Biodegradable nanofiber coated human umbilical cord as nerve scaffold for sciatic nerve regeneration in albino Wistar rats.

BACKGROUND: Human umbilical cord (hUC) is encompassed by a mucoid connective tissue called Wharton's jelly (WJ), made of hyaluronic acid, collagen, and stromal cells to support the blood vessels of hUC. This study was aimed to determine the in vitro neuronal differentiation of WJ-derived mesenchymal stem cells (WJMSCs), and in vivo axonal regeneration potential of nanofiber coated human Wharton's jelly as a neuronal graft after sciatic nerve injury in immunosuppressed albino Wistar rats. MATERIALS AND METHODS: Wharton's jelly-derived mesenchymal stem cells could be differentiated to neuron-like cells by inducing with neuronic supplementing media. The test animal's axotomized nerves were implanted with trimmed human umbilical cord devoid of vascularity and nanocoated with electro-spun poly-l-lactic acid nanofibers. The control animals were bridged with native sciatic nerve reversed and sutured. Post-surgical functional recovery was studied by walking track, pinprick, muscle weight, and sweating quantification. At the end of the 4th week, the animals were euthanized, and magnetoneurography was performed. The explanted grafts were quantified by immunohistochemistry for immuno-rejection, neural scarring, neural adhesion axon regeneration, fibre diameter, myelin thickness, and G-ratio. The sciatic function index values were similar by walking track analysis for both the test and control groups. RESULTS: The animals had functional and sensation recovery by the end of 2 weeks. No mortality, signs of inflammation, and acute immune rejection were observed post-surgery. CONCLUSIONS: The hUCWJ devoid of vascular elements can be a perfect peripheral nerve graft, and we hypothesis that the cryopreserved hUC could be an ideal resource for axonal regeneration in the future.

Deep sequencing unveils altered cardiac miRNome in congenital heart disease.

Congenital heart disease (CHD) surges from fetal cardiac dysmorphogenesis and chiefly contributes to perinatal morbidity and cardiovascular disease mortality. A continual rise in prevalence and prerequisite postoperative disease management creates need for better understanding and new strategies to control the disease. The interaction between genetic and non-genetic factors roots the multifactorial status of this disease, which remains incompletely explored. The small non-coding microRNAs (miRs, miRNAs) regulate several biological processes via post-transcriptional regulation of gene expression. Abnormal expression of miRs in developing and adult heart is associated with anomalous cardiac cell differentiation, cardiac dysfunction, and cardiovascular diseases. Here, we attempt to discover the changes in cardiac miRNA transcriptome in CHD patients over those without CHD (non-CHD) and find its role in CHD through functional annotation. This study explores the miRNome in three most commonly occurring CHD subtypes, namely atrial septal defect (ASD), ventricular septal defect (VSD), and tetralogy of fallot (TOF). We found 295 dysregulated miRNAs through high-throughput sequencing of the cardiac tissues. The bioinformatically predicted targets of these differentially expressed miRs were functionally annotated to know they were entailed in cell signal regulatory pathways, profoundly responsible for cell proliferation, survival, angiogenesis, migration and cell cycle regulation. Selective miRs (hsa-miR-221-3p, hsa-miR-218-5p, hsa-miR-873-5p) whose expression was validated by qRT-PCR, have been reported for cardiogenesis, cardiomyocyte proliferation, cardioprotection and cardiac dysfunction. These results indicate that the altered miRNome to be responsible for the disease status in CHD patients. Our data expand the existing knowledge on the epigenetic changes in CHD. In future, characterization of these cardiac-specific miRs will add huge potential to understand cardiac development, function, and molecular pathogenesis of heart diseases with a prospect of epigenetic manipulation for cardiac repair.

Advanced imaging and digitization of preserved heart specimens using virtual reality - A primer.

