Colloids and Surfaces B: Biointerfaces 173 (2019) 599–606

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Colloids and Surfaces B: Biointerfaces

journal homepage: www.elsevier.com/locate/colsurfb

Delivery of MTH1 inhibitor (TH287) and MDR1 siRNA via hyaluronic acid- T
based mesoporous silica nanoparticles for oral cancers treatment
⁎
Xiao-Lei Shia,1, Yuan Lia,1, Lu-Ming Zhaoa, Lin-Wang Sub, Gang Dinga,
a
Department of Stomatology, Yidu Central Hospital of Weifang, Weifang, Shandong, 262500, China
b
Department of Stomatology, Liaocheng People's Hospital, Liaocheng, Shandong, 252000, China

A R T I C LE I N FO A B S T R A C T

Keywords: Oral cancer accounts for 95% of all maxillofacial malignant neoplasm. Presently, surgery and radiation (with and
Oral cancer without chemotherapy) are the major treatments for oral cancer; however it met with limited therapeutic
MDR1 siRNA outcome. Overcoming the multidrug resistance and finding new therapeutic agents for oral cancer treatment are
TH287 some of the serious challenges. Small molecule, TH287 potently and selectively inhibits the MTH1 protein in
Mesoporous silica nanoparticles
cells and could act as a new chemotherapeutic agent. The study reports the successful loading and delivery of
Hyaluronic acid
MTH1 inhibitor – TH287 and MDR1 siRNA in oral squamous cell carcinoma. Our results showed that HA-
assembled mesoporous silica nanoparticles were effective in controlling the drug release and internalization in
CAL27 cancer cells. Cytotoxicity and apoptosis assay showed that combination of TH287 + MDR1 siRNA was
significantly more effective in inducing the anticancer effect compared to that of TH287 (MTH1 inhibitor) alone.
SiTMSN and HA-siTMSN significantly reduced the tumor burden compared to that of untreated control and free
TH287. The study strongly supports the fact that the HA-siTMSN plays dual of inhibiting the MDR1 function and
enhancing the cell killing effect of TH287 in the cancer tissues. Overall, results suggest that the HA-siTMSN
platform is a promising vector the systemic delivery of MDR1 siRNA/TH287 and combinational therapeutics
could be a viable solution for treatment of cancer of oral cavity.

1. Introduction dGTP to prevent misincorporation of damaged nucleotides into DNA,

has been shown to prevent RAS-induced DNA damage and senescence
Oral squamous cell carcinoma (OSCC) attributes to the 95% of all and suppress the high mutation rate of mismatch repair-defective
maxillofacial malignant neoplasm and emerged as a critical health issue cancer cells. Down-regulation of MTH1 protein expression as a result of
across the globe [1]. At present, chemotherapy is prime treatment of exposure to particular small molecules can drastically reduce tumor cell
choice for the treatment of OSCC; however, it suffers from limited survival and replication in an increased level of oxidative stress situa-
therapeutic effect due to severe drug-related toxicity and multi-drug tion [9,10]. Among all MTH1 inhibitor, TH287 is shown to be most
resistance (MDR) [2–4]. To overcome such challenges, multiple stra- effective in suppressing the cancer cells [11]. However, as with all small
tegies need to be combined that could act by different mechanism in drugs, it undergo short half-life, poor stability and limited aqueous
cancer cells. The combination of therapeutic strategies is expected to solubility, warranting a therapeutic carrier.
improve the anticancer efficacy while at the same time it will minimize One of the important reasons for the failure of drug therapy is due to
the associated side effects. In this perspective, reactive oxygen species the multi-drug resistance (MDR) protein 1 (MDR1) or P-glycoprotein 1
(ROS)-based therapeutic strategy is increasingly getting more attention [12]. MDR1 is a cell membrane protein that plays an important role in
[5]. ROS can indiscriminately damage the biomolecules such as dNTP. pumping foreign substances out of cells. Although MDR1 in normal
The prevention of nucleotide accumulation and incorporation in DNA cells perform vital function; MDR1 in tumor cells results in drug re-
will result in tremendous cancer cell death [6,7]. The protein MutT sistance as the higher expression of this protein prevents the in-
homolog 1(MTH1) is gaining increasing importance due to its potent tracellular accumulation of anticancer drugs [13,14]. It is well docu-
role in inhibiting the cancer cell proliferation and enhance therapeutic mented that ∼60% of drug resistance relates to primary drug resistance
efficacy [8]. MTH1, which hydrolyzes the oxidized forms of dATP and and ∼35% relates to acquired drug resistance among patients.

