Home About us Editorial board Ahead of print Current issue Search Archives Submit article Instructions Subscribe Contacts Login 

 Table of Contents  
Year : 2019  |  Volume : 15  |  Issue : 3  |  Page : 480-490

Development of α-tocopherol surface-modified targeted delivery of 5-fluorouracil-loaded poly-D, L-lactic-co-glycolic acid nanoparticles against oral squamous cell carcinoma

1 Department of Oral and Maxillofacial Surgery, King George's Medical University, Lucknow, Uttar Pradesh, India
2 Department of Oral Pathology and Microbiology, King George's Medical University, Lucknow, Uttar Pradesh, India
3 Department of Clinical Laboratory Science, College of Applied Medical Sciences, King Khalid University; Research Centre for Advanced Materials Science, King Khalid University, Abha, Kingdom of Saudi Arabia

Date of Web Publication29-May-2019

Correspondence Address:
Dr. Saurabh Srivastava
Department of Oral and Maxillofacial Surgery, King George's Medical University, Lucknow, Uttar Pradesh
Login to access the Email id

Source of Support: None, Conflict of Interest: None

DOI: 10.4103/jcrt.JCRT_263_18

Rights and Permissions
 > Abstract 

Objective: The aim of the study to develop surface modified targeted moiety α-tocopherol (α-t) encapsulated with 5-fluorouracil (5-FU)-poly-D, L-lactic-co-glycolic acid nanoparticles (PLGA NPs) toward the anticancer activity against oral squamous cell carcinoma (OSCC).
Materials and Methods: 5-FU was conjugated with the polymer, PLGA by ionic cross-linking and α-tocopherol use as a functionalized surface moiety. Characterization, drug entrapment efficiency, and in-vitro drug release system were optimized at different pH 7.4 and pH 4.5. The in-vitro cell was performed to optimize the anticancer activity through MTT assay and apoptotic staining assay was also performed by flow cytometry to evaluate the cellular apoptotic activity and cellular uptake.
Results: The particle size was distributed within an average range of 145–162 nm, the polydispersity index values lie 0.16–0.30, and the surface charge was at the negative side, –17mV to –23mV. The in vitro drug release system showed more sympathetic situation at pH 7.4 as compared to pH 4.5, for targeted NPs, approximately 86% and 69%, respectively. The non-targeted 5-FU-PLGA NPs showed drug release of 83% and 64% at pH 7.4 and 4.5 subsequently. In vitro anticancer activity confirmed the intense inhibition by α-t-FU-PLGA NPs of 79.98% after 96 h treatment of SCC15 cells and confirmed the steady-state inhibition of 83.74% after 160 h incubation in comparison to 5-FU-PLGA NPs. Subsequently, the early apoptosis, 27.98%, and 16.45%, and late apoptosis, 47.29%, and 32.57%, suggested the higher apoptosis rate in targeted NPs against OSCC.
Conclusions: The surface modified α-t-FU-PLGA NP was treated over SCC15 cells, and the oral cancer cells have shown the high intensity of cellular uptake, which confirmed that the target moiety has successfully invaded over the surface of cancer cells and shown advanced targeted delivery against OSCC.

Keywords: Anticancer, fluorouracil, nanoformulation, oral squamous cell carcinoma, poly-D,L-lactic-co-glycolic acid, squamous cell carcinoma-15 cells, targeted moiety

How to cite this article:
Srivastava S, Gupta S, Mohammad S, Ahmad I. Development of α-tocopherol surface-modified targeted delivery of 5-fluorouracil-loaded poly-D, L-lactic-co-glycolic acid nanoparticles against oral squamous cell carcinoma. J Can Res Ther 2019;15:480-90

How to cite this URL:
Srivastava S, Gupta S, Mohammad S, Ahmad I. Development of α-tocopherol surface-modified targeted delivery of 5-fluorouracil-loaded poly-D, L-lactic-co-glycolic acid nanoparticles against oral squamous cell carcinoma. J Can Res Ther [serial online] 2019 [cited 2022 Jan 28];15:480-90. Available from: https://www.cancerjournal.net/text.asp?2019/15/3/480/251400

 > Introduction Top

Cancer is the uncontrolled expansion of flawed cells to build into a pulpy mass called tumor, which affects surrounding healthy tissues to made collusion.[1] It is the topmost reason for the loss of life all around the world, the responsibility of new patients was increased around 14.5 million till 2012,[2] and it is rising at a high rate of approximately 50% up to the next upcoming decades.[3],[4] Specifically, in India, approximately 1.2 million malignant subjects were reported,[5] and it was estimated that the approximately 1 million new cancer patients will be raised by 2026.[6]

Oral cancer is the most frequent type of solid tumor globally [7] and comes under the top three most occurred types of cancer in India only.[8] Oral cancer could be seen in the inner part of mouths, such as tongue, lips,[9] inside part of the cheek, tooth gum parts, and all the related part.[10] Annually, 25 billion fresh subjects have been reported, and from them, 0.55 million cases were excluded from India till 2014,[11] with maximum cases within an average age of 45–55 years.[12] The rate of survival of oral cancer is 5 years.[13] The prime risk factor for oral cancer is tobacco, about 75%, including any type of tobacco use.[14],[15]

Squamous cell carcinoma (SCC),[16] established within the squamous cells, flat cells, underline the entire human body. The dissimilar forms of SCC were determined, and the most traditional forms are verrucous and basaloid.[17],[18] SCC over the oral cavity calls oral SCC (OSCC) and about 80% of oral cancer is SCC.[19],[20] OSCC may also the outcome of one of the continual irritations, such as dental cavity, much use of mouthwash, betel nuts, smoking and smokeless,[21],[22] and alcoholism, including other aspects, immunosuppression, catastrophic metabolism, deficiency in the enzyme, which regulates DNA, human genetic perceptivity,[23] human papillomavirus,[24],[25] and diet.[26] OSCC evoked on mucosa and premalignant condition, mostly leukoplakia, erythroplakia, dysplasia, lichen planus, and oral submucous fibrosis.[27],[28],[29],[30] Approximately 45% of OSCC proposed from the tongue surface.[31] In vitro cells studies with drug treatment quantify and explain biological characteristics of the destructive infirmity and provide a provisional mechanism to understand oral cancer.[32],[33]

OSCC was primarily treated by surgical incision additionally chemo-radio-therapy,[34] and the distinctive class of chemodrugs may be introduced against OSCC, which may be alone or in combination. There are different classes of chemotherapeutic drugs, which are used to treat oral cancer such as alkylating, nucleotide, anthracylines, alkaloids, and others.[35],[36] These agents have the competence to kill the tumor tissue through distraction in cell proliferation.[37]

