scholarly journals Hepato(Geno)Toxicity Assessment of Nanoparticles in a HepG2 Liver Spheroid Model

Nanomaterials ◽  
2020 ◽  
Vol 10 (3) ◽  
pp. 545 ◽  
Author(s):  
Elisabeth Elje ◽  
Espen Mariussen ◽  
Oscar H. Moriones ◽  
Neus G. Bastús ◽  
Victor Puntes ◽  
...  
Keyword(s):  
Laser Doppler Anemometry ◽  
Animal Experiments ◽  
Toxicity Assessment ◽  
In Vitro Models ◽  
Substantial Part ◽  
Ag Nps ◽  
Zno Nps ◽  
Vis Spectroscopy ◽  
Vitro Model

(1) In compliance with the 3Rs policy to reduce, refine and replace animal experiments, the development of advanced in vitro models is needed for nanotoxicity assessment. Cells cultivated in 3D resemble organ structures better than 2D cultures. This study aims to compare cytotoxic and genotoxic responses induced by titanium dioxide (TiO2), silver (Ag) and zinc oxide (ZnO) nanoparticles (NPs) in 2D monolayer and 3D spheroid cultures of HepG2 human liver cells. (2) NPs were characterized by electron microscopy, dynamic light scattering, laser Doppler anemometry, UV-vis spectroscopy and mass spectrometry. Cytotoxicity was investigated by the alamarBlue assay and confocal microscopy in HepG2 monolayer and spheroid cultures after 24 h of NP exposure. DNA damage (strand breaks and oxidized base lesions) was measured by the comet assay. (3) Ag-NPs were aggregated at 24 h, and a substantial part of the ZnO-NPs was dissolved in culture medium. Ag-NPs induced stronger cytotoxicity in 2D cultures (EC50 3.8 µg/cm2) than in 3D cultures (EC50 > 30 µg/cm2), and ZnO-NPs induced cytotoxicity to a similar extent in both models (EC50 10.1–16.2 µg/cm2). Ag- and ZnO-NPs showed a concentration-dependent genotoxic effect, but the effect was not statistically significant. TiO2-NPs showed no toxicity (EC50 > 75 µg/cm2). (4) This study shows that the HepG2 spheroid model is a promising advanced in vitro model for toxicity assessment of NPs.

Velocity Measurements Inside a Hemodialysis Graft In Vitro Model

2004 ◽  
Author(s):  
David S. Smith ◽  
Francis Loth ◽  
Hisham S. Bassiouny ◽  
Paul Fischer ◽  
Jennifer K. Grogan ◽  
...  
Keyword(s):  
In Vitro Model ◽  
Laser Doppler Anemometry ◽  
In Vitro Models ◽  
Venous Anastomosis ◽  
Rate Environment ◽  
Plastic Cast ◽  
Computational Fluid Dynamics Cfd ◽  
Vitro Model ◽  
Good Agreement

Arteriovenous (AV) grafts, which provide an access site for hemodialysis, typically produce a high flow rate environment with pressure and velocity fluctuations; high and low wall shear stress, and vibration. Laser Doppler anemometry (LDA) was performed at Reynolds number (Re) 1200 on an in vitro model, which was constructed from computerized tomography (CT) images of a perfusion fixed plastic cast of a canine venous anastomosis. The results obtained were compared to numerical results and to results previously obtained with idealized in vitro models. This study showed the importance of an accurate geometry in characterizing the flow environment inside an AV graft. Good agreement between the computational fluid dynamics (CFD) and LDA was observed although differences were clearly present.

scholarly journals Nanoparticle induced barrier function assessment at liquid–liquid and air–liquid interface in novel human lung epithelia cell lines

2019 ◽  
Vol 8 (6) ◽  
pp. 1016-1027 ◽  
Author(s):  
Lars Leibrock ◽  
Sandra Wagener ◽  
Ajay Vikram Singh ◽  
Peter Laux ◽  
Andreas Luch
Keyword(s):  
Cell Lines ◽  
Barrier Function ◽  
Human Lung ◽  
Animal Experiments ◽  
A549 Cells ◽  
In Vitro Models ◽  
Cell Models ◽  
Zno Nps

