Silver Nanoparticles Phytofabricated through <i>Azadirachta indica</i>: Anticancer, Apoptotic, and Wound-Healing Properties

Silver nanoparticles (AgNPs) have unlocked numerous novel disciplines in nanobiotechnological protocols due to their larger surface area-to-volume ratios, which are attributed to the marked reactivity of nanosilver, and due to their extremely small size, which enables AgNPs to enter cells, interact...

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Main Authors: Yogesh Dutt, Ramendra Pati Pandey, Mamta Dutt, Archana Gupta, Arpana Vibhuti, V. Samuel Raj, Chung-Ming Chang, Anjali Priyadarshini
Format: Article
Language:English
Published: MDPI AG 2023-01-01
Series:Antibiotics
Subjects:
Online Access:https://www.mdpi.com/2079-6382/12/1/121
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author Yogesh Dutt
Ramendra Pati Pandey
Mamta Dutt
Archana Gupta
Arpana Vibhuti
V. Samuel Raj
Chung-Ming Chang
Anjali Priyadarshini
author_facet Yogesh Dutt
Ramendra Pati Pandey
Mamta Dutt
Archana Gupta
Arpana Vibhuti
V. Samuel Raj
Chung-Ming Chang
Anjali Priyadarshini
author_sort Yogesh Dutt
collection DOAJ
description Silver nanoparticles (AgNPs) have unlocked numerous novel disciplines in nanobiotechnological protocols due to their larger surface area-to-volume ratios, which are attributed to the marked reactivity of nanosilver, and due to their extremely small size, which enables AgNPs to enter cells, interact with organelles, and yield distinct biological effects. AgNPs are capable of bypassing immune cells, staying in the system for longer periods and with a higher distribution, reaching target tissues at higher concentrations, avoiding diffusion to adjacent tissues, releasing therapeutic agents or drugs for specific stimuli to achieve a longer duration at a specific rate, and yielding desired effects. The phytofabrication of AgNPs is a cost-effective, one-step, environmentally friendly, and easy method that harnesses sustainable resources and naturally available components of plant extracts (PEs). In addition, it processes various catalytic activities for the degradation of various organic pollutants. For the phytofabrication of AgNPs, plant products can be used in a multifunctional manner as a reducing agent, a stabilizing agent, and a functionalizing agent. In addition, they can be used to curtail the requirements for any additional stabilizing agents and to help the reaction stages subside. <i>Azadirachta indica</i>, a very common and prominent medicinal plant grown throughout the Indian subcontinent, possesses free radical scavenging and other pharmaceutical properties via the regulation of proinflammatory enzymes, such as COX and TOX. It also demonstrates anticancer activities through cell-signaling pathways, modulating tumor-suppressing genes such as p53 and pTEN, transcriptional factors, angiogenesis, and apoptosis via bcl2 and bax. In addition, it possesses antibacterial activities. Phytofabricated AgNPs have been applied in the areas of drug delivery, bioimaging, biosensing, cancer treatment, cosmetics, and cell biology. Such pharmaceutical and biological activities of phytofabricated AgNPs are attributed to more than 300 phytochemicals found in <i>Azadirachta indica</i>, and are especially abundant in flavonoids, polyphenols, diterpenoids, triterpenoids, limonoids, tannins, coumarin, nimbolide, azadirachtin, azadirone, azadiradione, and gedunin. Parts of <i>Azadirachta indica</i>, including the leaves in various forms, have been used for wound healing or as a repellent. This study was aimed at examining previously biosynthesized (from <i>Azadirachta indica</i>) AgNPs for anticancer, wound-healing, and antimicrobial actions (through MTT reduction assay, scratch assay, and microbroth dilution methods, respectively). Additionally, apoptosis in cancer cells and the antibiofilm capabilities of AgNPs were examined through caspase-3 expression, dentine block, and crystal violet methods. We found that biogenic silver nanoparticles are capable of inducing cytotoxicity in HCT-116 colon carcinoma cells (IC<sub>50</sub> of 744.23 µg/mL, R<sup>2</sup>: 0.94), but are ineffective against MCF-7 breast cancer cells (IC<sub>50</sub> >> 1000 µg/mL, R<sup>2</sup>: 0.86). AgNPs (IC<sub>50</sub> value) induced a significant increase in caspase-3 expression (a 1.5-fold increase) in HCT-116, as compared with control cells. FITC-MFI was 1936 in HCT-116-treated cells, as compared to being 4551 in cisplatin and 1297 in untreated cells. AgNPs (6.26 µg/mL and 62.5 µg/mL) induced the cellular migration (40.2% and 33.23%, respectively) of V79 Chinese hamster lung fibroblasts; however, the improvement in wound healing was not significant as it was for the controls. AgNPs (MIC of 10 µg/mL) were very effective against MDR <i>Enterococcus faecalis</i> in the planktonic mode as well as in the biofilm mode. AgNPs (10 µg/mL and 320 µg/mL) reduced the <i>E. faecalis</i> biofilm by >50% and >80%, respectively. Natural products, such as <i>Syzygium aromaticum</i> (clove) oil (MIC of 312.5 µg/mL) and eugenol (MIC of 625 µg/mL), showed significant antimicrobial effects against <i>A. indica</i>. Our findings indicate that <i>A. indica</i>-functionalized AgNPs are effective against cancer cells and can induce apoptosis in HCT-116 colon carcinoma cells; however, the anticancer properties of AgNPs can also be upgraded through active targeting (functionalized with enzymes, antibiotics, photosensitizers, or antibodies) in immunotherapy, photothermal therapy, and photodynamic therapy. Our findings also suggest that functionalized AgNPs could be pivotal in the development of a novel, non-cytotoxic, biocompatible therapeutic agent for infected chronic wounds, ulcers, and skin lesions involving MDR pathogens via their incorporation into scaffolds, composites, patches, microgels, or formulations for microneedles, dressings, bandages, gels, or other drug-delivery systems.