5-Fluorouracil (5-FU),[38] nucleotide analog, has the consequential efficacy against OSCC through anticipating chromosomal cloning to suppress thymidylate synthesis exertion and ultimately block the transfiguration of deoxyuridine monophosphate to deoxythymidine monophosphate, resulting in inhibition of DNA replication through deoxythymidine triphosphate and further cell proliferation.[39],[40],[41] The conventional chemotherapy of 5-FU has utmost restriction over nonspecificity, limited half-life, less bioavailability, and limited therapeutic index, besides less unyielding to OSCC.[42] It was suggested that the drawback of 5-FU conventional dosage may be reduced through the nanoparticles (NPs)-targeted therapy specifically against OSCC.[43] The targeted drug delivery has the ability to directly inflate over the affected region.[44] It could interim cancer cells by inhibiting the protein, which is susceptible to tumor proliferation. The efficiency of 5-FU as a chemotherapeutic agent could be enhanced through a chambered drug–polymer nanoparticulate system that targeted over a specific region.[45] The 5-FU being designed to engulf in a bucket of polymer and surface would be protected through the target moiety, which has the capability to bind with a surface protein of the OSCC cells and made a way to transfer the drug directly on specific site and antiproliferate OSCC cells exclusively and may not affect the normal cells.[46],[47] In addition, the nanoformulation drug delivery has the competence of enhanced permeability and retention (EPR) effect,[48],[49] which assists aggregation of the drug-loaded-nanoparticulate system on leaky tumor tissues as compared to normal tissues.[50],[51] Poly-D, L-lactic-co-glycolic acid (PLGA) polymer extensively used to prepare the nanoparticulate control-release system for the drug, which is enclosed within the PLGA complex and specifically exaggerates the bioavailability of poor absorbed low lipophilic drug like 5-FU.[52],[53],[54] The targeted drug delivery of 5-FU NPs formulation was specifically design to attach over OSCC cells, according to the biochemical behavior of tumor site [55],[56] and thus provide an advanced drug delivery platform to minimize the side effects, toxicity,[57] and nonspecific inhibition over surrounding normal cells.[58] This advancement may improve the drug efficacy, bioavailability, prolonged release, and targeted over affected region.[59],[60]

α-Tocopherol (α-t), a fellow of Vitamin E group,[61] acquires phenol hydroxyl group having a maximal biological activity along with nontoxic characteristics and vigorously absorbed by humans.[62] The concussion of α-t in the inhibition of chronic diseases affirmed to be concord with oxidative stress and influences apoptosis in OSCC cells and tempers the accessibility of cancer cell surface that enhances the penetration of 5-FU.[63],[64] The targeting moiety, α-t, has the ability to bind with a specific class of receptors over the surface of cancer cells and fabricate the path to deliver the anticancer drug 5-FU. Infrequent target moieties such as peptides, folic acid, vitamins, and antibodies were composite within a network for targeted drug delivery system.[65],[66]

The goal of the study to formulate α-t surface-modified targeted PLGA NPs engulfed 5-FU and their evaluation on the basis of control-release system and OSCC cell treatment. The targeted α-t-FU-PLGA NPs were compared with nontargeted 5-FU nanoformulation, 5-FU-PLGA NPs on SCC15 cells, followed by in vitro assays, which were performed to assess the result through inhibitory cytotoxicity study and the affinity of in vitro cellular uptake and cell targeting and cell apoptosis rate [Figure 1].
Figure 1: Schematic representation for formulation of targeted α-tocopherol-5-fluorouracil-poly-D,L-lactic-co-glycolic acid nanoparticles and nontargeted 5-fluorouracil-poly-D,L-lactic-co-glycolic acid nanoparticles and its squamous cell carcinoma-15 cell internalization

Click here to view

 > Materials and Methods Top


5-FU 99% was purchased from (Cas No. 51.21.9) Sigma-Aldrich, India, and α-t was purchased from TCI Chemicals Pvt. Ltd. (India) (Cas No. 59-02-8). PLGA was purchased from Sigma-Aldrich, India (Product No. 808482-5G), SCC15, cancer cells were obtained from National Centre for Cell Sciences, Pune, India, and the rest chemicals used for experiments, standard analytical grade, were acquired from Sigma-Aldrich Chemicals Pvt. Ltd., Bangalore, India.

Preparation of targeted α-tocopherol-5-fluorouracil-poly-D, L-lactic-co-glycolic acid nanoparticles/5-fluorouracil-poly-D, L-lactic-co-glycolic acid nanoparticles

5-FU was conjugated to the polymer PLGA by ionic cross-linking and α-t used as a functionalized surface moiety,[67] for the preparation of α-T-FU-PLGA NPs. PLGA 34.50 mg was dissolved in acetic acid 1% w/v (10 ml), and the pH was maintained at 4.8. The drug 5-FU was added to that solution. The solution was added in 0.5% polyvinyl alcohol solution and allowed for magnetic stirring for 1 h. The 5-FU-PLGA solution was allowed to ultrasonicated for 12 min at 20% amplitude to facilitate the solubility and retrieved a homogeneous amalgamation followed by washing with deionized water, then lyophilized, and stored at 4°C.

Surface functionalized of α-tocopherol as targeted moiety on 5-fluorouracil-poly-D, L-lactic-co-glycolic acid nanoparticles

30.25 mg of α-t was added in pH 7.4 phosphate-buffered saline (PBS). Subsequently, 17.5 ml 0.1% (w/v) 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide aqueous solution was added drop-wise, under low-velocity magnetic stirring condition about 2 h to form cross-links. The formations of NPs have been formed impulsively under the carbodiimide reaction. The prepared 5-FU-PLGA NPs were activated by adding in 7.4 PBS and 250 μl of N-hydroxysuccinimide (1 mg/ml) under magnetic stirring for 3 h after that unreacted chemical has been washed with PBS. Both the solutions were added under magnetic stirring further for 3 h followed by overnight incubation and ultrasonicated [68] for 15 min, and the pellets were collected after washed with PBS. Eventually, the targeted α-T-FU-PLGA NPS were obtained and used for furthermore experiments.

Characterization of α-tocopherol-5-fluorouracil-poly-D, L-lactic-co-glycolic acid/5-fluorouracil-poly-D, L-lactic-co-glycolic acid nanoparticles

The particle size, zeta potential, and polydispersity index (PDI) of targeted α-T-FU-PLGA and nontargeted 5-FU-PLGA NPs were optimized through the instrument Malvern Zetasizer ver. 7.12 (Malvern Instrument, UK.), and morphology of the NPs was obtained through scanning electron microscope (SEM) (Quanta 450 FEG, the Netherlands).

In vitro drug-release system and drug entrapment efficiency

To study the release system and entrapment efficiency (EE) of targeted NPS and nontargeted NPs, a standard curve has been plotted between 5-FU concentration (μg/ml) and absorbance (nm) [Figure 2]. The absorbance of the solution of 5-FU was established by the ultraviolet (UV) spectrophotometer (PerkinElmer, Lambda 25, USA) at absorption maxima 267 nm. The α-t-FU-PLGA/5-FU-PLGA NPs were accommodated into a dialysis bag and engrossed at different pH 7.4 and pH 4.5 in PBS. The dialysis bag (Sigma-Aldrich, India) procured as per the protocol. The USP dissolution appliance grade 1 basket type was used to perform the analysis, and the speed of the apparatus was 100 rpm, 37°C. The dialysis bag was poured into the dissolution solution (gastric fluid maintained pH 7.4 and 4.5), and the 5 ml supernatant at definite time interval in h (0, 20, 40, 60, 80, 120, 160) was withdrawn and further analyzed for drug content through established standard calibration curve of the 5-FU solution using UV visible spectrophotometer and the in vitro drug release was calculated by the given formula. The EE value was monitored through high-speed centrifugation of 5-FU-PLGA/α-t-FU-PLGA NPs at 12,000 rpm for 45 min. The supernatant was further filtered with 0.2 μm membrane filter. The collected sample was further allowed to evaluate the absorbance value through UV spectroscopy and compare with earlier plotted graph of 5-FU solution, and percentage of EE was calculated by given formula.
Figure 2: Standard calibration curve of the 5-fluorouracil solution in between concentration (μg/ml) versus absorbance (nm)

Click here to view

  • In vitro drug release (%): Concentration of drug at different time interval/absolute bulk of the drug × 100
  • EE (%): The absolute bulk of the drug − The unbound drug particles in supernatant/the absolute bulk of the drug × 100.

