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American Journal of  
Life Science and Innovation (AJLSI)

Impact of  Bulky & Liposomal Secukinumab on Lymphocytes of  Psoriatic Patients, to 
Determine the Improvement in Bioavailability of  Secukinumab Nanolipo

Gazala Layas1*, Mohammad Isreb2, Nader Ghaderi3, Fanila Shahzad1, Pouria Akhbari4, Diana Anderson1, Andrew Wright3,
Mojgan Najafzadeh1

Volume 4 Issue 1, Year 2024
ISSN: 2833-1397 (Online)

DOI: https://doi.org/10.54536/ajlsi.v4i1.3268
https://journals.e-palli.com/home/index.php/ajlsi

Article Information ABSTRACT

Received: June 24, 2024

Accepted: July 30, 2024

Published: June 15, 2025

Psoriasis is an immune-mediated disease affecting the skin and joints. biologic treatments are 
often used in moderate to severe cases, administered as a monotherapy or in combination 
with other treatments. Liposomes are versatile and can enhance drug retention, reducing 
systemic side effects, and are used for therapy and research. This research study aimed to 
investigate the Geno toxicity of  Secukinumab, an ideal biologic treatment for psoriasis (a 
human IgG1k antibody, anti-IL17A), in bulk and liposome nanoparticles on the lymphocytes 
of  psoriatic patients in comparison with healthy persons. Geno toxicity of  Secukinumab in 
bulk and liposomal form was evaluated and compared by using the Comet and micronucleus 
assays. From assays, it was demonstrated that Secukinumab in both forms did not exhibit 
Geno toxicity and reduced DNA damage following the treatment of  psoriatic patient 
lymphocytes and healthy individual’s lymphocytes with Secukinumab bulk and liposomes 
format. Secukinumab used (2.1 and 2.8 µg/mL) with two different concentrations, and 
effectively decreased DNA damage induced by H2O2 in both groups to almost the negative 
control level. Secukinumab bulk and liposome form markedly reduced the H2O2-induced 
damage and proficiently diminished its adverse effects both in the Comet (p<0.0001) and 
micronucleus as-says (p<0.01). Overall, Secukinumab in both forms showed anti-genotoxic 
and protective effects by expressing its potential to reduce DNA damage produced by 
oxidative stress and it was observed that it would not induce any further damage in the 
lymphocytes of  healthy individuals and patients.

Keywords

Geno Toxicity, Micronucleus 
Assay, Oxidative Stress, Psoriasis, 
Secukinumab

INTRODUCTION 
Psoriasis was initially characterized as a dermatologic 
disorder primarily affecting epidermal keratinocytes. 
However, recent studies have shed light on its immune-
mediated pathogenesis, making it one of  the most prevalent 
immune-mediated conditions (Kadam et al., 2010). 
Psoriasis is a chronic autoimmune disease characterized 
by systemic manifestations and inflammatory changes 
that predominantly affect the skin and joints. 
The disease can significantly compromise the overall quality 
of  life of  those affected. Hyper proliferating body patches 
are primarily caused by autoimmune illness, a disorder 
marked by an overactive immune system. Psoriasis is a 
multifaceted disease, and its pathogenesis is not yet fully 
understood. Despite the availability of  several treatment 
options, psoriasis remains a challenging disease to manage.
Therefore, further research is necessary to understand 
the disease’s pathogenesis and develop more effective 
treatments to improve patients’ quality of  life (Rendon 
& Schäkel, 2018). Throughout the world population, 
up to 2.5% of  Psoriasis Vulgaris (chronic skin disorder) 
prevalence rate was reported (Elder, 2018). Various 
psoriatic clinical phenotypes are highly regarded as 
chronic plaque (Psoriasis Vulgaris) with an accountancy 
of  90% cases (Griffiths & Barker, 2007). 
Genetically mutated individuals are believed to have 

