Nghiên cứu điều chế hydrogel nanocomposite trên cơ sở chitosan và curcumin ứng dụng trong tái tạo mô

• Nghien cixu - Ky thu4t
de

dinh logng terpin hydrat trong cac the phern da
thanh phan vai dieu kien pic terpin va chuen nal phai
tech khoi cac thanh phan kink trong hen hqp.

Tai liOu tham khao
1.80 lite (2009), Ducyc di4n Vitt Nam IV, tr. 596-597 .
2. Ducec dion Mk (USP 38) 2015, pp. 5497-5498.
3. Douglas A. Skoog, James J. Leary (1992), "Principles

of instrumental analysis", A harcourt brace Jovanovich college
publisher, the United States of America, pp. 588, 592.
4. Reich U. A. (2011), "Illustrated Handbook of Toxicology,
Georg Thieme Verlag KG, Germany, pp. 106-107.
5. International conference on harmonization of technical
requirements for registration of Pharmaceuticals for human
use (2005), Validation of analytical procedure: Text and
Methodology, pp. 1-13.

(Ngay nhan bai: 22/02/2017 - Ngay phan Nan: 21/03/2017 - Ngay duyat clang: 03/04/2017)

Nghien cfru ditu the hydrogel nanocomposite tren
chitosan va curcumin *lig dung trong tai tao mo
Ding Thi

Huynh Nguyen Tuin Anil', Nguyen Dai Hai', TrAn Le Bao Ha'
Doan Nguyen Vie, Tran Ng9c Quyenv, Nguyen Thi Bich Tram3
iVien Khoa hoc Vat lieu Ong dung - Vien Han liim KHCN Viet Nam
'Khoo Sinh hoc, Dpi hoc Khoa hoc Tlf nhien Tp. Ha Chi Minh

3Khoa Hoa hoc, Dai hoc Thu D6u Mat, Thanh pha Thu Dau Mat, Tinh Binh arcmg
* E-mail:
Summary

A temperature-sensitive hydrogel based on chitosan-pluronic was prepared. Copolymers were characterized
by 1H-NMR spectroscopy. Sol-gel phase transition behavior of the obtained hydrogel was rheologically determined
indicating formation of gel when the temperature increased from 25°C to 46°C. The copolymer solution may served
as a dispersant system for preparing nanocurcumin by emulsification. Nanocurcumine distribution ranged from 7 to
10 nm in the thermosensitive copolymer solution and the hydrogel nanocomposite could form at 34°C and enhanced
second-degreed bum healing effect in rats.
Keywords: Nanocurcumin, chitosan, Pluronic F127, thermal sensitive CP hydrogel.

001 Vin de
Cho den nay nhieu loai hydrogel sinh hoc duvc
nghien civu va Ong dung trong y sinh. Cac hydrogel
ten ca sa chitosan cho they hieu qua vocrt tr6i trong
tai tao mo va den truyen thuoc [11. Trong nghien cOu
nay, chUng tdi dieu the hydrogel nhay nhiet ten ca so'
chitosan-pluronic mang nanocucurmin Ong dung china
lanh vet thuvng. Pluronic cb ceu tr)c PEO-PPO-PEO,
duvc sir dung nhu' mat to duvc lam tang da h6a tan
cua thu6c. Trong dO PPO le phan ky, nu'O'c &wig vai tr6
tao lien kat ngang vat 19 tao hydrgel khi tang nhiet da
dung dich copolymer 12]. Bien tinh chitosan vai pluronic
tao deh xuet chitosan hem tan va nhpy cam nhiet de
mang va nha cham cac host chat. Curcumin cO tac
dyng he trg china lanh vet &Krung nh6 cac dac tinh nhu,
chang viem, chOng nhiem trong, chang oxy h6a, tang
hinh thanh m6 hat, kich thich nguyen bao sgi phat trien
tao collagen. Tuy nhien, curcumin rat kern tan va da

hep thu thep nen hien nay nano h6a curcumin de tang
sinh khd dung duvc quan tam [31. Ben canh hopt tinh
cO Ich cua chitosan va nanocurcumin, vai dac tinh cua
cac hydrogel, he nanocomposite hydrogel giOp bao phei
vet thirang ngan can cac vi khuen xem nhap, but dich
TAP CHI Dtroc HQC - 4/2017 (SO 492 NAM 57)

kr vet thirang va gidp hO trg lanh h6a vet throng (4] •

Vat li0u, phu'o'ng phi!" va thyv nghiOm
H6a chOt

Chitosan (MW-300 kD, 95% acetyl hoe), pluronic
F127 (Mw: 12.500 g/mol, Acros Organics), p-nitrophenyl
chloroformat (NPC) (BASF Corp), tetrahydrofuran
(THF), dimethylformamid (DMF), 3-amino-1-propanol
(99%, Acros Organic).
TOng hqp copolymer chitosan- F127
0,25 g chitosan hoe tan trong dung dich HCI
khuey trong 24 gite, sau dO dieu chinh pH ye 5,5. NPC-