Abstract Inhalation is the most relevant entry point for nanoparticles (NPs) into the human body. To date, toxicity testing of nanomaterials in respect to oral, dermal and inhalative application is mainly based on animal experiments. The development of alternative test methods is the subject of current research. In vitro models can help to investigate mechanistic aspects, as e.g. cellular uptake or genotoxicity and might help to reduce in vivo testing. Lung cell lines are proper in vitro tools to assess NP toxicity. In respect to this, various cell models have been developed during the recent years, but often lack in a proper intact barrier function. However, besides other important in vivo criteria which are still missing like e.g. circulation, this is one basic prerequisite to come closer to the in vivo situation in certain mechanistic aspects such as particle translocation which is an important task for risk assessment of nanomaterials. Novel developed in vitro models may help to investigate the translocation of nanomaterials from the lung. We investigated the barrier function of the recently developed human lung cell lines CI-hAELVi and CI-huAEC. The cells were further exposed to CeO2 NPs and ZnO NPs, and their suitability as in vitro models for toxicological investigations was proven. The obtained data were compared with data generated with the A549 cell line. Measurement of transepithelial resistance and immunohistochemical examination of tight junctions confirmed the formation of a functional barrier for both cell lines for submerged and air–liquid cultivation. For particle exposure, hAELVi and huAEC cells showed comparable results to A549 cells without losing the barrier function. CeO2 NP exposure revealed no toxicity for all cell lines. In contrast, ZnO NPs was toxic for all cell lines at a concentration between 10–50 μg ml−1. Due to the comparable results to A549 cells CI-hAELVi and CI-huAEC offer new opportunities to investigate nanoparticle cell interactions more realistic than recent 2D cell models.

scholarly journals Advanced In Vitro Models for Replacement of Animal Experiments

Small ◽  
2021 ◽  
Vol 17 (15) ◽  
pp. 2101474
Author(s):  
Martin J. D. Clift ◽  
Shareen H. Doak
Keyword(s):  
Animal Experiments ◽  
In Vitro Models

Μελέτη της τοξικής, κυτταροτοξικής και γενοτοξικής δράσης νεοσυντιθέμενων ενώσεων και νανοϋλικών σε in vitro συνθήκες

2021 ◽  
Author(s):  
Ιωάννα Ευθυμίου
Keyword(s):  
Humic Acid ◽  
Vibrio Fischeri ◽  
Anodic Stripping Voltammetry ◽  
Micronucleus Assay ◽  
Stripping Voltammetry ◽  
Neutral Red ◽  
Flame Spray ◽  
Ag Nps ◽  
Zno Nps