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spelling doaj.art-89449a9694f2451c9556708074fc88ad2023-11-30T20:56:02ZengMDPI AGAntibiotics2079-63822023-01-0112112110.3390/antibiotics12010121Silver Nanoparticles Phytofabricated through <i>Azadirachta indica</i>: Anticancer, Apoptotic, and Wound-Healing PropertiesYogesh Dutt0Ramendra Pati Pandey1Mamta Dutt2Archana Gupta3Arpana Vibhuti4V. Samuel Raj5Chung-Ming Chang6Anjali Priyadarshini7Department of Microbiology, SRM University, 39, Rajiv Gandhi Education City, Post Office P.S. Rai, Sonepat 131029, Haryana, IndiaDepartment of Microbiology, SRM University, 39, Rajiv Gandhi Education City, Post Office P.S. Rai, Sonepat 131029, Haryana, IndiaMamta Dental Clinic, Opposite Sector 29, Main Badkhal Road, Faridabad 121002, Haryana, IndiaDepartment of Biotechnology, SRM University, 39, Rajiv Gandhi Education City, Post Office P.S. Rai, Sonepat 131029, Haryana, IndiaDepartment of Biotechnology, SRM University, 39, Rajiv Gandhi Education City, Post Office P.S. Rai, Sonepat 131029, Haryana, IndiaDepartment of Microbiology, SRM University, 39, Rajiv Gandhi Education City, Post Office P.S. Rai, Sonepat 131029, Haryana, IndiaMaster & Ph.D. Program in Biotechnology Industry, Chang Gung University, No. 259, Wenhua 1st Rd., Guishan District, Taoyuan City 33302, TaiwanDepartment of Microbiology, SRM University, 39, Rajiv Gandhi Education City, Post Office P.S. Rai, Sonepat 131029, Haryana, IndiaSilver nanoparticles (AgNPs) have unlocked numerous novel disciplines in nanobiotechnological protocols due to their larger surface area-to-volume ratios, which are attributed to the marked reactivity of nanosilver, and due to their extremely small size, which enables AgNPs to enter cells, interact with organelles, and yield distinct biological effects. AgNPs are capable of bypassing immune cells, staying in the system for longer periods and with a higher distribution, reaching target tissues at higher concentrations, avoiding diffusion to adjacent tissues, releasing therapeutic agents or drugs for specific stimuli to achieve a longer duration at a specific rate, and yielding desired effects. The phytofabrication of AgNPs is a cost-effective, one-step, environmentally friendly, and easy method that harnesses sustainable resources and naturally available components of plant extracts (PEs). In addition, it processes various catalytic activities for the degradation of various organic pollutants. For the phytofabrication of AgNPs, plant products can be used in a multifunctional manner as a reducing agent, a stabilizing agent, and a functionalizing agent. In addition, they can be used to curtail the requirements for any additional stabilizing agents and to help the reaction stages subside. <i>Azadirachta indica</i>, a very common and prominent medicinal plant grown throughout the Indian subcontinent, possesses free radical scavenging and other pharmaceutical properties via the regulation of proinflammatory enzymes, such as COX and TOX. It also demonstrates anticancer activities through cell-signaling pathways, modulating tumor-suppressing genes such as p53 and pTEN, transcriptional factors, angiogenesis, and apoptosis via bcl2 and bax. In addition, it possesses antibacterial activities. Phytofabricated AgNPs have been applied in the areas of drug delivery, bioimaging, biosensing, cancer treatment, cosmetics, and cell biology. Such pharmaceutical and biological activities of phytofabricated AgNPs are attributed to more than 300 phytochemicals found in <i>Azadirachta indica</i>, and are especially abundant in flavonoids, polyphenols, diterpenoids, triterpenoids, limonoids, tannins, coumarin, nimbolide, azadirachtin, azadirone, azadiradione, and gedunin. Parts of <i>Azadirachta indica</i>, including the leaves in various forms, have been used for wound healing or as a repellent. This study was aimed at examining previously