Lecithin soybean phospholipid nano‐transfersomes

as potential carriers for

Transdermal delivery of the human growth hormone
Maryam Kateh Shamshiri1 | Amir Abbas Momtazi ‐Borojeni2,3 |
Mahvash Khodabandeh Shahraky 4 | Fatemeh Rahimi4
Journal of Cellular Biochemistry

Mohammad Wildan Abiyyi

Research and Development Officer

PT. Daewoong Pharmaceutical Company Indonesia

BACKGROUND: CHRONIC WOUNDS
Chronic wounds usually found as a complication in
diabetic patients which caused by neuropathy disease
and peripheral
The prevalence was reach 34% and 10% should be
amputated1

Debridement
 General therapies for chronic
Topical antibiotic
wounds:
Metabolic therapy

(-) This general therapy were used only for prevent infectious disease, without enhancing skin
regeneration and scars healing

The suitable and optimum active ingredient is needed

Epidermal Growth Factor as therapeutic agent

 BACKGROUND: EPIDERMAL GROWTH FACTOR
Epidermal Growth Factor (EGF)1 :
 Protein berukuran 6200 Dalton
Menstimulasi proliferasi dan migrasi sel
Dapat menstimulasi epitelisasi dan regenerasi epidermis pada
1 2 3 4 luka

EGF topikal terbukti dapat membantu penyembuhan ulkus diabetes2

Regen® (Ointment) Easyef® (Spray)

EGF topikal memiliki stabilitas yang rendah akibat degradasi oleh protease pada luka dan
sulit dipenetrasikan ke dalam kulit sehingga menurunkan efektifitas penyembuhan
terhadap ulkus3
adanya delivery system yang tepat sangat dibutuhkan

(Tsang et al., 20031; Garcia Orue et al., 20172; Saghazadeh et al., 201
BACKGROUND :TRANSFERSOM
A Hidrofili
kAPI
Lipofilik API Advantages of transfersomes:
• Provide a protection for active ingredients
inside
Surfaktan Fosfolipi
• Optimum penetration by ultradeformable
d

Transfersom
To obtain transfersomes vesicle with have optimum physicochemical
B
characteristic, optimum formulation is needed:
Stratum corneum
 Suitable lipid type
Deformasi  Suitable surfactant type
Pori  Optimum concentration of lipid and surfactant
Reformasi Optimisasi formulasi EGF Transfersom belum pernah
dilakukan
Optimation study to obtain optimum transfersomes characteristic is needed
 IDENTIFICATION OF PROBLEM
• How to develop therapeutically superior hEGF formulation that can
enhance optimum efficacy for chronic wound healing

PURPOSE
• use a novel drug delivery system for noninvasive administration of drugs to
overcome the problem of transferring large protein molecules across the skin

HYPOTHESIS
• to make use of transfersome as flexible vesicles in the transdermal delivery of the
epidermal growth factor (EGF)
MATERIALS
o Soybean Phosphatidylcholine
o hEGF
o Methanol
o Acetic acid
o Sodium phosphate dibasic dihydrate
o Potassium phosphate monobasic
o Sodium chloride
o Polysorbate 20
o Stearylamine
o Sodium deoxycholate

(Ternullo et al., 2018)

INSTRUMENTS
o Bath sonicator (Branson UltraSonic Co)
o Centrifuged (L‐90K, Beckmun)
o Dynamic light scattering technique on a Zetasizer (Nano ‐ZS, Malvern, UK)
o Extruder (Avestin Co, Canada)
o Franz diffusion cell
o Hitachi S3200N transmission electron microscope (TEM; Hitachi, Tokyo, Japan).
o Rotary vacuum evaporator apparatus (Heidolph, Germany)

(Ternullo et al., 2018)

Preparation of Transferosom containing Method
hGH with thin film hydration method

Lecithin soybean phospholipids

• Add with surfactant (85:15) EGF transfersomes composition
• Dissolved with methanol
• Evaporated in a rotary vacuum evaporator at
Sample hEGF SPC Surfactants (ug)
45°C and 55 mbar for 1 hour
(ug) (ug)
EGF-NDLs 100 170 30 (PS 20)
Thin film layer EGF-CDLs 100 153 30 (PS 20) + 17 (SA)
• Add 5 mL sample hEGF solution buffer (pH
7,4) EGF-ADLs 100 170 30 (SDC)

