"Ficus johannis Boiss. leaves ethanolic extract ameliorate streptozotocin-induced diabetes in rats by upregulating the expressions of GCK, GLUT4, and IGF and downregulating G6P".

The leaves of Ficus johannis Boiss (F. johannis), commonly known as Fig tree, Anjir, and Teen, are used by the folk medicinal practitioners in Iran for controlling hyperglycemia in diabetic patients. This study investigated the pharmacological basis for antidiabetic effect of the ethanolic extract of F. johannis leaves using in vitro and in vivo experimental models. Qualitative screening of phytochemicals, estimation of total phenolic and flavonoid contents, and in vitro antioxidant and α-amylase inhibition assays were performed. Moreover, the High-performance liquid chromatography (HPLC) quantification, acute toxicity, glucose tolerance, and in vivo antidiabetic effect along with the evaluation of gene expressions involved in diabetes mellitus were carried out. Significant quantities of phenolic (71.208 ± 2.89 mgg-1 GAE) and flavonoid (26.38 ± 3.53 mgg-1 QE) were present. Inhibitory concentration (IC50) of the plant extract exhibited an excellent in vitro antioxidant (IC50 = 33.81 µg/mL) and α-amylase (IC50 = 12.18 µg/mL) inhibitory potential. The HPLC analysis confirmed the gallic acid (257.79 mgg-1) as main constituent of the extract followed by kaempferol (22.86 mgg-1), myricetin (0.16 mgg-1), and quercetin (3.22 mgg-1). Ethanolic extract displayed glucose tolerance in normo-glycemic rats. Streptozotocin-induced hyperglycemia declined dose dependently in the extract treated rats with improvement in lipid profile and liver and renal function biomarkers. The F. johannis-treated groups showed an increase in mRNA expressions of glucose transporter 4 (GLUT-4), glucokinase, insulin growth like factor 1 and peroxisomal proliferator activating receptor gamma in pancreas. However, the Glucose-6-phosphatase was downregulated. Present study suggests that the ethanolic extract of F. johannis leaves demonstrates a good anti-diabetic profile by improving insulin sensitivity, GLUT-4 translocation, and carbohydrate metabolism while inhibiting lipogenesis.

In vivo and in vitro toxicity profile of tetrabutylammonium bromide and alcohol-based deep eutectic solvents.

Deep eutectic solvents (DESs) have emerged as new promising solvents in the field of "green chemistry," which possess a broad range of potential applications. However, the ecotoxicological profile of these solvents is still poorly known. In this study, ammonium-based deep eutectic solutions with glycerol (2:2), ethylene glycol (1:2), and diethylene glycol (1:2) as hydrogen bond donors in 1:2 proportion were evaluated for their interaction with various biological systems, including gram-positive and negative bacteria, fungi, fish, and human fibroblast cell lines. The DES synthesis was confirmed by Fourier transform infrared spectroscopy analysis, which analyses the interactions between DES precursors for their synthesis. The antimicrobial activity of tetrabutylammonium bromide: ethylene glycol was the most potent, while tetrabutylammonium bromide: diethylene glycol had a higher LC50 against C. carpio fish. Tetrabutylammonium bromide: glycerol was supposed to be the most suitable DES in terms of cell viability percentage (118%) and 2,2-diphenyl-1-picrylhydrazyl scavenging activity (93%). Finally, tetrabutylammonium bromide in glycerol can be considered an eco-friendly solvent due to its lower toxicity in both in vivo and in vitro environments.

The frequency and pattern of deranged lipid profile in patients with ischemic stroke: a retrospective study.

