Optimization of silver nanoparticles synthesis via Plackett-Burman experimental design: in vitro assessment of their efficacy against oxidative stress-induced disorders.

Nanoparticles possess remarkable biological activities owing to their small size and large surface-to-volume ratio. Given the increasing adoption of environmentally sustainable practices in silver nanoparticle (AgNP) fabrication, this study presents a simple lab-scale green synthesis of AgNPs using banana peels. Large amounts of banana peels are disposed off in Pakistan every day. As the fruit is available throughout the year and contains many active components with potent biological activities, we aimed to synthesize silver nanoparticles using its peel, through an energy-efficient and inexpensive route. The synthesis was optimized according to the Plackett-Burman design (PDB) of experiments, which helped identify significant factors and saved time and resources. For characterization, UV-Vis spectroscopy and SEM-EDX analysis were performed, revealing spherical particles in the 45-65 nm size range. To investigate functional groups, FT-IR analysis was performed, revealing the presence of N-C[double bond, length as m-dash]O amide I bonds of proteins, C-H bonds of tannins and C-O bonds involved in the capping and stabilization of nanoparticles. The free radical scavenging property of banana peel-mediated silver nanoparticles (BP-AgNPs) was studied against 2,2-diphenyl-1-picrylhydrazyl (DPPH), and the antioxidant potential was found to be 79% at 500 µg mL-1 concentration. The efficacy of BP-AgNPs with respect to certain biological activities were studied through anti-inflammatory assays, which demonstrated better results compared to a standard drug, and an anti-glycation assay, wherein only 4% of AGEs were formed, demonstrating 96% of AGE inhibition in vitro. The findings not only demonstrated the effectiveness of the PBD approach but also highlighted the potent property of BP-AgNPs against disorders associated with oxidative stress.

Bio-inspired synthesis of silver nanoparticles usingSalsola imbricataand its application as antibacterial additive in glass ionomer cement.

Nanotechnology has gained immense popularity and observed rapid development due to the remarkable physio-chemical properties of nanoparticles (NPs) and related nanomaterials. The green production of NPs has many benefits over traditional techniques because the current procedures are expensive, time-consuming, and involve harmful substances that limit their applicability. This study aimed to use a novel green source, theSalsola imbricata(SI) plant, which is commonly found in Central Asia and known for its medicinal properties as a reducing and stabilizing agent for the synthesis of AgNPs. The current study also utilized efficient statistical design, the Plackett-Burman Design (PBD) of Experiment method to synthesize the NPs. The characterization of NPs was carried out using UV-Vis spectroscopy, Fourier transform infrared spectroscopy, and scanning electron microscopy (SEM). The PBD results showed that only two out of four factorsi.e.AgNO3concentration and incubation time, were significant for the synthesis of SI-AgNPs. While remaining factors, incubation temperature and plant extract: AgNO3ratio were non-significant. The SEM analysis result showed that SI-AgNPs had a size of 20-50 nm. The SI-AgNPs demonstrated strong antibacterial activity against oral pathogens such asS. mutans and Lactobacillus acidophilus, with the highest efficacy observed at a concentration of 2 mg ml-1. The addition of SI-AgNPs in glass ionomer cement significantly increased the antibacterial activity of GIC againstS. mutans. Based on the results of the current study, the plant based AgNPs can be further evaluated in detail as alternate antimicrobial agent either alone or in combination with other antimicrobial agents for different dental applications.

Biofabrication of silver nanoparticles using Spirulina platensis: In vitro anti-coagulant, thrombolytic and catalytic dye degradation activity.

