Fathy et al.

BMC Pharmacology
BMC Pharmacology and Toxicology (2023) 24:21
https://doi.org/10.1186/s40360-023-00662-1 and Toxicology

RESEARCH Open Access

Controlled release of silica‑coated

insulin‑loaded chitosan nanoparticles
as a promising oral administration system
Mohamed M. Fathy, Asmaa A. Hassan, Anwar A. Elsayed and Heba M. Fahmy*

Abstract
Background Oral insulin administration has recently become one of the most exciting research subjects. Different
approaches have been carried out to get an effective oral insulin delivery system using nanotechnology. The devel-
opment of a delivery system that overcomes the difficulties of oral insulin administration, achieving high stability
and minimal side effects, is still an urgent need. Therefore, this study is considered one of the efforts to design a new
prospective drug delivery nano-composite (silica-coated chitosan-dextran sulfate nanoparticles).
Methods Chitosan-dextran sulfate nanoparticles (CS-DS NPs) were prepared via a complex coacervation method
and then coated with silica. Uncoated and silica-coated CS-DS NPs were physically characterized via different tech-
niques. Transmission electron microscopy (TEM), scanning electron microscopy (SEM), energy-dispersive X-ray (EDX)
analysis, and atomic force microscopy (AFM) have been used to investigate the chemical elements, size, morphology,
and surface properties of the prepared formulations. Differential scanning calorimetry (DSC) to assess the thermal
properties of formed nano-formulations. Fourier transform infrared (FT-IR) spectroscopy investigated the silica coat
and chitosan interaction. The encapsulation efficiency was evaluated using high-performance liquid chromatography
(HPLC) analysis. The insulin release profile of nano-formulations was performed with and without silica coat at two
different pHs (5.5,7), nearly simulating the environment of the gastrointestinal tract (GIT).
Results The silica-coated CS-DS NPs revealed interesting physicochemical properties exemplified by suitable core
particle size obtained by TEM images (145.31 ± 33.15 nm), hydrodynamic diameter (210 ± 21 nm), high stability
indicated by their zeta potential value (-32 ± 3.2 mV), and adequate surface roughness assessed by AFM. The encap-
sulation efficiency of insulin-loaded chitosan nanoparticles (ICN) was (66.5%) higher than that of insulin-chitosan
complex nanoparticles (ICCN). The silica-coated ICN demonstrated a controlled insulin release profile at pHs (5.5 and
7) compared with uncoated ICN.
Conclusion The silica-coated ICN can be an efficient candidate as a desired oral delivery system, overcoming the
common obstacles of peptides and proteins delivery and achieving high stability and controlled release for further
applications.
Keywords Oral administration, Silica coat, Insulin-loaded chitosan nanoparticles, Drug release test

*Correspondence:
Heba M. Fahmy
hfahmy@sci.cu.edu.eg
Biophysics Department, Faculty of Science, Cairo University, Giza, Egypt

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Fathy et al. BMC Pharmacology and Toxicology (2023) 24:21 Page 2 of 11