Cell culture

Human tongue SCC15, as oral cancer cell lines, cultured (DMEM/F12) and enriched with 10% heat-inactivated fetal bovine serum followed by addition of 1% concentration ratio of penicillin and streptomycin, and further, SCC15 cells were preserved in typical humidified incubator supplied with 5% CO2, 95% air at 37°C ± 0.5°C.

Cytotoxicity of α-tocopherol-5-fluorouracil-poly-D, L-lactic-co-glycolic acid/5-fluorouracil-poly-D, L-lactic-co-glycolic acid nanoparticles by methyl thiazolyl tetrazolium assay

SCC15 cells were fixed with a frequency of 10,000 cells into 96-well plates and acquiesced to abide by 24 h. The SCC15 cells were exhibited to 10 μL of α-t-FU-PLGA/5-FU-PLGA NPs at predetermined time intervals (24, 48, and 72 h) in a different dose; methyl thiazolyl tetrazolium (MTT) (0.2 mg/ml) was composite to all the well plate and sustenance for 4–6 h. 250 μL DMSO was mixed after the removal of medium and further vibrated for 12 min.

Cytotoxicity of α-tocopherol-5-fluorouracil-poly-D, L-lactic-co-glycolic acid/5-fluorouracil-poly-D, L-lactic-co-glycolic acid nanoparticles in drug-resistant squamous cell carcinoma 15 cells

The drug-resistant SCC15 cell lines were placed in into 96-well plates with a density of 10,000 cells per plates for 24 h, centrifuged, and then procured; the drug-resistant cells were again incubated different drug concentration of α-t-FU-PLGA/5-FU-PLGA NPs at dose 0, 0.25, 1.50, 3.0, 4.5, 6.0, and 7.5 μg/ml.

Therapeutic productivity of α-tocopherol-5-fluorouracil-poly-D, L-lactic-co-glycolic acid/5- fluorouracil-poly-D, L-lactic-co-glycolic acid nanoparticles against squamous cell carcinoma 15 cells

The therapeutic productivity of targeted α-t-FU-PLGA and nontargeted 5-FU-PLGA NPs as antiproliferating agent established against SCC15 cell lines through MTT method. The malignant cells, SCC15, were fixed with the frequency of 5000 cells into 96-well plates for the overnight incubation, the cells were treated with different concentrations 1.0, 0.5, and 0.1 mg/ml, and the antiproliferating effect of the α-t-FU-PLGA/5-FU-PLGA NPs was examined. The cell viability of the NPs was quantified in 96 h and indirectly the cytotoxicity effects of the targeted and nontargeted NPs.

In vitro cellular uptake of α-tocopherol-5-fluorouracil-poly-D, L-lactic-co-glycolic acid/5-fluorouracil-poly-D, L-lactic-co-glycolic acid nanoparticles

SCC15 cell lines were seeded into six-well plates and allowed to treat with for 1 day; after that, the cells were exhibited to formulated α-t-FU-PLGA and nontargeted 5-FU-PLGA NPa labeled with distinct concentration of fluorescein isothiocyanate (FITC) for 4–6 h, and the cellular uptake and targeting were observed by fluorescent microscopy (Carl Zeiss, Germany) using 485 nm excitation for FITC and fluorescence intensity within the treated SCC15 cells being quantified by a microplate reader.

Cell apoptosis by α-tocopherol-5-fluorouracil-poly-D, L-lactic-co-glycolic acid/5-fluorouracil-poly-D, L-lactic-co-glycolic acid nanoparticles

AV-FITC/PI (Invitrogen, Thermo Fisher Scientific, India) apoptotic staining assay was performed by flow cytometry analysis to obtain the programmed cell death.[69] The SCC15 cells were incubated with α-t-FU-PLGA/5-FU-PLGA NPs formulation to attain the rate of cell apoptosis.[70] The SCC15 cells treated with α-t-FU-PLGA/5-FU-PLGA NPs were matured in six-well plates at the density 5 × 103 cells. The treated cells were washed with PBS, and 10 μL AV-FITC conjugate and PI staining solution were poured to each cells solution and incubated at 25°c for 15 min and defended from light, and the fluorescence of the SCC15 cells was examined through flow cytometry (BD Influx Model, USA).

Data analysis

All the scientific data were expressed as in the standard deviation and mean and one-way ANOVA analysis and least significance difference test applied for comparison of the groups. Prism 7.0 (GraphPad, Fay Avenue, Suite 230, La Jalla, CA 92037, USA) statistical software, was used for data analysis (P > 0.05).

 > Results Top

Characterization of α-tocopherol-5-fluorouracil-poly-D, L-lactic-co-glycolic acid/5-fluorouracil-poly-D, L-lactic-co-glycolic acid nanoparticles

The surface conjugation of α-t as a targeted moiety on 5-FU-PLGA NPs eventually increased in average particle size ranges from 145 nm to 160 nm as the nontargeted NPs formulation having the negative surface charge about −17 mV and increased in α-t functionalized α-t-FU-PLGA nanoformulation as shown in [Figure 3]a and [Figure 3]b. The PDI, particle size, and zeta potential are summarized in [Table 1]. The SEM of the α-t-FU-PLGA/5-FU-PLGA nanoformulation confirmed that as a spherical shape of the nanoparticles with the easy surface as shown in [Figure 4].
Figure 3: Zetasizer peak report for particle size, zeta potential, and polydispersity index of α-tocopherol-5-fluorouracil-poly-D,L-lactic- co-glycolic acid/5-fluorouracil-poly-D,L-lactic-co-glycolic acid nanoparticles. (a) Particle size and polydispersity index distribution of the nanoparticles; (b) zeta potential distribution of nanoparticles

Click here to view
Table 1: Particle size, polydispersity index, and zeta potential of targeted a-tocopherol-5-fluorouracil-poly-D,L-lactic-co-glycolic acid nanoparticles/nontargeted 5-fluorouracil-poly-D,L-lactic-co-glycolic acid nanoparticles (n=3; mean±standard deviation)

Click here to view
Figure 4: Scanning electron microscope showing spherical nanoparticles, easy surface, no adherence between the particles in α-tocopherol-5-fluorouracil-poly-D,L-lactic-co-glycolic acid nanoparticles/5-fluorouracil-poly-D,L-lactic-co-glycolic acid nanoparticles

Click here to view

In vitro drug-release system and entrapment efficiency

The in vitro release study and EE of α-t-FU-PLGA/5-FU-PLGA NPs were standardized through a standard calibration curve of the 5-FU solution, which was plotted a linear regression ranging from 0.1 to 10 μg/ml; a straight line was found between 5-FU concentration (μg/ml) and absorbance (nm) through the UV spectrophotometer at absorption maxima 267 nm. In vitro drug release of NPs also depends upon particle size, when size is less, the surface area of NPs increases, and more surface area comes in contact with the medium, resulting in a faster release. The cumulative in vitro drug release at pH 7.4 and pH 4.5 to identify the pH-susceptible difference in a 5-FU release from the nanoparticulate system and the EE of 5-FU loaded in α-t-FU-PLGA/5-FU-PLGA NPs were 68% and 63%, respectively, for both the nanoformulations. The total amount of commutative in vitro drug release at the pH 4.5 was increased 70% at the time range of up to 160 h; the results of 5-FU-PLGA NPs showed steady-state release approximately at the time 140–160 h and 63% in α-t-FU-PLGA NPs which had targeted moiety. In vitro drug-release data for α-t-FU-PLGA and 5-FU-PLGA NPs in pH 7.4 showed rapid releases at time range 40–60 h about 35%–60% and 25%–50%, respectively, which was followed by the cumulative drug release up to 160 h about 85% and 82%, respectively. The slope of the graph for in vitro release system for the α-t-FU-PLGA and 5-FU-PLGA NPs at different pH (7.4 and 4.5) ranges confirm the optimum drug release at different time intervals up to 160 h, as shown in [Figure 5], and the release kinetic at the pH 7.4 and 4.5 confirms that cumulative drug release strongly influences the pH of the dissolution solution. The data for drug release are summarized at a different time interval in [Table 2].
Figure 5: In vitro releases of α-tocopherol-5- fluorouracil-poly-D,L-lactic-co-glycolic acid nanoparticles and 5-fluorouracil-poly-D,L-lactic-co-glycolic acid nanoparticles in phosphate-buffered saline at pH 7.4 and 4.5