BACKGROUND AND AIM: Ischemic Stroke has been recognized as the principal cause of disability and the third leading cause of mortality worldwide. The aim of this study was to determine the frequency and pattern of dyslipidemia in patients presenting with ischemic stroke at a tertiary care hospital in Karachi, Pakistan and to evaluate the effect of demographic and clinical factors on the frequency and pattern of dyslipidemia in ischemic stroke subjects.   Methods: A cross-sectional study carried on a sample size of 235 patients presenting to the out-patient clinic with paralysis, difficulty in speech, and/or loss of consciousness lasting for one hour or more. Blood samples were analyzed for total cholesterol (TC), triglycerides (TG), low-density lipoproteins (LDL), very low-density lipoproteins (VLDL) and high-density lipoproteins (HDL) by the enzymatic colorimetric methodology. These values were recorded on the pre-defined proforma by the investigators. All analysis was performed using SPSS version 23.0.   Results: The average age of the patients was 50.84±11.51 years and 62.1% of them were males. The frequency of dyslipidemia was observed in more than half (n=134/235, 57.02%) of ischemic stroke patients. Regarding the dyslipidemia pattern, TC, VLDL-C and TG levels were deranged in more than 50% of the cases. The most commonly deranged values were of TC and VLDL-C, followed by TG levels. It was observed that patients with a previous history of DM (73.9%, p=0.002) and HTN (81.3%, p=0.001) had significantly higher rates of deranged lipid profiles. Lipid values were found to be more deranged in patients aged 41-50 years (p=0.002) however, no statistically significant differences were observed with respect to BMI (p=0.192) and symptoms duration (p=0.334).   Conclusions: Dyslipidemia is an important risk factor for ischemic stroke, and elevated LDL-C is usually the lipid fraction implicated in the pathologic mechanism of stroke.

Intensivist and COVID-19 in the United States of America: a narrative review of clinical roles, current workforce, and future direction.

COVID-19 continues to spread across borders and has proven to be a challenge for the existing healthcare system. The demand for intensivists has dramatically increased in the United States, in the backdrop of an expected lack of intensivists in many States even before the pandemic. One proposal has been to organize multidisciplinary teams functioning under one intensivist, as this approach would make use of the existing healthcare force and lessen the burden on intensivists. Another recommendation is the adaptation of Tele-ICUs, which have demonstrated constructive outcomes in the past. Moreover, ensuring the provision of all types of personal protective equipment, adequate testing and, other provisions such as mental health support, financial incentives for intensivists should be prioritized. More intensivists should be trained for the future, for which better institutional policies are essential.

Role and Perspectives of Inflammation and C-Reactive Protein (CRP) in Psychosis: An Economic and Widespread Tool for Assessing the Disease.

Schizophrenia is a major psychotic disorder affecting nearly 23.6 million people globally and greatly impacting the cognitive and social functioning of individuals. Multiple risk factors, including genetic, environmental, and epigenetic factors have been identified. However, the exact mechanism by which some factors aid in the development of schizophrenia is still uncertain. Acute and/or long-standing inflammation has been implicated as both a cause and effect of schizophrenia. Heightened immune responses have been documented in large cohorts of individuals with schizophrenia. While not completely known, multiple hypotheses, such as disruption of the blood-brain barrier, alterations in the kynurenine/tryptophan pathway, and increased microglial activation, have been presented to correlate inflammation with schizophrenic symptoms. Measurement of C-reactive protein (CRP) is a commonly performed and inexpensive test on patients' serum to determine levels of systemic inflammation in the body. Multiple studies have reported an elevated CRP level in different stages of schizophrenia, indicating its potential to be used as a viable biomarker in the diagnosis and monitoring of schizophrenia along with assessing treatment response to conventional and non-conventional treatment regimens. This review aims to evaluate the role of inflammation, in general, and CRP, in particular, in the pathogenesis of schizophrenia and its potential significance in diagnostic, therapeutic, and preventative approaches towards schizophrenia and psychosis.

Cardiac manifestations of the coronavirus disease-19: a review of pathogenesis, clinical manifestations, diagnosis, and treatment.