Background study aims to improve the release properties of CS-DS

One of the most significant nano-medical applica- NPs using a biocompatible and non-toxic silica coat.
tions is designing a smart drug delivery nano-carrier. The modified Stober method achieved the coating tech-
The administration of drugs orally has been considered nique [16] to enhance the pH stability of CS-DS NPs and
the most preferable over other routes of drug delivery increase their therapeutic effect.
regarding the pain and possible infections caused by
them [1, 2]. However, the oral bioavailability of some
Materials and methods
therapeutic agents is limited due to incomplete absorp-
Materials
tion, the influence of pH alteration through GIT on the
Human insulin (100 IU/ml) was brought from VACSERA,
drug and drug carriers, and the enzymatic degradation
Egypt. Chitosan (medium molecular weight, degree of
of many medications, particularly peptides and proteins,
deacetylation 96%), tetraethyl orthosilicate (TEOS), and
leading to their poor stability [3, 4].
sodium hydroxide (NaOH) were obtained from Sigma
Several approaches have been developed to overcome
Aldrich, Germany. Dextran sulfate sodium salt was pro-
these problems and enhance the therapeutic effect of
vided from Applichem, Germany. Acetic acid (99.7%) and
orally administrated protein and peptide pharmaceuti-
Ammonium hydroxide solution ­(NH4OH) were obtained
cals like insulin [5]. Recently, oral insulin administration
from Piochem for Egypt’s laboratory chemicals. Ethanol
for managing type 2 diabetes developed as an alterna-
(96%) was purchased from Diachem chemicals. Hydro-
tive method to the traditional delivery of insulin through
chloric acid (HCL) was obtained from SDFCL, India.
injection [6].
For oral insulin administration, many drug nano-carrier
have been developed to protect insulin from biodegra- Methods of preparation
dation and enhance transmucosal absorption [7, 8]. Bio- Preparation of Chitosan‑ dextran sulfate nanoparticles
degradable polymeric nano-carrier, such as the cationic (CS‑DS NPs)
polysaccharide chitosan (CS), was introduced to beat CS-DS nano-carriers were formed using a complex coac-
some difficulties of drug stability in GIT and control the ervation method [17]. Chitosan (0.1 g) was dissolved in
drug release due to its favorable properties, such as much- 100 ml glacial acetic acid solution (1%) and was left on
adhesiveness, permeation-enhancing capability, biocom- a magnetic stirrer overnight at room temperature. DS
patibility, and low toxicity [9, 10]. Mukhopadhyay et al. solution was prepared by dissolving 0.1 g DS in 100 ml
reported the self-assembly of chitosan/insulin nanoparti- deionized water. Then, 50 ml of DS solution was added
cles for oral insulin delivery. The prepared nano-formu- dropwise to 75 ml of CS solution under stirring, so the
lation average size range was about 200—550 nm with ratio of CS to DS becomes 1.5:1. After nanoparticle for-
encapsulation of about 85% [11]. mation, the solution was centrifuged at a temperature of
However, chitosan nano-carriers encounter some 25 °C and 10,000 rpm for 20 min to collect the formed
obstacles, such as their high solubility at low pH, caus- CS-DS NPs that were washed using deionized water
ing earlier leakage of insulin within the stomach [12]. To three times to remove any chemical residuals.
overcome this problem, chitosan nano-composites have
been developed using the ionic gelation method with an
oppositely charged ionic polymer such as dextran sul- Encapsulation of insulin
fate (DS) to provide a controlled released and stable drug Human insulin (100 IU/ml) was encapsulated to the pre-
delivery system due to better crosslinking of CS with DS pared CS-DS NPs at two different pHs, which conse-
during the formation of nano-carrier [13]. Pechenkin quently affects the encapsulation efficiency of insulin.
et al. formed chitosan-dextran sulfate nano-formulation
for oral insulin delivery [14]. This preparation achieved (A) Preparation of insulin‑chitosan complex nanoparticles
a relatively high encapsulation efficiency (65%), and the (ICCN)
in vitro release was studied at different pH values simu- Chitosan was dissolved in 1% glacial acetic acid solution,
lating the GIT tract. Additionally, mesoporous silica and the resulting solution’s pH was adjusted to 5.8 using
nanoparticles coated with Chitosan have been designed NaOH solution. Then, 1 ml of insulin solution (with a
as an injectable insulin delivery system. The interaction concentration of 6.94 mg/ml and pH 5.8) was added to
of insulin with CS and silica was indicated by surface ten- the CS solution under magnetic stirring for 1 h at room
sion studies showing high encapsulation efficiency, which temperature. Finally, DS solution with pH adjusted at
reflects favorable obtained results [15]. 5.8 was added dropwise to the mixture of CS and insulin
Accordingly, administering insulin orally is considered under stirring (CS to DS final mass ratio is to be 1.5:1),
a significant challenge in drug delivery. Therefore, this and ICCN was collected using centrifugation [18].
Fathy et al. BMC Pharmacology and Toxicology (2023) 24:21 Page 3 of 11