Click here to view
Table 2: In vitro drug release of targeted a-tocopherol-5-fluorouracil-poly-D,L-lactic-co-glycolic acid nanoparticles and nontargeted 5-fluorouracil-poly-D,L-lactic-co-glycolic acid nanoparticles

Click here to view

Cytotoxicity of α-tocopherol-5-fluorouracil-poly-D, L-lactic-co-glycolic acid/5-fluorouracil-poly-D, L-lactic-co-glycolic acid nanoparticles in dose-time dependent on squamous cell carcinoma 15 cells

The cytotoxicity study was performed in reference to time-dose dependent manner. 5-FU-PLGA/α-t-FU-PLGA NPs have inhibited the growth of SCC15 cells at different time interval and α-t-FU-PLGA NPs have shown high inhibition rate as compare to 5-FU-PLGA NPs. The targeted α-t-FU-PLGA NPs (8.0 μg/ml) have confirmed the high percentage of inhibition rate 79.39%, 56.93%, and 46.63 at different time interval (24, 48 and 72 h respectively). In other hand non-targeted 5-FU-PLGA NPs (8.0 μg/ml) have shown comparatively low inhibition rate 69.45%, 48.96% and 36.28 at different time interval (24, 48 and 72 h respectively) in SCC15 cells the results confirmed the intense inhibition of OSCC by α-t-FU-PLGA NPs as shown in the graph of [Figure 6]a and [Figure 6]b. The inhibition rate for α-t-FU-PLGA NPs was increased as nontargeted 5-FU-PLGA NPs approximately in lesser intense 45.29% and 39.58%, respectively, for both at time 24 h in SCC15, as the time increases the cytotoxicity effects of α-t-FU-PLGA NPs in SCC15 showed higher percentage of inhibition of 79.98% at time 96 h and confirmed approximately steady-state inhibition of 83.74% up to 160 h, as 5-FU-PLGA NPs showed lower inhibition rate up to 59.25% at the time 160 h as plotted graph in [Figure 7].
Figure 6: Cytotoxicity study, inhibition rate of α-tocopherol-5-fluorouracil-poly-D,L-lactic-co-glycolic acid nanoparticles and 5-fluorouracil-poly-D,L-lactic-co-glycolic acid nanoparticles in squamous cell carcinoma-15 cells detected by methyl thiazolyl tetrazolium assay. (a) Inhibition rate of different dose-concentration-time (24, 48, 72 h) of 5-fluorouracil-poly-D,L-lactic-co-glycolic acid nanoparticles on squamous cell carcinoma-15 cells. (b) Inhibition rate of different dose-concentration-time (24, 48, 72 h) of α-tocopherol-5-fluorouracil-poly-D,L-lactic-co-glycolic acid nanoparticles on squamous cell carcinoma-15 cells

Click here to view
Figure 7: Cytotoxicity study, inhibition rate of α-tocopherol-5- fluorouracil-poly-D,L-lactic-co-glycolic acid nanoparticles and 5-fluorouracil-poly-D,L-lactic-co-glycolic acid nanoparticles on squamous cell carcinoma-15 cells up to time 160 h

Click here to view

Cytotoxicity of α-tocopherol-5-fluorouracil-poly-D, L-lactic-co-glycolic acid/5-fluorouracil-poly-D, L-lactic-co-glycolicacid nanoparticles in drug-resistant squamous cell carcinoma 15 cells

The drug-resistant cell line was fixed to optimize the inhibitory activity of α-t-FU-PLGA/5-FU-PLGA NPs in drug-resistant SCC15 cells. IC50 values of the inhibitory activity of α-t-FU-PLGA/5-FU-PLGA NPs on drug-resistant SCC-15 cell were 13.19 and 23.25 μg/ml. The drug resistance for the cytotoxicity of α-t-FU-PLGA/5-FU-PLGA NPs in respect to dose-dependent manner, inhibition rate was remarkably high in α-t-FU-PLGA approximately 80% at the concentration 8.0 μg/ml with a comparison to inhibition rate for 5-FU-PLGA NPs, 65% at the concentration of 8.0 μg/ml and inhibition rate with respect to time-dependent [Table 3], cytotoxicity in α-t-FU-PLGA nanoparticles was higher (58%) in comparison to 5-FU-PLGA nanoparticles (45%), shown in [Figure 8]a and [Figure 8]b.
Table 3: Inhibitory effects of targeted a-tocopherol-5-fluorouracil-poly-D,L-lactic-co-glycolic acid nanoparticles and nontargeted 5-fluorouracil-poly-D,L-lactic-co-glycolic acid nanoparticles on the proliferation of the squamous cell carcinoma-15 cells, 5-fluorouracil resistance cell line (n=3; mean±standard deviation)

Click here to view
Figure 8: Inhibition of α-tocopherol-5-fluorouracil-poly-D,L-lactic-co-glycolic acid nanoparticles and 5-fluorouracil-poly-D,L-lactic-co-glycolic acid nanoparticles on drug-resistant squamous cell carcinoma-15 cells detected by methyl thiazolyl tetrazolium assay. (a) Dose-concentration-dependent inhibition rate; (b) time-dependent inhibition rate

Click here to view

Therapeutic productivity of α-tocopherol-5-fluorouracil-poly-D, L-lactic-co-glycolic acid/5-fluorouracil-poly-D, L-lactic-co-glycolic acid nanoparticles in squamous cell carcinoma 15 cells

The therapeutic productivity of α-t-FU-PLGA/5-FU-PLGA NPs was examined on the basis of the cellular update, cytotoxicity investigation of targeted nanoformulation, α-t-FU-PLGA NPs and nontargeted 5-FU-PLGA NPs on the SCC15 cell line; the nanoformulations produced expected level of toxicity to the cell at different concentration (0.1, 0.5, and 1.0 mg/ml). The α-t-FU-PLGA NPs produced higher percentage of cell viability, 83.59%, 67.82%, and 41.25% in all concentration levels 1, 0.5, and 0.1 mg/ml, respectively, confirmed the therapeutic productivity, and the internalization of 5-FU and nontargeted 5-FU-PLGA NPs produced less cell viability 65.23%, 39.82%, 29% at the concentration of 1.0, 0.5, and 0.1 mg/ml, respectively, which confirmed the comparatively less therapeutic productivity of 5-FU, as shown in [Figure 9].
Figure 9: Cell viability assays of α-tocopherol-5-fluorouracil-poly-D,L-lactic-co-glycolic acid nanoparticles and 5-fluorouracil-poly-D,L-lactic-co-glycolic acid nanoparticles in squamous cell carcinoma-15 cells at the concentration of 1.0 mg/ml, 0.05 mg/ml, and 0.1 mg/ml

