derived from only PSCs). This co-culture system mimics the natural conditions of uterine implantation and the role of the endometrial fibroblasts in the induction of further embryonic development. The fibroblast co-cultured iPSC EBs had better roundness scores than the normal iPSC EBs and had a higher expression of lineage-specific markers.

The Human Neonatal Skin Fibroblast, an Available Cell Source for Tissue Production and Transplantation, Exhibits Low Risk of Immunogenicity In Vitro.

The immunogenicity of allogeneic skin fibroblasts in transplantation has been controversial. Whether this controversy comes from a natural heterogeneity among fibroblast subsets or species-specific differences between human and mouse remains to be addressed. In this study, we sought to investigate whether fibroblasts derived from either adult or neonatal human skin tissues could induce different immune responses toward phagocytosis and T cell activation using in vitro co-culture models. Our results indicate that both phagocytosis and T cell proliferation are reduced in the presence of neonatal skin fibroblasts compared to adult skin fibroblasts. We also show that neonatal skin fibroblasts secrete paracrine factors that are responsible for reduced T cell proliferation. In addition, we show that neonatal skin fibroblasts express less class II human leukocyte antigen (HLA) molecules than adult skin fibroblasts after interferon gamma priming, which might also contribute to reduced T cell proliferation. In conclusion, this study supports the use of allogeneic neonatal skin fibroblasts as a readily available cell source for tissue production and transplantation to treat patients with severe injuries.

Molecular Evidence of Breast Cancer Cell Proliferation Inhibition by a Combination of Selected Qatari Medicinal Plants Crude Extracts.

Breast cancer (BC) is the most common malignancy, and conventional medicine has failed to establish efficient treatment modalities. Conventional medicine failed due to lack of knowledge of the mechanisms that underpin the onset and metastasis of tumors, as well as resistance to treatment regimen. However, Complementary and Alternative medicine (CAM) modalities are currently drawing the attention of both the public and health professionals. Our study examined the effect of a super-combination (SC) of crude extracts, which were isolated from three selected Qatari medicinal plants, on the proliferation, motility and death of BC cells. Our results revealed that SC attenuated cell growth and caused the cell death of MDA-MB-231 cancer cells when compared to human normal neonatal fibroblast cells. On the other hand, functional assays showed that SC reduced BC cell migration and invasion, respectively. SC-inhibited cell cycle and SC-regulated apoptosis was most likely mediated by p53/p21 pathway and p53-regulated Bax/BCL-2/Caspace-3 pathway. Our ongoing experiments aim to validate these in vitro findings in vivo using a BC-Xenograft mouse model. These findings support our hypothesis that SC inhibited BC cell proliferation and induced apoptosis. These findings lay the foundation for further experiments, aiming to validate SC as an effective chemoprevention and/or chemotherapeutic strategy that can ultimately pave the way towards translational research/clinical trials for the eradication of BC.

Neonatal hyperoxia induces activated pulmonary cellular states and sex-dependent transcriptomic changes in a model of experimental bronchopulmonary dysplasia.

Hyperoxia disrupts lung development in mice and causes bronchopulmonary dysplasia (BPD) in neonates. To investigate sex-dependent molecular and cellular programming involved in hyperoxia, we surveyed the mouse lung using single cell RNA sequencing (scRNA-seq), and validated our findings in human neonatal lung cells in vitro. Hyperoxia-induced inflammation in alveolar type (AT) 2 cells gave rise to damage-associated transient progenitors (DATPs). It also induced a new subpopulation of AT1 cells with reduced expression of growth factors normally secreted by AT1 cells, but increased mitochondrial gene expression. Female alveolar epithelial cells had less EMT and pulmonary fibrosis signaling in hyperoxia. In the endothelium, expansion of Car4+ EC (Cap2) was seen in hyperoxia along with an emergent subpopulation of Cap2 with repressed VEGF signaling. This regenerative response was increased in females exposed to hyperoxia. Mesenchymal cells had inflammatory signatures in hyperoxia, with a new distal interstitial fibroblast subcluster characterized by repressed lipid biosynthesis and a transcriptomic signature resembling myofibroblasts. Hyperoxia-induced gene expression signatures in human neonatal fibroblasts and alveolar epithelial cells in vitro resembled mouse scRNA-seq data. These findings suggest that neonatal exposure to hyperoxia programs distinct sex-specific stem cell progenitor and cellular reparative responses that underpin lung remodeling in BPD.