(B) Preparation of insulin‑loaded chitosan nanoparticles space on the order of tens to hundreds of angstroms away
(ICN) from it and, therefore, cannot disturb or destroy the sam-
Chitosan was dissolved in a 1% glacial acetic acid solu- ple. The surface topography in non-contact AFM mode
tion. The pH value of the solution was confirmed to be is measured by utilizing the attractive atomic force in the
4. Then, 1 ml of insulin solution (with a concentration distance between the tip and a sample surface.
of 6.94 mg/ml and pH 4) was mixed with the DS solu-
tion; its pH was adjusted to 4. This mixture was then Differential Scanning Calorimetry (DSC)
added to the CS solution under stirring. After that, The Differential Scanning Calorimeter DSC131 Evo
the prepared nano-formulations were centrifuged and (SETARAM Inc., France) was used to evaluate the ther-
washed with deionized water. mal behavior of the two prepared formulations (uncoated
and silica-coated CS-DS NPs). The DSC analysis was per-
Silica coating of the prepared formulations formed at a heating range from 25 °C to 350 °C with a
By adding NH4OH to prepared formulations, silica coat- heating rate of 10 °C / min. The samples were weighed in
ing was provided to adjust their pH to 10. Then a solu- an Aluminum crucible 120 ul and presented to the DSC.
tion of TEOS (15 µl) and ethanol (0.5 ml) was then slowly The results of the thermo-gram were processed using
added at a rate of 10µ/min under sonication at 40 °C [19]. (CALISTO Data processing software v.149).
The Silica-coated NPs collected via centrifugation.
Fourier Transform Infrared (FTIR) spectroscopy
The FTIR spectrometer (Edwards High Vaccum, Crae-
Characterization of the prepared nano‑formulations ley Sussex, England) was used to assess (for the pellet
The size and morphology of the prepared formulations of uncoated and silica-coated CS-DS NPs) the interac-
(uncoated and silica-coated CS-DS NPs) were obtained tion between the silica coat and the surface of CS-DS
using TEM (JEM 1230 electron microscope Jeol, Tokyo, NPs. FTIR scanning speed was 2 mm/sec, and the reso-
Japan). A drop of each diluted formulation was applied to lution was 4 ­cm−1. The resulting different spectra pat-
a copper grid and left to dry for 15 min. The two sam- terns are recorded as a relation between wave number
ples were negatively stained using 1% phosphotung- and transmittance %
stic acid and left to dry. Then, the images were captured
with a high-resolution transmission electron microscope Insulin encapsulation efficiency using HPLC
TEM. The surface structure and quantitative chemical The two prepared formulations (ICCN and ICN) were
analysis of the uncoated and silica-coated CS-DS NPs centrifuged (VS-18000 M, Korea, power 220 V/ 50 HZ) at
were determined using SEM (Quanta TM 250 FEG, FEI; 10,000 RPM for 20 min. The supernatant of the two formu-
USA), equipped with an Energy-Dispersive X-ray (EDX) lations was then collected to estimate the insulin concen-
spectrometer to measure the elemental concentrations tration (the free drug) using the high-performance liquid
in the formulations. The accelerating voltage was 20 kV. chromatography technique (HPLC) (Young Lin Instru-
The hydrodynamic size distribution and zeta potential of ment, Korea). The flow rate was maintained at 1.0 ml/min,
nano-formulations (uncoated and silica-coated CS-DS and the injection volume was 20 μL. The drug (insulin) was
NPs) were determined using Zetasizer (Nano ZS90, monitored by its UV absorbance at 270 nm. The run time
Malvern Instruments, UK) at 25 °C. The formulations was 9 min, and the insulin had a retention time of ~ 6 min.
were appropriately diluted with deionized water for the The insulin encapsulation efficiency was calculated for
analysis. The Dynamic Light Scattering (DLS) technique each formulation from the following equation:

Initialconcentration − Supernatantconcentration
EncapsulationEfficiency% = ×100%
Initialconcentration

measured the hydrodynamic size. And the zeta poten- In vitro drug release
tial was directly measured by the migration of particles The insulin release profiles from the prepared formula-
in an electric field [20]. Each measurement was analyzed tions (ICCN and ICN) were performed using the dialy-
three times to calculate the mean values and the standard sis bag method (MWCO 12,000 g/mole; Sigma-Aldrich)
errors. The prepared formulations’ topographic proper- in two different pHs (5.5, 7) to ensure the stability and
ties were investigated by well-resolved non-contact AFM controlled release of the insulin within media like those
(Wet – SPM9600, Shimadzu, Japan). In non-contact of human GIT [16]. A volume of 20 ml of phosphate
AFM, the tip mounted on the end of a cantilever for sam- buffer saline (pH 7) was prepared at 3 ­ 7o C and intro-
ple surface scanning never contacts the sample leaving a duced to each falcon tube for each formulation as a ready
Fathy et al. BMC Pharmacology and Toxicology (2023) 24:21 Page 4 of 11