F127-OH Ong hqp then quy trinh 15 '61, duvc hoe tan
trong nutec cat lanh 4°C. Dung dich NPC-F127-OH
duvc nhO tin tin vao dung dich chitosan lanh va khuey
qua dem. Dung dich duvc them tech vai mang cellulose
trong 4 ngay de loai bo cac tap chat va dong kh6 de
thu chitosan-pluronic F127 cho danh gia ceu trUc va thi
nghiem.
With gia dac tinh nhay nhiet cue copolymer
Chitosan-pluronic duvc hob tan trong nutec cat

(5, 8, 10, 12, 15 va 20%) duvc khao sat kha nang

47


• Nghien aim - Kg thuot
nhay nhiet a cac nhiet de: 4, 25, 30, 37 va 40°C bang
phu'ong phap deo ngt.rac dung djch copolymer. Diem
nhiet de chuyen pha tao gel cua dung djch copolymer
theo nhiet de duvc xac Binh tren may do lu'u bien ten
may HAAKE RheoStress 6000.
Dieu the nanocomposite hydrogel
Curcumin trong con (100 mM) nho vao dung djch
15% copolymer (chitosan:pluronic = 1:15; dung djch
nu*c va ethanol (9:1 v/v) sjeu am trong 15 phOt roj ce•
quay loci bo dung m6i, dang kh6 de thu nanocomposite
do kith thulac
va sau da haa tan trong mac cat lanh
hat nanocurcumin, diem nhiet tao gel va danh gia hieu
qua china bong.
Danh gid china lanh vet bong
Chi.* nhet trang (Mus muscuius var. albino) co
trong luvng 28 - 30 g mua ter Vier) Pasteur Tp. HO
Chi Minh, duvc nuOi On dinh 1 tuhn, can such !Ong a
vOng lung va gay me bang h6n hap ketamin va ntrac
(1:1) vai lieu luvng 0,02 ml/g the tong, rOi g8y bong
nhiet vat de 2. Tien hanh thi nghiem vai 4 nhOm: d6i
chang (kh6ng XeY 19), boi chitosan-pluronic hydrogel,
boi nCur-CS-F127 va kern trj bang de 2 (Biafine,
Filar)). Theo doi tien trinh lanh h6a, danh gia m6 hoc

sau 14 ngay.

a

Ket qua thuv nghiem va ban luen
Ket qua teng hop copolymer nhay nhidt
chitosan-pluronic
Copolymer &vac tong hap lien ket giCra nhOm
amin ten chitosan va NPC-pluronic. Hinh 1 cho they
khong con tin hieu pic cua p-NPC (5 - 7,4 va 8,3
ppm). Cac tin hieu the hien Ong Wang cua proton
methylen cuai mach pluronic da ghep vao chitosan
xu6t hien b 4,25 ppm. Cac proton dec tru'ng cua
pluronic va chitosan cOng dirqc ghi nhen. Ket qua thu
&vac cho they de tong hap thanh citing copolymer
ghep chitosan-pluronic.

12

I

F

a

,a

U- 45`A-1A1°101ti
S

1S 11 11 111 •

111

7

•

11

I

11

I

PM

Hinh 1: PM 'H-NMR cua copolymer ghep chitosanpluronic F127
Khao sat ddc tinh nhay nhidt cue copolymer
ghep chitosan-pluronic F127
Khi khao sat cac nong do tao gel bang phirang phap
deo nguvc, m6u chitosan-pluronic (t9 le 1:1 theo kin&
luvng ghep) kh6ng the to gel. Khi tang t9 le pluronic
len, kha nang tao gel duvc hinh thanh. Cac chitosanpluronic 6 t9 ghep 1:15 va 1:20 co gel hinh thanh tot 6
nhiet de 30-35°C. Mk chitosan-pluronic 1:20 cho they
gel c.6 the hinh thanh 6 nhiet de 25°C. NW vey, nhiet de
chuyen gel tang tiler) khi tang ham luvrig pluronic trong
meu chitosan-pluronic. Phirang phap do lu'u bien hoc
cOng cho they nhiet de chuyen gel cua dung djch 15%

chitosan-pluronic xay ra a 35°C vai ghep a t9 le 1:15.
Diem chuyen nhiet nay rat thich hap cho cac Ong dvng
y sinh vi gen vai nhiet de ca the.
Ddnh gia hg hydrogel nanocomposite
Phuvng phap dieu the he hydrogel china
nanocurcumin trong nghien thu cho they nanocurcumin
duvc dieu the trong dung djch chitosan-pluronic vai
&rang kinh di..rai 10 nm. He phan tan tren c6 the chuyen
thanh nanocomposite hydrogel china 15% chitosanpluronic F127 khi tang nhiet len 35°C (hinh 2).