Green synthesis of nanoparticles is an emerging field due to it's biosafety and promising results. Biological systems due to their biodiversity are employed in different production processes. In this study Spirulina platensis mediated silver nanoparticles (S-AgNPs) production was done. The Uv spectra, FTIR and SEM analysis was performed for the characterization of biosynthesized S-AgNPs. The biocompatibility evaluation of S-AgNPs was done through hemolysis analysis. S-AgNPs were also evaluated for anticoagulant and thrombolytic potential. In addition to the medical applications of S-AgNPs, silver nanoparticles have been known to show potential industrial applications among which one application is the utilization of silver nanoparticles in the degradation of toxic industrial dyes. Therefore, degradation assay of Eosin Y and Methylene Blue dyes was estimated. The SEM analysis of S-AgNPs showed the particle size of 50-65 nm, whereas the biocompatibility analysis showed that these S-AgNPs are biocompatible at ≤400 µM concentration. The S-AgNPs showed good anticoagulant potential and thrombolytic potential and were able to degrade 44% of the thrombus. The S-AgNPs significantly degraded 76% of Eosin Y within 30 min, whereas Methylene Blue was 80% degraded within 20 min (P-value ≤ .001). To the best of our knowledge, the dye degradation of Eosin Y, thrombolytic activity and anticoagulant activity of S-AgNPs produced from the biomass of Spirulina platensis has been reported for the first time. In the current study, we conclude that our biosynthesized S-AgNPs showed promising medical and industrial applications and these nanoparticles can be further evaluated and upscaled for large scale applications.

Translation and validation of the Urdu version of the neck pain and disability scale.

PURPOSE: To translate and cross-culturally adapt the neck pain and disability scale (NPDS) into Urdu language (NPDS-U), and to investigate the NPDS-U's psychometric properties in patients with non-specific neck pain (NSNP). METHODS: The NPDS was translated and cross-culturally adapted into Urdu in accordance with the previously described guidelines. The study included 200 NSNP patients and 50 healthy participants. The NPDS-U, Urdu version of neck disability index (NDI-U), neck Bournemouth questionnaire (NBQ), and numerical pain rating scale (NPRS) were completed by all participants. After three weeks of physiotherapy, the patients completed all of the above-mentioned questionnaires, along with the global rating of change scale. Reliability, factor analysis, validity, and responsiveness were all tested. RESULTS: The NPDS-U demonstrated excellent test-retest reliability (ICC2,1=0.92) and high internal consistency (Cronbach's alpha = 0.96). There were no floor or ceiling effects. A three-factor structure was extracted, which explained 70.42% of the total variance. The NPDS-U showed moderate to strong correlations with NPRS, NDI-U, and NBQ (r = 0.67-0.76, p < 0.001). A significant difference in the NPDS-U change scores between the stable and the improved groups (p < 0.001) confirmed its responsiveness. CONCLUSION: The NPDS-U is a reliable, valid, and responsive scale for assessing neck pain and disability in Urdu-speaking patients with NSNP.

The Urdu version of Neck Pain and Disability Scale (NPDS-U) is a reliable and valid tool for assessing neck pain and disability in Urdu-speaking patients with non-specific neck pain (NSNP).The NPDS-U can be used to distinguish between people who have neck pain and disability and those who do not.NPDS-U exhibited a 3-factor structure.The NPDS-U is a responsive instrument that can be used to assess treatment efficacy in NSNP patients.The NPDS-U is a simple and easy to use in clinical and research settings.

Extraction, Partial Purification and Characterization of Bromelain from Pineapple (Ananas Comosus) Crown, Core and Peel Waste

Abstract Ananas Comosus (also known as pineapple) is a part of Bromeliaceae family and it is consumed as food as well as folk medicine for the treatment of various diseases. It is reported that pineapple is a rich source of bromelain, a cysteine protease and it is considered as an important enzyme in different industries due to its significant therapeutic and industrial applications such as anticancer, anti-inflammatory and meat tenderizing. Bromelain is mostly present in fruit and stem of pineapple, but it is reported that crown, core, and peels, which constitute the waste of the pineapple plant, also contain bromelain but limited data is available. Therefore, the proposed study aimed at utilizing pineapple waste for the extraction and characterization of bromelain. Firstly, crude bromelain was extracted with phosphate buffer (pH 7), then it was subjected to partial purification using different fractions of ammonium sulphate (NH4)2SO4 such as 30, 40, 50 and 60% followed by desalting and concentration. Enzyme activity was calculated by using casein digesting unit (CDU) method. The results demonstrated that the crown bromelain showed highest purification of 4.34-fold at 30% (NH4)2SO4 saturation, whereas core and peel bromelain showed highest purification of 2.75 and 2.59-fold at 40% (NH4)2SO4 saturation. The molecular weight of crude and partially purified bromelain was determined by SDS-PAGE analysis and found to be 26 KDa. The pH and thermal stability of all the parts of pineapple showed maximum stability at pH 7 and at 35oC temperature.

