Accepted Manuscript

Title: Highly selective nicotinohydrazide based ‘turn-on’

chemosensor for the detection of bioactive zinc(II): its
biocompitability and bioimaging application in cancer cells

Authors: Manohar Patil, Shilpa Bothra, Suban K. Sahoo, Hilal

Ahmad Rather, Rajesh Vasita, Ratnamala Bendre, Anil Kuwar

PII: S0925-4005(18)30923-7
DOI: https://doi.org/10.1016/j.snb.2018.05.022
Reference: SNB 24678

To appear in: Sensors and Actuators B

Received date: 19-1-2018

Revised date: 4-5-2018
Accepted date: 7-5-2018

Please cite this article as: Manohar Patil, Shilpa Bothra, Suban K.Sahoo, Hilal
Ahmad Rather, Rajesh Vasita, Ratnamala Bendre, Anil Kuwar, Highly selective
nicotinohydrazide based ‘turn-on’ chemosensor for the detection of bioactive zinc(II):
its biocompitability and bioimaging application in cancer cells, Sensors and Actuators
B: Chemical https://doi.org/10.1016/j.snb.2018.05.022

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 Highly selective nicotinohydrazide based ‘turn-on’ chemosensor for the detection of

bioactive zinc(II): its biocompitability and bioimaging application in cancer cells

Manohar Patila, Shilpa Bothrab, Suban K. Sahoob*, Hilal Ahmad Ratherc, Rajesh Vasitac,

Ratnamala Bendrea*, Anil Kuwara*

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School of Chemical Science, North Maharashtra University, Jalgaon-425001, India.

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b
Department of Applied Chemistry, SV National Institute of Technology, Surat-395007 India.

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c
School of Life Sciences, Central University of Gujarat, Gandhinagar-382030 India

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*Corresponding author: E-mail: kuwaras@gmail.com (Dr Kuwar),

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suban_sahoo@rediffmail.com (Dr Sahoo); Phone No. 0257-2257431

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Highlights:
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 A new Sciff base receptor 3 was develoed for the detection of Zn2+.
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 Receptor 3 showed both colorimetric and turn-on fluorescence response.

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 Receptor 3 exhibit good selectivity and lower detection limit in semi-aqueous medium.

 The receptor 3 was applied for in-vitro detection of Zn2+ in live A549 cells.
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Abstract
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A novel optically active (E)-N’-(5-allyl-2-hydroxy-3-methoxybenzylidene)

nicotinohydrazide (receptor 3) was synthesized by simple Schiff base condensation of 5-allyl-

2-hydroxy-3-methoxybenzaldehyde with nicotinohydrazide. The ability of receptor 3 was

investigated for the selective sensing of cations by chromogenic and fluorogenic technique.

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Receptor 3 showed highly selective response for Zn2+ ions in the binary mixture of

acetonitrile/water (50:50, v/v) and possessed no interference from any of the potentially

competing cations. Upon additions of Zn2+, the fluorescence intensity of the receptor 3 showed

significant enhancement at 517 nm due to the chelation-enhanced fluorescence (CHEF) effect.

The receptor 3 showed a good linearity for Zn2+ ion with the detection limit down to 4.35 nM.

With negligible toxicity towards live cells, the receptor 3 could be successfully employed to

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monitor the Zn2+ ions in live A549 cells.

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Keywords: Fluorescent sensor; Zn2+; Schiff base; Live cells imaging; CHEF effect.