Η συνεχώς αυξανόμενη παραγωγή προϊόντων της Nανοτεχνολογίας εγείρει στις μέρες μας σημαντικά ερωτήματα σχετικά με τον περιβαλλοντικό αντίκτυπό τους. Συγκεκριμένα, νανοσωματίδια (Nanoparticles, NPs) όπως το οξείδιο του ψευδαργύρου (ZnO) και αργύρου (Ag), παρουσιάζουν ευρεία εξάπλωση και εφαρμογή, γεγονός που αυξάνει τις πιθανότητες να βρεθούν βιοδιαθέσιμα στο περιβάλλον. Παρόλο που υπάρχει εκτεταμένη βιβλιογραφία αναφορικά με τα προαναφερθέντα NPs, συνήθως κάθε μελέτη εστιάζει σε συγκεκριμένο παράγοντα κάθε φορά όπως για παράδειγμα στη σύνθεση των NPs, τα βελτιωμένα χαρακτηριστικά τους, τις πιθανές εφαρμογές τους ή τις τοξικές τους επιδράσεις σε συγκεκριμένο οργανισμό ή κυτταρική σειρά. Αν και κάθε έρευνα ενισχύει το μέχρι τώρα γνωσιακό υπόβαθρο και συμπληρώνει κενά αναφορικά με τη Νανοτεχνολογία, με την παρούσα μελέτη έγινε η προσπάθεια πραγματοποίησης μιας ολοκληρωμένης εργασίας. Συγκεκριμένα, το ερευνητικό πλάνο περιλαμβάνει (α) τη σύνθεση των ZnO, Ag και ZnO-Ag NPs, (β) το χαρακτηρισμό τους (μεμονωμένα και σε διασπορά μέσα σε υδατικά διαλύματα), (γ) την αξιολόγηση των πιθανών τοξικών, κυτταροτοξικών και γενοτοξικών τους επιδράσεων σε διαφορετικά βιολογικά συστήματα (ανθρώπινα λεμφοκύτταρα, βακτήρια και αιμοκύτταρα δίθυρου μαλακίου) σε in vitro συνθήκες, (δ) τη μελέτη της αλληλεπίδρασης των εξεταζόμενων NPs παρουσία χουμικών οξέων (Humic Acids, HAs), προσομοιάζοντας έτσι τις πραγματικές περιβαλλοντικές συνθήκες. Αναλυτικότερα, τα NPs (ZnO, Ag και ZnO-Ag NPs) παρασκευάστηκαν με την καινοτόμο τεχνική πυρόλυσης ψεκασμού φλόγας (Flame Spray Pyrolysis, FSP) που χρησιμοποιείται για τη σύνθεση μεμονωμένων και σύνθετων NPs με υψηλή καθαρότητα και βελτιωμένα μορφολογικά και φυσικοχημικά χαρακτηριστικά. Ακολούθησε χαρακτηρισμός των NPs με περίθλαση ακτίνων Χ (powder X ray Diffraction, pXRD), με ηλεκτρονική μικροσκοπία διέλευσης (Transmission Electron Microscopy, TEM) και με δυναμική σκέδαση φωτός (Dynamic Light Scattering, DLS). Στη συνέχεια, διερευνήθηκε η ενδεχόμενη γενοτοξική και κυτταροτοξική δράση των NPs παρουσία και απουσία δύο χαρακτηρισμένων HAs (Humic acid-like-polycondensate, HALP; Leonardite Humic Acid, LHA) σε ανθρώπινα λεμφοκύτταρα με την εφαρμογή της τεχνικής των μικροπυρήνων με χρήση της κυτταροχαλασίνης-Β (Cytokinesis Block Micronucleus assay, CBMN assay). Η έκπλυση ιόντων Zn2+ προσδιορίστηκε στα ιζήματα των λεμφοκυττάρων που προέκυψαν από την τεχνική CBMN, μέσω της ανοδικής αναδιαλυτικής βολταμμετρίας (Anodic Stripping Voltammetry, ASV). Έπειτα, οι τοξικές επιδράσεις των NPs παρουσία και απουσία των δύο HAs μελετήθηκαν στο βακτήριο Vibrio fischeri με τη χρήση του συστήματος Microtox. Τέλος, εξετάστηκαν οι κυτταροτοξικές και οξειδωτικές επιδράσεις των NPs σε αιμοκύτταρα του μυδιού Mytilus galloprovincialis, μέσω προσδιορισμού (α) της λυσοσωμικής αποσταθεροποίησης (Τεχνική ουδέτερου ερυθρού/Neutral Red Retention