The emerging role of exosomes in innate immunity, diagnosis and therapy

Exosomes, which are nano-sized transport bio-vehicles, play a pivotal role in maintaining homeostasis by exchanging genetic or metabolic information between different cells. Exosomes can also play a vital role in transferring virulent factors between the host and parasite, thereby regulating host gene expression and the immune interphase. The association of inflammation with disease development and the potential of exosomes to enhance or mitigate inflammatory pathways support the notion that exosomes have the potential to alter the course of a disease. Clinical trials exploring the role of exosomes in cancer, osteoporosis, and renal, neurological, and pulmonary disorders are currently underway. Notably, the information available on the signatory efficacy of exosomes in immune-related disorders remains elusive and sporadic. In this review, we discuss immune cell-derived exosomes and their application in immunotherapy, including those against autoimmune connective tissue diseases. Further, we have elucidated our views on the major issues in immune-related pathophysiological processes. Therefore, the information presented in this review highlights the role of exosomes as promising strategies and clinical tools for immune regulation

Nanoceutical Adjuvants as Wound Healing Material: Precepts and Prospects

Dermal wound healing describes the progressive repair and recalcitrant mechanism of 12 damaged skin, and eventually, reformatting and reshaping the skin. Many probiotics, nutritional supplements, metal nanoparticles, composites, skin constructs, polymers, and so forth have been associated with the improved healing process of wounds. The exact mechanism of material-cellular interaction is a point of immense importance, particularly in pathological conditions such as diabetes. Bioengineered alternative agents will likely continue to dominate the outpatient and perioperative management of chronic, recalcitrant wounds as new products continue to cut costs and improve the wound healing process. This review article provides an update on the various remedies with confirmed wound healing activities of metal-based nanoceutical adjuvanted agents and also other nano-based counterparts from previous experiments conducted by various researchers

Nuclear factor-kappa B and JNK mediate macrophage polarization shift induced by C-phycocyanin

381-389C-phycocyanin (C-pc), a fluorescent protein purified from the blue-green algae, Spirulina fusiformis, is reported to possess anti-inflammatory activity. Employing in vitro experiments and in vivo mouse models, we had earlier demonstrated the wound healing property of C-pc. In the current study, we evaluated the influence of C-pc on M1/M2 polarization of resting (M0) macrophages, using RAW 264.7 macrophage cells. Incubation of macrophages with C-pc resulted in upregulation of M1 markers (iNOS, IL-1β, IL-6 and TNF-α), and generation of reactive oxygen species and nitric oxide. In contrast, M2 markers, IL-10 and Arginase-1, exhibited reduced expressions. Immunoblotting and immunofluorescence studies revealed the activation and nuclear translocation of p65- NF-ĸB and JNK proteins. Inhibition studies showed that NF-ĸB and JNK were instrumental in shifting the polarization of M0 macrophages towards pro-inflammatory phenotype. NF-ĸB was responsible for the increase in TNF-α, IL-1β, and iNOS. JNK was responsible for the effect on IL-1β, IL-6 and Arg-1. This is the first report on the influence of NF-ĸB and JNK in mediating the pro-inflammatory action of C-pc, which could help towards phagocytosis and antimicrobial activity, processes required during the initial phases of wound healing. This study also exposes the biphasic action of C-pc towards modulating inflammatory processes

Biosynthesis and characterization of nano magnetic hydroxyapatite (nMHAp): An accelerated approach using simulated body fluid for biomedical applications