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1. Introduction

Design and development of optically active molecular receptors for the selective and

distinct detection of biochemical and environmental significant analytes have gained rapid

attention in the field of supramolecular chemistry and chemosensing. In the present scenario,

the design of fluorescent based sensors which possesses the ability for the recognition of various

clinically and biologically essential ions have emerged as one of the most thrust area of research

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because of their potential applications in various fields like material, medical and environmental

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analyses [1,2]. Several approaches to chemosensors development have been explored, but the

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most universal approach involves the coordination to the lone pair of a nitrogen atom in the

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close proximity of fluorophore such that fluorescence emission gets altered upon ions binding

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[3]. The receptor unit could be bind with the cations/anions and can cause associated change in
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one or more optical or photophysical properties of the system, that arises from the
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complexation, substitution, or ring transformation [4,5].
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Zinc is the second most trace element widely essential in cellular

functions of the body such as intracellular signalling molecule, capable of communicating

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between cells, converting extracellular stimuli to intracellular signals, and controlling

intracellular events. It also plays vital role in various biological processes, such as peptide
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synthesis, DNA synthesis, RNA transcription, metabolism of cells, metalloenzyme regulation

and neurophysiology, and also induces the formation of β-amyloid, which is related to the
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neurological function [6]. However, the presence of this ion at low concentration may result in

decrease in the ability of the islet cells of the pancreas to produce and secrete insulin,
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acrodermatitis enteropathica, impaired cognition, immune dysfunction, night blindness and

diarrhoea while an excessive dosage of the same may cause superficial skin diseases, prostate

cancer, and diabetes and brain diseases [7-8]. Numerous techniques developed for the detection

of zinc ions are atomic absorption spectrometry (AAS) [9], inductively coupled plasma mass

spectrometry (ICP-MS) [10] and voltammetry [11], but are expensive and time-consuming.
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Thus, many alternate techniques have been adopted for the selective and sensitive detection of

Zn2+ ions, but due to the closed shell 3d10 electronic structure without oxidation–reduction

activity in biological environments makes the detection of Zn2+ difficult [12]. Also, the

detection of Zn2+ ion have received tremendous attention, but is challenging to develop selective

and sensitive chemosensor for Zn2+ ion that can distinguish it from Cd2+ ion as the two belongs

to same group in the periodic table and have similar characteristic properties [13]. However,

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the fluorescent sensors for the sensing of Zn2+ have found preferential merits owing to the high

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sensitivity, low cost, ease of handling, robustness and the possibility of real-time monitoring

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with fast response time [14]. Many fluorescent chemosensors for Zn2+ recognition over other

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metal ion has successfully reported [15-26].

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Thus, as a part of our ongoing research on optical sensor development with high
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selectivity and sensitivity [27], we have designed and synthesized the receptor 3 i.e. (E)-N’-(5-
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allyl-2-hydroxy-3-methoxybenzylidene) nicotinohydrazide (Scheme 1). The developed
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receptor possess extensive substrate for nucleophilic addition reaction; its carbonyl group can

be activated intramolecular by the neighbouring phenolic hydroxyl group through

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intramolecular hydrogen bonding which function as a sensor toward Zn2+ ions comparatively

lower detection limits in biologically competing solvents.

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Scheme 1. Synthesis of receptor 3.

2. Experimental

2.1. Chemicals and Instruments

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 All reactions were carried out using oven-dried glasswares. All the chemicals were

purchased from Sigma Aldrich, India and 5-allyl-2-hydroxy-3-methoxybenzaldehyde was

prepared in our laboratory by using our previously reported method [28]. All reactions were

monitored using thin-layer chromatography (TLC) to confirm the successful formation of

product. 1H-NMR (400 MHz) and 13C-NMR (100 MHz) spectra were determined on a Bruker

Advance II 400 spectrometer. Chemical shifts for 1H NMR were reported in parts per million

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(ppm), calibrated to the solvent peak set. Fluorescence measurements were carried out on a

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Horiba Jobin Yvon, Fluoromax-4 Spectrofluorometer equipped with a xenon lamp. UV-Vis

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absorption spectra were recorded on a Shimadzu UV-2450 spectrophotometer.

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2.2. Synthesis of the receptor 3

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Receptor 3 was prepared using the reported literature procedures [28]: solution of 5-allyl-
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2-hydroxy-3-methoxybenzaldehyde (0.21 g, 1 mmol) in ethanol (10.0 mL) was added drop-
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wise to a solution of nicotinohydrazide (0.15 g, 1 mmol) in ethanol (10.0 mL) at room
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temperature. The reaction mixture was stirred and refluxed for 4 hours. Progress of the reaction

was monitored by TLC. After the reaction solution was stirred for additional three hours at
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room temperature gave yellow coloured powder was filtered and recrystallized with ethanol.