The coronavirus disease-19 (COVID-19), first appearing in Wuhan, China, and later declared as a pandemic, has caused serious morbidity and mortality worldwide. Severe cases usually present with acute respiratory distress syndrome (ARDS), pneumonia, acute kidney injury (AKI), liver damage, or septic shock. However, with recent advances in severe acute respiratory syndrome-coronavirus-2 (SARS-CoV-2) research, the virus´s effect on cardiac tissues has become evident. Reportedly, an increased number of COVID-19 patients manifested serious cardiac complications such as heart failure, increased troponin, and N-terminal pro-B-type natriuretic peptide levels (NT-proBNP), cardiomyopathies, and myocarditis. These cardiac complications initially present as chest tightness, chest pain, and heart palpitations. Diagnostic investigations such as telemetry, electrocardiogram (ECG), cardiac biomarkers (troponin, NT-proBNP), and inflammatory markers (D-dimer, fibrinogen, PT, PTT), must be performed according to the patient´s condition. The best available options for treatment are the provision of supportive care, anti-viral therapy, hemodynamic monitoring, IL-6 blockers, statins, thrombolytic, and anti-hypertensive drugs. Cardiovascular disease (CVD) healthcare workers should be well-informed about the evolving research regarding COVID-19 and approach as a multi-disciplinary team to devise effective strategies for challenging situations to reduce cardiac complications.

Generalized Anxiety Disorder and Depressive Symptoms among Pakistani Population during the Second Wave of the COVID-19 Pandemic: A Regression Analysis.

Over a span of 1 year, with millions infected, COVID-19 has spread to every part of the world and now poses a health threat to each and every one of us. The outbreak has consequently resulted in multiple health problems such as stress, anxiety, depressive symptoms, insomnia, panic, and denial globally. Several factors have contributed to this rising number of psychiatric consults all over the world. The primary objective of this study was to investigate the impact of COVID-19 pandemic on the mental health of Pakistani population during the second wave of the pandemic in this region. We conducted an online web-based cross-sectional survey comprising 500 participants. The questionnaire assessed the demographic information, attitude, and knowledge concerning COVID-19 outbreak in addition to generalized anxiety disorder (GAD) utilizing the GAD-7 scale and depressive symptoms using the Center for Epidemiology Scale for Depression (CES-D) scale. The response rate of the study was 90.9%. The results of the survey indicated a prevalence of 25.4% of GAD, and 18.8% of depressive symptoms. Furthermore, nearly 34.8% of participants feared contracting COVID-19, 62.8% obtained constant critical updates regarding COVID-19, while 17.6% did not understand the knowledge regarding COVID-19. In the multivariate regression models, GAD was significantly associated with gender, age, and checking constantly of critical updates regarding COVID-19. Similarly, participants under 30 years had a higher risk of developing depressive symptoms than those above (> 30 years). Lastly, participants with no formal education were also found to be more prone to developing depression. We identified a potential threat to mental health during the pandemic.

Synergistic Effect of Barbadensis miller and Marsdenia Condurango Extracts Induces Apoptosis Promotes Oxidative Stress by Limiting Proliferation of Cervical Cancer and Liver Cancer Cells.

BACKGROUND: Drug synergy is the combine effect of drug efficacy. Synergistic combinations of active ingredients have proven to be highly effective and more useful in therapeutics. In contrast, the individual effect of drug is usually undesirable and mostly used for selecting drug-resistant mutations. Purpose of this study was to check synergistic effects of both plants (Barbadensis miller and Marsdenia condurango) against liver and cervical cancer. METHODOLOGY: Culturing of HeLa (cervical cancer cell line) and HepG2 (liver cancer cell line) cells, IC50 evaluation, viability assays (trypan blue, crystal violet), p53 ELISA and immunocytochemistry, MUSE analysis (count and viability), antioxidants (GSH, SOD, CAT), at the end RT-PCR was performed. RESULTS: IC50 evaluation was done of each plant individually and with combination for synergistic effects, IC50 with plants combination (synergism) was applied on further viability assays (trypan blue, crystal violet, MUSE analysis via count and viability kit) p53 ELISA and immunocytochemistry for evaluation of cellular apoptosis, antioxidants assays (GSH, SOD, CAT), and RT-PCR with proliferative and apoptotic markers along with internal control. CONCLUSION: According to current study it was observed that synergistic effect of these plants has more anticancer properties with minimum effective dose. It was also observed that extracts possess the ability to induce apoptosis, restrict proliferation and enhanced oxidative stress.