There are several metal ions including iron oxide nanoparticles, which has been used for doping hydroxyapatite to overcome the existing shortcoming associated with poor mechanical strength, less bioactivity and inert properties. Iron oxide nanoparticles, commonly known as magnetite, has shown a great potential in tissue engineering and various other biomedical applications. It has been proven that small quantity of iron increases the mechanical properties as well antioxidative property of the material. Magnetic hydroxyapatite is synthesized by many different synthesis routes such as hydrothermal, spray drying, ultrasonic irradiation etc. These synthesis routes are time consuming and requires sophisticated process parameters. Hence, to overcome these complexities, this study aims to synthesize and characterize nano magnetic hydroxyapatite (nMHAp) in a shorter duration than the conventionally used duration (over 24 h) through a novel bio-mimetic approach. The physicochemical properties were checked using XRD, FTIR, TGA, XPS, DLS, SEM, TEM and VSM. The results suggest that synthesized nMHAp contains magnetic properties and resembles carbonated apatite with the traces of Na, Mg, K, Cl and Fe. SEM and TEM analysis confirmed the spherical nature of nMHAp with average size between 10 and 200 nm. The optimized average time for nMHAp synthesis was found to be 3 h. Cellular biocompatibility and the measurement of reactive oxygen species using L929 cells confirmed the biocompatible and antioxidative nature of nMHAp. Hence, Our studies show that the nMHAp can be synthesized biomimetically in 3 h and potentially could be used for bone tissue engineering and other biomedical applications

Cloning and expression of MPT83 gene from Mycobacterium tuberculosis in E. coli BL21 as vaccine candidate of tuberculosis: A preliminary study

The appearance of Mycobacterium tuberculosis strains leading to drug resistance has caused new problems in TB treatment in various parts of the world and forces WHO to declare TB as a global emergency. With the increase of TB drug resistance, it is convinced that a more effective vaccine development will stop the epidemic of TB. Some M. tuberculosis antigens, one of which is MPT83, have been examined as TB vaccine candidate. MPT83 antigen, which is very immunogenic in lipoprotein micro bacteria, is identified as surface cell interrelated to antigen with cytometry circulation. Having TB resistance from BCG vaccine, MPT83 is considered TB vaccine candidate that can protect people against TB at adult age. The purpose of this research is to conduct amplification of MPT83 antigen cloning, and expression of its antigen on E. coli bacteria. From the result of the research, it is expected that raw material to produce TB vaccine as well as a high-quality antigen can be obtained. The band of DNA in PCR product is 660 bp, while the one in pGEMT-Easy-Mpt83 recombinant plasmid is 3678 bp. This is expressed in E. coli BL21 strain and produces 48 kDa protein as well as GST-MPT83 fusion protein. Keywords: Tuberculosis, Cloning, GST-MPT8

Insight into OroxylinA-7-O-β-d-Glucuronide-Enriched Oroxylum indicum Bark Extract in Oral Cancer HSC-3 Cell Apoptotic Mechanism: Role of Mitochondrial Microenvironment

Oroxylum indicum, of the Bignoniaceae family, has various ethnomedical uses such as an astringent, anti-inflammatory, anti-bronchitis, anti-helminthic and anti-microbial, including anticancer properties. The druggability of OI stem bark extract was determined by its molecular docking interactions with PARP and Caspase-3, two proteins involved in cell survival and death. Note that 50 µg/mL of Oroxylum indicum extract (OIE) showed a significant (p < 0.05%) toxicity to HSC-3 cells. MTT aided cell viability and proliferation assay demonstrated that 50 µg/mL of OIE displayed significant (p < 0.5%) reduction in cell number at 4 h of incubation time. Cell elongation and spindle formation was noticed when HSC-3 cells were treated with 50 µg/mL of OIE. OIE initiated DNA breakage and apoptosis in HSC-3 cells, as evident from DNA ladder assay and calcein/EB staining. Apoptosis potential of OIE is confirmed by flow cytometer and triple-staining (live cell/apoptosis/necrosis) assay. Caspase-3/7 fluorescence quenching (LANCE) assay demonstrated that 50 µg/mL of OIE significantly enhanced the RFU of caspases-3/7, indicating that the apoptosis potential of OIE is probably through the activation of caspases. Immuno-cytochemistry of HSC-3 cells treated with 50 µg/mL of OIE showed a significant reduction in mitochondrial bodies as well as a reduction in RFU in 60 min of incubation time. Immunoblotting studies clearly showed that treatment of HSC-3 cells with OI extract caused caspase-3 activation and PARP deactivation, resulting in apoptotic cell death. Overall, our data indicate that OIE is an effective apoptotic agent for human squamous carcinoma cells and it could be a future cancer chemotherapeutic target