Yield: 90%. 1H NMR (DMSO-d6, δ ppm) : 12.18 (s, 1H, -OH), 10.9 (s, 1H, -NH), 9.12 (s, 1H,
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-CH), 8.76-8.75 (d, 1H, Ar), 8.59 (s, 1H, Ar), 8.29-8.28 (d, 1H, Ar), 7.53-7.49 (q, 1H, Ar), 6.89

(s, 1H, Ar), 6.80-6.80 (s, 1H, Ar), 6.01-5.59 (m, 1H), 5.05-5.05 (d, 2H), 3.86 (s, 3H), 3.36-3.33
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(d, 2H). 13C NMR: (DMSO-d6, δ ppm) 161, 152, 149, 148, 147, 145, 137, 135, 130,128, 123,

120, 117, 115, 114, 55, 29. LC-MS (M + Na) = Calcd: 334.00; Found: 334.05.
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2.3. UV-Vis and fluorescence spectral measurements

All the stock and working solutions were prepared in double distilled water and

spectroscopic grade acetonitrile. The stock solution of receptor 3 (1 × 10-5 M) was prepared in

binary solvent i.e. acetonitrile: water (50:50) and cations (1 × 10-4 M) were prepared in double

distilled water. The UV-Visible absorption and fluorescence experiments were carried out at
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room temperatures (298 K) with the aim of determining the selectivity of the receptor 3 towards

a series of cations (Cr3+, Fe3+, Co2+, Ni2+, Cu2+, Zn2+, Cd2+, Hg2+, Pb2+, Na+, K+, Mg2+, Ca2+,

Ba2+, Cs2+ and Al3+).

The absorption and fluorescence titrations with the selectively detected analyte Zn2+

showed satisfactory linear relationship between the added concentrations and the absorbance

intensity. The titration was accomplished through successive addition of Zn2+ (0.02 ml) in water

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to a solution of receptor 3 (2.0 ml) in a binary solvent mixture of acetonitrile: water (50:50; v/v)

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system. The collected data were processed using the HypSpec program to calculate the

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association constant (Ka) of detected analyte Zn2+ ion. The fluorescence intensity of emission

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peak maxima at 517 nm was used to calculate stoichiometry ratio for Zn2+ ion. The plot of [HG]

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versus Xi was used to determine the stoichiometry of the complex formed. The concentration

of [HG] was calculated by the equation of [HG] = (∆F/F0) [H] and Xi (i.e. mole fraction) =
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[H]v/ [H]v + [G]v. The fluorescence spectra were recorded at an excitation wavelength of 293
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nm. The excitation and emission slits were both set to 5.0 nm.

2.4. Cell culture studies

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The cytotoxicity assay (Alamar blue assay) and bioimaging of receptor 3, Zn2+ and

receptor 3-Zn was performed using A549 (lung cancer cell line). The A549 cells were procured
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from National Centre for Cell Science, Pune, India. RPMI 1640 media, foetal bovine serum

(FBS) and trypsin was procured from Gibco, USA. Resazurin was purchased from Sigma
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Aldrich, India. Cell culture plates were procured from Falcon, Fischer Scientific, USA and glass

cover slips were purchased from Himedia, India. The A549 cells were grown in a RPMI 1640
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media supplemented with 10% FBS in humidified incubator at 37°C and 5% CO2. Cells were

trypsinized for seeding once the cell confluency was 70-90%.