MiR-124- and let-7-Mediated Reprogram of Human Fibroblasts into SST Interneurons.

Many neurological disorders stem from defects in or the loss of specific neurons. Dysfunction of γ-aminobutyric acid (GABA)ergic interneurons may cause a variety of neurological and psychiatric disorders such as epilepsy, autism, Alzheimer's disease, and depression. Unlike other types of neurons, which can be generated relatively easily by direct reprogramming, it is difficult to generate GABAergic neurons by traditional methods. Neuronal transdifferentiation of fibroblasts mediated by nongenomic-integrated adenovirus has many advantages, but the efficiency is low, and there is a lack of studies using human cells as the initial materials. In this study, we explored the feasibility of the conversion of human fibroblasts into neurons through adenovirus-mediated gene expression and found that by introducing two microRNAs, miR-124 and let-7, together with several small chemical compounds, they can effectively generate GABAergic neuron-like cells from human neonatal fibroblasts without reverting to a progenitor cell stage. Most of these cells expressed neuronal markers and were all somatostatin (SST)-positive cells. Therefore, our study provides a relatively safe and efficient method to generate SST interneurons.

Surfactant-Free Chitosan/Cellulose Acetate Phthalate Nanoparticles: An Attempt to Solve the Needs of Captopril Administration in Paediatrics.

The Paediatric Committee of the European Medicines Agency encourages research into medicinal products for children, in particular, the development of an age-appropriate formulation of captopril is required in the cardiovascular therapeutic area. The aim of this study was the development of a liquid formulation using nanoparticles based only on chitosan and cellulose acetate phthalate containing captopril for the treatment of hypertension, heart failure and diabetic nephropathy in paediatric patients. Nanoparticles were prepared by a nanoprecipitation method/dropping technique without using surfactants, whose use can be associated with toxicity. A range of different cellulose to chitosan weight ratios were tested. A good encapsulation efficiency (61.0 ± 6.5%) was obtained when a high chitosan concentration was used (1:3 ratio); these nanoparticles (named NP-C) were spherical with a mean diameter of 427.1 ± 32.7 nm, 0.17 ± 0.09 PDI and +53.30 ± 0.95 mV zeta potential. NP-C dispersion remained stable for 28 days in terms of size and drug content and no captopril degradation was observed. NP-C dispersion released 70% of captopril after 2 h in pH 7.4 phosphate buffer and NP-C dispersion did not have a cytotoxicity effect on neonatal human fibroblasts except at the highest dose tested after 48 h. As a result, chitosan/cellulose nanoparticles could be considered a suitable platform for captopril delivery in paediatrics for preparing solid/liquid dosage forms.

Homo Sapiens (Hsa)-microRNA (miR)-6727-5p Contributes to the Impact of High-Density Lipoproteins on Fibroblast Wound Healing In Vitro.

Chronic, non-healing wounds are a significant cause of global morbidity and mortality, and strategies to improve delayed wound closure represent an unmet clinical need. High-density lipoproteins (HDL) can enhance wound healing, but exploitation of this finding is challenging due to the complexity and instability of these heterogeneous lipoproteins. The responsiveness of primary human neonatal keratinocytes, and neonatal and human dermal fibroblasts (HDF) to HDL was confirmed by cholesterol efflux, but promotion of ‘scrape’ wound healing occurred only in primary human neonatal (HDFn) and adult fibroblasts (HDFa). Treatment of human fibroblasts with HDL induced multiple changes in the expression of small non-coding microRNA sequences, determined by microchip array, including hsa-miR-6727-5p. Intriguingly, levels of hsa-miR-6727-5p increased in HDFn, but decreased in HDFa, after exposure to HDL. Delivery of a hsa-miR-6727-5p mimic elicited repression of different target genes in HDFn (ZNF584) and HDFa (EDEM3, KRAS), and promoted wound closure in HDFn. By contrast, a hsa-miR-6727-5p inhibitor promoted wound closure in HDFa. We conclude that HDL treatment exerts distinct effects on the expression of hsa-miR-6727-5p in neonatal and adult fibroblasts, and that this is a sequence which plays differential roles in wound healing in these cell types, but cannot replicate the myriad effects of HDL.