Click here to view

In vitro cellular uptake and targeting of α-tocopherol-5-fluorouracil-poly-D, L-lactic-co-glycolic acid/5-fluorouracil-poly-D, L-lactic-co-glycolic acid nanoparticles

In vitro cellular uptake analysis of targeted drug-loaded α-t-FU-PLGA NPs and nontargeted 5-FU-PLGA NPs was examined in SCC15 cancer cells by fluorescent microscopy. FITC-labeled α-t-FU-PLGA NPs crucially inflate the acquisition of drug into SCC15 as compared nontargeted NPs. In [Figure 10], FITC labeled α-t-FU-PLGA/5-FU-PLGA NPs were assembled in the cytoplasm when treated with SCC15. The robust color fluorescence was distinguished in SCC15 cells, designed the endocytosis of a vast number of α-t-FU-PLGA NPs, and ascribed to extent of the targeting moiety. The exposure of SCC15-5-FU-PLGA NPs showed the modest strength fluorescence. The higher intensity of cellular uptake for α-t-FU-PLGA NPs described that target moiety of NPs successfully invaded the cancer cells via receptor-mediated endocytosis in OSCC.
Figure: 10: Cellular uptake of fluorescein isothiocyanate labeled α-tocopherol-5-fluorouracil-poly-D,L-lactic-co-glycolic acid nanoparticles and 5-fluorouracil-poly-D,L-lactic-co-glycolic acid nanoparticles. (a) α-Tocopherol-5-fluorouracil-poly-D,L-lactic-co-glycolic acid nanoparticles, (b) 5-fluorouracil-poly-D,L-lactic-co-glycolic acid nanoparticles in squamous cell carcinoma-15 cells by fluorescent microscopy analysis on the scale bar is 50 μm

Click here to view

Cell apoptosis by α-tocopherol-5-fluorouracil-poly-D, L-lactic-co-glycolic acid/5-fluorouracil-poly-D, L-lactic-co-glycolic acid nanoparticles

Cell apoptosis assay was performed to investigate the cell death rate in SCC15, which was investigated by nontargeted NPs (5-FU-PLGA NPs) and targeted NPs (α-t-FU-PLGA NPs). The NPs have incubated with SCC15 cells approximately 96 h, with the exposure of AV-FITC/PI staining protocol by flow cytometry. The data confirm the apoptosis for early and late phase as shown in [Figure 11], and flow cytometry investigation for the ratio of AV/PI-positive cells was treated with α-t-FU-PLGA/5-FU-PLGA NPs. The observation for early apoptosis, 27.98% and 16.45%, and late apoptosis, 74.29% and 61.13%, of SCC15 cells for α-t-FU-PLGA/5-FU-PLGA NPs, respectively.
Figure 11: Flow cytometry analysis of quantitative apoptosis and apoptosis of squamous cell carcinoma-15 cells and the corresponding cell (%) in early and late apoptosis for α-tocopherol-5-fluorouracil-poly-D,L-lactic-co-glycolic acid nanoparticles and 5-fluorouracil-poly-D,L-lactic-co-glycolic acid nanoparticles

Click here to view

 > Discussion Top

In current scenario, advancement in the treatment system against cancer takes multiple levels up to increase the survival rate of affected population.[71] Cancer is a massive health problem which effects almost every part of the world. Mostly, Asian countries are reported higher rate cancer death, approximately 3 million.[2] In that, oral tumor is highly reported and underlined tumor because of ignorance and consumption of tobacco.[72] The prime treatment of oral cancer is surgery with chemotherapy and radiotherapy.[73] Chemotherapy is the treatment which is used to treat oral cancer by killing or inhibiting the growth of tumor cells.[74] The anticancer drug, 5-FU, was used as a chemotherapeutic agent against oral cancer treatment. Although 5-FU has high efficacy, it may be also restricted with limited therapeutic index and reduced bioavailability because of its conventional dosage formulation.[75] 5-FU may also cause some side effects and toxicity because of overdose and nonspecific treatment. Hence, these drawbacks were always demands to overcome nonspecific activity of 5-FU along with minimum toxicity.[43] Nanotechnology may enhanced the effectiveness and biological activity of 5-FU with targeted delivery as surface attachment of functional moiety, α-t along with PLGA NPs engulfed 5-FU drug molecules. The earlier report showed very less effective release of drug as this study enhances the release up to 80% with control release of delivery up to 5 days over the specific region.[76],[77] The polymeric NPs, as PLGA nanoparticulate system,[78] loaded 5-FU was prepared through ultrasonication method and α-t, tagged upon the surface of PLGA-FU NPs to bind with the cancer cell surface and release the drug. The targeted α-t-FU-PLGA NPs showed higher cytotoxicity in comparison to nontargeted 5-FU-PLGA NPs because of surface modification with the tagging of α-t. The targeted moiety α-t acted to increase the endocytosis toward SCC15 cells. α-t-modified PLGA-FU NPs showed enhanced cellular permeation and enhanced cancer cell accumulation.[79] The extracellular fluid may activate the α-t tagging and influx the accumulation within the cytosol and adhere over SCC15 cells and accurately followed the EPR effects over tumor site. On the other hand, nontargeted 5-FU-PLGA NPs showed limited cell uptake due to nonmodified surface. The over-treated damaged cells may produce drug resistance. The results of cytotoxicity of drug-resistant SCC15 cells may produce high inhibition with concentration of NPs. The IC50 values of α-t-FU-PLGA NPs showed high regulation of growth inhibition and reduced level of cell survival via growth inhibition in G1 phase. The PLGA polymer nanoparticulate system provided a parallel advancement as surface modification by α-t to the drug delivery of 5-FU.[48] This system may sort the issue which limited the dissolution, solubility, therapeutic efficacy, and biological activity against OSCC. The in vitro encapsulation of cytotoxicity may enhance the 5-FU efficacy about 5-fold and cellular uptake was increased as time-dependent manner. The apoptosis study confirmed the cell internalization and produced thymineless cell death affecting the cellular pathways. Flow cytometry revealed the early apoptosis through targeted drug delivery of 5-FU. The in vitro drug assessment plots a significant result data for formulated α-t-modified PLGA-FU NPs against OSCC, SCC15 cells.

 > Conclusion Top

Farthermost, the objectives of the current work were to formulate the targeted NPs specifically on oral cancer region and uncollision with beside healthy cells. In this study, targeted α-t-functionalized surface moiety successfully attached within 5-FU-loaded PLGA nanoformulation and comparatively nontargeted 5-FU-PLGA nanoformulation was also synthesized. The characterization studies of blended NPs in both formulations were confirmed the uniformed dispersion, particle size, and negative charges. The cumulative in vitro drug-release pattern of the α-t-FU-PLGA/5-FU-PLGA NPs gradually increased with concentration-time-dependent manner. The assessment of cytotoxicity in dose-time-dependent and drug-resistant in SCC15 cells confirmed the ability of α-t-FU-PLGA/5-FU-PLGA NPs to inhibit the growth of dividing cancer cells. The physiology of SCC15 cells and on the basis of result data of increased cellular uptake, cellular augmentation, specific cellular targeting, cell viability, and cellular toxicity confirmed the successful formulation of targeted α-t-FU-PLGA NPs. It is concluded that the attractive-targeted drug within the polymeric nanoformulation provides a possible platform for delivering the drug to the specific site for trigged therapeutic action toward oral cancer.


Authors are thankful to Indian Council of Medical Research (ICMR), New Delhi, India for providing financial support.