2.5. Cytotoxicity assay and bioimaging

The A549 cells were seeded in 96 well plates as 10,000 cells per well. After 24 hours,

receptor 3, Zn2+ ion and receptor 3-Zn ion (1 μM and 10 μM) in media was added to the wells
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and incubated for another 24 hours. Control wells were treated with equal volume of dimethyl

sulfoxide (DMSO) as the channel for compounds. Further, after 24 hours, the media was

discarded and replaced with fresh media containing 10% v/v of the Alamar blue solution and

incubated for an hour at 37°C in the dark. The plates were taken out and the absorbance was

recorded at 570 nm to evaluate the cell viability using multi-plate reader (Synergy H1,

BioTek®). The experiment were performed as per reported procedure in literature [29]. All the

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experiments were performed in triplicates. For data analysis, Graphpad Prism software was

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used to plot the graph and Student’s t-test (Two-tailed) was performed by comparing each group

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individually with control.

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For bioimaging, the A549 cells were seeded separately on 14 mm coverslips and allowed

to grow till 24 hours. For bio imaging of each compound, 0.5 μM of Zn 2+ ion and 5 μM of
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receptor was incubated separately for 30 minutes at 37°C and 5% CO2 in dark. However, for

bioimaging of receptor 3-Zn. 0.5 μM of Zn2+ ion were incubated separately for 30 minutes.
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Then, media was replaced with 5 μM of receptor 3 and incubated for additional 30 minutes.
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Further, the media was removed and the cells were washed with PBS followed by fixing with
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2% para-formaldehyde for 30 minutes. Thereafter, imaging was performed under florescent

microscope (Zeiss-Scope A1, Germany). The excitation and emission wavelengths were set
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from 390 to 650 nm using filter set 25 (488028-0000).

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3. Results and discussion

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3.1. Synthesis of 3

Receptor 3 was synthesized by simple Schiff base condensation reaction of one mole of
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nicotinic hydrazide with one mole of 5-allyl-2-hydroxy-3-methoxybenzaldehyde in ethanolic

medium as shown in scheme 1. The obtained receptor 3 was in good quantitative yield with

bright yellow coloured powder. The structure of receptor 3 was successfully characterized by

various spectroscopic techniques such as 1H NMR, 13C NMR and mass (Fig. S1-S3).

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3.2. Receptor 3 as a sensory probe for Zn2+ ion

The binding behaviour of receptor 3 towards different cations was monitored using UV-

Vis absorption and fluorescence spectroscopy. The absorption spectral properties of receptor 3

was studied in binary mixture of acetonitrile: water (50:50; v/v) upon addition of various cations

such as Cr3+, Fe3+, Co2+, Ni2+, Cu2+, Zn2+, Cd2+, Hg2+, Pb2+, Na+, K+, Mg2+, Ca2+, Ba2+, Cs2+ and

Al3+. The absorption spectra of receptor 3 contains two maxima band at 300 nm and 347 nm

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arising due to the π-π* and n- π* transitions, respectively (Fig. S4). Addition of Zn2+ ion to the

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receptor 3, the band at 300 nm shows red-shift to 323 nm along with the appearance of a new

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band at 406 nm. However, other tested cations with receptor 3 showed no significant changes

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in the absorbance spectra, which clearly indicates that the receptor 3 binds to Zn2+ ions

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selectively in the ground state which perturbs the electronic properties of receptor 3. The UV-

Vis titration of receptor 3 (c = 1 × 10-5 M) was performed by incremental addition of Zn2+ (Fig.
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1). Upon gradual addition of Zn2+, the receptor band at 300 nm shifted to 323 nm with the
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formation of two isosbestic points at 267 nm and 316 nm and a new band formed at 406 nm.

The presence of isosbestic points in the spectra indicates the formation of new species upon
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interaction Zn2+ with the receptor 3. Also, the receptor band at 347 nm disappeared upon

addition of Zn2+, which clearly shows the coordinative interaction between receptor 3 and Zn2+
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ions. It may be assumed that the imine-N, carbonyl-O and phenolate-O of receptor 3 binds with

Zn2+ ions and forms a 3-Zn complex in solution, which accelerates the intramolecular charge
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transfer (ICT) process between the receptor 3 and Zn2+ ion and also brings the lowest excited

state closer to the highest ground state [13].