Perspective on the development of a bioengineered patch to treat heart failure: rationale and proposed design of phase I clinical trial

This perspective focuses on the development of tissue engineered (TE) cell-based therapies to treat left ventricular (LV) dysfunction and chronic heart failure (CHF). The development of induced pluripotent stem cells enabled investigators to seed or co-culture human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs) alone and in combination with other cells onto bioengineered scaffolds applied to the epicardial surface of the damaged left ventricle. Using our work as an example, we show how a xenograft implant of a bioengineered scaffold embedded with human neonatal fibroblasts and seeded with hiPSC-CMs partially reversed maladaptive LV remodeling and improved LV systolic/diastolic function in an immune-competent rat model of CHF. The fibroblasts lay down an extracellular matrix and secrete growth factors that increase myocardial blood flow. This approach provides an improved cell payload that covers a larger area of the damaged left ventricle as opposed to direct cell injections into the heart or down the coronary arteries. These studies combined with ongoing studies in immune-competent Yucatan mini swine treated with the same xenograft led to the preliminary design of a proposed Phase I clinical trial that will be presented to the Federal Drug Administration. For the proposed Phase I clinical, this TE patch will be implanted onto the epicardial surface of non-immunosuppressed patients undergoing elective Coronary Artery Bypass Grafting with Ejection Fractions ≥ 20% and ≤ 45%. The primary endpoints will be adverse events/severe adverse events associated with placing the TE patch on the heart. While Phase I trials are primarily safety trials, this proposed trial is designed to obtain some potential efficacy endpoints to help with the design of future Phase II/III clinical trials. These endpoints include changes in LV remodeling that were seen in the pre-clinical animal models as well as including endpoints that focus on patient well-being.

Abstract MP205: Human Induced Pluripotent Stem Cell Derived Cardiomyocyte And Fibroblast Patch To Treat Heart Failure

Background: We tested a tissue engineered (TE) patch composed of a biodegradable mesh embedded with human neonatal fibroblasts and seeded with human induced pluripotent stem cell derived cardiomyocytes (hiPSC-CMs) to treat heart failure in Yucatan mini swine receiving no immune suppression. Methods: Swine (N=12) underwent a 90-minute balloon occlusion/reperfusion of the left anterior descending coronary artery to create a myocardial infarction (MI). Following a 4-week recovery, the TE patch was implanted via a mini median sternotomy. The following were obtained: Cardiac Magnetic Resonance (CMR) imaging, cardiac catheterization, activity monitoring with FitBark collars, treadmill testing, 24/7 ECGs with implanted loop recorders. Results: At 4 weeks after MI, swine had increased left ventricular (LV) volumes, decreased end-systolic elastance (Ees), a shift of the diastolic pressure/volume (P/V) to the right of baseline and an increase in the LV mass/volume. After 6 months of treatment, the TE treated swine (N=7) compared to inert tissue treated swine (N=5): End-systolic volume (2% decrease vs 18% increase); End-diastolic volume (7% decrease vs 26% increase): Ees (1.0±0.2 vs 1.9±0.2 mmHg/mL, P=0.006); the diastolic P/V loops shifted back toward baseline with no change in slope, and LV mass decreased. There was no mortality related to treatment; the TE patch was well tolerated as assessed by CMR and histology. The loop recorders showed TE treated animals remained in sinus rhythm throughout with no ventricular arrhythmias, no change in heart rate and a 20% increase in daily activity levels and a 20% increase in exercise tolerance. Conclusions: This TE patch with human neonatal fibroblasts and hiPSC-CMs improves LV function, partially reverses LV remodeling and improves exercise in non-immune suppressed swine with heart failure after 6 months of treatment.