Time), (β) της παραγωγής σουπεροξειδικών ανιόντων, (γ) της παραγωγής οξειδίων του αζώτου (υπό μορφή νιτρωδών) και (δ) των επιπέδων λιπιδικής υπεροξείδωσης. Σύμφωνα με τα αποτελέσματα, στην περίπτωση της τεχνικής CBMN, δεν υπήρξε εκδήλωση γενοτοξικών φαινομένων σε καμία περίπτωση, για κανένα από τα υπό μελέτη NPs τόσο παρουσία όσο και απουσία των δύο HAs. Από την άλλη πλευρά, όλα τα NPs εκδήλωσαν κυτταροτοξική δράση. Ωστόσο, αν και τα Ag και ZnO-Ag NPs οδήγησαν σε επαγωγή κυτταροτοξικότητας παρουσία και απουσία των δύο HAs, τα μίγματα των ZnO NPs με τα δύο HAs ελάττωσαν την κυτταροτοξικότητα που προκλήθηκε από τα μεμονωμένα ZnO NPs. Αναφορικά με τα ποσοστά έκπλυσης ιόντων Zn2+, παρατηρήθηκε ότι οι μεγαλύτερες συγκεντρώσεις των μιγμάτων (ZnO, ZnO-Ag)NPs-HAs διατήρησαν ένα μικρό ποσοστό ιόντων, ενώ στις υπόλοιπες περιπτώσεις το ποσοστό ήταν αμελητέο. Στην περίπτωση προσδιορισμού της τοξικότητας των NPs έναντι του βακτηρίου Vibrio fischeri, διαπιστώθηκε ενισχυμένη τοξικότητα των ZnO και ZnO-Ag NPs ενώ τα Ag NPs εμφάνισαν τη μικρότερη τοξική δράση. Ο συνδυασμός με τα δύο HAs δεν οδήγησε σε κάποια σημαντική αλλαγή της τοξικότητας σε σύγκριση με τα μεμονωμένα NPs, στην περίπτωση των ZnO και ZnO-Ag NPs. Αντιθέτως, τα μίγματα Ag NPs-HAs ελάττωσαν την τοξικότητα που προκλήθηκε από τα μεμονωμένα Ag NPs. Όσον αφορά τις επιδράσεις των NPs στα αιμοκύτταρα των μυδιών, διαπιστώθηκε ότι κάθε NP διέθετε διαφορετικό μηχανισμό εκδήλωσης τοξικότητας. Συγκεκριμένα, τα ZnO NPs - μέσω του επιφανειακού τους φορτίου και της σωματιδιακής συσσωμάτωσης - ενδέχεται να εισέλθουν στα κύτταρα πριν την εκδήλωση κυτταρικής θνησιμότητας, ενώ η απελευθέρωση των ιόντων Zn2+ μπορούσε να οδηγήσει στην παραγωγή ριζών μέσω διέγερσης της διαδικασίας της αναπνευστικής έκρηξης. Αντιθέτως, η παρατηρούμενη κυτταρική και οξειδωτική καταπόνηση που προκλήθηκε από τα Ag NPs, πιθανότατα λόγω της απελευθέρωσης ιόντων Ag+, δε φάνηκε να σχετίζεται με την αναπνευστική έκρηξη. Ομοίως, οι κυτταρικές και οξειδωτικές βλάβες που προκλήθηκαν από τα ZnO-Ag NPs, υπέδειξαν την παρουσία ανταγωνιστικής/συνεργιστικής δράσης μεταξύ των μεταλλικών ιόντων (Zn2+, Ag+) που ανάλογα με τις εκάστοτε συνθήκες μπορούν να ρυθμίσουν τη συμπεριφορά και τις βιολογικές επιδράσεις του σύνθετου NP. Συμπερασματικά, τα αποτελέσματα υποδεικνύουν τη δυνατότητα των νεοσυντιθέμενων ZnO, Ag και ZnO-Ag NPs να προκαλούν τοξικές, κυτταροτοξικές και οξειδωτικές επιδράσεις. Παρόλα αυτά, διαπιστώθηκε ότι οι επιδράσεις των NPs ποικίλουν τόσο μεταξύ των ίδιων των NPs, όσο και μεταξύ των τεχνικών και των οργανισμών μοντέλων και/ή κυττάρων που χρησιμοποιήθηκαν. Κατά συνέπεια, είναι εμφανές ότι είναι αναγκαίο να αξιολογείται το τοξικολογικό προφίλ των NPs με τη χρήση ενός εύρους τεχνικών, βιοδεικτών και περιβαλλοντικών σεναρίων, καθώς και να γίνεται παράλληλα ένας ολοκληρωμένος και αξιόπιστος χαρακτηρισμός αυτών σε κάθε περίπτωση.