Microwave-Assisted Synthesis of (Piperidin-1-yl)quinolin-3-yl)methylene)hydrazinecarbothioamides as Potent Inhibitors of Cholinesterases: A Biochemical and In Silico Approach.

Alzheimer's disease (AD), a progressive neurodegenerative disorder, characterized by central cognitive dysfunction, memory loss, and intellectual decline poses a major public health problem affecting millions of people around the globe. Despite several clinically approved drugs and development of anti-Alzheimer's heterocyclic structural leads, the treatment of AD requires safer hybrid therapeutics with characteristic structural and biochemical properties. In this endeavor, we herein report a microwave-assisted synthesis of a library of quinoline thiosemicarbazones endowed with a piperidine moiety, achieved via the condensation of 6/8-methyl-2-(piperidin-1-yl)quinoline-3-carbaldehydes and (un)substituted thiosemicarbazides. The target N-heterocyclic products were isolated in excellent yields. The structures of all the synthesized compounds were fully established using readily available spectroscopic techniques (FTIR, 1H- and 13C-NMR). Anti-Alzheimer potential of the synthesized heterocyclic compounds was evaluated using acetylcholinesterase (AChE) and butyrylcholinesterase (BChE) enzymes. The in vitro biochemical assay results revealed several compounds as potent inhibitors of both enzymes. Among them, five compounds exhibited IC50 values less than 20 µM. N-(3-chlorophenyl)-2-((8-methyl-2-(piperidin-1-yl)quinolin-3-yl)methylene)hydrazine carbothioamide emerged as the most potent dual inhibitor of AChE and BChE with IC50 values of 9.68 and 11.59 µM, respectively. Various informative structure-activity relationship (SAR) analyses were also concluded indicating the critical role of substitution pattern on the inhibitory efficacy of the tested derivatives. In vitro results were further validated through molecular docking analysis where interactive behavior of the potent inhibitors within the active pocket of enzymes was established. Quinoline thiosemicarbazones were also tested for their cytotoxicity using MTT assay against HepG2 cells. Among the 26 novel compounds, there were five cytotoxical and 18 showed proliferative properties.

Suicide in Healthcare Workers: Determinants, Challenges, and the Impact of COVID-19.

The Coronavirus disease-19 (COVID-19), which first appeared in Wuhan, China, and was later declared a pandemic, has caused significant morbidity and mortality worldwide. Numerous efforts have been made worldwide to understand the disease's physical manifestation. However, less emphasis has been placed on the pandemic's mental health challenges for healthcare workers (HCWs) who played a critical role in fighting the disease. Existing literature shows the detrimental psychological impact and increased incidence of depression and anxiety among HCWs. It is expected that the mental health crisis will become a serious issue affecting HCWs, with long-term negative consequences following COVID. Physicians and nurses already represent the highest risk groups of suicide among the general population, and suicide can be regarded as an occupational hazard in the healthcare industry. Increased workload, burnout and fatigue, multifaceted challenges women HCWs, and increased substance abuse are contributing factors to suicide ideation. In this article, we identify the risk factors of suicide among HCWs, discuss mental health challenges exacerbated by the pandemic and its impact on suicide ideation.

Effect of COVID-19 on Mental Health Rehabilitation Centers.

The SARS-CoV-2 outbreak is an unprecedented event in modern history worldwide. To facilitate speedy and smooth recovery during this time period, an added responsibility is placed upon rehabilitation center counselors, staff and management. In this paper we expose the role that psychiatric rehabilitation plays during the COVID-19 crisis. Since COVID-19 pandemic has remarkably raised mental health concerns, one of the high risks and possibly neglected groups includes individuals undergoing mental health rehabilitation, the impact on which can be significant as compared to the rest of the population. In order to ensure that individuals on the road to recovery are not alone and forgotten during the times of coronavirus crisis they should be provided with adequate resources, such as online meetings and group therapy sessions, assessment, treatment and support by telephone and video consultation and getting the medications they need to recover. This article highlights how the coronavirus pandemic, that has hit hard the health care and all categories of patients affected by mental disorders, is likely to be particularly dramatic for mental health rehabilitation centers. Indeed, the COVID-19 crisis has severely tested the health systems worldwide, reducing sometimes the quality and quantity of care offered to psychiatric patients both hospitalized and not in rehabilitation centers. The main goal of this paper is to raise awareness of the importance, often underestimated, of mental health rehabilitation centers, on which one should invest for the both future of psychological and psychiatric rehabilitation and the current crisis as well.