⁎
Corresponding author.
E-mail address: [email protected] (G. Ding).
1
These two authors contribute to this work equally.

https://doi.org/10.1016/j.colsurfb.2018.09.076
Received 29 March 2018; Received in revised form 11 August 2018; Accepted 28 September 2018
Available online 29 September 2018
0927-7765/ © 2018 Elsevier B.V. All rights reserved.
X.-L. Shi et al. Colloids and Surfaces B: Biointerfaces 173 (2019) 599–606

Therefore, in order to improve the treatment efficiency, new strategies by centrifugation.

needs to be outlaid to counter the MDR. One such promising strategy Aqueous solution of HA was prepared in a concentration of 1 mg/
would be to employ RNA interference (RNAi) such as small interfere mL and filtered using a 200 nm filter to remove the bulkier part. The HA
RNA (siRNA). SiRNA is a 20–25 base pairs of RNA molecules that ac- solution (2.5 parts) was mixed with MSN in a weight ratio of 2.5:10 (w/
tively involves in the regulation of specific gene expression by con- w) and incubated for 1 h under gentle rotation. The negatively charged
trolling the gene silencing activity [15–17]. Therefore, in this study, we HA will bind to the positively charged MSN via electrostatic interaction.
have employed MDR1 siRNA to inhibit or suppress the gene expression The unbound HA was removed by centrifuging at 1800 rpm for 10 min.
of MDR1 in the cancer cells which is expected to improve the efficacy of The pellet was collected and re-dispersed in ultrapure water.
TH287 or any other small molecules. However, systemic stability and
target specificity of siRNA in the blood circulation is still questionable 4. Particle size and morphology
and remains a major obstacle in the clinical development.
Among all the carriers, mesoporous silica nanoparticles (MSN) of- The particle size of nanoparticles was determined by dynamic light
fers interesting features such as tunable pore structure, surface func- scattering (DLS) method using ZetaSizer Nano ZS (Malvern Zetasizer,
tionalization, excellent cellular internalization which makes them as an UK). The particles were adequately diluted and performed the experi-
ideal carrier system. The pore structure will allow the maximal loading ments in triplicate. The morphology of particles was determined by
of the small molecule while high surface area will allow the higher transmission electron microscopy (TEM) using JEOL 2100 F at an ac-
oligos loading to the particle [18,19]. The negatively charged siRNA celerating voltage of 120 kV.
will adsorb on the positive surface charge of MSN particles. However,
gene delivery based on physical adsorption in the blood compartment is 5. Gel electrophoresis
arguable as the DNA/RNA will not be protected from the nucleases.
Therefore, we have employed hyaluronic acid (HA) - a anionic glyco- The binding and loading of siRNA into the nanoparticle was con-
saminoglycan to assemble on top of siRNA/MSN and act as a protective firmed by gel electrophoresis using 2% agarose. For this purpose, 2%
layer [20]. Most importantly, HA has been actively taken up by the agarose gel was prepared in tris-acetate-ethylenediaminetetraacetic
most cancer cells owing to the overexpression of CD44 receptors [21]. acid (EDTA) buffer that consist of 0.5 mg of ethidium bromide (EtBr).
The presence of HA is expected to serve 3 purpose (i) act as a barrier for The samples were run for 15 min for electrophoretic mobility at 110 V
the drug release from pores of MSN (ii) protects the siRNA from the for 10 min. The gel was photographed using Alpha Innotech gel image
harsh environment of blood and (iii) act as a targeting agent to cancer system.
cells [22].
The main aim of present study is to study the combinational effect of 6. In vitro drug release study
TH287 and MDR1 siRNA towards the therapeutic efficacy in oral
squamous cell carcinoma. For this purpose, TH287 and siRNA was The drug release of TH287 from siTMSN and HA-siTMSN was stu-
loaded in a HA-based MSN and the biological effect was studied in died using a dialysis tube (Spectra/Pore, MWCO 3500). The study was
CAL27 cancer cells. The in vivo therapeutic efficacy was studied in a performed at 37 °C in a regular shaker (100 rpm). 1 mL of nanoparticle
xenograft model and histochemical analysis was performed. dispersions containing 2 mg equivalent of TH287 was used for the re-
lease study. The drug-loaded nanoparticle dispersions were sealed in a
2. Materials and method dialysis tube and placed in a 25 mL of phosphate buffered saline (PBS,
pH 7.4) in a conical tube. The tube was placed in a shaker bath
TH287 hydrochloride was purchased from Sigma-Aldrich, China. maintained at 37 °C. Art regular intervals, aliquots of samples were
Hyaluronic acid, Tetraethyl orthosilicate (TEOS), cetyl- collected and replaced with equal volume of the fresh sample to
trimethylammonium chloride (CTAB), and 3 aminopropyltriethox- maintain the sink conditions. The amount of drug released in the buffer
ysilane (APTES) was purchased from Sigma-Aldrich, China. The target was calculated using HPLC method. Briefly, Waters LC Module 1 HPLC
sequence of MDR1-siRNA is 5′-GAGCTTAACACCCGACTT ACATT-3′ system connected to a C18 column (Uptisphere C18, 3 m, 4 mm ×
and purchased from QIAGEN GmbH (Germany). The lipofectamine was 150 mm) was used to analyze the drug concentration. The mobile
purchased from Life Technologies Corp (Carlsbad, CA). All other che- phase comprised of a mixture of 0.05 M trichloro-acetic acid (TCA) and
micals are of reagent grade and used without additional purifications. acetonitrile (60/40, v/v) and pH 2.5 was maintained. Fluorimetric
detection was performed with a Waters 470 Scanning Fluorescence
3. Preparation of hyaluronic acid modified mesoporous silica Detector at a wavelength of 480 nm (excitation). The amount of drug
nanoparticles release was plotted against the time interval.