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The calibration plot at 406 nm was plotted at varying concentration of Zn2+ ions by

using the titration data (Fig. S5), and the limit of detection (LOD) was estimated by adopting

the IUPAC approved equation, LOD = 3σ/slope (where, σ represents the standard deviation of

ten blank samples) [30]. The estimated LOD was obtained down to 10.75 nm. Further, the

titration data was fitted in the least-square fitting program HypSpec to determine the binding
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constant between receptor 3 and Zn2+ ions. The best fit model was obtained for the 1:1 binding

stoichiometry with the binding constant of log  = 6.19 (0.04) (Fig. S6) [31].

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Fig. 1. UV-Vis absorption titration of receptor 3 (c = 1 × 10-5 M) in acetonitrile: water (50:50;

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v/v) after the successive addition of Zn2+ (0 to 200 μL, c = 1 × 10-4 M). Inset shows the colour

change of vials from colourless to yellow.

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Considering the 1:1 binding stoichiometry between the receptor 3 and the Zn2+ ions, the

possible 3D structure was calculated by applying the density functional theory (DFT) method.
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All calculations were performed in Gaussian 09W by applying the exchange-correlation

functional B3LYP and the basis sets 6-31G(d,p) for the C, H, N, O, atoms whereas LANL2DZ
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for the Zn atom [32]. The DFT computed structure of the receptor 3 and its receptor 3-Zn

complex is shown in Fig. 2a. The optimized structure of the receptor 3 showed an

intramolecular hydrogen bonding of length 1.814 Å, which complements well with the peak

observed for the phenolic-OH in the downfield region of 1H NMR at 12.18 ppm (Fig. S1). The

receptor 3 provides a suitable cavity to accommodate the Zn2+ ions through the imine-N,
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carbonyl-O and phenolate-O donor atoms. Upon coordination with Zn2+, the interaction energy

(Eint = Ecomplex-Ereceptor-EZn2+) is lowered by -109.03 kcal/mol which indicates the formation of

a stable complex. Further, the formation of new band upon complexation was complemented

from the lowering in the band gap between the highest occupied molecular orbital (HOMO)

and lowest unoccupied molecular orbital (LUMO) of receptor 3 (Fig. 2b-c).

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Fig. 2. (a) The DFT computed optimized structure of receptor 3 and its complex with Zn2+. The

LUMO’s (b) and HOMO’s (c) diagrams of receptor 3 and its complex with Zn2+.

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 The metal ions sensing ability of the receptors 3 (1×10-5 M) was investigated by

fluorescence method in similar mixed solvents medium by adding the nitrate salt of series of

cations like Cr3+, Fe3+, Fe2+, Co2+, Ni2+, Cu2+, Zn2+, Cd2+, Hg2+, Pb2+, Na+, K+, Mg2+, Ca2+,

Ba2+, Cs2+and Al3+ (Fig. 3a). The receptor 3 showed weak fluorescence upon excitation at 293

nm due to the conformational flexibility upon isomerization of -C=N double bond along with

the photo-induced electron transfer (PET) process occurred at the excited state. Among the

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various metal ions examined (1 equiv.), the receptor 3 showed highly selective ‘Off–On’

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fluorescence enhancement at 517 nm in the presence of Zn2+ (Fig. 3a). The fluorescence

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enhancement of receptor 3 in the presence of Zn2+ at 517 nm can be explained due to the

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inhibition of the -C=N isomerisation at the excited state that caused a large chelation-enhanced