Knowledge, attitude and practices of medical undergraduates of Rawalpindi Medical University regarding potential organ donation.

OBJECTIVE: To determine the knowledge, attitude and practices of medical undergraduates regarding potential organ donation. METHODS: The descriptive cross-sectional study was conducted from April to July 2017 at Rawalpindi Medical University, Rawalpindi, Pakistan and comprised medical students of either gender selected from each of the five academic years. Data was collected using a structured questionnaire on knowledge, attitude and practices. Data was analysed using SPSS 22. RESULTS: Of the 400 subjects, there were 40(10%) male and 40(10%) female students from each of the five academic years. The mean age of the sample was 20.98+/-1.63 years (range: 18-27 years). Overall, 363(90.8%) students had knowledge of organ donation and 96(24%) knew about potential organ donation. Of the 184(46%) subjects willing for live donation, 86(46.7%) were males and 98(53.2%) were females. Of the 245(61.5%) subjects willing for potential organ donation, 62(77.5%) were from the 5th year and 30(37.5%) were from the 1st year. CONCLUSIONS: Medical students were found to have good knowledge regarding live organ donation, but they had very little information about potential organ donation.

Knowledge, attitude and practice of medical students towards HIV patients in their pre-clinical and post-clinical years in Karachi, Pakistan: a dual-center cross-sectional study.

The human immunodeficiency virus (HIV) is currently a global threat with an estimated 38.6 million people affected with HIV worldwide. According to the Joint United Nations Program on HIV/AIDS (UNAIDS), since 2004 the total number of cases of HIV in Pakistan has risen from 2700 to 130,000. In light of the rising burden of HIV/AIDS across the country, it is essential that medical students possess appropriate knowledge regarding the subject. Therefore, we aimed to assess the knowledge, attitude and practice of medical students towards HIV patients in their pre-clinical and post-clinical years in Karachi, Pakistan. A cross-sectional study was conducted among 518 pre-clinical (year 1 and 2) and post-clinical (year 3, 4, and 5) medical students from two medical schools in Karachi during the months of October - December 2019. Similar numbers of participants were taken from each year. Data were analyzed using SPSS. Descriptive statistics were used to report frequencies and proportions for categorical responses. Chi-square and Kruskal-Wallis tests were used as the primary statistical tests. About 55% of participants were female, and most belonged to the Islamic faith. More than half of the participants learned about HIV from books (315/518), followed by medical personnel (287/518). A quarter (134/518) of the participants believed HIV could be transmitted by sharing saliva, more than half of whom consisted of pre-clinical year students. Over half the participants (60.4%) knew that there was a difference between HIV and AIDs, most of whom belonged to the 5th year group. When detecting HIV, only about 30% of participants knew about indirect fluorescent antibody. Regarding attitudes, one-third would not be friends with a person diagnosed with HIV/AIDS. As a medical officer, a large majority (76.6%; n=397/518) of the participants would be anxious or somewhat anxious. Two-thirds believed that treating an HIV patient can make them contract HIV, and a majority of participants (333/518) did not feel adequately prepared to deal with the psycho-social problems of an HIV/AIDS patient. Finally, regarding practice, only one-third of the participants were willing to treat an HIV/AIDS patient, most of whom belonged to the pre-clinical 2nd year group and fewer to the post-clinical 3rd year group. Knowledge amongst medical students regarding HIV/AIDS was generally high, although there are some knowledge inadequacies which require more emphasis in the medical school curriculum. However, contrasting with the level of knowledge, in terms of attitude the majority were anxious or somewhat anxious when treating an HIV patient, and only one-third were willing to treat a patient with HIV.

Delonix regia a Folklore Remedy for Diabetes; Attenuates Oxidative Stress and Modulates Type II Diabetes Mellitus.