LB811 Exosomes from human neonatal fibroblasts conditioned media play an important role in skin rejuvenation

Extracellular vesicles (EVs), including exosomes, are lipid bilayer delimited particles that are naturally released from a cell and play a major role in the communication of various cells by serving as vehicles to transfer cytosolic proteins, lipids, and RNAs between cells. Cell culture conditioned media (CCM) from human neonatal fibroblasts cultured under hypoxic conditions have demonstrated potential to stimulate wound healing and skin regeneration. EVs are known to secrete into medium from cultured cells; therefore we characterized the content of exosomes from CCM to investigate its potential role in skin.

No mitigation of IFN-β and HLA class I expression in early sub-cultured human neonatal skin fibroblasts but both molecules are overexpressed in starved cells

Allogeneic fibroblasts containing-dermal substitutes are known as neutral allografts. There is evidence that successive subculturing of fibroblasts leads to a decrease in HLA class I expression. The aim of this study was to investigate this downregulation and its correlation with the expression of interferon-beta (IFN- β) as a HLA class I regulating cytokine. This correlation was also measured under serum starvation that is prevalent in wounds, especially in burn wounds, and that is also used in pre-transplantation fibroblasts culture to manufacture bioengineered skin substitutes on an industrial scale. Fibroblasts isolated from neonatal foreskin were cultured for eight successive passages in DMEM supplemented with bovine serum (n = 3). Those cells were also cultured for different time points in DMEM without serum (n = 4). Flow cytometry, Elisa, and real-time PCR methods were used to determine the levels of HLA class I expression, IFN-β protein and IFN-β mRNA respectively. The expression of HLA class I molecules was not significantly altered by cell subculturing (p > 0.05). Interestingly, these cells also produced substantial levels of IFN-β -mRNA and -protein with non-significant alterations upon successive sub-culturing (p > 0.05). However, HLA expression in starved fibroblasts was significantly raised in a timely-dependent manner (r2 = 0.84, p < 0.001), with strong correlation with IFN-β concentrations (r2 = 0.86, p < 0.0001).These data reveal that sub-cultured allogeneic fibroblasts are not HLA negative cells supported by cells spontaneous production of IFN-β, a multifunctional immunoregulatory cytokine. Even the levels of this antigen can be increased in association with IFN-β up-regulation in certain stressful conditions, such as growth factor deprivation.These findings open a new window in transplantation immunology for immuneregulatory roles of metabolic stress- IFN-β-HLA axis leading to successful allogeneic HLA-unmatched organ transplantation in immunocompetence recipients. Moreover these data can be applied in the optimization of conditions for pre-clinical fibroblasts culture to the better designation of human dermal substitutes.

Integration of choline geranate into electrospun protein scaffolds affords antimicrobial activity to biomaterials used for cutaneous wound healing.

Wound healing attempts to maintain homeostasis in the wound while minimizing the risk of infection to the tissue by foreign agents, such as opportunistic bacterial pathogens. Biofilms established by these pathogens are a common cause of chronic infections that slow the healing process. Preparation of skin wound healing devices comprised of electrospun proteins associated with skin have been shown to accelerate the healing process relative to conventional wound dressings. In this work, we have developed electrospinning methods to incorporate the antimicrobial ionic liquid/deep eutectic solvent choline geranate (CAGE) into these devices. Integration of CAGE into the dressing material was verified via 1 H nuclear magnetic resonance spectrometry, and the effect on the material property of the resultant devices were assessed using scanning electron microscopy. CAGE-containing devices demonstrate a concentration-dependent inactivation of exogenously applied solutions of both gram-positive and gram-negative pathogens (Enterococcus sp and Pseudomonas aeruginosa, respectively), but maintain their ability to serve as a compatible platform for proliferation of human dermal neonatal fibroblasts.