At first, 200 mg of CTAB was dissolved in 100 mL of 0.125 M 7. In vitro cytotoxicity assay
NH4OH solution and the solution was heated to 80 °C. Under gentle
stirring, 2.85 mL of 0.45 M TEOS solution was added. The mechanical The cytotoxic potential of TH287 in combination with siRNA
stirring was continued for 2 h and the solution was heated at 80 °C (siTMSN and HA-siTMSN) against CAL27 cell was studied by 3-(4,5-
continuously. The surfactant template was removed by adding 100 mL dimethylthiazol-2-yl)-2,5-diphenyl tetrazoliumbromide (MTT) assay
of ethanol (100%) and re-fluxed at 60 °C for 24 h. The resulting solution protocol. The CAL27 cells were maintained in DMEM media supple-
was dialyzed against distilled water for 24 h. The as-synthesized MSN mented with 10% fetal bovine serum (FBS) and antibiotics penicillin
was collected after centrifugation. The dried MSN was dissolved in to- (100 IU/mL) and streptomycin (100 mg/mL). The cells were grown in
luene and sonicated for 5 min. APTES was added to the MSN solution in standard conditions of 5% CO2 at 37 °C. Briefly, cells were seeded in a
a molar ratio of 1:5 of APTES to MSN. The mixture was allowed to react 96-well plate (0.8 × 104 cells/well) using the media and incubated for
for 12 h and then filtered and refluxed under nitrogen atmosphere. The 24 h. Next old media was aspirated and replaced with new media
TH287-loaded MSN was prepared by incubating 10% w/w of TH287 containing free TH287, siTMSN and HA-siTMSN. Each of the formula-
with the MSN particles for 12 h under gentle shaking. Followed by, tions contain a fixed concentration of 50 nM of MDR1 siRNA. TH287
particles were centrifuged and supernatant containing the free drug was was used in an increasing concentration while a fixed concentration of
removed and the drug-loaded MSN was collected. The siRNA was siRNA (50 ng) was used for the study. Blank nanoparticles without drug
loaded into MSN by mixing the siRNA and MSN in MES buffer and were also added to estimate the cytotoxicity of carrier itself. The cell
allowed to incubate for 60 min. The siRNA-loaded MSN was collected was incubated for 24 h and then carefully supernatant was removed and

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Fig. 1. Schematic illustration of preparation of TH287 and siRNA-loaded Hyaluronic acid assembled mesoporous silica nanoparticles.