Mesothelial Cells

2007 ◽  
Vol 27 (2_suppl) ◽  
pp. 110-115 ◽  
Author(s):  
Susan Yung ◽  
Chan Tak Mao
Keyword(s):  
Mesothelial Cell ◽  
In Vitro Models ◽  
Mesothelial Cells ◽  
Dynamic Membrane ◽  
Peritoneal Mesothelial Cells ◽  
Immunohistochemical Markers ◽  
Vitro Model ◽  
And Function

♦ Background The introduction of peritoneal dialysis (PD) as a modality of renal replacement therapy has provoked much interest in the biology of the peritoneal mesothelial cell. Mesothelial cells isolated from omental tissue have immunohistochemical markers that are identical to those of mesothelial stem cells, and omental mesothelial cells can be cultivated in vitro to study changes to their biologic functions in the setting of PD. ♦ Method The present article describes the structure and function of mesothelial cells in the normal peritoneum and details the morphologic changes that occur after the introduction of PD. Furthermore, this article reviews the literature of mesothelial cell culture and the limitations of in vitro studies. ♦ Results The mesothelium is now considered to be a dynamic membrane that plays a pivotal role in the homeostasis of the peritoneal cavity, contributing to the control of fluid and solute transport, inflammation, and wound healing. These functional properties of the mesothelium are compromised in the setting of PD. Cultures of peritoneal mesothelial cells from omental tissue provide a relevant in vitro model that allows researchers to assess specific molecular pathways of disease in a distinct population of cells. Structural and functional attributes of mesothelial cells are discussed in relation to long-term culture, proliferation potential, age of tissue donor, use of human or animal in vitro models, and how the foregoing factors may influence in vitro data. ♦ Conclusions The ability to propagate mesothelial cells in culture has resulted, over the past two decades, in an explosion of mesothelial cell research pertaining to PD and peritoneal disorders. Independent researchers have highlighted the potential use of mesothelial cells as targets for gene therapy or transplantation in the search to provide therapeutic strategies for the preservation of the mesothelium during chemical or bacterial injury.

scholarly journals Mesenchymal Stem Cells as a Promising Cell Source for Integration in Novel In Vitro Models

Biomolecules ◽  
2020 ◽  
Vol 10 (9) ◽  
pp. 1306
Author(s):  
Ann-Kristin Afflerbach ◽  
Mark D. Kiri ◽  
Tahir Detinis ◽  
Ben M. Maoz
Keyword(s):  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Cell Types ◽  
In Vitro Models ◽  
Cell Source ◽  
Fundamental Research ◽  
Multiple Cell ◽  
Vitro Model

The human-relevance of an in vitro model is dependent on two main factors—(i) an appropriate human cell source and (ii) a modeling platform that recapitulates human in vivo conditions. Recent years have brought substantial advancements in both these aspects. In particular, mesenchymal stem cells (MSCs) have emerged as a promising cell source, as these cells can differentiate into multiple cell types, yet do not raise the ethical and practical concerns associated with other types of stem cells. In turn, advanced bioengineered in vitro models such as microfluidics, Organs-on-a-Chip, scaffolds, bioprinting and organoids are bringing researchers ever closer to mimicking complex in vivo environments, thereby overcoming some of the limitations of traditional 2D cell cultures. This review covers each of these advancements separately and discusses how the integration of MSCs into novel in vitro platforms may contribute enormously to clinical and fundamental research.