BACKGROUND: The folkloric profile of Delonix regia demonstrates that it can be used in the management of diabetes. OBJECTIVE: The present study was conducted to evaluate the safety profile of the aerial part extracts of Delonix regia and their antidiabetic potential along with improvement in oxidative stress. MATERIALS AND METHODS: Phytochemical screening, total phenolic, and flavonoid contents along with in-vitro antioxidant and alpha-amylase inhibitory activities were determined. HPLC analysis, acute toxicity, glucose tolerance, in-vivo antidiabetic effect along with the influence on biochemical, oxidative stress parameters, and comet assay of the active extract were performed and assessed. RESULTS: Total phenolic (831.6±0.002 mg/g GAE) and flavonoid (361.4±0.002 mg/g QE) contents were found to be higher in the methanolic extract. Inhibitory concentration IC50 indicated better results for the methanolic extract in DPPH (47.6µg/mL) and alpha-amylase inhibitory (14.61µg/mL) assays. HPLC analysis of the methanolic extract confirmed the presence of quercetin, gallic acid, caffeic acid, cinnamic acid, ferulic acid, and p-coumaric acid. Acute oral toxicity exhibited no mortality and morbidity during the 24h period. The methanolic extract showed better tolerance to glucose. Streptozotocin- nicotinamide (55-110 mg/kg) induced hyperglycemia declined along with improvement in hematological, biochemical parameters and oxidative stress markers (SOD, CAT, H202) in a dose-dependent manner. The maximum effect was recorded at 500mg/kg dose. Comet assay was performed for genotoxic studies and it was observed that the methanolic extract of Delonix regia showed the maximum genoprotective effect at 100µg/mL. CONCLUSION: The findings suggest that the methanolic aerial part extract of Delonix regia exhibited hypoglycemic, antioxidant, and hypolipidemic activities. The antidiabetic effect was comparable to glibenclamide suggesting its therapeutic use as a natural anti-diabetic remedy.

Hepato-protective role of itraconazole mediated cytochrome p450 pathway inhibition in liver fibrosis.

Liver is a vital organ and is routinely exposed to toxins. Carbon tetrachloride is one such noxious agent which cause toxicity in liver when CYP450 enzyme bio-activates it. Many hepatoprotective agents are available in market with severe side effects. Appropriate agent is required to combat such liver problems. Azole compounds have much therapeutic values in many diseases. Based upon this fact, present study is aimed to evaluate the repurposing of Itraconazole in the prevention of hepatic fibrosis via inhibition of cytochrome P450 pathway. For in-vitro evaluation of cyto-protective effects in HepG2 cells (untreated and treated groups), cell viability assays, antioxidant evaluation, enzyme linked immunosorbent assay (ELISA) and immunocytochemistry was used. For in-vivo evaluation, CCl4 induced liver fibrotic rat model was used and post treated evaluation was done by blood biochemistry, hematoxylin and eosin (H&E) staining and gene expression profiling. Results of the current study indicated hepatoprotective role of itraconazole via inhibition of CYP450 pathway inhibition. Therefore, Itraconazole use could be a potential therapeutic approach to prevent liver fibrosis.

In-Vitro Anti-Proliferative, Apoptotic and Antioxidative Activities of Medicinal Herb Kalonji (Nigella sativa).

BACKGROUND: Natural product with apoptotic activity could serve as a potential new source for anti-cancer medicine. Numerous phytochemicals from plants have shown to exert antineoplastic effects via programmed cell death (apoptosis). Cancer is one of the leading causes of death in prosperous countries. The subject study was intended to evaluate the anticancer properties of Kalonji extracts against cancer cell lines HeLa and HepG2 and normal cell lines BHK and VERO were used as normal controls. MATERIALS & METHODS: For the evaluation of anti-proliferative effects, cell viability and cell death in all groups of cells were evaluated via MTT, crystal violet and trypan blue assays. For the evaluation of angiogenesis, Immunocytochemistry and ELISA of VEGF were done. Immunocytochemistry and ELISA of Annexin-V and p53 were performed for the estimation of apoptosis in all groups of cells. Furthermore, LDH assay, antioxidant enzymes activity (GSH, APOX, CAT and SOD) and RT-PCR with proliferative and apoptotic markers along with internal control were also performed. Cancer cells of both cell lines HepG2 and HeLa cells showed reduced viability, angiogenesis and proliferation with increased apoptosis when treated with Kalonji extracts. Whereas anti-oxidative enzymes show enhanced levels in treated cancer cells as compared to untreated ones. CONCLUSION: It was observed that Kalonji extracts have the ability to induce apoptosis and improve the antioxidant status of HeLa and HepG2 cells. They can also inhibit the proliferation and angiogenesis in both these cancer cell lines.