Rutin and rutin-conjugated gold nanoparticles ameliorate collagen-induced arthritis in rats through inhibition of NF-κB and iNOS activation.

Numerous studies have suggested that nuclear factor-κB (NF-κB) and inducible nitric oxide synthase (iNOS) are important mediators of inflammatory response in human and animal models of arthritis. Besides, oxidative stress markers, nitric oxide (NO) and peroxide (PO) are also major contributors in the pathogenesis of rheumatoid arthritis (RA). Over expression of these inflammatory mediators leads to the extracellular matrix degradation, and excessive cartilage and bone resorption, ultimately leading to the irreversible damage to joints. The aim of the present study was to investigate the anti-arthritic mechanism of bioflavonoids, rutin and rutin-conjugated gold nanoparticles (R-AuNPs) by determining their role in the modulation of NF-κB and iNOS expression in collagen-induced arthritis (CIA) model of rats. Arthritis was induced by the subcutaneous administration of bovine type II collagen. Treatment was started with rutin, indomethacin + rutin (I + R) and R-AuNPs on the day of CIA induction. The severity of arthritis was determined by measuring the arthritic score on alternate days until mean arthritic score of 4 was observed. The NO and PO levels were also analyzed in serum samples. NF-κB and iNOS expression levels were determined in spleen tissue samples by real time RT-PCR and immunohistochemistry. Marked reduction in the arthritic score as well as in the NO and PO levels was observed in the treated groups. A significant downregulation in the NF-κB and iNOS expression levels was also observed in the treatment groups compared to the arthritic control group. Collectively, the findings suggest potential clinical role of rutin and R-AuNPs in the treatment of rheumatoid arthritis.

N-(2-Hydroxyphenyl)acetamide: a Novel Suppressor of RANK/RANKL Pathway in Collagen-Induced Arthritis Model in Rats.

RANKL and RANK are potential contributors of inflammatory cascade in human and animal model of arthritis. The current study aims to investigate the effect of N-(2-hydroxyphenyl)acetamide (NA-2) on regulation of RANKL pathway in collagen-induced arthritis (CIA) model in rats. CIA was induced using bovine type II collagen in female Wistar rats. The clinical parameters, level of pro-inflammatory and oxidative stress markers were measured to determine the progression of the disease. The mRNA level of RANKL and RANK and downstream mediators of inflammation i.e. c-fos, c-jun, NF-κB and Akt were analysed in spleen tissue using real-time PCR. Immunohistochemical analysis of iNOS, pAkt and c-Fos was also done in spleen tissue. Treatment with NA-2 and indomethacin showed increase in body weight and significant reduction in paw volume and arthritic score (p < 0.0001). Marked reduction in the level of oxidative stress markers, NO, PO and GSH (p < 0.0001), and pro-inflammatory markers, IL-1ß (p < 0.0001) and TNF-α (p < 0.01), was also observed. Likewise, NA-2 and indomethacin treatment also significantly suppressed the mRNA expression of RANKL, RANK, c-fos, c-jun, NF-κB (p < 0.0001) and Akt (p < 0.01) and protein expression of iNOS, pAkt and c-Fos (p < 0.0001) compared to the arthritic control group. Our findings suggest that NA-2 is an antiarthritic agent acting in a pleiotropic manner in CIA rats by not only reducing the clinical signs of arthritis, inflammatory cytokines and free radical production but also attenuating the RANK/RANKL signaling pathway.