10 μL of MTT solution (5 mg/ml) was added and incubated for 4 h. After sacrificing the mice and fixed in 10% formalin. The tumor was paraffin-
4 h incubation, supernatant was removed and 100 μL of DMSO was embedded, sectioned and stained with H&E staining agents and ob-
added and kept aside for 15 min. The cell viability was then determined served under fluorescence microscope.
by measuring the absorbance of formazan salt at 570 nm. The cell
viability was calculated with relative to that of untreated control. A plot 11. Statistical analysis
of concentration vs % cell viability was drawn.
The data are presented as means ± standard deviation. The one-
8. Cellular uptake – CLSM way ANOVA followed by the post-Turkey comparison tests was used for
statistical analysis. A difference of p < 0.05 was considered statisti-
The cellular uptake of nanoparticle was evaluated by means of cally significant.
confocal laser scanning microscope (CLSM). Briefly, cells were platted
in 6-well plate (0.8 × 104 cells/well) using DMEM media and incubated 12. Results and discussion
for 24 h. Rhodamine B was used as a fluorescence agent which is loaded
in siTMSN and HA-siTMSN in the same concentration. The cells were Oral squamous cell carcinoma (OSCC) accounts for the highest
incubated with siTMSN and HA-siTMSN (2 μg of rhodamine B) for 3 h. number oral cavity cancers. Despite the improvement in the treatment
The culture medium was removed and the cells were washed thrice options, 5-year survival rate of OSCC remains at 30% necessitating
carefully. The cells were further fixed with 4% paraformaldehyde for novel strategy in addition to surgery or radiation. Chemotherapy re-
15 min followed washed with PBS twice to get rid of all the residues. mains one of the viable options for the treatment of OSCC; however,
The images were obtained from CLSM microscope (TCS SP2). multidrug resistance to many drugs limits the therapeutic effectiveness
of present drug-therapies [22]. The combination of therapeutic strate-
9. Flow cytometer-based apoptosis assay gies is expected to improve the anticancer efficacy while at the same
time it will minimize the associated side effects. In this perspective,
The apoptosis potential of formulation was evaluated by Annexin V- MutT homolog 1(MTH1) is gaining increasing importance due to its
FITC and PI-based double staining using flow cytometer (BD FACS). potent role in inhibiting the cancer cell proliferation and enhance
Briefly, cells were seeded in a 12-well plate (2 × 105 cells/well) using therapeutic efficacy. Among all MTH1 inhibitor, TH287 is shown to be
the media and incubated for 24 h. Next, old media was aspirated and most effective in suppressing the cancer cells. Similarly, MDR1 gene
replaced with new media containing free TH287, siTMSN and HA- plays key roles in multidrug resistance phenomenon. MDR1-siRNA is
siTMSN and incubated for 24 h. A fixed concentration of TH287 (1 μg/ emerging as an important tool to control the expression of MDR1 gene
mL) and siRNA (50 nM/mL) was used for the study. After 24 h, cells in multiple cancers [23]. However, TH287 and siRNA undergoes short
were collected, washed with cold PBS, centrifuged. The pellet cells were half-life and poor stability in blood stream. In this study, we have
re-dispersed in 100 μL of binding buffer and stained with 3 μL of employed mesoporous silica nanoparticles (MSN) whose pore structure
Annexin V-FITC and PI each and incubated for 15 min. Following in- will allow the maximal loading of the small molecule while high surface
cubation, volume was made up to 800 μL using 1X binding buffer and area will allow the higher oligos loading to the particle (Fig. 1).
analyzed using flow cytometer.
13. Particle characterization
10. In vivo antitumor efficacy
The dynamic light scattering analysis (DLS) showed that particle
The therapeutic supremacy of individual formulations was eval- size of siTMSN was 140.25 ± 1.23 nm while the particle size increased
uated by antitumor efficacy analysis in male Balb/c mice (6 weeks old). to 184.25 ± 2.14 nm after the assembly of HA on top of the particle
The tumor model was generated by subcutaneously injecting 2 × 106 surface (HA-siTMSN) (Fig. 2A). A slight increase in particle size was
CAL27 cells in the right flank of mice. After the tumor attains the size of mainly attributed to the coating of hydrophilic polymer on the particle.
50-80 mm3, mice were randomly divided into 5 groups with 8 mice in It has been believed that negatively charged HA will bind to the posi-
each group. The group includes blank NP, free TMSN, siTMSN and HA- tively charged MSN surface via electrostatic interaction. Nevertheless,
siTMSN. Untreated mice were considered as an appropriate control. The overall size of particle is < 200 nm indicating its suitability for cancer
TH287 was administered at a fixed dose of 6 mg/kg and siRNA as targeting applications. The successful assembly of HA on MSN was
18 μg/mice and formulations were administered 3 times with a span of further proved by surface charge analysis. The surface charge of
3 days. The tumor size was measured every 3 days once until 18th day. siTMSN was 27.2 ± 1.46 mV while it reversed to -18.4 ± 2.41 mV
The tumor was size was measured using Vernier Caliper and tumor after the HA assembly indicating the presence of anionic polymer layer
volume was calculated using a × b×0.5; whereas ‘a’ and ‘b’ are short on the particles.
and long diameter of tumor, respectively. The tumor was isolated after The morphology of HA-siTMSN was evaluated by transmission