Bi-layered α-tocopherol acetate loaded membranes for potential wound healing and skin regeneration.

With an increase in the demand for skin regeneration products, there is a noticeable increase in developing materials that encourage, wound healing and skin regeneration. It has been reported that antioxidants play an important role in anti-inflammatory reactions, cellular proliferation and remodeling phase of wound healing. While consideration all these factors, a novel α-tocopherol acetate (vitamin E) (VE) loaded bi-layered electrospun membrane, based on lower polycaprolactone (PCL) layer and upper polylactic acid (PLA) layer, was fabricated through electrospinning. Fourier-transform infrared spectroscopy (FTIR), scanning electron microscopy (SEM), in-vitro degradation studies, swelling studies and VE release studies were performed to evaluate structural, physical and in-vitro behavior of membranes. Biological properties of membranes were evaluated through cell proliferation assay, cell adhesion studies, live/dead cell assay and CAM assay. SEM images showed that the average diameter of nanofibers ranged from 1 to 6 µm, while addition of VE changed the diameter and morphology of fibers. Bi-layered membranes showed significant swelling behavior through water uptake, membranes loaded with 30% VE showed 8.7% and 6.8% degradation in lysozyme and H2O2 respectively. 20% and 30% VE loaded membranes followed Korsmeyer-Peppas and first order drug release kinetics followed by non-fickian drug release kinetics. Membranes showed non-toxic behavior and supported cell proliferation via alamar blue assay, cell adhesion via SEM, cell viability via live/dead assay and wound healing by scratch assay. CAM assay showed that membranes having VE supported angiogenesis and showed significant formation of blood vessels making it suitable for skin regeneration and wound healing. Results showed that large surface area of nanofibers, porous structure and biocompatible nature are suitable for targeted clinical applications.

Human neonatal stem cell-derived skin substitute improves healing of severe burn wounds in a rat model.

Conventional approaches can repair minor skin injuries; however, severe burn injuries require innovative approaches for efficient and better wound repair. Recent studies indicate that stem cell-based regenerative therapies can restore severe damaged skin both structurally and functionally. The current study aims to evaluate the wound healing potential of skin substitute derived from human neonatal stem cells (hNSCs) using a severe burn injury rat model. Amniotic epithelial cells (AECs) and mesenchymal stem cells (MSCs) were isolated from placenta (a source of neonatal stem cells) by explant culture method. After characterization, AECs and umbilical cord-MSCs were differentiated into keratinocyte and fibroblasts, respectively. Morphological changes, and expression of corresponding keratinocyte and fibroblast specific markers were used to verify differentiation into respective lineage. A skin substitute was developed by mixing hNSCs-derived skin cells (hNSCs-SCs) in plasma for transplantation in a rat model of severe burn injury. Results indicated that placenta-derived AECs and MSCs were efficiently differentiated into skin cells, that is, keratinocytes and fibroblasts, respectively, as indicated by morphological changes, immunostaining, and polymerase chain reaction analysis. Further, transplantation of hNSCs-SCs seeded in plasma significantly improved basic skin architecture, re-epithelization rate, and wound healing concurrent with reduced apoptosis. In conclusion, neonatal stem cell-derived skin substitute efficiently improved severe burn wounds in a rat model of burn injury. Unique properties of placenta-derived stem cells make them superlative candidates for the development of "off-the-shelf" artificial skin substitutes for future use.