state for the in vitro release experiment. Equivalent vol- Results

umes of each formulation (5 ml) were filled in the dialy- Physical characterization of the prepared formulations
sis bags and immersed in 20 ml buffer (pH 7). The same Figure 1 (A & B) demonstrates transmission electron
steps were repeated for the buffer (pH 5.5). At a discreet micrographs for uncoated and silica-coated CS-DS
time interval, a sample of 2 ml was withdrawn from NPs. TEM images showed the successful preparation of
each buffer solution (to measure the amount of insulin uncoated and silica-coated CS-DS NPs with homogenous
released) and replaced with fresh buffers. The absorbance sizes. The dominant morphology of the resulting particles
of the samples was then measured using a UV–Vis spec- of the two formulations was almost spherical, with few
trophotometer at a wavelength of 270 nm. And the con- aggregations. TEM image (Fig. 1B) revealed the dense layer
centrations of released insulin were calculated. covered with CS-DS NPs that proved the successful forma-
tion of silica coat (arrow indicated). The resulting average
Statistical analysis nano-size of the two prepared formulations was 117 nm
DLS, zeta potential, and drug release data were meas- for uncoated CS-DS NPs and 145 nm for silica-coated
ured as three replicates. The results were expressed as CS-DS NPs. Figure. 2 shows SEM images for uncoated
mean ± S.E.M. Statistical Package for Social Sciences ori- and silica-coated CS-DS NPs. SEM images revealed a
gin software (version 94E) used for all data. Data analysis near-coherent structure for uncoated CS-DS NPs where
was implemented on a compatible computer. the nature of CS dominates the surface. In contrast,

Fig. 1 TEM images for (A) uncoated CS-DS NPs and (B) silica-coated CS-DS NPs

Fig. 2 SEM images for (A) uncoated CS-DS NPs and (B) silica-coated CS-DS NPs
Fathy et al. BMC Pharmacology and Toxicology (2023) 24:21 Page 5 of 11

Fig. 3 EDX spectra for (A) uncoated CS-DS NPs and (B) silica-coated CS-DS NPs

silica-coated CS-DS NPs showed a compact surface incor-

porating two clear phases and a porous structure. The
prominent elements composing the prepared uncoated
and silica-coated CS-DS NPs were determined through the
EDX spectra shown in Fig. 3 (A & B). The silica (Si) absorp-
tion peaks at 1.8 keV, as seen in Fig. 3 (B). The hydrody-
namic size distribution results showed that the average
hydrodynamic size of silica-coated CS-DS NPs was about
210 ± 21 nm, which is larger than that of uncoated CS-DS
NPs (190 ± 19 nm) (Fig. 4). The average zeta potential value
of uncoated CS-DS NPs was 48.75 ± 4.88 mV, whereas that
of silica-coated CS-DS NPs was -32 ± 3.2 mV (Fig. 5). The
topographic images obtained by AFM for uncoated and
silica-coated CS-DS NPs are demonstrated in Fig. 6 A &
B, respectively. The nanoscale features of the two for-
mulations can be observed, portraying the roughness of
Fig. 4 Particle size distribution for uncoated and silica-coated CS-DS
their materials. The average roughness values of uncoated NPs
CS-DS NPs were (7.72 ± 2.75) larger than that of silica-
coated CS-DS NPs (2.21 ± 0.81).
Differential Scanning Calorimetry (DSC).
As seen in Fig. 7A, the DSC thermogram of uncoated shifted to a higher temperature at 257.324 ºC, and the
CS-DS NPs showed two endothermic peaks, the first at exothermic peak disappeared.
90.747 ºC and the second at 254.5ºC, in addition to an
exothermic peak at 230.8ºC. While the DSC thermo- Fourier transform infrared (FT‑IR) spectroscopy
gram of silica-coated CS-DS NPs (Fig. 7B) showed a shift FTIR spectra of uncoated and silica-coated CS-DS NPs
in the first endothermic peak to a lower temperature at are shown in Fig. 8. For the uncoated CS-DS NPs spec-
64.772ºC, whereas the second endothermic peak was trum, a peak corresponding to asymmetric stretching of
Fathy et al. BMC Pharmacology and Toxicology (2023) 24:21 Page 6 of 11

Fig. 5 Zeta potential analysis for (A) uncoated CS-DS NPs and (B) silica-coated CS-DS NPs

Fig. 6 AFM images for (A) uncoated CS-DS NPs and (B) silica-coated CS-DS NPs

S˭O was found at around 1233 ­cm−1[22]. An intense peak tiny adjacent bands appeared in the spectrum of polysac-
was observed at about 3114 ­cm−1, characteristic of O–H charides and were noticed at 2490 and 2564 ­cm−1, cor-
stretching and intramolecular hydrogen bonds[23]. Two responding to the C-H symmetric and C-H asymmetric
Fathy et al. BMC Pharmacology and Toxicology (2023) 24:21 Page 7 of 11