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fluorescence (CHEF) effect [28]. One important criteria for a chemosensor is to give a specific
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response for the selective detection of target analytes over a wide range of potentially competing
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analytes, and to avoid the cross sensitivity. The competitive experiment was performed by

recording the fluorescence spectra of receptor 3 in the presence of 1 equiv. of Zn2+ ion mixed
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with 1 equiv. of other potent cations mentioned above (Fig. 3b). No significant variation in the
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fluorescence spectra of receptor 3 was found in comparison with the same amounts of Zn2+

solution in the presence and absence of other cations, and showed relative error below ± 5%.
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These results indicates that the recognition of Zn2+ by receptor 3 is not notably interfered by

other potentially competing cations. Therefore, the receptor 3 exhibits a high selectivity and
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specificity toward Zn2+, and opens a new door for the analytical application of receptor 3 in real

sample analysis.
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Fig 3. (a) Fluorescence spectra (λexc= 293 nm) of receptor 3 (1×10−5 M) in the absence and

presence of metal ions (1 equiv.) in acetonitrile: water (50:50; v/v) and (b) competitive study
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of receptor 3 towards Zn2+ in the presence of one equivalent of other interfering metals ions.

Further, the fluorescence titration of receptor 3 (c = 1 × 10-5 M) was performed by

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successive incremental addition of Zn2+ ion (Fig. 4a). The fluorescence band at 517 nm of
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receptor 3 increased continuously with the increasing concentration of Zn2+ due to the CHEF
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effect. Using the fluorescence titration data, the binding constant was determined by non-linear

curve fitting of the fluorescence titration data of receptor 3 for Zn2+ (Fig. S7) and was found to

be log  = 5.63 (0.15) for the best fit model of 1:1 binding stoichiometry. The binding

stoichiometry of receptor 3-Zn complex was determined using the Job’s plot experiment [33].

The fluorescence intensity was plotted against the molar fraction of the chemosensor under a
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constant total concentration. A maximum intensity was observed when the molar fraction was

~0.5, which indicates a 1:1 ratio for the receptor 3-Zn complex (Fig. 4b). Further, the LOD of

receptor 3 for the detection of Zn2+ using fluorescence approach was estimated down to 4.35

nM with regression coefficient of 0.959 (Fig. S8). The sensing ability of the developed receptor

3 for Zn2+ ion clearly delineated that this fluorescence assay was found to be sensitive, facile

and highly selective in compared to the various reported methods (Table 1).

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Fig. 4. Fluorescence spectra (λexc = 293 nm) of receptor 3 (c = 1 × 10-5 M) after the successive

addition of Zn2+ (c = 1 × 10-4 M) in acetonitrile: water (50:50; v/v). (b) Host guest relationship

from Job’s plot obtained for the complexation of receptor 3 with Zn2+.
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Table 1. Brief account of various fluorescent sensing systems reported for Zn2+ detection.

Sr. No. Developed Systems LOD, M Real samples References

1. N,N′- 1.5 × 10−7 water samples [34]
phenylenebis(salicylideaminato) and waste water
2. Pyridoxal-2-hydrazinopyridine 70× 10−9 live cells [35]
Schiff-base
3. Furan-2-carbohydrazide-5-(2- 77 × 10−9 - [36]
thienyl)-salicylaldehyde
4. Diarylethene derivative 8.10 × 10−8 - [37]

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5. Tetra-peptides conjugated with 32 × 10−9 live cells [38]

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dansyl groups
6. Receptor 3 4.35 × 10−9 lung cancer cell This study

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line

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3.3. Cytotoxicity study and bio-imaging application of receptor 3

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To examine the biological applications of receptor 3, the receptor was used for the
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detection of Zn2+ in living A549 cells (Fig. 5). The cells showed non-significant fluorescence
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upon incubation only with Zn2+ ions (0.5 μM) and a very weak fluorescence was observed when
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they were incubated only with receptor 3. However, an increase in the brightness of the

fluorescence was observed when the cells were incubated with receptor 3 in the presence of
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Zn2+. Thus, receptor 3 showed the potential to detect Zn2+ in an in vitro cellular system.

Alamar blue assay was performed to evaluate the cytotoxicity of receptor 3, Zn2+ and
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receptor 3-Zn after exposure of cells to a concentration range of 1 to 10 μM for 24 hours. The
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results shows that the percent cell growth in each group compared to the control, which were
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treated with equal volume of vehicle. We observed no significant cell death even after 24 hours

of treatment at any concentration of the receptor 3, Zn2+ ions and receptor 3-Zn individually or
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in combination as shown in Fig. 6. These results clearly indicates the excellent

cytocompatibility of receptor 3, Zn2+ and receptor 3-Zn making it useful for biosensors

applications.