Introduction: Preserved congenital heart specimens are an important component of training professionals working with children and adults with congenital heart disease. They are curated in few institutions worldwide and not freely accessible. This was a proof-of-concept project to explore the use of advanced cardiac imaging modalities (computed tomography [CT] and magnetic resonance imaging [MRI]) and virtual reality (VR) simulation to assess the feasibility and identify the best method of imaging curated cardiac pathology specimens. Methods: Seven specimens in glass jars with formalin, with varied anatomic lesions, from a curated collection were imaged using MRI and high-dose CT to compare the fidelity of models created via each modality. Three-dimensional (3D) models were created and loaded into a VR headset and viewed in virtual space. Two independent physicians performed a "virtual dissection" and scored the resultant models. Results: The highest fidelity and tissue characterization of more delicate structures was achieved with T2 spoiled gradient-echo sequences on MRI (median score of 4 out of 5). CT (median score of 3), while excellent for external anatomy, lost some fidelity with delicate internal anatomy, even at high-radiation doses. No specimens were damaged. Conclusions: We believe that in vitro heart specimens can be easily scanned with high fidelity at a relatively low cost, without causing damage, using high-dose CT and MRI. The ability to "walk through" different chambers of the heart makes the understanding of anatomy easy and intuitive. VR and 3D printing are technologies that could be easily adapted to digitize preserved heart specimens, making it globally accessible for teaching and training purposes.

Fabrication and Electrospinning of 3D Biodegradable Poly-l-Lactic Acid (PLLA) Nanofibers for Clinical Application.

Poly-L-lactic acid (PLLA) is a biodegradable synthetic polyester synthesized by polymerization or polycondensation. PLLA hydrolytically degrades into lactic acid, a biocompatible metabolic by-product, making it suitable for clinical applications. PLLA scaffolds or nanofibers have been used in various regenerative medicine and drug delivery applications. These scaffolds impart biocompatible properties of high surface area, hydrophobicity, native extracellular properties, and mechanical strength for an organ system. Moreover, PLLA nanofibers hold great promise as drug delivery systems, where fabrication parameters and drug-PLA compatibility greatly affect the drug release kinetics. In this chapter, we present the protocols to fabricate, electrospinning, and validation of 3D PLLA nanofibrous scaffolds for tissue engineering application and offer perspectives on their future use.

3D Printing to Model Surgical Repair of Complex Congenitally Corrected Transposition of the Great Arteries.

We report the use of three-dimensional (3D) modeling to plan surgery for physiologic repair of congenitally corrected transposition of the great arteries with pulmonary atresia, dextrocardia, and complex intra cardiac anatomy. Based on measurements made from the 3D printed model of the actual patient's anatomy, we anticipated using a composite valved conduit (Dacron tube graft, decellularized bovine jugular vein, and aortic homograft) to establish left ventricle-to-pulmonary artery continuity with relief of stenosis involving the pulmonary artery confluence and bilateral branch pulmonary arteries.

Right ventricular outflow tract obstruction: a quest for ideal management.

Management of right ventricular outflow tract obstruction has undergone much change over the last century. Techniques described in the literature include anatomical repairs and the use of various patches, conduits, and innovative grafts. However, many of these approaches require reoperations or catheter-based interventions, leading to increased morbidity, mortality, and cost. The search for the ideal long-lasting conduit continues and there are new techniques on the horizon, using genetic engineering and nanotechnology. This review discusses the evolution of various techniques for repair of right ventricular outflow tract obstruction, past and current conduits, as well as ongoing research.

GSK-3 Inhibitors: A Double-Edged Sword? - An Update on Tideglusib.

GSK-3 inhibitors are an emerging tool for clinical interventions in human diseases and represent a niche area in combinational therapy. They possess diverse facets in applications of nervous system disorders, Type 2 diabetes, regenerative medicine and cancer. However, conflicting reports suggest the controversial role of GSK-3 inhibitors in cancers. This review aims to highlight the rise of GSK-3 inhibitors as tools for molecular-targeted research and its shift to a promising drug candidate. The review also focuses on key GSK-3 inhibitors and their roles in cancer and regenerative medicine with special emphasis to tideglusib. In addition, the decisive roles of GSK-3 in various molecular pathways will be concisely reviewed. Finally, this review concludes the emergence of GSK-3 inhibitors as a 'double-edged sword' in the treatment against human diseases cautioning researchers about the potential ramifications of off-target pharmacological effects.