high rate of  susceptibility of  Psoriasis vulgaris through 
environmental factors, as a result of  which adaptive 
and innate immune pathways activates inflammatory 
responses. Lesions are highly visible, directing to a 
substantial proportion of  physical, emotional, social and 
psychological disability in the patients (Nestle et al., 1994; 
Chen et al., 2018).
The cause of  psoriasis is still unknown. According to a 
study, erythrocyte-superoxide dismutase activity was lower 
in psoriasis patients and blood levels of  malondialdehyde 
and nitric oxide end products were greater. The 
pathogenesis of  psoriasis is not fully elucidated; three 
factors are responsible for the disease pathogenicity 
including; T-cells, Tumor necrosis factor-alpha (TNFα), 
and Dendritic cells (Matsuzaki & Umemura, 2018) .
While its major clinical findings are the presence of  
psoriatic patches on epidermal layer and deep down 
histologically to the dermis, which revealed that it is not 
restricted to the epidermal layer. The main member of  
the IL-17 group, IL-17, interacts with several immune 
cell types and keratinocytes in the dermal layer of  the 
skin, making psoriasis more pathogenic. There are six 
components of  IL 17 (A-F). 
Consequently, the human T cell has number of  subsets, 
and one of  these cells is Th17, which produces the 
cytokine IL-17. The research findings from animal 

1 School of  Life Sciences, University of  Bradford, Bradford, BD7 1DP, UK
2 School of  Pharmacy, University of  Bradford, Richmond Road, Bradford, BD7 1DP, UK
3 Bradford Teaching Hospitals NHS Foundation Trust, St Luke’s Hospital, Little Horton Lane, BD5 0NA, UK
4 Institute of  Biomedical and Clinical Science, College of  Medicine and Health, University of  Exeter, EX2 5DW, UK
* Corresponding author’s e-mail: layas698@outlook.com



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knockout models proposed that IL-17 and Th17 play a 
major role to cause the pathogenesis of  this disease. By 
stimulating keratinocyte multiplication in the epidermis, 
Th17 cytokines IL-17, IL-21, and IL-22, which are 
released by activated T cells, the adaptive immune system 
is maintained and psoriatic inflammation is sustained.
IL-17A has been found to be used in the treatment of  
moderate to severe psoriasis plaque with an improved 
efficacy. The inhibitory mechanism of  IL-17A works 
swiftly and induces sustained responses with a significant 
safety assurance profile. Through the clinical severity 
of  the pathological condition, treating parameters of  
psoriasis are determined, which are based on numerous 
factors, for instance the dimensions of  affected area, the 
PASI grades, and the Life Quality Index of  Dermatology.
Mild conditions can be addressed with topical therapies, 
whereas moderate to severe cases may necessitate 
phototherapy or systemic therapy, comprising biological 
or non-biological interventions or a combination of  both. 
Secukinumab, also known as AIN457, is a biological 
medication that functions as an anti- IL17A monoclonal 
antibody (mAb). It is employed to treat moderate and 
severe psoriasis, as well as psoriatic arthritis (Wagener et 
al., 2013). Chinese Hamster Ovary (CHO) cells were used 
to manufacture Secukinumab, an IgG1/K-class property 
(Molden et al., 2021).
Secukinumab has been shown to have a superiority over 
other biological treatments in terms of  achieving severity 
index PASI 75 (Bagel et al., 2021). Also, it has the ability to 
sustain its effectiveness over time contrasted to additional 
biological treatments (Bissonnette et al., 2018). Neutralized 
and binding aspects of  the cytokine interleukin-17A (IL-
17A) are responsible to improve the Secukinumab work 
efficiency for the treatment of  psoriasis.
Secukinumab is a fully human mono-clonal antibody that 
selectively binds and neutralizes the pro-inflammatory 
cytokine inter-leukin-17A (IL-17A31009130) without 

