Synthesis of PLGA Nanoparticles of Tea Polyphenols and Their Strong in Vivo Protective Effect Against Chemically Induced DNA Damage

8/10/2019 Synthesis of PLGA Nanoparticles of Tea Polyphenols and Their Strong in Vivo Protective Effect Against Chemically I

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International Journal of Nanomedicine Dovepressopen access to scientific and medical research

Open Access Full Text Article O r i g i n a l R e s e a r c h

Synthesis of PLGA nanoparticles of

tea polyphenols and their strong invivo protective eect againstchemically induced !A damage

This article "as pu#lished in the follo"ing ove Press $ournal%

&nternational 'ournal of

!anomedicine () April *+()

!um#er of times this article has #een vie"ed

Amit ,umar Srivastava(

Priyan-a .hatnagar*

/adhuli-a Singh(

San$ay /ishra(

Pradeep ,umar*

0ogesh"er Shu-la(

,ailash 1hand Gupta(2*

(Proteomics La#oratory2 &ndian&nstitute of Toxicology Research

31S&R42 Luc-no"2 &ndia5 *!ucleic AcidResearch La#oratory2 &nstitute ofGenomics and &ntegrative .iology31S&R42 elhi 6niversity 1ampus2 &ndia

Abstract: In

spite of proficient

results of severalphytochemicals in

preclinical

settings, the

conversion rate

from bench to

bedside is not very

encouraging.

Many reasons are

attributed to this

limited success,

including

inefficient

systemic delivery

and bioavailability

under in vivo

conditions. To

achieve improved

efficacy,

polyphenolic

constituents of

black (theaflavin

[TF] and green

(epigallocatechin!

"!gallate [#$%$]

tea in poly(lactide!

co!glycolide

nanoparticles

(&'$!)&s *ere

entrapped *ith

entrapment efficacy of 7+- and /-,

respectively. Further, their preventive potential

against 0,+!dimethylben1anthracene (2M3!induced 2) damage in mouse skin using

2) alkaline un*inding assay *as evaluated.

&retreatment (topically of mouse skin *ith

either TF or #$%$ (+44 g5mouse doses

e6hibits protection of 78."7- and ."-,

respectively, against 2M3!induced 2)

damage. 9o*ever, pretreatment *ith TF!loaded

&'$!)&s protects against 2) damage

/7.7+- by +54th dose of bulk, 0+.0:- by

+5+4th dose of bulk, and 0.7/- by +58th dose

of bulk. ;imilarly, 8+.- (+54th of bulk,

80./"- (+5+4th of bulk, and /".+7- (+58th of

bulk prevention *as noted using #$%$!loaded

&'$!)& doses. These results sho*ed that tea

polyphenol!loaded &'$!)&s have 7"4!fold

dose!advantage than bulk TF or #$%$ doses.

dditionally, TF! or #$%$!loaded &'$!)&s

sho*ed significant potential for induction of

2) repair genes (XRCC1, XRCC3, and

ERCC3 and suppression of 2) damage

responsive genes (p53, p21, MDM2,

GADD45, and COX-2 as compared *ith

respective bulk TF or #$%$ doses. Taken

together, TF! or #$%$!loaded &'$!)&s

sho*ed a superior ability to prevent 2M3!

induced 2) damage at much lo*er

concentrations, thus opening a ne* dimension

in chemoprevention research.

Keywords: 2) alkaline un*indingassay, mouse skin, 2) repair

1orrespon

dence%

0ogesh"er

Shu-laProteomic

s

La#oratory2 &ndian &nstitute

of Toxicology Research

31S&R42

PO .ox 8+2 /G/arg2 Luc-no"6P **9++(2 &ndia

Tel +:( ;**

*:9)8*84 has gained

importance for the encapsulation of a *ide variety of drugs as it

is biodegradable, biocompatible, capable of controlling the

release of the incorporated entity, and less to6ic.+/,+0

The choice

of &'$ *as also supported by the fact that it can be used to

form stable nanoparticles ()&s, and has already been approved

for use in humans by the B; Food and 2rug dministration.+

Thus, it has been chosen as the polymer scaffold to fabricate

the )&s used in the present study.

Mammalian skin forms a

hurdle bet*een the host and

surrounding

environments.+:,4

9o*ever,

it is also a ma=or route of

entry for several to6ic agents

and a potential target of

environmental ha1ards, *hich

can cause several skin!related

abnormalities including skin

cancer.+

#pidemiological

stud!ies suggest that almost

one!third of all ne* cancers

diagnosed annually

*orld*ide originate from

skin.