Transferosomal suspension

(Ternullo et al., 2018)

EGF-Transfersom Analysis Case Study

Physical characterization by zeta sizer and transmission electron microscope

Determination of drug entrapment efficiency by dialysis-ELISA method

In vitro drug release by dialysis-ELISA

Stability study at 4°C and 25°C for 30 days

Ex vivo hormone penetration using rat skin by Franz diffusion cell

(Shamsiri et al., 2018)

Size, PDI, and zeta potential of the hGH‐ RESULT

loaded nanoTFs

(Cevc et al., 1992) has synthesized nanoTFs with a size range of 90 ‐110 nm
(Ahad et al., 2012) has proofed that nanoTFs in a size range of 72 to 176 nm could easily
penetrate across rat's skin
Accordance with other ex vivo study that showed nanoTFs (60 ‐200 nm) posing
significant penetration power and flexibility for transdermal penetration of drug
(Chaudmary et al., 2013).
vesicles with a size of less than 500 nm can pass through the skin, considering that
TFs penetrate across pores that are five times smaller than their own size (Cevc et
al., 2004)

(Shamsiri et al., 2018)

Size, PDI, and zeta potential of the hGH‐ RESULT

loaded nanoTFs

phosphatidylcholine is neutral and its isoelectric point is between 6 and 7, the pH

value in this experiment was 7.4, which is higher than its isoelectric point, and thereby
vesicles were found to obtain a negative charge.
the using surfactants were ionic and, hence, at pH 7.4, both F1 and F2 TFs have a
negative charge (Guo et al., 2000).

Negative charge has been considered favorable for the stability of the TFs and the
enhancement of transdermal permeation of drug in account of the electrostatic
repulsion between the same charge of the TFs and the skin surface
(Shuwaili et al., 2016; Malakar et al., 2012).
(Shamsiri et al., 2018)
 RESULT
Port analit

Wadah donor
Wadah donor

Membran
Port sampling

Heater/ sirkulator

Wadah reseptor
Air Stirrer

(Shamsiri et al., 2018)

The stability of hGH‐TFs RESULT

Amount
F1 of hGH inwere
hGH‐nanoTFs F1 nanoTFs
enlargedwas dropped
by 17.7% by 18.9%
(43.33 ± 4.6 (−
nm)7.95
and±45.38%
0.1 ng·mL−1
(114.7)
± and 46.59%
3.2 nm) (− 19.43
at 4°C ± 0.15
and 25°C, ng·mL−1 ) The
respectively. at 4°C and 25°C,
particle respectively.
size of There
F2 hGH ‐nanoTFs
were
was 43.88%
found to be(−increased
17.76 ± 0.2 ng·mL−1(43
by 27.33% ) and
± 5 99.1%
nm) and (− 84.9%
39.91 ±(142.7
0.3 ng·mL−1
± 3.6 nm))
reduction in the hormone content of F2
at 4°C andhGH ‐nanoTFs stored at 4°C and 25°C
25°C
(Shamsiri et al., 2018)
Ex vivo transdermal penetration study RESULT

The highest amount of hGH transdermal passage

by the F1 (489.54 ± 8.301 ng·cm−2 ) and F2 TFs
(248.46 ± 4.019 ng·cm−2 ) occurred after the
eighth and fourth hour, respectively, while those
of the control, the aqueous solution of hGH was
only 15.43 ± 0.904 ng·cm−2

(Shamsiri et al., 2018)

 Conclusion

hGH theraphy using transfersome as an ultradeformable vesicle shows

enhanced permeation in TDDS from 15.43±0.904 ng·cm-2 become
489.54±8.301 ng·cm-2

Use sodium deoxycholate as an edge activator (surfactant) can enhanced

the permeation flux, cumulative drug release and entrapment efficiency for
hGH theraphy and the F1 formula that contain SDC has far greater
potential in future medical applications.
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Shamsiri M K, Amir A MB, Mahvash K S, Fatemeh R. Lecithin soybean phospholipid nano-transfersomes as potential
carriers for transdermal delivery of the human growth hormone. Journal of Cellular Biochemistry. 2018;1-11
Thank You