biosynthesized (from <i>Azadirachta indica</i>) AgNPs for anticancer, wound-healing, and antimicrobial actions (through MTT reduction assay, scratch assay, and microbroth dilution methods, respectively). Additionally, apoptosis in cancer cells and the antibiofilm capabilities of AgNPs were examined through caspase-3 expression, dentine block, and crystal violet methods. We found that biogenic silver nanoparticles are capable of inducing cytotoxicity in HCT-116 colon carcinoma cells (IC<sub>50</sub> of 744.23 µg/mL, R<sup>2</sup>: 0.94), but are ineffective against MCF-7 breast cancer cells (IC<sub>50</sub> >> 1000 µg/mL, R<sup>2</sup>: 0.86). AgNPs (IC<sub>50</sub> value) induced a significant increase in caspase-3 expression (a 1.5-fold increase) in HCT-116, as compared with control cells. FITC-MFI was 1936 in HCT-116-treated cells, as compared to being 4551 in cisplatin and 1297 in untreated cells. AgNPs (6.26 µg/mL and 62.5 µg/mL) induced the cellular migration (40.2% and 33.23%, respectively) of V79 Chinese hamster lung fibroblasts; however, the improvement in wound healing was not significant as it was for the controls. AgNPs (MIC of 10 µg/mL) were very effective against MDR <i>Enterococcus faecalis</i> in the planktonic mode as well as in the biofilm mode. AgNPs (10 µg/mL and 320 µg/mL) reduced the <i>E. faecalis</i> biofilm by >50% and >80%, respectively. Natural products, such as <i>Syzygium aromaticum</i> (clove) oil (MIC of 312.5 µg/mL) and eugenol (MIC of 625 µg/mL), showed significant antimicrobial effects against <i>A. indica</i>. Our findings indicate that <i>A. indica</i>-functionalized AgNPs are effective against cancer cells and can induce apoptosis in HCT-116 colon carcinoma cells; however, the anticancer properties of AgNPs can also be upgraded through active targeting (functionalized with enzymes, antibiotics, photosensitizers, or antibodies) in immunotherapy, photothermal therapy, and photodynamic therapy. Our findings also suggest that functionalized AgNPs could be pivotal in the development of a novel, non-cytotoxic, biocompatible therapeutic agent for infected chronic wounds, ulcers, and skin lesions involving MDR pathogens via their incorporation into scaffolds, composites, patches, microgels, or formulations for microneedles, dressings, bandages, gels, or other drug-delivery systems.https://www.mdpi.com/2079-6382/12/1/121<i>Azadirachta indica</i>phytofabricationsilver nanoparticlesanticancer drugscaspase-3 expressionwound healing
spellingShingle Yogesh Dutt
Ramendra Pati Pandey
Mamta Dutt
Archana Gupta
Arpana Vibhuti
V. Samuel Raj
Chung-Ming Chang
Anjali Priyadarshini
Silver Nanoparticles Phytofabricated through <i>Azadirachta indica</i>: Anticancer, Apoptotic, and Wound-Healing Properties
Antibiotics
<i>Azadirachta indica</i>
phytofabrication
silver nanoparticles
anticancer drugs
caspase-3 expression
wound healing
title Silver Nanoparticles Phytofabricated through <i>Azadirachta indica</i>: Anticancer, Apoptotic, and Wound-Healing Properties
title_full Silver Nanoparticles Phytofabricated through <i>Azadirachta indica</i>: Anticancer, Apoptotic, and Wound-Healing Properties
title_fullStr Silver Nanoparticles Phytofabricated through <i>Azadirachta indica</i>: Anticancer, Apoptotic, and Wound-Healing Properties
title_full_unstemmed Silver Nanoparticles Phytofabricated through <i>Azadirachta indica</i>: Anticancer, Apoptotic, and Wound-Healing Properties
title_short Silver Nanoparticles Phytofabricated through <i>Azadirachta indica</i>: Anticancer, Apoptotic, and Wound-Healing Properties
title_sort silver nanoparticles phytofabricated through i azadirachta indica i anticancer apoptotic and wound healing properties
topic <i>Azadirachta indica</i>
phytofabrication
silver nanoparticles
anticancer drugs
caspase-3 expression
wound healing
url https://www.mdpi.com/2079-6382/12/1/121
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