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Fig. 5. a, b, c and d shows the bright field images of control, cells incubated with Zn2+, 3 and

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3.Zn. Whereas, e, f, g and h shows their fluorescence images respectively. All the images were

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captured under fluorescent microscope (Zeiss-Scope A1, Germany).
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Fig. 6. Cytotoxicity assay of 3, Zn2+ and 3.Zn2+ on A549 cells after 24 hours. The data shows

no significant decrease in the % cell growth in the groups treated with 3, Zn2+ or in combination.

The control wells were treated with DMSO vehicle. The experiment was performed in

triplicates (n=3).

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4. Conclusions

In summary, an easy-to-prepare fluorescent ‘turn-om’ sensor for the detection of Zn2+

ions was developed. In acetonitrile/water (50:50; v/v), the weakly fluorescent receptor 3 showed

highly significant fluorescence enhancement at 517 nm due to the chelation-enhanced

fluorescence (CHEF) effect. The chelation occurred between the receptor 3 and Zn2+ in 1:1

binding stoichiometry and the structure was proposed by DFT calculation. With the nanomolar

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detection limit, the receptor 3 showed no interference from any of the potentially competing

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cations. Importantly, the receptor 3 showed excellent cytocompatibility, cell permeability and

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successfully employed to monitor the Zn2+ ions in live A549 cells.

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Figures
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Authors Biography
Manohar Patil is a Ph.D. student in North Maharashtra University, Jalgaon, India. He did his
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M.Sc degree from the same University. His current research is focused on design of
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fluorescent based chemosensors, Biologically active molecule synthesis, Pesticide

Formulation development.
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Shilpa Bothra has completed her PhD degree under the supervision of Dr Sahoo from SV
National Institute of Technology, Surat, India. She has published more than 20 research
articles in the fields of nanomaterials and optical sensors development using functionalized
nanoparticles.
Hilal Ahmad Rather is a graduate Ph.D student at Central University of Gujarat, India. His
research work is focusing on Mesenchymal Stem cells on 3D mimetic microenvironments for

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tissue regeneration. His expertise includes functionalization of polymeric biomaterials,
fluorescent imaging, hydrogel fabrication and 3D cell culture.
Suban K. Sahoo is working as an assistant professor in the Department of Applied
Chemistry, S.V. National Institute Technology, Surat, India. He has published more than 130
research papers in various international peer reviewed journals. His area of research includes
supramolecular chemistry and molecular recognition, Colorimetric and fluorescent sensors for
anions and cations, ISEs and Sensors based on functionalized nanoparticles and QDs.
Rajesh Vasita is a faculty member at School of Life Sciences, Central University of Gujarat

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and leading a research group in Biomaterials & Biomimetic Laboratory. The focus of his

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research group includes surface engineering of nano-biomaterials and investigation of

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structure-property-function relationships of these materials. The applications of these
materials include tissue regeneration in development and disease, stem cell fate regulation,

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and drug delivery. Current projects include creating in vitro niche for homing meshenchymal
stem cells for bone tissue engineering and designing scaffold for 3D tumor.

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Ratnamala Bendre is currently working as an Professor in North Maharashtra University,
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Jalgaon, India. She had completed his Ph.D. and M.Sc from Pune University, India. Her
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current main research is coordination and bioinorganic chemistry.
Anil Kuwar is currently working as an Assistant Professor in North Maharashtra University,
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Jalgaon India. He had completed his Ph.D. in 2007 from the North Maharashtra University.
He had completed his postdoctoral research in Sunchon National University, Sunchon, South
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Korea and University of Utsunomiya, Utsunomiya, Japan. His current main research is
Supramolecular Chemistry and Bioinorganic Chemistry.
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