Re-Cellularised Kidney Scaffold for Chikungunya Virus Propagation: A Novel Approach.

BACKGROUND: Re-emerging viral attacks are catastrophic for health and economy. It is crucial to grasp the viral life cycle, replication and mutation policies and attack strategies. It is also absolute to fathom the cost-efficient antiviral remedies earliest possible. METHODS: We propose to use a lab-grown organ (re-cellularized scaffold of sheep kidney) for viral culture and understand its interaction with extra-cellular matrices of the host tissue. RESULTS: Our findings showed that the chikungunya virus (CHIKV) could be better replicated in tissue-engineered bio models than cell culture. A decrease in ds-DNA levels emphasized that CHIKV propagates within the re-cellularized and cell culture models. There was an increase in the viral titres (pfu/ml) in re-cellularized scaffolds and control groups. The lipid peroxidation levels were increased as the infection was progressed in cell culture as well as re-cellularized and control groups. The onset and progress of the CHIKV attacks (cellular infection) lead to transmembrane domain fatty acid peroxidation and DNA breakdown, landing in cellular apoptosis. Simultaneously cell viability was inversely proportional to non-viability, and it decreased as the infection progressed in all infected groups. Histological findings and extracellular matrix evaluation showed the impairment in medullary, cortex regions due to propagation of CHIKV and plaques generations. CONCLUSION: This method will be a breakthrough for future virus culture, drug interaction and to study its effect on extracellular matrix alterations. This study will also allow us to investigate the correct role of any vaccine or antiviral drugs and their effects on re-engineered organ matrices before moving towards the animal models.

In vitro and in vivo studies reveal α-Mangostin, a xanthonoid from Garcinia mangostana, as a promising natural antiviral compound against chikungunya virus.

BACKGROUND: Chikungunya virus (CHIKV), a serious health problem in several tropical countries, is the causative agent of chikungunya fever. Approved antiviral therapies or vaccines for the treatment or prevention of CHIKV infections are not available. As diverse natural phenolic compounds have been shown to possess antiviral activities, we explored the antiviral activity of α-Mangostin, a xanthanoid, against CHIKV infection. METHODS: The in vitro prophylactic and therapeutic effects of α-Mangostin on CHIKV replication in Vero E6 cells were investigated by administering it under pre, post and cotreatment conditions. The antiviral activity was determined by foci forming unit assay, quantitative RT-PCR and cell-based immune-fluorescence assay. The molecular mechanism of inhibitory action was further proposed using in silico molecular docking studies. RESULTS: In vitro studies revealed that 8 µM α-Mangostin completely inhibited CHIKV infectivity under the cotreatment condition. CHIKV replication was also inhibited in virus-infected mice. This is the first in vivo study which clearly showed that α-Mangostin is effective in vivo by significantly reducing virus replication in serum and muscles. Molecular docking indicated that α-Mangostin can efficiently interact with the E2-E1 heterodimeric glycoprotein and the ADP-ribose binding cavity of the nsP3 macrodomain. CONCLUSIONS: The findings suggest that α-Mangostin can inhibit CHIKV infection and replication through possible interaction with multiple CHIKV target proteins and might act as a prophylactic/therapeutic agent against CHIKV.

Fabricating a pre-vascularized large-sized metabolically-supportive scaffold using Brassica oleracea leaf.

BACKGROUND: There is a significant pitfall in clinical translation of large-sized tissue-engineered grafts - a lack of vascularization. This study was carried out to find the answer in a plant leaf, as plants and animals share structural similarities. METHODS AND RESULTS: We fabricated a scaffold using Brassica oleracea leaves (10%SDS) and expanded the endothelial cells onto them. The vascularity was demarcated by angiography. The thermal decomposition confirmed that the oxidation resistance of the scaffold is parallel to the natural leaf. The acellularity of the scaffold as well as the presence of cellular establishment after culture on the scaffold was confirmed by histology, scanning electron microscopy, periodic acid-Schiff, and DNA quantification. Further, we estimated various biochemical markers like MDA, catalase, total proteins, and total nitric oxide for confirming their metabolic activities. Cell-specific markers like vWF, lectin established their phenotype. Cytotoxicity and live-dead assay showed the viability of cells. CONCLUSION: Our findings proved that the decellularized leaf scaffold preserves vascularity, exhibits non-toxicity, maintains the cell identity, and supports mammalian cells for their metabolic activities. The study gives a futuristic hope in combating the ever-growing issues of clinical applicability of large-sized grafts.

Fabrication of aortic bioprosthesis by decellularization, fibrin glue coating and re-endothelization: a cell scaffold approach.

Aortic dysfunctions (aneurysm, aortitis) lead to the most serious conditions related to aortic wall with life-threatening complications. The most common modality of management for such conditions is replacement (diseased part) of aorta by a larger diameter stent (reconstructive vascular surgery) which in itself is a big trial. The most natural way is to use a re-endothelized scaffold. Developing a scaffold with biomimetic properties is an experimental aim for most of the scientists and surgeons. We aim to structure a strategy to overcome the well-known problems associated with aorta. In this study, we plan to remold a larger diameter blood vessel such as aorta from xenogeneic origin using different protocols to decellularize and comparing them with normal aorta. The chemicals and enzymes used for bovine aorta decellularization are 1% SDS (group II), 70% ethanol + 0.25% trypsin (group III), 70% ethanol (group IV), and 0.25% trypsin (group V). Group I served as control (without decellularization). Histology and SEM study were conducted for cellular presence/absence in all scaffolds. Later, the scaffolds were coated with the fibrin glue (FG) and endothelial cells were proliferated over them. 3D images were taken showing the remolding of the endothelial cells on FG-coated surfaces. The re-endothelization was confirmed by lectin and vWF+/+ expression. Graft elasticity and burst pressure were confirmed by biomechanical tensile testing. Further, the absence of host tissue DNA and presence of cellular DNA after re-endothelialization were confirmed by PicoGreen assay. The acceptability for metabolically active cellular proliferation on scaffolds and its non-toxicity were proved by cell viability assay. Current findings accomplish that larger diameter aorta extracellular matrix scaffold (group II) can be fabricated and re-endothelialized to develop non-thrombotic surfaces with improved graft patency with promising results compared to other fabricated scaffold groups.