producing any effect on IL-17F and Th17 cells (Berg 
et al., 2021). Additionally, it has no direct impact on the 
Th1 cascade. Current studies have revealed that increased 
T-cell abnormalities and oxidative stress (OS) are central 
parameters that lead to the pathogenesis of  psoriasis.   
Reactive oxygen species (ROS) suppress regulatory 
T lymphocytes’ (Treg) ability to reduce inflammation 
while promoting the growth and specialization of  Th1/
Th17/Th22 cells. Successive secretions of  inflammatory 
cytokines, such as tumor necrosis factor-alpha (TNF-α), 
vascular endothelial growth factor (VEGF), interleukin 
(IL)-17, IL-22, and interferon-gamma (IFN-γ), stimulate 
keratinocyte number increasing and angiogenesis (Lai et 
al., 2018; Ciążyńska et al., 2021). 
Liposomes are vesicles that contain several thin layered 
membrane or shells comprised of  a phospholipid bilayer 
covering an insignificant amount of  aqueous liquid, 
used for biotechnological and biomedical roles. On the 
basis of  preparation and its use, they are different in size 
ranging from several specific nanometers to hundred 
micrometers (Lombardo & Kiselev, 2022). Bio-logically 
active compounds of  liposomes, such as genetic materials, 
chelating agents, proteins, enzymes, peptides, vaccines, 
hormones and antimicrobial agents have been evaluated 
for patient use. 
Because of  its superior mechanical, optical, and electrical 
capabilities, quantum mechanics, and nanoscale volume-
surface area ratio, liposomes are becoming more and more 
common in sophisticated technology (Sercombe et al., 
2015; Nsairat et al., 2022). Because they increase the flow 
of  drugs, they also aid in reducing treatment side effects.
This study aimed to determine that how liposomes 
and Secukinumab bulk form affected DNA damage 
in peripheral blood cells from both healthy people 
and psoriasis sufferers by using the Comet assay 
and Micronucleus (CBMN) assay. Additionally, the 
study aimed to determine the best concentration of  

Figure 1: Pathogenesis of  psoriasis, the cytokines were activated in psoriasis and their further cascades in the skin. In-
terleukin (IL)-23 produced by various IL-23- producing cells induces IL-17 production from various IL-17-producing 
cells such as T-helper (Th)17, IL-17 producing CD8+ T cells (Tc17), and innate lymphoid cells type 3 (ILC3). IL-17 
stimulates keratinocytes to produce inflammatory cytokines/chemokines, which further activate IL-23-producing cells 
and recruit IL-17-producing cells and neutrophils



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Secukinumab liposome to prepare topically for treating 
psoriasis in the future.
The cellular response to the bulk and liposome forms was 
investigated and the optimal dose for bulk and liposome 
forms was obtained. In the presented study, the Geno 
toxicity assays with different end-points were used to 
assess the impact of  Secukinumab bulk and liposome 
forms on lymphocytes from psoriasis patients contrasted 
to healthy controls (Yamanaka, 2021).

MATERIALS AND METHODS
Materials
Secukinumab was obtained from Novartis Pharmaceuticals 
(Cosentyx). 1, 2-Dipalmitoyl-sn-glycero-3-phospho-
rac-(1-glycerol) sodium salt (DPPG), 1,2-dioleoyl-sn-
glycero-3-phsphoethanolamine (DOPE), purity >99%, 
Cholesterol (Chol), Methanol, Dichloromethane (DCM) 
were obtained from Sigma-Aldrich Company Ltd. (UK). 
All other reagents used were derived from Sigma Aldrich 
U.K. unless otherwise stated. For the preparation of  
Secukinumab solutions, drugs were in distilled water 
(150mg/ml). A 75μg/ml stock solution was prepared. The 
prefilled Secukinumab syringes (150 mg) were used in this 
study to produce the liposome format of  this biological 
agent and investigate its comparison with bulk form. The 
compound was diluted to a suitable concentration for 
treating the cells in vitro.

Characterization and Production of  Peptide-Loaded 
Liposome
Liposomes were prepared by using the Bangham process, 
which involved dissolving methanol and Dichloro-
methane (DCM) and methanol (3:1 v/v) with1,2-
dipalmitoyl-sn-glycero-3-phospho-rac-(1-glycerol) 
sodium salt DPPG, and (2 mg), 1,2-dioleoyl-sn-glycero-
3-phsphoethanolamine DOPE, and cholesterol (2:2:1 
molar ratio). The solution was transmitted to a rotary 
evaporator flask and at 40 °C the respective organic 
solvent mixture and after six hours of  evaporation, the 
mixture was rehydrated with 10ml distilled water. 
After that, the films were placed within 1ml of  
Secukinumab for 1 h at 60 °C by employing a bath 
Sonicator (150 W), then the sample exposed through 
freeze-thaw cycles at -20°C and thawing at 60°C, and then 
diluted with 75 μg/ml of  distilled water. Centrifuging 
the finished mixture at 22,000 x g for 30 minutes at 4 
°C, helped to guarantee purification and eliminate any 
remaining non-encapsulated medicines.
The modest size and Poly Disparity index of  the liposome 
formulations were evaluated by means of  dynamic light 
scattering (DLS) with Zetasizer ZS-90 and Model ZEN 
3600. In triplicate form, all measurements were performed. 
Measures were found to be < 150 nanometers. 
 