16470 Clinical efficacy of a novel growth factor–based serum: A randomized, double-blind, placebo-controlled, cross-over study

Topical physiologically balanced growth factors have been shown to provide improvements in skin rejuvenation. Advancements in cell culture technology have enabled harvesting of a novel growth factor blend derived from neonatal human fibroblasts cultured under hypoxic conditions, leveraging unique properties exclusive to this particular environment. To assess the efficacy and tolerability of the novel growth factor–based serum (TNSA) on subjects presenting with moderate to severe facial photodamage, a 24-week, randomized, double-blind, vehicle placebo–controlled, cross-over study was conducted. 66 women aged 37-70 with Fitzpatrick skin types I-V completed the 24-week study (Active: n = 43, Control: n = 23). During phase 1 (baseline to week 12), TNSA (Active) and vehicle (Control) were applied to the face twice daily. Both groups also used a basic skincare regimen (cleanser and moisturizer with SPF). During phase 2 (week 12-24), Control crossed-over and received TNSA while Active continued with TNSA. TNSA provided significant improvements over Control in coarse lines/wrinkles at weeks 4, 8, and 12; overall photodamage, fine lines/wrinkles and overall hyperpigmentation at weeks 8 and 12; and sagging at week 12 (all P ≤ .01; Wilcoxon signed rank test). After Control cross-over to TNSA during phase 2, significant improvements in overall photodamage, sagging, coarse lines/wrinkles, and fine lines/wrinkles were achieved at weeks 18 and 24 compared with week 12 “baseline.” Results from the validated patient reported outcome tool Face-Q and histologic analysis of biopsies support improvements observed by the investigator. Study results suggest that the novel growth factor-based serum may provide a treatment option for patients seeking facial skin rejuvenation.

15998 Effects of a topical cosmetic growth factor serum in postmenopausal subjects with moderate to severe facial photodamage

Menopause, defined by the cessation of menses, is associated with hormonal alterations including decreased estrogen production. Because the skin contains numerous estrogen receptors, these hormonal changes can significantly affect the skin, including diminished collagen levels, turnover and synthesis. Visible manifestations of menopause-related skin changes may include dryness, lines/wrinkles and sagging. Limited studies have been conducted using topical cosmetic products on the postmenopausal patient population. Recently, a novel topical growth factor serum (test product) derived from neonatal human fibroblasts cultured under hypoxic conditions was clinically-proven to improve the appearance of moderate to severe facial photodamage. To explore the effects of the test product in postmenopausal subjects, a 12-week clinical study was conducted. 15 postmenopausal female subjects aged 46-65 years with Fitzpatrick skin types I-III and presenting with moderate to severe facial photodamage, completed the study. Subjects applied the test product twice daily, morning and evening for 12 weeks. Visits occurred at baseline, week 6 and week 12 and included investigator assessments, questionnaires and standardized photography. Significant improvements in investigator assessments for overall photodamage, skin tone evenness, perioral fine lines/wrinkles, periocular and cheeks coarse lines/wrinkles and tactile roughness were observed at all follow-up visits (all P ≤ .02; paired t test). At week 12, significant improvements were also observed for periocular, forehead and cheeks fine lines/wrinkles and perioral coarse lines/wrinkles (all P ≤ .04; paired t test). Study results suggest that the test product may help improve the appearance of facial skin quality in postmenopausal subjects. However additional studies in this unique population are needed.

Antimicrobial Properties of Gallium(III)- and Iron(III)-Loaded Polysaccharides Affecting the Growth of Escherichia coli, Staphylococcus aureus, and Pseudomonas aeruginosa, In Vitro