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Fig. 2. (A) Particle size analysis of HA-siTMSN via dynamic light scattering (DLS) methods; (B) Transmission electron microscopy (TEM) image of HA-siMSN.

electron microscope (TEM) (Fig. 2B). As seen, perfectly spherical

shaped particles were observed by TEM experiment. A slight greyish
layer at the outré circumference indicates the presence of polymer
layer. The particle maintained their size and stability for more than 30
days at 4 °C. The exact mechanism of the stability of the nanoparticle is
not known however it is believed that a protective coating of MSN with
a hydrophilic polymer will confer the stability. The HA act as a stabi-
lizer that protects the particle from destabilizing in the dispersion
medium. Moreover, TEM image clearly showed that the particles are
uniformly distribution with no sign of aggregation indicating its ex-
cellent stability in the dispersion medium.

14. Gel electrophoresis

Agarose gel electrophoresis was conducted to study the binding

pattern of siRNA to the positively charged MSN (Fig. 2C). As seen, free Fig. 3. In vitro drug release kinetics of TH287 from siTMSN and HA-siTMSN.
siRNA easily migrated to the opposite end whereas siRNA-loaded in The release study was performed in dialysis tube and the experiment was
performed in triplicate.
MSN was gradually retarded as the n/p ratio increased. At n/p ratio of 4
and 5, 100% of siRNA migration was retarded indicating the effective
loading in the MSN. Highly negative charged siRNA bind electro- and HA-siTMSN. For example, ∼40% of drug released from siTMSN
statically to the positively charged MSN. while only ∼25% of drug released from HA-siTMSN. The trend con-
tinued until the end of the study with ∼100% of drug released from
siTMSN compared to ∼70% of drug release from HA-siTMSN at the end
15. Drug loading and in vitro drug release study
of 80 h. The difference in release pattern was mainly attributed to the
HA coating on the MSN surface. The presence of polymer layer on the
The entrapment efficiency of TH287 was observed to 94.2 ± 1.35%
pores might obstruct its early release leading to a delayed pattern of
while the drug loading was 8.91 ± 1.24% w/w. The drug release of
release. Overall, a controlled release of drug from HA-siTMSN is ben-
TH287 from siTMSN and HA-siTMSN was studied by dialysis method
eficial for the cancer targeting applications.
(Fig. 3). Briefly, nanoparticle dispersions were loaded in dialysis tube,
sealed and placed in a shaker. The samples were collected at specific
time intervals. The TH287 released in a controlled manner from both 16. Cellular uptake of nanoparticles
the nanoparticle system throughout the length of the study period.
Significant differences in release percentage were seen between siTMSN The delivery efficacy of drug and siRNA was evaluated in terms of

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Fig. 4. (A) Cellular uptake analysis of siTMSN and HA-siTMSN in CAL27 cancer cells via confocal microscopy; (B) in vitro cytotoxicity analysis of free TH287,
siTMSN and HA-siTMSN in the cancer cells. The cell viability was analyzed by MTT assay protocol.

Fig. 5. Analysis of apoptosis cells determined by Annexin V-FITC/PI staining using flow cytometer. The cells were treated with formulations and after 24 h, cells were
stained with the respective apoptosis detecting agent and studied by flow cytometer.

cellular uptake (Fig. 4a). The CAL27 cells were treated with siTMSN 17. In vitro cytotoxicity assay
and HA-siTMSN loaded with Rhodamine B. Results clearly showed that
both the nanoparticles were internalized appreciably in the cancer cells. It is proven beyond any doubt that MTH1 is essential for cancer cell
As shown, free rhodamine B randomly diffused in the cancer cells survival. We have evaluated the cytotoxic effect of MTH1 inhibitor
whereas drug-loaded nanoparticles internalized via a specialized en- alone and in combination with siRNA in CAL27 cells (Fig. 4b). We can
docytosis mechanism. Marked uptake of HA-siTMSN in the cancer cells see that TH287 showed a potent anticancer effect in a concentration-
indicates its ability to deliver the encapsulated therapeutics in the in- dependent manner [24,25]. To be specific, cytotoxic effect was pro-
tracellular environment. nounced in combination with MDR1 siRNA. It is noted that cytotoxic
effect of siTMSN and HA-siTMSN was almost similar at the in vitro