Ethical Approval 
The study on the genetic and environmental effects 
on lymphocytes from different illness situations, 
such as inflammatory, malignant, and precancerous 
ones, employing different genetic endpoints, has been 
authorized by the Leeds East Research Ethics Committee 

(rec) (rec reference number: 12/YH/0464). Ethical 
approval has been given by the subcommittee for Ethics 
in Research with Human Subjects at the University of  
Bradford (Reference no.: 0405/8). The Research Support 
and Governance Office Bradford Teaching Hospital 
NHS Foundation granted the Re DA number: 1202.

Blood Sample Collection 
Blood samples were collected from 40 individuals (20 
healthy and 20 patients) using the vein-puncture technique, 
after obtaining consent and following safety measures. 
Basic information was gathered from individuals about 
health and lifestyle factors using a questionnaire, as 
shown in Table. 1, while healthy individual’s data was 
presented in Table. 2. Blood was collected from a patient 
with moderate Psoriasis under systemic treatment; non 
were taking any biological treatment.

Table 1:  List of  patients’ confounding factors

C
on

fo
un

di
ng

 
Fa

ct
or

s

C
at

eg
or

ie
s

Fr
eq

ue
nc

ie
s 

(n
=

20
)

Pe
rc

en
ta

ge
 

(n
=

20
)

Age 
 

21-40 6 30
41-60 7 35
60-80 7 35

Gender Male 9 45
Female 11 55

Ethnicity Asian 3 15
Caucasian 17 85

Smoking history Ex-Smoker 5 25
Non-Smoker 15 75

Table 2:  List of  healthy controls confounding factors

C
on

fo
un

di
ng

 
Fa

ct
or

s

C
at

eg
or

ie
s

Fr
eq

ue
nc

ie
s 

(n
=

20
)

Pe
rc

en
ta

ge
 

(n
=

20

Age 20-35 14 70
36-50 6 30

Gender Male 10 50
Female 10 50

Ethnicity Asian 10 50
Caucasian 7 35
African 1 5
Arab 2 10

Smoking history Non Smoker 19 95
Ex-Smoker 1 5

Cell Viability Determination 
Cell viability was determined using the cell counting kit 8 
(CCK8) from VWR, UK. Blood samples were added with 
the chemicals in Eppendorf  tubes as described earlier for 



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the Comet assay treatment step and RPMI-1640 medium 
for the preparation of  cell culture. After that tubes were 
subjected to centrifugation at 3000 rpm (705 g) and the 
remained precipitates were discarded, and the mass of  cells 
were suspended. Following the manufacturer’s procedure, 
the experimental strategy was carried out and the kit’s 
contents were applied to the cell suspension. Cell viability 
of  ≥80% was considered for use in all experiments.

Cell Treatment and the Comet Assay
Complete blood samples from 20 psoriasis patients and 
20 healthy persons were collected and supplemented with 
10% DMSO and RPMI medium and quickly stored at -80 
°C. Five groups were selected for treatment, in order to 
conduct the experiment, including an un-treated group 
(NC) containing the blood sample and RPMI 1640 medium. 
A positive control containing media, blood, and a total 
of  10μl, 75μM/ml Hydrogen Peroxide (H2O2), and two 
treatment groups with different doses of  “Secukinumab 
(2.8µg /ml, 2.1µg/ml) including or excluding H2O2”, and 
1000μl of  treatment volume were used in the experiment. 
It was then “centrifuged at 3000 rpm” and incubated at 
37°C (Tice et al., 2000; Najafzadeh & Anderson, 2016).
Using a computer terminal Komet 6 software and 
Kinetic Imaging (Andor Technology Ltd, Belfast) and 
fluorescence microscope (20X magnification) equipped 
with CCD camera, 100 cells from each treatment group 
were fixed using Comet assay parameters such as Olive 
tail moment and percentage tail DNA.