Fig. 7 DSC of (A) uncoated CS-DS NPs and (B) silica-coated CS-DS NPs

Fig. 8 FTIR of uncoated CS-DS NPs and silica-coated CS-DS NPs

Fathy et al. BMC Pharmacology and Toxicology (2023) 24:21 Page 8 of 11

stretching, respectively, as well as small adjacent peaks

were recorded at around 637, 848, and 1001 which are
corresponding to ­ SO4 asymmetric bending, S–O–S
vibration and C-O stretching, respectively[22, 24, 25].
The FTIR spectrum of silica-coated CS-DS NPs is nearly
similar to that of uncoated, but it’s observed that the
minor peaks at 637, 848, 2490, and 2564 almost dis-
appeared. So it was suggested that these groups were
engaged in the interaction with the silica coat.

HPLC assay of insulin (Drug encapsulation efficiency)

The drug encapsulation efficiency of ICN prepared at
pH 4 was 66.5%, whereas the encapsulation efficiency of
ICCN prepared at pH 5.8 was 60.177%.

In vitro drug release

The in-vitro release profile of insulin from uncoated
and silica-coated ICN was studied to evaluate their effi-
ciency as insulin nano-carrier. The release of insulin from
uncoated and silica coated ICN at 5.5 and 7 pH were
expressed in terms of cumulative release (%) versus time,
as shown in Fig. 9 A&B. For acidic pH, the release behav-
ior of insulin from uncoated ICN showed a burst release
state; approximately 40% of insulin was released through
the first 5 h at a relatively rapid rate, and most of the
insulin amount was released within 24 h. In contrast, the
release behavior of insulin from silica-coated ICN showed
a slow and sustained release state. For pH 7, the insulin
release from uncoated ICN was higher than that from
silica-coated ICN. Only about 10% of insulin was slowly
released from silica-coated ICN during the experiment.
Fig. 9 The in vitro release study of insulin from uncoated and
Discussion silica-coated ICN in (A) pH 5.5 and (B) pH 7
To overcome insulin’s oral drug delivery problems and
improve the properties of CS-DS NPs as an efficient insu-
lin carrier, this study focused on evaluating silica as a coat
for CS-DS NPs and studying the emerging properties of system was proposed to be ± 30 mV [27, 28] as nano-
adding the silica coat. Comprehensive analyses were exe- particles show poor colloidal stability at low positive or
cuted to show the impact of the silica coat on CS-DS NPs negative zeta potential and tend to agglomeration. There-
and investigate the validity of the prepared formulation fore, the measured zeta potential value of silica-coated
for further applications. Different physical techniques CS-DS NPs (-32 ± 3.2 mV) indicates higher colloidal
studied the properties of silica-coated CS-DS NPs com- stability than uncoated CS-DS NPs having a zeta poten-
pared to those of uncoated CS-DS NPs. Morphological tial value of 48.75 ± 4.88 mV [29, 30]. In addition, it was
characterization demonstrated by TEM, SEM, and AFM proved that the ability of the negatively charged NPs to
images confirmed a uniform layer of silica coating the penetrate the mucus layer (coverage of the epithelial cells
surface of CS-DS NPs. The larger size of silica-coated of the GIT) with less interaction than that emerges from
CS-DS NPs, compared to the size of uncoated CS-DS the high positive NPs (characterized by a mucoadhesive
NPs, was inferred by TEM and DLS results. property) [31]. Therefore, silica-coated CS-DS NPs are
The colloidal stability of silica-coated CS-DS NPs was not trapped in the mucus network and can deliver the
studied using the zeta potential technique, which is con- loaded drug to the underlying desired tissues. Moreover,
sidered a fundamental analysis to evaluate the colloidal the negative surface potential of silica-coated NPs mini-
stability of nanoparticles [26]. The required minimum mizes their tendency to adsorb plasma proteins, increas-
value for the stability assurance of a colloidal suspension ing their stability in blood circulation [32].
Fathy et al. BMC Pharmacology and Toxicology (2023) 24:21 Page 9 of 11