Flanocurcumin

Hinh 2: Chu* hoa sol-gel cua chitosan-F127 va hinh thai nanocurcumin
48

TAP CHI DUQC HQC - 4/2017 (SO 492 NAM 57)


• Nghien ciru - Kji thOt
Dinh gia Neu qua china lanh vet thuvng bong
de 2
Ket qua thin nghiem china lanh vet bong ten m6
hinh chuet cho they sau 14 ngey vet bong phi) vai
nanocomposite hydrogel china nanocurcumin lanh

gen nhu, hoan town co nhieu long mai mix tren mo tai
tao. D6i vai Ink thirang mai vet b6ng china lanh kha tot
vai dO co vet thuang ten 95%. Mau khong dieu tri thi
vet bong chi china lanh khoang 80% (hinh 3).

Hinh 3: Hinh anh qua trinh china lanh vet bang va d6 thi danh gia dO co vet Chuang
Ket qua danh gia m6 hoc cho they cac hydrogel vai thu6c thirang mai Biafine. Tuy nhien, trong meu mo
khong gay di Ong, khong lam ngan can qua trinh dap Crng tai tao vai hydrogel composite cho trot tu' hinh thanh
binh thirang cua CU the. Vet bong ph6 nanocomposite lap thuvng bi, ha bi va lap ca c6 trot tu' nhu, mau m6
hydrogel co tac dung day nhanh qua trinh tai to va lanh tu' nhien (hinh 4). Ket qua nghien ciru sa khai nay cho
has vet thirang bong vai mat de collagen tai tao c6 trat they tiern Wang cua hydrogel ten ca sa chitosan mang
ty,, nhieu mach mau va m6 hat hinh thanh twang &rang nanocurcumin trong china lanh yet thu,ang.
ket qua phan tich mo hoc oh vet bOng duvc x6, l'
Pr

Hinh 4: Ket qua nhu6m h6a m6 Hematoxyline&Eosin cua:
a) mo thutmg, b) m6 chug vcif Biafine va c) m6 chfia vOi hydrogel composite
Ket luan
Copolymer ghep nhay nhiet chitosan-pluronic duvc
tong hap thanh cong va sir dung nhu' chat nen cho tong
hap nanocomposite hydrogel. Dung dich copolymer
mang nanocurcumin phan tan co diem chuyen nhiet
giCra tang thai sol sang gel 0, 35°C, day la khoang
nhiat de 19 twang cho cac thin nghiem oh vat lieu
nanocomposite hydrogel trong china lanh vet thuvng
ngoai da. Ket qua thin nghiem butrc du cho they
nanocomposite hydrogel tang cirang hieu qua china
lanh vet bong de 2 ten m6 hinh chi*.
TAP CHI DEW HQC - 4/2017 (SO 492 NAM 57)

Lel cam an
Nghien cCru tai tra 1361 Vien Han lam KHCN Wet
Nam theo hap ding VAST03.08/17-18.
Tai liOu tham khao
1. Nguyen D. H., Tran N. Q., Nguyen C. K. (2013),

"Tetronic-grafted chitosan hydrogel as an injectable and
biocompatible scaffold for biomedical applications", Journal of
Biomaterials Science, 24 (14), 1636-1648.
2. Kamel A. H. (2002), "In vitro and in vivo evaluation
of pluronic F127-based ocular delivery system for timolol
maleate", Int. J. Pharm., 241, 47-55.

49


• Nghiên ciru - Ky thOt
3. Dania Akbika, Maliheh Ghadiria, Wojciech
Chrzanowskia, Ramin Rohanizadeha (2014), "Curcumin as a
wound healing agent", Life Sciences, 116,1-7.
4. Zhoua H. Y, Zhanga Y. P., Zhang W. F., Chen X. G.
(2011), "Biocompatibility and characteristics of injectable
chitosan-based thermosensitive hydrogel for drug delivery",
Carbohydrate Polymers, 83,1643-1651.
5. Nguyen Thi Tram Chau, Thi Hiep Nguyen, Tran
Ngoc Quyen, Nguyen Cuu Khoa (2017), "Highly lipophilic

pluronics-conjugated polyamidoamine dendrimer nanocarriers
as potential delivery system for hydrophobic drugs",
Materials Science and Engineering C: Materials for Biological
Applications, 70,992-999.
6. Choi J. H., Joung Y. K., Bae J. W., Choi J. W., Tran
N. Q., Park K. D. (2011), "Self-assembled nanogel of
pluronic-conjugated heparin as a versatile drug nanocanier",
Macromolecular Research, 19(2), 180-188.