Cytokinesis Block Micronucleus Assay (CBMN) 
Fresh blood sample (350 μl) and 130μl of  
phytohaemagglutinin (PHA) were added to 25cm3 
vented cap corning flasks containing “4.5 ml RPMI-1640 

medium augmented with 1% of  Penicillin-Streptomycin 
(Aminoglycoside antibiotic) and 15% Fetal Bovine 
Serum (FBS)”. Followed by incubation for 24h at room 
temperature i.e., 37⁰C in the existence of  5% CO2 and then 
the test chemicals Secukinumab bulk and liposome, H2O2 
were added. For the untreated negative control plain basic 
medium was used, and as positive controls Mitomycin C 
(MMC) (0.4μM) and H2O2, (75μM) were used. In each flask 
30μl of  Cytochalasin B (cyt B) (1mg/ml) was added at 44h 
and cultured under the same conditions for another 20h 
(Fenech et al., 2016). 
Using a 40X microscope to score 1000 cells per treatment 
group, including the Nuclear Division Index and the 
percentage of  binucleated cells, the frequency of  Micro 
Nuclei (MNi) was ascertained (Fenech, 2007). The NDI 
was calculated using the Equation 
Eq. A.1: NDI = M1 + 2(M2) + 3(M3) / N
Where, M1 = mononucleated cells, M2= binucleated 
cells, M3 = multinucleated cells, N = total number of  
viable cells scored (Fenech et al., 2016).

Statistical Analysis 
For the determination of  significant values, data was 
analyzed through one-way ANOVA (tofind the difference 
between results obtained from research) and t-tests to 
determine the significant values. A value P= <0.05 was 
considered statistically significant. Graph Pad Prism 8 
was used for statistical data analysis. 

RESULTS AND DISCUSSION
Results
When Secukinumab liposomes were TEM-analyzed, 
homogeneous, spherical bio particles with sizes between 
100 and 200 nm were discovered.

Figure 2: (a) TEM analysis of  the aqueous dispersion of  Secukinumab, (b) TEM imaging the Liposomal form of  
Secukinumab

Figure 3: Cytotoxicity of  Secukinumab on human peripheral lymphocytes after 24 hrs exposure to different 
concentrations (2.1, 2.8, and 3.751 μg/mL) by using CCK8



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The proliferation rate was computed by dividing the total 
number of  counted cells by the number of  living or viable 
cells. A cell counting kit was used to check the integrity 
and viability of  the lymphocytes, as shown in Figure 3.

The Effects of  Secukinumab and H2O2 on the 
Lymphocytes from Healthy 
The responses of  bulk and liposome forms of  
Secukinumab on lymphocytes of  DNA from healthy 
and psoriatic patients treated with H2O2 using “Olive 
Tail Moment” (OTM), standard errors and significance 
were displayed in the Figure 4. Using one-way ANOVA, 

the study showed that H2O2, treatments significantly 
increased DNA damage in cells by increasing OTM **p 
≤ 0.01in psoriatic and ***p≤0.001 in healthy individuals 
in comparison to untreated cells. However, Secukinumab 
treatments in the two different forms with or without 
H2O2 were not significant (ns). 
In comparison with the positive control treatments, a 
remarkable reduction in the DNA damage was observed 
after the addition of  Secukinumab ***p≤0.001 for the 
bulk in patients without the H2O2, and **p ≤ 0.01 for all 
the other treatments, as shown in Figure 4.