In-stent restenosis: surgical and histopathological perspective.

Background In-stent restenosis has been recognized as a distinct clinical entity that warrants a repeat procedure either in the form of percutaneous reintervention or coronary artery bypass grafting. Multivessel grafting with endarterectomy and explantation of the stent is rarely performed, with few cases reported in the literature. We aim to study the pathomorphology of the stent-vascular interface in coronary vessels undergoing in-stent restenosis. Methods Over a period of 3 years, 3 patients who had undergone angioplasty for diffuse coronary artery disease developed in-stent restenosis and were advised coronary artery bypass. The mean age was 53 years, the average time from the previous intervention was 77 months. Coronary endarterectomy with stent removal and concomitant multivessel coronary artery bypass was performed. Results Histology showed significant proliferation of the well-endothelialized intima as the reason for in-stent restenosis. There were no signs of local thrombus formation or increased inflammatory activity in any of the specimens. After coronary artery bypass, all patients were asymptomatic at a mean follow-up of 32 months. Conclusion Coronary endarterectomy with stent explantation and multivessel coronary artery bypass is a procedure that requires attention because the need is increasing due to the rise in the number of angioplasties. The complexity of this procedure increases to the extent that the adventitia is involved during stent explantation.

MitraClip and mitral annuloplasty device as a bridge to transplantation.

A 42-year-old man with dilated cardiomyopathy and endstage heart failure was evaluated for heart transplantation. He received a MitraClip and Carillon annuloplasty device for functional mitral regurgitation as palliation for his heart failure. Subsequently, he underwent successful heart transplantation.

Role of Antioxidants in Horse Serum-mediated Vasculitis in Swine: Potential Relevance to Early Treatment in Mitigation of Coronary Arteritis in Kawasaki Disease.

BACKGROUND: Horse serum-induced immune complex coronary vasculitis in swine is the first experimental model to mimic most of the pictures of Kawasaki disease. Immune complex mechanism has been implicated as one of the possible mechanisms in the pathogenesis of vasculitis in Kawasaki disease. Antioxidants have a significant role in the reduction of cardiovascular diseases in both human and animal studies. We tried giving vitamins A, E, and C to treat immune complex vasculitis, in the hope of mitigating coronary vasculitis in Kawasaki disease. METHODS: Our study group consisted of 30 pure bred male piglets of 2-3 months of age, and they were divided into test and control groups. The test (AEC) group (n = 20) received two doses of horse serum, 10 mL (0.65 g protein)/kg body weight at 5-day intervals, and oral vitamins A, E, and C once daily for 14 days. The control group (n = 10) was further divided into the saline group (n = 3) receiving two doses of normal saline and the horse serum group (n = 7) receiving two doses of horse serum at 5-day intervals. Piglets were observed for the rashes and coronary artery dimensions. RESULTS: Both the AEC and the control horse serum group developed rashes after horse serum infusions, but the AEC group developed significantly fewer rashes, and no rashes were seen in the saline group. The control horse serum group (mean ± standard deviation = 2.13 ± 0.72) showed significant coronary artery dilatation, whereas there was no significant dilatation in the AEC group (mean ± standard deviation = 0.81 ± 0.58) or the control saline group (p = 0.002). CONCLUSION: Serum sickness is a prototype of immune complex vasculitis, and the severity can be ameliorated with antioxidants. A trial of therapeutic dosages of vitamins A, E, and C in acute phase of Kawasaki disease, may be effective in mitigation of coronary artery lesion in addition to intravenous immunoglobulin and aspirin.