It was found that there is an actual correlation between silica coat, and two mechanisms suggested this: First,
the surface roughness of NPs and their cellular attach- It was proved that the silica coating could improve the
ment [33, 34]. Rough surfaces have a high ability for the chitosan resistance against pH denaturation [43]. Sec-
entrapment of glycosylated proteins (mucins) compos- ond, the presence of a silica coat decreases the surface
ing the mucus layer lining the GIT, affecting their per- porosity of the prepared nano-carrier (demonstrated by
meability through the epithelial cells and hindering their the decreasing surface roughness values). That reduces
accessibility to blood circulation [35, 36]. Therefore, the the insulin molecules to be released, suggesting that
interaction between highly rough surfaces of uncoated the coating of ICN provides a protective shield allow-
NPs and mucus cells decreases their in vivo stability. ing controlled release of the drug during its journey
Hence, the lower roughness value (2.21 ± 0.81) of silica- through pH varieties of GIT.
coated CS-DS NPs, indicated by AFM, confirmed their
higher bioavailability than uncoated nanoparticles with a
roughness value of 7.72 ± 2.75. Conclusion
For the DSC thermogram of silica-coated CS-DS This study presents silica-coated IC as an efficient prom-
NPs, the shift in the first endothermic peak to lower ising strategy for the oral administration of insulin. The
temperature indicated the decreased crystallinity of favorable characteristics results were obtained for silica-
CS due to its entrapment with silica in addition to the coated CS NPs compared to uncoated CS NPs, includ-
interaction between polymers and silica, causing dehy- ing the relevant nano-size indicated by TEM images and
dration associated with hydrophilic groups of polymers DLS results and the colloidal stability inferred from zeta
[37–39]. The second endothermic peak (of CS-DS NPs) potential, AFM, and DSC results. The higher encapsu-
was slightly shifted to a higher temperature when add- lation efficiency was obtained at, the lower pH loading
ing silica which can be explained by higher formula- method, silica-coated ICN, achieving sustained release
tion stability due to the formed physical crosslinking results for insulin at pHs (5.5,7), mimicking the media of
[40]. It was also noted that the intensity of endothermic GIT. This work demonstrates inclusive investigation of
peaks was decreased, which may suggest the reduced a potential insulin delivery system as a novel challenge
reorganization ability of the CS-DS matrix by adding overcoming the main problems of oral peptides and pro-
silica, causing a stiffer matrix [40]. At the same time, teins administration. It should be considered a pave the
the disappearance of the exothermic peak by adding way for further research supporting its possible medical
silica may be evidence that no degradation of polymers applications in biological systems.
has occurred [39]. FTIR also characterized the silica
coating, whereas the disappearance of the peaks at
Abbreviations
637 ­cm−1 and the small peaks at 2490 and 2564 ­cm−1, CS Chitosan
which correspond to the skeletal vibrations of poly- DS Dextran sulfate
saccharides, as well as the disappearance of the peak NPs Nanoparticles
TEOS Tetraethyl orthosilicate
­at848cm−1corresponding to sulfated polysaccharides NaOH Sodium hydroxide
and the formation of glycosidic linkages as a result of NH4OH Ammonium hydroxide
the interaction of silica coating [41]. The calculated HCL Hydrochloric acid
ICCN Insulin-Chitosan Complex Nanoparticles
encapsulation efficiency was higher for the lower pH ICN Insulin-loaded Chitosan Nanoparticles
method (insulin-loaded CS NPs). That may be due to ml Millieliter
the isoelectric point of insulin (PI = 5.3), meaning that mg Millie gram
w/v Weight per Volume
insulin in acidic media less than its PI, like that of the µl Microliter
ICN method with pH 4, has positive charges. So, when IU International unit
it is first mixed with negatively charged DS before add- C Degree Celsius
RPM Revolutions per minute
ing to CS, it is expected that an electrostatic attraction TEM Transmission electron microscopy
occurs between the two oppositely charged molecules SEM Scanning electron microscopy
in addition to the electrostatic attraction that may then EDX Energy-Dispersive X-ray
DLS Dynamic Light Scattering
emerge between the positively charged CS and DS mol- AFM Atomic Force Microscopy
ecules resulting in high encapsulation efficiency [15, DSC Differential Scanning Calorimetry
42]. The release profiles of ICN and silica coated ICN FTIR Fourier transform infrared spectroscopy
S.M.E. Standard error of the mean
at two different pHs simulating the media of the human HPLC High-Performance Liquid Chromatography technique
GIT were assessed. The slow and sustained release was UV–Vis Ultraviolet–visible
obtained in the case of silica-coated ICN at pHs (5.5, GIT Gastrointestinal tract
PI Isoelectric point of insulin
7), The low release of silica-coated ICN is due to the
Fathy et al. BMC Pharmacology and Toxicology (2023) 24:21 Page 10 of 11

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