Figure 4: The effects of  Secukinumab and H2O2, on the lymphocytes from healthy and patient groups measuring 
OTM (* com-paring the treatments to the without treatments or negative control or NC, ns (non-significant) 
comparing the groups with different treatments to the NC group, + comparing the groups with different treatments 
to the positive control or PC) ( N=20 in the healthy control group and 20 in the patient group), ), N/C stands for the 
negative control, P/C; positive control, Bulk; Secukinumab bulk, liposome (liposome form of  Secukinumab), empty; 
liposomes with no drug, hydrogen peroxide (H2O2)

Results presented in Figure 4 illustrate the responses of  
bulk and liposome forms of  Secukinumab on lymphocyte 
DNA from healthy and psoriatic individuals treated 
with H2O2 using Tail DNA %, standard errors SE and 
significance. DNA damage observed in the patient group 
was higher compared to the healthy as predicted. The PC 

revealed a significant rise in the Tail DNA control cells, 
****p ≤ 0.0001. In comparison to the NC, Secukinumab 
treatments—liposomal and bulk, with or without 
H2O2, were not very important. When compared to PC 
therapies, the inclusion of  Secukinumab ++++p ≤ 0.000, 
considerably reduced the damage, as shown in Figure 5.

Figure 5: The effects of  Secukinumab and H2O2 on the lymphocytes from 20 healthy and 20 patient groups measuring 
Tail DNA% (* comparing the treatments to the NC, ns (non-significant) comparing the treatments to the NC, + 
comparing the treat-ments to the PC) ( N=20 in the healthy control group and 20 in the patient group), ), N/C stands 
for the negative control, P/C; positive control, Bulk; secukinumab bulk, liposome (liposome form of  secukinumab), 
empty; liposomes with no drug, hydrogen peroxide (H2O2).



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Cytokinesis Block Micronucleus Assay (CBMN) 
The micronucleus assay reveals higher levels of  MNis 
in patients who are not receiving therapy, making it a 
dependable and appropriate approach for examining 

DNA damage at chromosomal levels. Secukinumab 
2.1µg/ml in bulk and liposome forms has shown some 
attenuation of  the consequences caused by H2O2, as 
illustrated in Figure 6.

Figure 6: The average of  MNi in BiNC scored per 1000 cells from 5 healthy individuals and 5 psoriasis patients, 
n=1000. Data are expressed as means ± standard error (SE).Seven treatment groups included the negative control, 
two positive control groups (0.4µM MMC) (75µМ of  H2O2), 2.1µg/ml of  Secukinumab, 2.1µg/ml of  Secukinumab 
with H2O2 and the liposome form of  both (*** represents P< 0.0001, **p<0.001, *p<0.02, *p<0.018, ns=not 
significant). ( N=5 in the healthy control group and 5in the patient group), ), N/C stands for the negative control, 
P/C; positive control, Bulk; Secukinumab bulk, liposome (liposome form of  Secukinumab), empty; liposomes with 
no drug, hydrogen peroxide (H2O2)

Discussion 
Secukinumab currently is one of  the best choices with 
proper efficacy on psoriasis conditions, even on the 
increased level of  IL-23 and JAK inhibitors. However, 
in the case of  Secukinumab failure, combined therapy 
would be ideal (Damiani et al., 2022). Also, Secukinumab 
is a well-known biological therapy considered the most 
cost-effective bio-logical agent (Schweikert et al., 2020) .
Moreover, a range of  side effects linked with these 
biological compounds, such as paradoxical reactions 
(PRs), which are the result of  worsening of  immune-
mediated inflammatory disease (Miyagawa, 2022) and 
increased eosinophilic disorders (Bridgewood et al.,, 2022). 
It is therefore desirable to reduce the con-centration of  
the antibody in the blood and increase its residency in 
the psoriatic dermal layer by encapsulation in liposomes. 
Using the Comet and CBMN assays, the study examined 
the effects of  “Secukinumab bulk and liposome forms” 
on human peripheral lymphocytes from 20 psoriasis 
patients and 20 healthy persons.
The research examined Secukinumab’s possible protective 
effects on H2O2-induced DNA damage in lymphocytes, 
demonstrating the anti-oxidant properties of  both forms 
and damage that was similar to that of  patients who 
were not treated. Our results showed that Secukinumab 
significantly de-creased the percentage of  DNA damage 
in lymphocytes in the Comet assay (+++ p < 0.001). 
Both groups had considerable levels of  DNA damage, 
according to the Comet assay, but Secukinumab 
dramatically reduced the effects of  H2O2 in both liposomal 
and bulk forms. The investigation, which used both 