Correlative Studies Unravelling the Possible Mechanism of Cell Death in Tideglusib-Treated Human Ovarian Teratocarcinoma-Derived PA-1 Cells.

This study aims to unravel the use of GSK-3 inhibitors as viable apoptotic inducers for teratocarcinoma-derived ovarian PA-1 cells. MTT assay was carried out to assess inhibitory concentrations of LiCl and TDG. AO/EB staining and Hoechst 33258 staining were employed to assess the damage. Mitochondrial membrane potential (ΔΨm) and ROS generation were assessed with IC50 concentrations of LiCl and TDG. Tumor-related genes (p53, p21, IL-8, TNF-α, MMP-2, Fas-L, Cox-2, and caspase-3) were assessed with 1/4 IC50, 1/2 IC50, IC50 concentrations by semi-quantitative RT- PCR. Cell cycle analysis was performed with IC50 concentration of LiCl and TDG. Western blot analysis was performed for caspase-3, caspase-7, caspase-9, PARP to estimate the possible damage induced by GSK-3 inhibitors and regulation of GSK-3ß, pGSK-3ß, Cox-2. GSK-3 inhibitors demonstrated a concentration and time-dependent reduction in cell viability, exhibiting significant ROS generation and reduced ΔΨm at their IC50 values. Substantial concentration-dependent gene expression changes with significant upregulation of P21, Cox-2, TNF-α, caspase-3, Fas-L were observed. Protein expression of caspase-3 caspase-7, caspase-9, PARP exhibited significant cleavage in LiCl and TDG-treated cells. Protein expression of Cox-2 was significantly increased in IC50 concentration of TDG. Cell cycle analysis showed significant accumulation of cells at sub-G0-G1.

Tideglusib induces apoptosis in human neuroblastoma IMR32 cells, provoking sub-G0/G1 accumulation and ROS generation.

Neuroblastoma is the most common tumor amongst children amounting to nearly 15% of cancer deaths. This cancer is peculiar in its characteristics, exhibiting differentiation, maturation and metastatic transformation leading to poor prognosis and low survival rates among children. Chemotherapy, though toxic to normal cells, has shown to improve the survival of the patient with emphasis given more towards targeting angiogenesis. Recently, Tideglusib was designed as an 'Orphan Drug' to target the neurodegenerative Alzheimer's disease and gained significant momentum in its function during clinical trials. Duffy et al. recently reported a reduction in cell viability of human IMR32 neuroblastoma cells when treated with Tideglusib at varying concentrations. We investigated the effects of Tideglusib, at various concentrations, compared to Lithium chloride at various concentrations, on IMR32 cells. Lithium, a known GSK-3 inhibitor, was used as a standard to compare the efficiency of Tideglusib in a dose-dependent manner. Cell viability was assessed by MTT assay. The stages of apoptosis were evaluated by AO/EB staining and nuclear damage was determined by Hoechst 33258 staining. Reactive oxygen species (ROS) and mitochondrial membrane potential (ΔΨm) were assessed by DCFDA dye and Rhodamine-123 dye, respectively. Tideglusib reported a significant dose-dependent increase in pro-apoptotic proteins (PARP, Caspase-9, Caspase-7, Caspase-3) and tumor-related genes (FasL, TNF-α, Cox-2, IL-8, Caspase-3). Anti-GSK3 ß, pGSK3 ß, Bcl-2, Akt-1, p-Akt1 protein levels were observed with cells exposed to Tideglusib and Lithium chloride. No significant dose-dependent changes were observed for the mRNA expression of collagenase MMP-2, the tumor suppressor p53, or the cell cycle protein p21. Our study also reports Tideglusib reducing colony formation and increasing the level of sub-G0/G1 population in IMR32 cells. Our investigations report the significance of Tideglusib as a promising apoptotic inducer in human neuroblastoma IMR32 cells. Our study also reports that LiCl reduced cell viability in IMR32 cells inducing apoptosis mediated by ROS generation.