Financial support and sponsorship

Indian Council of Medical Research (ICMR), New Delhi, India (Project id-45/21/2013-NAN-BMS).

Conflicts of interest

There are no conflicts of interest.

 > References Top

Iorio F, Knijnenburg TA, Vis DJ, Bignell GR, Menden MP, Schubert M, et al. A landscape of pharmacogenomic interactions in cancer. Cell 2016;166:740-54.  Back to cited text no. 1
Torre LA, Sauer AM, Chen MS Jr., Kagawa-Singer M, Jemal A, Siegel RL, et al. Cancer statistics for Asian Americans, Native Hawaiians, and Pacific Islanders, 2016: Converging incidence in males and females. CA Cancer J Clin 2016;66:182-202.  Back to cited text no. 2
American Society of Clinical Oncology. The state of cancer care in America, 2016: A report by the American Society of Clinical Oncology. J Oncol Pract 2016;12:339-83.  Back to cited text no. 3
Stewart BW, Bray F, Forman D, Ohgaki H, Straif K, Ullrich A, et al. Cancer prevention as part of precision medicine: 'Plenty to be done'. Carcinogenesis 2016;37:2-9.  Back to cited text no. 4
D'Souza ND, Murthy NS, Aras RY. Projection of cancer incident cases for India -till 2026. Asian Pac J Cancer Prev 2013;14:4379-86.  Back to cited text no. 5
Allemani C, Weir HK, Carreira H, Harewood R, Spika D, Wang XS, et al. Global surveillance of cancer survival 1995-2009: Analysis of individual data for 25,676,887 patients from 279 population-based registries in 67 countries (CONCORD-2). Lancet 2015;385:977-1010.  Back to cited text no. 6
Cho JH, Lee MH, Cho YJ, Park BS, Kim S, Kim GC, et al. The bacterial protein azurin enhances sensitivity of oral squamous carcinoma cells to anticancer drugs. Yonsei Med J 2011;52:773-8.  Back to cited text no. 7
Coelho KR. Challenges of the oral cancer burden in India. J Cancer Epidemiol 2012;2012:701932.  Back to cited text no. 8
Yang JS, Lin CW, Hsieh YH, Chien MH, Chuang CY, Yang SF, et al. Overexpression of carbonic anhydrase IX induces cell motility by activating matrix metalloproteinase-9 in human oral squamous cell carcinoma cells. Oncotarget 2017;8:83088-99.  Back to cited text no. 9
Chinn SB, Myers JN. Oral cavity carcinoma: Current management, controversies, and future directions. J Clin Oncol 2015;33:3269-76.  Back to cited text no. 10
Tandon P, Dadhich A, Saluja H, Bawane S, Sachdeva S. The prevalence of squamous cell carcinoma in different sites of oral cavity at our rural health care centre in Loni, Maharashtra – A retrospective 10-year study. Contemp Oncol (Pozn) 2017;21:178-83.  Back to cited text no. 11
Harada K, Ferdous T, Ueyama Y. Therapeutic strategies with oral fluoropyrimidine anticancer agent, S-1 against oral cancer. Jpn Dent Sci Rev 2017;53:61-77.  Back to cited text no. 12
Gavish A, Krayzler E, Nagler R. Tumor growth and cell proliferation rate in human oral cancer. Arch Med Res 2016;47:271-4.  Back to cited text no. 13
Curado MP, Hashibe M. Recent changes in the epidemiology of head and neck cancer. Curr Opin Oncol 2009;21:194-200.  Back to cited text no. 14
Gupta B, Johnson NW. Systematic review and meta-analysis of association of smokeless tobacco and of betel quid without tobacco with incidence of oral cancer in South Asia and the Pacific. PLoS One 2014;9:e113385.  Back to cited text no. 15
Narang S, Kanungo N, Jain R. Squamous cell carcinoma: Morphological & topographical spectrum: A two year analysis. Indian J Surg 2014;76:104-10.  Back to cited text no. 16
Fan L, Hu X, Lin S, Zhou W, Fu S, Lv H, et al. Concurrent preoperative chemotherapy and three-dimensional conformal radiotherapy followed by surgery for oral squamous cell carcinoma: A retrospective analysis of 104 cases. Oncotarget 2017;8:75557-67.  Back to cited text no. 17
Pires FR, Ramos AB, Oliveira JB, Tavares AS, Luz PS, Santos TC, et al. Oral squamous cell carcinoma: Clinicopathological features from 346 cases from a single oral pathology service during an 8-year period. J Appl Oral Sci 2013;21:460-7.  Back to cited text no. 18
Marcinkiewicz KM, Gudas LJ. Altered epigenetic regulation of homeobox genes in human oral squamous cell carcinoma cells. Exp Cell Res 2014;320:128-43.  Back to cited text no. 19
Sommer G, Rossa C, Chi AC, Neville BW, Heise T. Implication of RNA-binding protein la in proliferation, migration and invasion of lymph node-metastasized hypopharyngeal SCC cells. PLoS One 2011;6:e25402.  Back to cited text no. 20
Ariyawardana A, Athukorala AD, Arulanandam A. Effect of betel chewing, tobacco smoking and alcohol consumption on oral submucous fibrosis: A case-control study in Sri Lanka. J Oral Pathol Med 2006;35:197-201.  Back to cited text no. 21
Awojobi O, Scott SE, Newton T. Patients' perceptions of oral cancer screening in dental practice: A cross-sectional study. BMC Oral Health 2012;12:55.  Back to cited text no. 22
Niaz K, Maqbool F, Khan F, Bahadar H, Ismail Hassan F, Abdollahi M, et al. Smokeless tobacco (paan and gutkha) consumption, prevalence, and contribution to oral cancer. Epidemiol Health 2017;39:e2017009.  Back to cited text no. 23
Eckert AW, Wickenhauser C, Salins PC, Kappler M, Bukur J, Seliger B, et al. Clinical relevance of the tumor microenvironment and immune escape of oral squamous cell carcinoma. J Transl Med 2016;14:85.  Back to cited text no. 24
Hübbers CU, Akgül B. HPV and cancer of the oral cavity. Virulence 2015;6:244-8.  Back to cited text no. 25
Grimm M, Cetindis M, Lehmann M, Biegner T, Munz A, Teriete P, et al. Association of cancer metabolism-related proteins with oral carcinogenesis – Indications for chemoprevention and metabolic sensitizing of oral squamous cell carcinoma? J Transl Med 2014;12:208.  Back to cited text no. 26
Lin CW, Chuang CY, Tang CH, Chang JL, Lee LM, Lee WJ, et al. Combined effects of icam-1 single-nucleotide polymorphisms and environmental carcinogens on oral cancer susceptibility and clinicopathologic development. PLoS One 2013;8:e72940.  Back to cited text no. 27
Messadi DV. Diagnostic aids for detection of oral precancerous conditions. Int J Oral Sci 2013;5:59-65.  Back to cited text no. 28
Mondal R, Ghosh SK, Choudhury JH, Seram A, Sinha K, Hussain M, et al. Mitochondrial DNA copy number and risk of oral cancer: A report from Northeast India. PLoS One 2013;8:e57771.  Back to cited text no. 29
Montero PH, Patel SG. Cancer of the oral cavity. Surg Oncol Clin N Am 2015;24:491-508.  Back to cited text no. 30
Yardimci G, Kutlubay Z, Engin B, Tuzun Y. Precancerous lesions of oral mucosa. World J Clin Cases 2014;2:866-72.  Back to cited text no. 31
Fu XJ, Li HX, Yang K, Chen D, Tang H. The important tumor suppressor role of PER1 in regulating the cyclin-CDK-CKI network in SCC15 human oral squamous cell carcinoma cells. Onco Targets Ther 2016;9:2237-45.  Back to cited text no. 32
Zhang F, Li Y, Zhang H, Huang E, Gao L, Luo W, et al. Anthelmintic mebendazole enhances cisplatin's effect on suppressing cell proliferation and promotes differentiation of head and neck squamous cell carcinoma (HNSCC). Oncotarget 2017;8:12968-82.  Back to cited text no. 33
Petti S, Masood M, Scully C. The magnitude of tobacco smoking-betel quid chewing-alcohol drinking interaction effect on oral cancer in South-East Asia. A meta-analysis of observational studies. PLoS One 2013;8:e78999.  Back to cited text no. 34
Bundela S, Sharma A, Bisen PS. Potential therapeutic targets for oral cancer: ADM, TP53, EGFR, LYN, CTLA4, SKIL, CTGF, CD70. PLoS One 2014;9:e102610.  Back to cited text no. 35
Hwang-Bo J, Park JH, Chung IS. Tumstatin induces apoptosis mediated by fas signaling pathway in oral squamous cell carcinoma SCC-VII cells. Oncol Lett 2015;10:1016-22.  Back to cited text no. 36
Alaufi OM, Noorwali A, Zahran F, Al-Abd AM, Al-Attas S. Cytotoxicity of thymoquinone alone or in combination with cisplatin (CDDP) against oral squamous cell carcinoma in vitro. Sci Rep 2017;7:13131.  Back to cited text no. 37
Kalantarian P, Najafabadi AR, Haririan I, Vatanara A, Yamini Y, Darabi M, et al. Preparation of 5-fluorouracil nanoparticles by supercritical antisolvents for pulmonary delivery. Int J Nanomedicine 2010;5:763-70.  Back to cited text no. 38
Focaccetti C, Bruno A, Magnani E, Bartolini D, Principi E, Dallaglio K, et al. Effects of 5-fluorouracil on morphology, cell cycle, proliferation, apoptosis, autophagy and ROS production in endothelial cells and cardiomyocytes. PLoS One 2015;10:e0115686.  Back to cited text no. 39
Nazim UM, Rasheduzzaman M, Lee YJ, Seol DW, Park SY. Enhancement of TRAIL-induced apoptosis by 5-fluorouracil requires activating bax and p53 pathways in TRAIL-resistant lung cancers. Oncotarget 2017;8:18095-105.  Back to cited text no. 40
Sun J, Wei Q, Zhou Y, Wang J, Liu Q, Xu H, et al. A systematic analysis of FDA-approved anticancer drugs. BMC Syst Biol 2017;11:87.  Back to cited text no. 41