;everal studies sho*

an association bet*een

different human cancers and

lifestyle5dietary habitsA

additionally, detailed studiesof muta!tional events in

human cancers have provided

evidence for a direct

involvement of

environmental

mutagens5carcinogens in the

progression of different types

of cancer.",7

In the present study, a

&'$!based drug

development strategy *asfollo*ed to encapsulate TF

and #$%$. The authors

envisioned that )&s!

mediated delivery of

bioactive catechins from

black tea (TF and green tea

(#$%$ could be useful in

enhancing their protective

potential against 0,+!

dimethylben1[a]anthracene

(2M3!induced 2)damage in mouse skin

tissue.

&ateria's andmethods1hemicals&'$ 84>84 (molecular *eight

[MC] 74D08 k2a, polyvinyl

alcohol (MC "4 k2a, #$%$,

TF, and 2M3 *ere purchased

from ;igma!ldrich (;t 'ouis,

M?, B;. 9ydro6ylapatite

and ),)!dimethylformamide

*ere purchased from ;isco

Eesearch 'aboratory (Mumbai,

India. ll other chemicals

used *ere of analytical grade

purity and procured locally.

Formulationof TFE or=G1GEloadedPLGAE!PsTF! or #$%$!loaded &'$!

)&s *ere prepared using the

standard solvent evaporation

method,8

*ith minor

modifications. 3riefly, a 4.8m' a


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Dovepress Protection against !A damage #y nanotied tea polyphenols

by *ashing *ith *ater, and finally, the )&s *ere free1e!dried

*ith a Bnivapo +849 Bnicryo free1e dryer (Bnie


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The mean particle si1e and distribution, as *ell as the

=valuation of TF or =G1Grelease

1eta potential of the TF! or #$%$!loaded &'$!)&s *as from !Psdetermined by a dynamic light scattering techni


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,ab'e 1 1haracteristics of TFJ=G1G loadedE!Ps

"anopartic'es - ie'd #i)e nm eta potentia' rug 3oading ncapsu'ation

+I m - 6ciency -

=G1GEPLGA :9?:+ (*


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9igure 1 Particle sie distri#ution of epigallocatechinE)Egallate 3A4 andtheaMavin4nanoparticles? 3

"ote: Nalues are represented as meanstandard deviation?

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Srivastava et al Dovepress

having all the properties re


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A 100 EGCG PLGA NPs90

80

release 70

60

50

%d

rug

40

30

20

100

0 1 2 3 4 5 6 7 8 9 10

Numer !" days

B #0 $% PLGA NPs80

70

release

60

&0

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40

30

20

'0

0

0 1 2 3 4 5 6 7 8 9 10

Numer !" days

9igure % lnEvitro releaseA4and TFprole34PLGAE!Ps?of =G1G 3

"ote: Nalues are represented as meanstandard deviation?

Abbre7iations: =G1G2epigallocatechinE)Egallate5 PLGAE!P2 poly3lactideEcoEglycolide4 nan

time interval of 7 hours follo*ing the doses of TF or #$%$. ;ignificant

protection (78."7- *as observed at the dose of +44 g5mouse of bulk TF

in 7 hours (Figure 8, *hile TF!loaded &'$!)&s at doses 8, +4, and 4g5mouse provided protection of /7.74-, 0+.+:-, and 0.7/-,

respectively, *hich clearly sho*s about a "4!fold dose!advantage over

the bulk counterpart. ;imilarly,

*ith bulk #$%$ (+44 g5

mouse, .- protection *as

observed, *hile #$%$!loaded

&'$!)&s at 8, +4, and 4

g5mouse dose provided

8+.-, 80./"-, an

protection, res

sho*ing about a "!f

advantage over bulk

9o*ever, &'$!lo

($roup KI and #$%$

25

rea(s

20

s

trand

15

!"

10

Numer

5

0

II

9igure 4 Reduction in /.AEinduced strand #rea-s#y #ul- and poly3lactideEcoEglycolide4Eloaded teapolyphenols?

"otes: ata sho"n is the meanstandard deviationof all the animals5

indicatesP,+?+;5Qindicatessignifigniccantrof nanotied teapolyphenol over its#ul-2 P,+?+;?

Abbre7iation:/.A2


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Srivastava et al Dovepress

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9igure $ =ect of

#ul- and PLGAE

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polyphenols on the

transcription of !A

d"as uantied #y

densitometryIE

actinand? expressedas a fold dierence

against

"otes: atasho"nis the result of threedierentexperimentsP,+?+;incomparisonto"ithGroup&&5Qsimindilcartesresultsignic?Teect ofnanotied teapolyphenol over its#ul-2 P,+?+;? Lanes%&2 1ontrol5 &&2 /.A3;* gJmouse45 &&&2TF 3(++ gJmouse4+/.A5 &N2 TFE

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Synthesis of PLGA Nanoparticles of Tea Polyphenols and Their Strong in Vivo Protective Effect Against Chemically Induced DNA Damage