Development of an antidiabetic formulation (ADJ6) and its inhibitory activity against α-amylase and α-glucosidase.

There has recently been much advancement in the diagnosis, treatment, and research of metabolic disorders, especially diabetes. Current research around the world is focused on finding an alternative source of treatment from natural resources for diabetic management, apart from the available synthetic medicines. The present study is a preliminary study of a polyherbal formulation using edible natural resources and an assessment of its antidiabetic activity. The formulation was screened for its phytochemical constituents, total phenols, flavonoids, and vitamin C content. It was also analyzed for its inhibitory effect against the digestive enzymes α-amylase and α-glucosidase, compared with the standard drug acarbose. The formulation showed the presence of major constituents such as steroids, cardiac glycosides, phenols, flavonoids, and saponins. It also had a high level of phenols (340 ± 2.5 mg/g), flavonoids (235.4 ± 8.3 mg/g), and vitamin C (470.8 ± 16.6 mg/g), and showed a half-maximal inhibitory concentration (IC50) value of 0.41 ± 0.03 mg/mL and 0.51 ± 0.01 mg/mL for amylase and glucosidase, respectively. The results showed that ADJ6 had a significant inhibitory activity on α-amylase and α-glucosidase; however, its inhibitory activity was less than that of acarbose. The plants that are formulated in ADJ6 possess potent antidiabetic activity. Thus, we found that ADJ6 is a potent lead for effective diabetic management; however, an evaluation of the formulation must be illustrated using an in vivo model.

Development of an antidiabetic polyherbal formulation (ADPHF6) and assessment of its antioxidant activity against ROS-induced damage in pUC19 and human lymphocytes - an in vitro study.

BACKGROUND: Polyherbalism, an alternative natural-based therapy for various disorders, has been quoted about 1,300 years before in Sharangdhar Samhita. Herbal-based combination therapy stages a vital role for the treatment of type 2 diabetes mellitus (T2DM) and associated complications. The present study aims at developing an Ayurvedic-based polyherbal formulation (ADPHF6) and the assessing its antidiabetic and antioxidant property. METHODS: ADPHF6 polyherbal formulation was measured for phytochemical components by qualitative methods. The polyherbal formulation was quantitatively estimated for its phytochemical constituents, i. e. total phenol and flavonoid content. Further, the antioxidant property of ADPHF6 formulation was evaluated by 2,2-diphenyl-1-picryl-hydrazyl-hydrate (DPPH) radical scavenging assay, hydrogen peroxide radical scavenging assay and metal chelating assay. α-Amylase and α-glucosidase inhibitory activities of polyherbal formulation were also assessed. ADPHF6 was further analysed for its protective antioxidant property against reactive oxygen species (ROS‾)-induced damage in human lymphocyte DNA and pUC19 plasmid. RESULTS: ADPHF6 polyherbal formulation revealed the presence of phytochemical constituents such as alkaloids, flavonoids, phenols, tannins, terpenoids, saponins and cardiac glycosides in significant levels. Further, it also measured the higher levels of total phenols (473.3±3.05 mg/g) and flavonoid (664±5.29 mg/g) content. Polyherbal formulation also exhibited IC50 values of 49.9±0.15, 65.1±0.10 and 60.1±0.05 mg/mL for 2,2- diphenyl-1-picryl-hydrazyl-hydrate (DPPH), hydrogen peroxide (H2O2) and Fe2+ radical scavenging activities, respectively. ADPHF6 revealed an inhibitory activity (IC50) of 0.67±0.01 and 0.81±0.01 mg/mL for α-amylase and glucosidase, respectively. Pre-treated human peripheral blood lymphocytes with ADPHF6 aqueous extract illustrated enhanced protection against ROS-mediated damage as compared with post-treated groups. DNA nicking assay rendered protective activity against the OH¯ radical-induced DNA damage in supercoiled pUC19 plasmid. CONCLUSIONS: Our present study demonstrates that ADPHF6 offers potent inhibitory activity against free radicals as well as digestive enzymes. However, studies should be conducted using in vivo model to further elucidate the effect against free radicals and its anti-hyperglycaemic activity in the management of non-insulin-dependent diabetes.