versions of  Secukinumab, supplemented with or without 
H2O2, to treat patient groups and healthy individuals, did 
not find any statistically significant differences in DNA 
damage.
Empty liposomes had no significant damage on the cell, 
which shows that it in itself  is safe on cells. According 
to the study, Secukinumab may be able to both protect 
and lessen the damage that H2O2 causes to DNA in both 
healthy people and psoriasis patients. 
Oxidative stress-induced synthesis and amplification 
of  pro-inflammatory cytokines can be inhibited by the 
anti-IL-17 drug Secukinumab (Zou & Meng, 2021). 
Increased reactive oxygen species (ROS) damage DNA, 
lipids, and proteins, as well as signaling molecules and 
other physiological activities, leading to diseases through 
oxidative stress.
When compared to the negative control, the oxidative 
stress therapy Secukinumab did not significantly damage 
DNA in lymphocytes from either patient group or from 
healthy individuals. The outcomes were very similar to 
those of  the negative control. This suggests that the 
concentration of  2.1μg/ml of  Secukinumab is non-
genotoxic to the cells in both groups. 
Hydrogen peroxide is an oxidative stress inducer 
compound which causes significant amounts of  oxidative 
stress-related DNA damage in peripheral lymphocyte cells 
(Najafzadeh et al., 2009; Stanić et al., 2016). Our results 
are in accordance with these studies as H2O2 significantly 
induced DNA damage in both subjective groups; healthy 
individuals and psoriasis patients. When compared to 
the positive control alone, the administration of  various 



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forms of  Secukinumab, such as liposome and bulk form, 
dramatically decreased DNA damage caused by H2O2.
Secukinumab, with antioxidant defense playing a part, 
reduced damage to a level comparable to the untreated 
control group. The Psoriasis Area and Severity Index and 
oxidative stress indicators were found to be positively 
correlated.
Using CCK8, the study examined the cytotoxic effects 
of  Secukinumab on healthy peripheral lymphocytes, 
and the results showed that these cells were cytotoxic 
in a concentration-dependent manner (Lin & Huang, 
2016). It was observed that, increased concentration of  
Secukinumab (3.75µg/ml) was found to be cytotoxic, 
while the two lower concentrations of  the drugs were 
tested 2.1µg/ml and 2.8µg/ml, had increased cell survival 
rates of  over 75%. 
The CBMN assay is an effective test to study the 
capability of  genotoxic agents to cause various clastogenic 
(chromosome breakage) and an eugenic effects 
(causing daughter cells to have an abnormal number of  
chromosomes during cell division) (Fenech, 2009). It 
was utilized in the current study to describe the effects 
of  Secukinumab on healthy individuals versus psoriasis 
patients. Also, to determine the protective potential of  
the drug against H2O2-induced Geno toxicity. This assay 
assesses several parameters, including micronuclei (MNi), 
which form during anaphase and indicate chromosomal 
remains or lost chromosomes generated during nuclear 
division.
To enhance the sensitivity and reliability of  the assay, 
cytokinesis is blocked using cytochalasin B, which facilitates 
the accumulation of  BiNC. The presence of  MNi in BiNC 
only reflects damage induced after treatment, reducing the 
possibility of  scoring pre-existing damage. This approach 
effectively determines the effects of  the test chemicals. 
By implementing this methodology, the sensitivity and 
reliability of  the assay are increased, and the effects of  the 
test chemicals are accurately determined (Magdolenova et 
al., 2012; Co-operation OfE, 2010).
In comparison with the untreated cells, no effects on 
MNi frequency were reported when the lymphocytes of  
healthy patients receiving both kinds of  Secukinumab 
treatment. 
“The MNi frequency in lymphocytes(***p<0.001) was 
considerably elevated by the MMC and 75 μM H2O2, yet 
Secukinumab (bulk or liposome co-supplemented with 
H2O2) exhibited significant reduction in the number of  
MNi in lymphocytes (*p<0.05)”. 
“It was manifested that liposomal Secukinumab caused a 
significant reduction in the number of  MNis in patients 
(***p<0.001 as compared to un-treated cells). Moreover, 
the inclusion of  MMC and 75 μM H2O2 revealed a 
significant increase (***p<0.001) in the MNi number in 
lymphocyte count in comparison to untreated cells”. This 
all confirms the safety of  Secukinumab in both forms, 
bulk and liposome, on lymphocytes. This could be the first 
step in using the liposome form as a topical treatment.
Therefore, the results demonstrate that all the treatment 