Antimicrobial resistance (AMR) has become a global concern as many bacterial species have developed resistance to commonly prescribed antibiotics, making them ineffective to treatments. One type of antibiotics, gallium(III) compounds, stands out as possible candidates due to their unique "Trojan horse" mechanism to tackle bacterial growth, by substituting iron(III) in the metabolic cycles of bacteria. In this study, we tested three polysaccharides (carboxymethyl cellulose (CMC), alginate, and pectin) as the binding and delivery agent for gallium on three bacteria (Pseudomonas aeruginosa, Escherichia coli, and Staphylococcus aureus) with a potential bioresponsive delivery mode. Two types of analysis on bacterial growth (minimum inhibitory concentrations (MIC) and minimum bactericidal concentrations (MBC)) were carried out while iron(III)-loaded polysaccharide samples were also tested for comparison. The results suggested that gallium showed an improved inhibitory activity on bacterial growth, in particular gallium(III)-loaded carboxymethyl cellulose (Ga-CMC) sample showing an inhibiting effect on growth for all three tested bacteria. At the MIC for all three bacteria, Ga-CMC showed no cytotoxicity effect on human dermal neonatal fibroblasts (HDNF). Therefore, these bioresponsive gallium(III) polysaccharide compounds show significant potential to be developed as the next-generation antibacterial agents with controlled release capability.

Calreticulin exploits TGF-β for extracellular matrix induction engineering a tissue regenerative process.

Topical application of extracellular calreticulin (eCRT), an ER chaperone protein, in animal models enhances wound healing and induces tissue regeneration evidenced by epidermal appendage neogenesis and lack of scarring. In addition to chemoattraction of cells critical to the wound healing process, eCRT induces abundant neo-dermal extracellular matrix (ECM) formation by 3days post-wounding. The purpose of this study was to determine the mechanisms involved in eCRT induction of ECM. In vitro, eCRT strongly induces collagen I, fibronectin, elastin, α-smooth muscle actin in human adult dermal (HDFs) and neonatal fibroblasts (HFFs) mainly via TGF-β canonical signaling and Smad2/3 activation; RAP, an inhibitor of LRP1 blocked eCRT ECM induction. Conversely, eCRT induction of α5 and β1 integrins was not mediated by TGF-β signaling nor inhibited by RAP. Whereas eCRT strongly induces ECM and integrin α5 proteins in K41 wild-type mouse embryo fibroblasts (MEFs), CRT null MEFs were unresponsive. The data show that eCRT induces the synthesis and release of TGF-β3 first via LRP1 or other receptor signaling and later induces ECM proteins via LRP1 signaling subsequently initiating TGF-β receptor signaling for intracellular CRT (iCRT)-dependent induction of TGF-β1 and ECM proteins. In addition, TGF-β1 induces 2-3-fold higher level of ECM proteins than eCRT. Whereas eCRT and iCRT converge for ECM induction, we propose that eCRT attenuates TGF-β-mediated fibrosis/scarring to achieve tissue regeneration.

De novo production of human extracellular matrix supports increased throughput and cellular complexity in 3D skin equivalent model.

Three-dimensional (3D) tissue models of human skin are being developed to better understand disease phenotypes and to screen new drugs for potential therapies. Several factors will increase the value of these in vitro 3D skin tissues for these purposes. These include the need for human-derived extracellular matrix (ECM), higher throughput tissue formats, and greater cellular complexity. Here, we present an approach for the fabrication of 3D skin-like tissues as a platform that addresses these three considerations. We demonstrate that human adult and neonatal fibroblasts deposit an endogenous ECM de novo that serves as an effective stroma for full epithelial tissue development and differentiation. We have miniaturized these tissues to a 24-well format to adapt them for eventual higher throughput drug screening. We have shown that monocytes from the peripheral blood can be incorporated into this model as macrophages to increase tissue complexity. This humanized skin-like tissue decreases dependency on animal-derived ECM while increasing cellular complexity that can enable screening inflammatory responses in tissue models of human skin.