The Effect of Angle and Flow Rate Upon Hemodynamics in Distal Vascular Graft Anastomoses: An In Vitro Model Study

1991 ◽  
Vol 113 (4) ◽  
pp. 458-463 ◽  
Author(s):  
R. S. Keynton ◽  
S. E. Rittgers ◽  
M. C. S. Shu
Keyword(s):  
Stagnation Point ◽  
Flow Rate ◽  
In Vitro Model ◽  
Laser Doppler Anemometry ◽  
Bypass Graft ◽  
Outer Wall ◽  
Model Study ◽  
Vitro Model ◽  
Junction Angle

A steady flow, in vitro model of distal arterial bypass graft junctions was used to examine the effects of junction angle and flow rate on the local velocity field. Three test sections were fabricated from Plexiglas™ tubing having anastomotic junction angles of either 30, 45, or 60 deg. Flow visualization revealed velocity profiles skewed toward the outer wall with a flow split around a clear stagnation point along the outer wall. Laser Doppler anemometry [LDA] measurements confirmed a distinct stagnation point at the outer wall and both reverse and forward shear were detected immediately upstream and downstream, respectively, of this site. Axial velocities and shear rates along the outer wall were higher than along the inner wall and occurred in the junction angle order: 45, 60, and 30 deg. This study clearly identified changes in wall shear which varied with the anastomotic angle and flow rate.

scholarly journals Encapsulated multicellular tumor spheroids as a novel in vitro model to study small size tumors

2003 ◽  
Vol 57 (12) ◽  
pp. 585-588 ◽  
Author(s):  
Elena Markvicheva ◽  
Lina Bezdetnaya ◽  
Artur Bartkowiak ◽  
Annie Marc ◽  
Jean-Louis Gorgen ◽  
...  
Keyword(s):  
Tumor Cells ◽  
Tumor Biology ◽  
Cell Encapsulation ◽  
Large Cell ◽  
In Vitro Models ◽  
Multicellular Tumor Spheroids ◽  
Tumor Spheroids ◽  
Vitro Model

Presently multicellular tumor spheroids (MTS) are being widely used in various aspects of tumor biology, including studies in biology and photodynamic therapy. The cellular organization of spheroids allows the recreation of in vivo small tumors much better than all common two-dimensional in vitro models. The cell encapsulation method could be proposed as a novel technique to quickly and easily prepare a large number of spheroids with narrow size distribution within a desirable diameter range. Moreover, the proposed technique for spheroid generation using encapsulated growing tumor cells could provide entirely new avenues to develop a novel spheroid co-culture model (for instance, the in vitro co-cultvation of tumor cells and monocytes, or epithelial cells, or fibroblasts etc). The current research was aimed at developing a simple and reliable method to encapsulate tumor cells and to cultivate them in vitro. In order to generate spheroids, MCF-7 cells were encapsulated and cultivated in 200 ml T-flasks in a 5% CO2 atmosphere at 37?C for 4-5 weeks. The cell proliferation was easily observed using a light microscope. The cells grew in aggregates increasing in size with time. The cell growth resulted in the formation of large cell clusters (spheroids) which filled the whole microcapsule volume in 4-5 weeks.

scholarly journals Biosynthesized ZnO-NPs from Morus indica Attenuates Methylglyoxal-Induced Protein Glycation and RBC Damage: In-Vitro, In-Vivo and Molecular Docking Study

Biomolecules ◽  
2019 ◽  
Vol 9 (12) ◽  
pp. 882 ◽  
Author(s):  
Satish Anandan ◽  
Murali Mahadevamurthy ◽  
Mohammad Azam Ansari ◽  
Mohammad A. Alzohairy ◽  
Mohammad N. Alomary ◽  
...  
Keyword(s):  
Molecular Docking ◽  
Therapeutic Potential ◽  
Morphological Changes ◽  
Diabetic Rats ◽  
Protein Glycation ◽  
Zno Nps ◽  
Vis Spectroscopy ◽  
Arginine Residues