Cardiac surgery-associated acute kidney injury in a developing country: Prevalence, risk factors and outcome.

Little is known about cardiac surgery-associated acute kidney injury (CS-AKI) in children in developing regions of the world. The study aimed to determine the prevalence of CSAKI, associated factors and its impact on mortality and utilization of hospital services. The hospital records of children aged 0-17 years who underwent CS at an Indian hospital were reviewed. CS-AKI was defined as a rise in serum creatinine of ≥0.3 mg/dL in any 48 h and or by urine output <0.5 mL/kg/h for an 8-h period in the first five days after CS. The study included 323 children with a median age of one year (0.04-17), of whom 22 (6.8%) were neonates and 18.3% had a single ventricle. About 60% of the children had Risk Adjusted Congenital Heart Surgery-I category 1 or 2 interventions. CS-AKI occurred in 39 children (12.1%). Factors associated with CS-AKI were sepsis and intraand post-operative hypotension. In-hospital mortality was six-fold higher in children who developed CS-AKI. CS-AKI was associated with two to three days more of mechanical ventilation and Intensive care unit stay. CS-AKI occurs in children in developing countries, but at a lower frequency mainly due to the predominance of post-neonatal children undergoing less-complex CSs. CS-AKI was associated with higher in-hospital mortality and increased utilization of hospital services. Factors associated with CS-AKI included intraand post-operative hypotension and sepsis.

The warden procedure through a posterolateral thoracotomy approach.

The Warden procedure for the correction of a right-sided partial anomalous pulmonary venous connection to the high superior vena cava is well established. It has the advantages of avoiding sinoatrial node dysfunction and pulmonary and systemic venous obstruction. In the case related here, a 3-year-old girl presented with a superior vena cava type of sinus venosus atrial septal defect and an anomalously draining right upper pulmonary vein, with bilateral superior venae cavae. Our approach to the Warden procedure was through a right posterolateral thoracotomy, which provided additional advantages.

Low frequency magnetic force augments hepatic differentiation of mesenchymal stem cells on a biomagnetic nanofibrous scaffold.

Liver tissue engineering using polymeric nanofibrous scaffold and stem cells holds great promises for treating end-stage liver failures. The aim of this study was to evaluate hepatic trans-differentiation potential of human mesenchymal stem cells (hMSCs) on a biomagnetic electrospun nanofibrous scaffold fabricated from a blend of poly-L-lactide (PLLA), collagen and fibrin-rich blood clot, under the influence of a low frequency magnetic field. The scaffold was characterized for surface properties, biochemical and biomechanical parameters and bio-magnetic behaviour. Cell proliferation assay revealed that the scaffold was suitable for hMSCs adhesion and proliferation. Hepatic trans-differentiation potential of hMSCs was augmented on nanofibrous scaffold in magnetic field exposure group compared to control groups, as evident by strong expression of hepatocyte specific markers, albumin release, urea synthesis and presence of an inducible cytochrome P450 system. Our results conclude that biomagnetic scaffold of PLLA/collagen/blood clot augments hepatic trans-differentiation of hMSCs under magnetic field influence.

Cruciate Fenestration in Ventricular Septal Defect Patch for High-Risk Patients With High Pulmonary Vascular Resistance.

Late presentation of patients with large ventricular septal defect (VSD) and elevated pulmonary vascular resistance (PVR) is not uncommon in developing countries. Surgical VSD closure in these patients carries risks of persistent pulmonary hypertension, right ventricular failure, and mortality. Several techniques for creation of valved patches or fenestrated patches have been developed to address these issues. We have successfully used a simple and easily reproducible technique in which a cruciate fenestration is created in the patch used for VSD closure.