Liu B, Han L, Liu J, Han S, Chen Z, Jiang L, et al. Co-delivery of paclitaxel and TOS-cisplatin via TAT-targeted solid lipid nanoparticles with synergistic antitumor activity against cervical cancer. Int J Nanomedicine 2017;12:955-68.  Back to cited text no. 42
Gao Z, Li Z, Yan J, Wang P. Irinotecan and 5-fluorouracil-co-loaded, hyaluronic acid-modified layer-by-layer nanoparticles for targeted gastric carcinoma therapy. Drug Des Devel Ther 2017;11:2595-604.  Back to cited text no. 43
Tawfik E, Ahamed M, Almalik A, Alfaqeeh M, Alshamsan A. Prolonged exposure of colon cancer cells to 5-fluorouracil nanoparticles improves its anticancer activity. Saudi Pharm J 2017;25:206-13.  Back to cited text no. 44
Balasubramanian S, Girija AR, Nagaoka Y, Iwai S, Suzuki M, Kizhikkilot V, et al. Curcumin and 5-fluorouracil-loaded, folate- and transferrin-decorated polymeric magnetic nanoformulation: A synergistic cancer therapeutic approach, accelerated by magnetic hyperthermia. Int J Nanomedicine 2014;9:437-59.  Back to cited text no. 45
Bae YH, Park K. Targeted drug delivery to tumors: Myths, reality and possibility. J Control Release 2011;153:198-205.  Back to cited text no. 46
Singh R, Lillard JW Jr. Nanoparticle-based targeted drug delivery. Exp Mol Pathol 2009;86:215-23.  Back to cited text no. 47
Esfandyari-Manesh M, Mostafavi SH, Majidi RF, Koopaei MN, Ravari NS, Amini M, et al. Improved anticancer delivery of paclitaxel by albumin surface modification of PLGA nanoparticles. Daru 2015;23:28.  Back to cited text no. 48
Li B, Li Q, Mo J, Dai H. Drug-loaded polymeric nanoparticles for cancer stem cell targeting. Front Pharmacol 2017;8:51.  Back to cited text no. 49
Butt AM, Mohd Amin MC, Katas H. Synergistic effect of pH-responsive folate-functionalized poloxamer 407-TPGS-mixed micelles on targeted delivery of anticancer drugs. Int J Nanomedicine 2015;10:1321-34.  Back to cited text no. 50
Nair KL, Jagadeeshan S, Nair SA, Kumar GS. Biological evaluation of 5-fluorouracil nanoparticles for cancer chemotherapy and its dependence on the carrier, PLGA. Int J Nanomedicine 2011;6:1685-97.  Back to cited text no. 51
Jain SK, Haider T, Kumar A, Jain A. Lectin-conjugated clarithromycin and acetohydroxamic acid-loaded PLGA nanoparticles: A novel approach for effective treatment of H. pylori. AAPS PharmSciTech 2016;17:1131-40.  Back to cited text no. 52
Taghavi S, Ramezani M, Alibolandi M, Abnous K, Taghdisi SM. Chitosan-modified PLGA nanoparticles tagged with 5TR1 aptamer for in vivo tumor-targeted drug delivery. Cancer Lett 2017;400:1-8.  Back to cited text no. 53
Pelaz B, Alexiou C, Alvarez-Puebla RA, Alves F, Andrews AM, Ashraf S, et al. Diverse applications of nanomedicine. ACS Nano 2017;11:2313-81.  Back to cited text no. 54
Fadaeian G, Shojaosadati SA, Kouchakzadeh H, Shokri F, Soleimani M. Targeted delivery of 5-fluorouracil with monoclonal antibody modified bovine serum albumin nanoparticles. Iran J Pharm Res 2015;14:395-405.  Back to cited text no. 55
Hrynyk M, Ellis JP, Haxho F, Allison S, Steele JA, Abdulkhalek S, et al. Therapeutic designed poly (lactic-co-glycolic acid) cylindrical oseltamivir phosphate-loaded implants impede tumor neovascularization, growth and metastasis in mouse model of human pancreatic carcinoma. Drug Des Devel Ther 2015;9:4573-86.  Back to cited text no. 56
Ragab DM, Rohani S, Consta S. Controlled release of 5-fluorouracil and progesterone from magnetic nanoaggregates. Int J Nanomedicine 2012;7:3167-89.  Back to cited text no. 57
Singh S, Kotla NG, Tomar S, Maddiboyina B, Webster TJ, Sharma D, et al. A nanomedicine-promising approach to provide an appropriate colon-targeted drug delivery system for 5-fluorouracil. Int J Nanomedicine 2015;10:7175-82.  Back to cited text no. 58
He X, Li J, Guo W, Liu W, Yu J, Song W, et al. Targeting the microRNA-21/AP1 axis by 5-fluorouracil and pirarubicin in human hepatocellular carcinoma. Oncotarget 2015;6:2302-14.  Back to cited text no. 59
Steichen SD, Caldorera-Moore M, Peppas NA. A review of current nanoparticle and targeting moieties for the delivery of cancer therapeutics. Eur J Pharm Sci 2013;48:416-27.  Back to cited text no. 60
Mah E, Sapper TN, Chitchumroonchokchai C, Failla ML, Schill KE, Clinton SK, et al. A-tocopherol bioavailability is lower in adults with metabolic syndrome regardless of dairy fat co-ingestion: A randomized, double-blind, crossover trial. Am J Clin Nutr 2015;102:1070-80.  Back to cited text no. 61
Horák D, Pustovyy VI, Babinskyi AV, Palyvoda OM, Chekhun VF, Todor IN, et al. Enhanced antitumor activity of surface-modified iron oxide nanoparticles and an α-tocopherol derivative in a rat model of mammary gland carcinosarcoma. Int J Nanomedicine 2017;12:4257-68.  Back to cited text no. 62
Pedeboscq S, Rey C, Petit M, Harpey C, De Giorgi F, Ichas F, et al. Non-antioxidant properties of alpha-tocopherol reduce the anticancer activity of several protein kinase inhibitors in vitro. PloS one 2012;7:e36811.  Back to cited text no. 63
Yu J, Zhou Y, Chen W, Ren J, Zhang L, Lu L, et al. Preparation, characterization and evaluation of α-tocopherol succinate-modified dextran micelles as potential drug carriers. Materials (Basel) 2015;8:6685-96.  Back to cited text no. 64
Abbad S, Wang C, Waddad AY, Lv H, Zhou J. Preparation, in vitro and in vivo evaluation of polymeric nanoparticles based on hyaluronic acid-poly(butyl cyanoacrylate) and D-alpha-tocopheryl polyethylene glycol 1000 succinate for tumor-targeted delivery of morin hydrate. Int J Nanomedicine 2015;10:305-20.  Back to cited text no. 65
Neophytou CM, Constantinou AI. Drug delivery innovations for enhancing the anticancer potential of Vitamin E isoforms and their derivatives. Biomed Res Int 2015;2015:584862.  Back to cited text no. 66
Harfouche R, Basu S, Soni S, Hentschel DM, Mashelkar RA, Sengupta S, et al. Nanoparticle-mediated targeting of phosphatidylinositol-3-kinase signaling inhibits angiogenesis. Angiogenesis 2009;12:325-38.  Back to cited text no. 67
Watson C, Ge J, Cohen J, Pyrgiotakis G, Engelward BP, Demokritou P, et al. High-throughput screening platform for engineered nanoparticle-mediated genotoxicity using CometChip technology. ACS Nano 2014;8:2118-33.  Back to cited text no. 68
Wlodkowic D, Skommer J, Darzynkiewicz Z. Flow cytometry-based apoptosis detection. Methods Mol Biol 2009;559:19-32.  Back to cited text no. 69
Alphonse G, Maalouf M, Battiston-Montagne P, Ardail D, Beuve M, Rousson R, et al. P53-independent early and late apoptosis is mediated by ceramide after exposure of tumor cells to photon or carbon ion irradiation. BMC Cancer 2013;13:151.  Back to cited text no. 70
Arnold M, Pandeya N, Byrnes G, Renehan PA, Stevens GA, Ezzati PM, et al. Global burden of cancer attributable to high body-mass index in 2012: A population-based study. Lancet Oncol 2015;16:36-46.  Back to cited text no. 71
Rivera C. Essentials of oral cancer. Int J Clin Exp Pathol 2015;8:11884-94.  Back to cited text no. 72
Block KI, Gyllenhaal C, Lowe L, Amedei A, Amin AR, Amin A, et al. Designing a broad-spectrum integrative approach for cancer prevention and treatment. Semin Cancer Biol 2015;35 Suppl: S276-304.  Back to cited text no. 73
Huang CY, Ju DT, Chang CF, Muralidhar Reddy P, Velmurugan BK. A review on the effects of current chemotherapy drugs and natural agents in treating non-small cell lung cancer. Biomedicine (Taipei) 2017;7:23.  Back to cited text no. 74
Oertel K, Spiegel K, Schmalenberg H, Dietz A, Maschmeyer G, Kuhnt T, et al. Phase I trial of split-dose induction docetaxel, cisplatin, and 5-fluorouracil (TPF) chemotherapy followed by curative surgery combined with postoperative radiotherapy in patients with locally advanced oral and oropharyngeal squamous cell cancer (TISOC-1). BMC Cancer 2012;12:483.  Back to cited text no. 75
Makadia HK, Siegel SJ. Poly lactic-co-glycolic acid (PLGA) as biodegradable controlled drug delivery carrier. Polymers (Basel) 2011;3:1377-97.  Back to cited text no. 76
Calixto G, Bernegossi J, Fonseca-Santos B, Chorilli M. Nanotechnology-based drug delivery systems for treatment of oral cancer: A review. Int J Nanomedicine 2014;9:3719-35.  Back to cited text no. 77
Piktel E, Niemirowicz K, Wątek M, Wollny T, Deptuła P, Bucki R, et al. Recent insights in nanotechnology-based drugs and formulations designed for effective anti-cancer therapy. J Nanobiotechnology 2016;14:39.  Back to cited text no. 78
Subudhi MB, Jain A, Jain A, Hurkat P, Shilpi S, Gulbake A, et al. Eudragit S100 coated citrus pectin nanoparticles for colon targeting of 5-fluorouracil. Materials (Basel) 2015;8:832-49.  Back to cited text no. 79