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2.4 μg/mL whereas IC50 value of siTMSN and HA-siTMSN were in the
range between 0.65 to 0.82 μg/mL indicating the therapeutic benefit of
small drug + siRNA combination. It has been earlier reported that
human MDR1 gene encodes an integral membrane protein, Pgp. In this
regard, MDR1-siRNA has been reported to counter the expression of
MDR1 gene in cancer cells and could potential inhibit the p-gp ex-
pression. We have analyzed the effect of MDR1 siRNA and scrambled
siRNA on the p-gp expression. As expected, scrambled siRNA did not
show any appreciable effect on the p-gp expression whereas MDR1
siRNA showed significant downregulation of p-gp expression (> 60%)
compared to that of scrambled siRNA. The marked effect of MDR1
siRNA on the p-gp silencing will enhance the anticancer effect of the co-
administered small molecule. Results in the present study clearly
Fig. 6. In vivo antitumor efficacy study – Tumor volume assessment in in- highlight the fact that MDR1-siRNA, to block gene expression of MDR1
dividual mice group after the treatment period. The antitumor effect of for- and potentiate the therapeutic efficacy of MTH1 inhibitor in OSCC. We
mulations and free drug was studied in CAL27 xenograft model and tumor size can conclude that the mesoporous silica nanoparticle could increase the
was measured using vernier caliper. intracellular levels of combination therapeutics (siRNA/TH287) in the
cancer cells. It is worth mentioning that blank nanoparticles did not
conditions. The sequence specificity of siRNA was confirmed from the show any cell killing effect indicating that nanoparticles are very safe
fact that HA-siTMSN with scrambled siRNA did not result in synergistic for the in vivo administration and therapeutic targeting.
cytotoxic effect whereas HA-siTMSN with MDR1 siRNA resulted in su-
perior cell killing effect. The IC50 value of TH287 was significantly
reduced in combination with siRNA. The IC50 value of TH287 was

Fig. 7. H&E staining of tumor tissues. The tumor was extracted and subjected to fixing and paraffin embedding. The tumor slice was stained with H&E agent and
observed by light microscope.

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18. Apoptosis assay CAL27 cancer cells. Cytotoxicity and apoptosis assay showed that
combination of TH287 + MDR1 siRNA was significantly more effective
Additionally, we have determined the anticancer effect of combi- in inducing the anticancer effect compared to that of TH287 (MTH1
nation therapeutics in terms of apoptosis assay via flow cytometer using inhibitor) alone. SiTMSN and HA-siTMSN significantly reduced the
an Annexin-V and PI staining protocol (Fig. 5). Untreated cells did not tumor burden compared to that of untreated control and free TH287.
exhibit any apoptosis and blank nanoparticle showed fewer than 8% of The study strongly supports the fact that the HA-siTMSN plays dual of
necrotic cells. Similarly, scrambled siRNA was not effective in inducing inhibiting the MDR1 function and enhancing the cell killing effect of
the apoptosis indicating the non-specific nature of siRNA. MDR1 siRNA TH287 in the cancer tissues. Overall, results suggest that the HA-
showed ∼22% of apoptosis and TH287 showed 15% of apoptosis cells siTMSN platform is a promising vector the systemic delivery of MDR1
consistent with its low cytotoxicity effect as mentioned above. As ex- siRNA/TH287 and combinational therapeutics could be a viable solu-
pected, TH287 in combination with MDR1 siRNA significantly elevated tion for treatment of cancer of oral cavity.
the apoptosis cells by 35% further portraying the anticancer effect of
MDR1 siRNA in the cancer cells. Finally, HA-siTMSN showed around Acknowledgement
45% of cancer cell apoptosis. Slight higher apoptosis effect of HA-
siTMSN compared to that of siTMSN might be attributed to better cel- The study was supported from the Research Grant of Yidu Central
lular uptake and controlled release of the encapsulated therapeutics. Hospital of Weifang, Weifang, China.

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