(Ngay nhan bai: 21/01/2017 - Ngay ph& bian: 15/02/2017 - Ngay duyat clang: 03/04/2017)

Nghien cfru xay dtrng vithim dinh phuvng phap,
LC-MS/MS dinh luyng dong thoi curcumin va chat
chuytn hoa tetrahydrocurcumin trong huytt tuvng chu(it
Hwang Thj Hong Anh'*, Hoing Van Dime, Le Thj Thu Huyen2
Nguyen Vin Long', Nguyen TrAn Linh'
Truing Dui hoc Amy Ha Nal
Vien Kie'm nghiOm thuoc Trung trong
E-mail:

Summary
A specific and sensitive LC-MS/MS method was developed for the determination of curcumin (CUR) and
tetrahydrocurcumin (THC) in mouse plasma. Curcumin, tetrahydrocurcumin and internal standard (IS) were extracted
from mouse plasma with tert-butyl methyl ether. The analytes were chromatographed as follows: Kinetex column (100
mm x 2.1 mm, i.d; 1.7 pm); Mobile phase: acetonitrile-formic acid 0.05% (60:40, v/v); Detector: positive ion electrospray
ionization (ESI) mass spectrometry with multi-reaction monitoring (MRM). Good linearity covered the concentration
range of 0.5-500 ng/ml, with a limit of quantification of 0.5 ng/mL for both CUR and THC, each. The intra- and inter-day
precisions (CV) for either CUR and THC were both less than 15.0% and the accuracy was from 91.3 to 112.6%. On
practical application, the method proved suitable for pharmacokinetic study of CUR and THC in experimental mice and
can be applicable to future bioavailablity study in human.
Keywords: Curcumin, tetrahydrocurcumin, LC-MS/MS, ESL

Dot van
Trong nhinng nam On day, curcumin (CUR) - hoot
chat duvc chiet mat tu, cay ngha yang (Curcuma longa
L.), dang thu htit duvc sy, quan tam Gila nganh Y hyc
ten the g161 do nhOng tac dyng sinh hyc vuvt trOi nhu,
tinh khang khuan, khang virus, chang viem, tang cutmg
mien

Tuy nhien, CUR ft tan trong nuttc [21, sinh
kha dung thap, bi chuyen haa manh qua gan, to ra mOt
so san pham chuyen h6a trong (16 tetrahydrocurcumin
(THC) la met chat c6 hoat tinh manh
Tren the gi&i, phan Ian cac nghien aru danh gia
SKD cilia CUR chi lien hanh dinh lityng CUR trong
huyet twang [4, 51. Met s6 It nghien c&u dinh luvng ca
CUR va chit chuyen h6a trong huyet tu,ang Ti Viet
Nam, cho den nay, cac thong s6 duvc dOng hoc dm
the pham chin CUR van chin duvc nghien dm day
du. Vi vey, bai bao nay trinh bay nghien cipu vai myc
tieu xay Ong va tham dinh phuvng phap LC-MS/MS
CO c10 dO nhey, dac hiau va chInh xac de dinh luvng
50

ding theyi CUR va chat chuyen h6a THC trong huyet
twang chuOt thf nghiem.
Nguyen lieu va phu'ong phap nghien cuu
Nguyen lieu
Chit chuin
+ Curcumin (CUR) — Chun phong thi nghiem, ham
luvng: 95,51% (nguyen tang).
+ Tetrahydrocurcumin (THC) — Chuan nha san xuat,
ham luvng: 95,0%; 0 am 0,09%
+ Glibenclamid (GBC) - Chun phong thi nghiem,
ham luvng 100,1%, dO am 0,09%. SO dyng lam chat
nei chuan (IS) trong phuvng phap phan tfch.
Dung m6i, h6a chit: Dat tau chi& tinh khiet dung
cho HPLC va LC/MS.
Huyit tuong treing: Lay tin cac chuOt nhat thf

nghiem khang sip dying thuoc.
Phuvng phap nghien cu'u
Khao sat dieu kien sac ky, khoi phi va phirang phap
TAP CHI DUVC HQC - 4/2017 (SO 492 NAM 57)