sets from both investigative groups have shown the BiNC 
% and the NDI within the normal ranges: a typical NDI 
value represents a successful division. Evaluation of  
the assays shows that treatment of  healthy and patient 
groups with Secukinumab in both forms did not induce 
a significant number of  MNi in BiNC at basal levels. 
However, when 2.1μg/ml of  the drug was co-treated 
with H2O2, Secukinumab provided substantial protection 
against H2O2-induced damage and reduced the frequency 
of  MNi in healthy BiNC.
A significantly raised number of  MNi were observed 
in Mono NC of  the patient group compared to healthy 
individuals, which indicates pre-existence of  DNA 
and chromosomal damage due to the disease state, 
medications or other confounding factors. Secukinumab 
2.1μg/ml re-duced the frequency of  MNi in MonoNC 
of  the patient group at a basal level. However, the results 
were not statistically significant. The consistency of  the 
data suggests that con-founding factors were not an issue.

Limitations 
➢ Due to differing degrees of  safety and efficacy, 

standardizing liposomal and bulk Secukinumab 
formulations and dosages is essential to maintaining 
similar results throughout investigations.
➢ The liposomal and bulk medication Secukinumab 

may protect against oxidative stress, but more research 
is necessary to assess any possible off-target effects and 
determine the treatment’s overall risk-benefit profile.

CONCLUSION 
The aim of  this study was to investigate the effect of  
Secukinumab, in both liposomal and non-liposomal 
forms, on DNA damage in healthy individuals and 
psoriasis patients. Despite higher levels of  damage in the 
sick group compared to the healthy group, Secukinumab 
significantly reduced H2O2-induced DNA damage in the 
presence of  H2O2, attenuating its harmful effects.
The results were consistent across the Comet and 
micronucleus assays, with Secukinumab 2.1μg/ml 
demonstrating enhanced efficacy against H2O2-mediated 
DNA damage. According to the study, both types of  
Secukinumab have the ability to protect against oxidative 
stress caused by H2O2, which may help with repair 
without having a major negative impact on Geno toxicity.
Furthermore, the liposomal form of  the drug was found to 
be safe for localized treatment via subcutaneous injection 
or transdermal drug delivery preparation. However, 
additional investigations are required to elucidate other 
possible molecular mechanisms involved.
In conclusion, this study demonstrated the ability to 
effectively deliver Secukinumab via liposomes, which 
may have clinical applications. These results highlight the 
potential of  Secukinumab to serve as a protective agent 
against oxidative stress-induced DNA dam-age. Overall, 
this study provides valuable insight into the effects of  
Secukinumab on DNA damage and its potential clinical 
applications.



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Recommendations
➢ Extensive research ought to appraise Secukinumab’s 

enduring impacts on oxidative stress and Geno toxicity in 
order to appraise its resilience and possible unfavorable 
consequences in the long run.
➢ Large-scale clinical trials involving human 

participants are required for further research studies to 
evaluate Secukinumab’s efficacy, safety, and tolerability in 
a range of  patient populations.

Acknowledgement 
Authors are thankful to University of  Bradford, for their 
continued support in this research work.

Authors Contribution
All authors contributed to the study conception and 
design. G. Layas & M. Isreb N. Ghaderi & Fanila: 
Conceptualization, Methodology including ethica 
approval, samples collections and statistical analysis; 
P. Akhbari, D. Andreson: Analytical interpretation; A. 
Wright & M. Najafzadeh: Research discussion and review.

Abbreviations
CHO Chinese Hamster Ovary cells
Chol Cholesterol
CBMN cytokinesis-block micronucleus
DCM methanol dichloromethane
DOPE Dioleoyl phsphoethanolamine
DPPG 1,2-Dipalmitoyl-sn-glycerol sodium salt
IL-17 Interleukin 17
mAb Monoclonal antibody
PSAI Psoriasis Area and Severity Index
Th T-Helper cells
TNFa Tumor necrosis factor-alpha
RPMI Mammalian cell culture media/ Roswell Park 
Memorial Institute

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