Fluro-Protein C-Phycocyanin Docked Silver Nanocomposite Accelerates Cell Migration through NFĸB Signaling Pathway

Currently, there is a great demand for the development of nanomedicine aided wound tissue regeneration via silver doped nanoceuticals. Unfortunately, very little research is being carried out on antioxidants-doped silver nanometals and their interaction on the signaling axis during the bio-interface mechanism. In this study, c-phycocyanin primed silver nano hybrids (AgcPCNP) were prepared and analyzed for properties such as cytotoxicity, metal decay, nanoconjugate stability, size expansion, and antioxidant features. Fluctuations in the expression of marker genes during cell migration phenomena in in vitro wound healing scenarios were also validated. Studies revealed that physiologically relevant ionic solutions did not exhibit any adverse effects on the nanoconjugate stability. However, acidic, alkali, and ethanol solutions completely denatured the AgcPCNP conjugates. Signal transduction RT2PCR array demonstrated that genes associated with NFĸB- and PI3K-pathways were significantly (p < 0.5%) altered between AgcPCNP and AgNP groups. Specific inhibitors of NFĸB (Nfi) and PI3K (LY294002) pathways confirmed the involvement of NFĸB signaling axes. In vitro wound healing assay demonstrated that NFĸB pathway plays a prime role in the fibroblast cell migration. In conclusion, the present investigation revealed that surface functionalized AgcPCNP accelerated the fibroblast cell migration and can be further explored for wound healing biomedical applications

c-Phycocyanin primed silver nano conjugates: Studies on red blood cell stress resilience mechanism

In this study, we have synthesized cPC functionalized hybrid Ag nanoparticles (AgcPCNPs) by employing a green chemistry approach. This study also entails an in-depth understanding of different biochemical interaction between synthesized AgcPCNPs with whole blood and hemoglobin (Hbc). The cPC primed on the surface of the AgNPs reduced the hemolysis and lipid oxidation. A variation in the K+ concentrations and were observed. Additionally, molecular docking technique did not indicate significant changes in the protein structures. Finally, the AgcPCNPS enhanced the cellular migration of fibroblast. [Display omitted] •Synthesis and characterization of silver core CPC capped nanoparticles.•cPC conjugation reverses blood cell stress phenomenon by AgNPs.•AgcPC nanoparticles enhanced migration of fibroblasts. Green synthesis of metal-encased nutraceutical nano-hybrids has been a target for research over the last few years. In the present investigation, we have reported temperature dependent facile synthesis of silver nanoparticles using FDA approved c phycocyanin (cPC). The cPC conjugated silver nanoparticles (AgcPCNPs) were characterized by TEM, Zeta Potential, UV–vis, XPS, FTIR, and CD Spectroscopy. The temperature optimization studies suggested the synthesis of stable AgcPCNPs at 40 °C while at higher temperature system shows aggregated appearance. Molecular docking studies predicted the exclusive interaction of C, D, I, and J chains of cPC with the surface of AgNPs. Moreover, AgcPCNPs significantly (p < 0.1 %) counteract the toxic nature of AgNPs on red blood cell by measuring parameters like total RBC count, % hemolysis, % hematocrit, coagulation time, pH, electrolyte concentrations and degree of blood cell lipid peroxidation by the anti-oxidation mechanism. Skin fibroblast in vitro cell migration result suggeststhat AgcPCNPs enhanced the degree of cell movement towards the wound area. Data obtained collectively demonstrate that AgcPCNPs can be a better agent in the dermal wound healing with reduced toxicity with the bi-phasic advantage of cPC as a wound healer and Ag nano-metal as an anti-bacterial agent