  [Figure 1], [Figure 2], [Figure 3], [Figure 4], [Figure 5], [Figure 6], [Figure 7], [Figure 8], [Figure 9], [Figure 10], [Figure 11]

  [Table 1], [Table 2], [Table 3]

This article has been cited by
1 5-Fluorouracil (5-FU) resistance and the new strategy to enhance the sensitivity against cancer: Implication of DNA repair inhibition
Chinmayee Sethy, Chanakya Nath Kundu
Biomedicine & Pharmacotherapy. 2021; 137: 111285
[Pubmed] | [DOI]
2 The prospects of nanotherapeutic approaches for targeting tumor-associated macrophages in oral cancer
Dwaipayan Bhattacharya, Kalyani Sakhare, Kumar Pranav Narayan, Rajkumar Banerjee
Nanomedicine: Nanotechnology, Biology and Medicine. 2021; 34: 102371
[Pubmed] | [DOI]
3 Nanoparticles in Dentistry: A Comprehensive Review
Gustavo Moraes, Carolina Zambom, Walter L. Siqueira
Pharmaceuticals. 2021; 14(8): 752
[Pubmed] | [DOI]
4 Sulfasalazine Microparticles Targeting Macrophages for the Treatment of Inflammatory Diseases Affecting the Synovial Cavity
Monica-Carolina Villa-Hermosilla, Ana Fernández-Carballido, Carolina Hurtado, Emilia Barcia, Consuelo Montejo, Mario Alonso, Sofia Negro
Pharmaceutics. 2021; 13(7): 951
[Pubmed] | [DOI]


Similar in PUBMED
   Search Pubmed for
   Search in Google Scholar for
 Related articles
Access Statistics
Email Alert *
Add to My List *
* Registration required (free)

  >Abstract>Introduction>Materials and Me...>Results>Discussion>Conclusion>Article Figures>Article Tables
  In this article

 Article Access Statistics
    PDF Downloaded144    
    Comments [Add]    
    Cited by others 4    

Recommend this journal