The development of advanced glycation end-products (AGEs) inhibitors is considered to have therapeutic potential in diabetic complications inhibiting the loss of the biomolecular function. In the present study, zinc oxide nanoparticles (ZnO-NPs) were synthesized from aqueous leaf extract of Morus indica and were characterized by various techniques such as ultraviolet (UV)-Vis spectroscopy, Powder X-Ray Diffraction (PXRD), Fourier Transform Infrared Spectroscopy (FT-IR), Scanning electron microscopy (SEM), and energy dispersive spectroscopy (EDS). Further, the inhibition of AGEs formation after exposure to ZnO-NPs was investigated by in-vitro, in-vivo, and molecular docking studies. Biochemical and histopathological changes after exposure to ZnO-NPs were also studied in streptozotocin-induced diabetic rats. ZnO-NPs showed an absorption peak at 359 nm with a purity of 92.62% and ~6–12 nm in size, which is characteristic of nanoparticles. The images of SEM showed agglomeration of smaller ZnO-NPs and EDS authenticating that the synthesized nanoparticles were without impurities. The biosynthesized ZnO-NPs showed significant inhibition in the formation of AGEs. The particles were effective against methylglyoxal (MGO) mediated glycation of bovine serum albumin (BSA) by inhibiting the formation of AGEs, which was dose-dependent. Further, the presence of MGO resulted in complete damage of biconcave red blood corpuscles (RBCs) to an irregular shape, whereas the morphological changes were prevented when they were treated with ZnO-NPs leading to the prevention of complications caused due to glycation. The administration of ZnO-NPs (100 mg Kg−1) in streptozotocin(STZ)-induced diabetic rats reversed hyperglycemia and significantly improved hepatic enzymes level and renal functionality, also the histopathological studies revealed restoration of kidney and liver damage nearer to normal conditions. Molecular docking of BSA with ZnO-NPs confirms that masking of lysine and arginine residues is one of the possible mechanisms responsible for the potent antiglycation activity of ZnO-NPs. The findings strongly suggest scope for exploring the therapeutic potential of diabetes-related complications.

Green Synthesis of Gold Nanoparticles Using Different Leaf Extracts of Ocimum gratissimum Linn for Anti-tubercular Activity

2019 ◽  
Vol 9 (2) ◽  
pp. 146-157
Author(s):  
Arti Gupta ◽  
Sonia Pandey ◽  
Bharat Variya ◽  
Shailesh Shah ◽  
Jitendra Singh Yadav
Keyword(s):  
Gold Nanoparticles ◽  
Mycobacterium Tuberculosis ◽  
Ethanolic Extract ◽  
Leaf Extracts ◽  
Ocimum Gratissimum ◽  
Mycobacterium Tuberculosis H37rv ◽  
Vis Spectroscopy ◽  
Lowenstein Jensen ◽  
Vitro Model

Background: Tuberculosis is a greatest threat to human health. It requires urgent need to seek new devise alternate strategies and ant-tubercular compounds. In the present scenario, Nonmaterias, have opened new avenues in medicine, diagnosis and therapeutics. Objective: In view of this, the current study aims to synthesize gold nanoparticles and determine its efficacy to inhibit Mycobacterium tuberculosis. Methods: Gold nanoparticles (GNPs) synthesized from medicinal plant, such as Ocimum gratissimum linn, were tested against Mycobacterium tuberculosis (H37RV strain). Gold nanoparticles were characterized by UV-Vis spectrophotometer, FTIR, SEM and TEM. TEM results revealed that the GNPs were found spherical in structure and around 10-25 nm in diameter. UV-Vis spectroscopy exhibited an absorption peak at 348 nm. Fourier transform infra-red spectroscopy showed the GNPs have coated with phytoconstituents (terpenoids) that indicate the role of bio-molecules responsible for efficient stabilization and capping of the gold nanoparticles. In vitro model was designed to determine minimum inhibitory concentration (MIC) of each sample by Lowenstein Jensen (LJ) slope method. Results: The results showed that the presence of ursolic acid in ethanolic and hydroalcoholic extracts was found to be 2.89% and 1.97%, respectively. GNPs of ethanolic and hydroalcoholic exhibited anti-tubercular activity, with MIC 2.5 µg/ml and 20 µg/ml, respectively. While ethanolic and hydroalcoholic extracts showed such activity at concentrations 50 µg/ml and 75 µg/ml, respectively. Conclusion: GNPs synthesized from ethanolic extract showed profound efficiency to kill mycobacteria. As in this method no chemical reagents were used, the synthesized gold nanoparticles have potential for biological applications. There is an urgent need to further development of nano-antibiotic for tuberculosis.

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