Human Induced Pluripotent Stem Cell–Derived Cardiomyocyte Patch in Rats With Heart Failure

To treat chronic heart failure (CHF), we developed a robust, easy to handle bioabsorbable tissue-engineered patch embedded with human neonatal fibroblasts and human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs). This patch was implanted on the epicardial surface of the heart covering the previously infarcted tissue. Sprague-Dawley rats (6-8 weeks old) underwent sham surgery (n= 12) or left coronary artery ligation (n= 45). CHF rats were randomized 3 weeks after ligation to CHF control with sham thoracotomy (n= 21), or a fibroblasts/hiPSC-CMs patch (n= 24) was implanted. All sham surgery rats also underwent a sham thoracotomy. At 3 weeks after randomization, hemodynamics, echocardiography, electrophysiologic, and cell survival studies were performed. Patch-treated rats had decreased (P < .05) left ventricular-end diastolic pressure and the time constant of left ventricular relaxation (Tau), increased anterior wall thickness in diastole, and improved echocardiography-derived indices of diastolic function (E/e' [ratio of early peak flow velocity to early peak LV velocity] and e'/a' [ratio of early to late peak left ventricular velocity]). All rats remained in normal sinus rhythm, with no dysrhythmias. Rats treated with the patch showed improved electrical activity. Transplanted hiPSC-CMs were present at 7 days but not detected at 21 days after implantation. The patch increased (P < .05) gene expression of vascular endothelial growth factor, angiopoietin 1, gap junction α-1 protein (connexin 43), β-myosin heavy 7, and insulin growth factor-1 expression in the infarcted heart. Epicardial implantation of a fibroblasts/hiPSC-CMs patch electrically enhanced conduction, lowered left ventricular end-diastolic pressure, and improved diastolic function in rats with CHF. These changes were associated with increases in cytokine expression.

Cytoprotective effect of a functional antipollutant blend through reducing B[a] P-induced intracellular oxidative stress and UVA exposure.

Benzo[a]pyrene (B[a]P) is a ubiquitous environmental pollutant that reacts with skin and induces intracellular oxidative stress through reactive oxygen species (ROS) accumulation. The antipollution properties of natural extracts, especially including antioxidants, for inhibiting ROS in cells are gaining importance, in addition to the anticancer effects attributed to them. In this study, a commercial functional antipollutant blend of plant extracts consisting of ellagic acid standardized Punica granatum peel extract, Sambucus nigra fruit extract, Prunus cerasus seed extract, and hydrolyzed wheat protein with high antioxidant properties and UV damage-protective properties attributed to each one was investigated. The cytoprotective effect of this functional antipollutant blend was determined by ROS assay through reducing the level of intracellular ROS induced by B[a]P as an oxidative stress factor in human neonatal keratinocytes and fibroblast cells. In addition, the cytoprotective effect of the functional antipollutant blend after UVA exposure was also determined. It is shown that the oxidative damage induced by B[a]P and UVA, which are the most abundant factors of chemical and physical pollution, would be prevented by the functional antipollutant blend. Thus, it can be concluded that this antipollutant functional blend may offer a promising ingredient for the cosmetic industry's skincare products.

The Role of Serotonin during Skin Healing in Post-Thermal Injury.

Post-burn trauma significantly raises tissue serotonin concentration at the initial stages of injury, which leads us to investigate its possible role in post burn wound healing. Therefore, we planned this study to examine the role of serotonin in wound healing through in vitro and in vivo models of burn injuries. Results from in vitro analysis revealed that serotonin decreased apoptosis and increased cell survival significantly in human fibroblasts and neonatal keratinocytes. Cellular proliferation also increased significantly in both cell types. Moreover, serotonin stimulation significantly accelerated the cell migration, resulting in narrowing of the scratch zone in human neonatal keratinocytes and fibroblasts cultures. Whereas, fluoxetine (a selective serotonin reuptake inhibitor) and ketanserin (serotonin receptor 2A inhibitor) reversed these effects. Scald burn mice model (20% total body surface area) showed that endogenous serotonin improved wound healing process in control group, whereas fluoxetine and ketanserin treatments (disruptors of endogenous serotonin stimulation), resulted in poor reepithelization, bigger wound size and high alpha smooth muscle actin (α-SMA) count. All of these signs refer a prolonged differentiation state, which ultimately exhibits poor wound healing outcomes. Collectively, data showed that the endogenous serotonin pathway contributes to regulating the skin wound healing process. Hence, the results of this study signify the importance of serotonin as a potential therapeutic candidate for enhancing skin healing in burn patients.