New Nanotechnology-Based Drug Delivery System Targets Lung Cancer with Precision

Editor:Nina

Scientists develop a folate-bovine serum albumin-coated ethoniosome-based drug delivery system using pterostilbene to target lung cancer cells, demonstrating enhanced tumor targeting and therapeutic effectiveness.

Key Preview

• Research Question: Can a new formulation of pterostilbene (PTS) using folate-bovine serum albumin-coated ethoniosomes improve targeted drug delivery for lung cancer treatment?
• Research Design and Strategy: The study employed a factorial design to optimize ethoniosomal formulations, followed by in vitro and in vivo evaluations to assess efficacy in targeting lung cancer cells.
• Method: PTS-loaded ethoniosomes were developed using a proniosome formulation method, characterized through various physicochemical analyses, and evaluated for cytotoxicity in A549 lung cancer cell lines and a mouse model.
• Key Results: The optimized formulation (F4) displayed an entrapment efficiency of 93.19% and a drug release rate of 75.10% after 24 hours. In vivo studies showed significant tumor reduction in treated mice.
• Significance of the Research: This innovative drug delivery system may enhance the therapeutic efficacy of PTS while minimizing side effects, offering a potential new avenue for lung cancer treatment.

Introduction

Lung cancer is a significant global health challenge and remains one of the leading causes of cancer-related mortality. Characterized by its aggressive nature and late-stage diagnosis, lung cancer has an alarming incidence rate, particularly in regions with high risk factors such as smoking, environmental pollutants, and genetic predispositions. The disease is primarily classified into two main types: small cell lung carcinoma (SCLC) and non-small cell lung carcinoma (NSCLC), with NSCLC accounting for approximately 85% of cases. The complexity of lung cancer, coupled with its resistance to treatment, underscores the urgent need for more effective therapeutic strategies.

Traditional treatment approaches for lung cancer typically involve chemotherapy, radiation therapy, and surgical intervention. These methods aim to eliminate cancer cells or reduce tumor size but often suffer from significant limitations. A common strategy in drug delivery involves systemic administration, where chemotherapy agents are distributed throughout the body. While this can be effective in targeting tumors, it frequently leads to systemic toxicity and adverse side effects, as healthy cells are also affected by the drugs. Moreover, the issue of multidrug resistance often arises, where cancer cells adapt to withstand the effects of chemotherapy, leading to treatment failure and poor patient outcomes.

The challenges inherent in current treatment modalities result in a high rate of recurrence and low progression-free survival among lung cancer patients. Many patients experience debilitating side effects, which can severely impact their quality of life and willingness to continue treatment. As a result, there is a pressing need for innovative strategies that can improve the efficacy of drug delivery while minimizing systemic toxicity.

In light of these challenges, the study introduces a novel drug delivery strategy utilizing pterostilbene (PTS) formulated into folate-bovine serum albumin-coated ethoniosomes. This innovative approach aims to enhance the solubility and targeted delivery of PTS specifically to lung cancer cells that overexpress folate receptors. By leveraging the principles of targeted drug delivery, this strategy seeks to improve therapeutic outcomes and reduce side effects, representing a promising advancement in the fight against lung cancer.

Research Team and Aim

The research team was led by Dr. Nemany A. N. Hanafy, along with co-authors Reham H. Abdelbadea, Abdelaziz E. Abdelaziz, and Eman A. Mazyed. This study was conducted in 2023 at the Institute of Nanoscience and Nanotechnology, Kafrelsheikh University, Egypt. The findings were published in the journal Cancer Nanotechnology under the title “Formulation and optimization of folate-bovine serum albumin-coated ethoniosomes of pterostilbene as a targeted drug delivery system for lung cancer: In vitro and in vivo demonstrations.”

The aim of the research, as articulated by Dr. Hanafy, was to formulate and optimize a novel drug delivery system using folate-bovine serum albumin-coated ethoniosomes loaded with pterostilbene (PTS). This formulation is intended to enhance the targeted delivery of PTS to lung cancer cells that overexpress folate receptors, ultimately improving therapeutic effectiveness while minimizing side effects.

Experimental Process

Primary Technique
The primary technique employed in this study was the development of pterostilbene (PTS)-loaded ethoniosomes using the proniosome formulation method. This innovative approach was designed to enhance drug delivery specifically targeting lung cancer cells, addressing the limitations of traditional drug delivery systems.

Key Steps of Each Experiment

1. Formulation Preparation: PTS-loaded ethoniosomes were prepared by dissolving PTS, surfactants (such as Span 60), and cholesterol in ethanol. This mixture was then injected into preheated aqueous media, leading to the formation of a milky ethoniosomal dispersion. The resultant dispersion was subjected to bath sonication to ensure uniformity and then stored overnight at 4°C for maturation.
2. Optimization and Characterization: The formulations were optimized through a 2^3 factorial design, varying the amounts of Span 60, ethanol, and cholesterol. The key performance indicators, including entrapment efficiency (EE%) and drug release percentage after 24 hours (Q24h), were measured. The optimized formulation was characterized for size, zeta potential, and elasticity using techniques like dynamic light scattering and extrusion methods.
   

Table 1. Composition of PTS-loaded EN according to 23 Factorial design

Table 2. Obtained data of the 23 factorial design of PTS-loaded ENs

3. In Vitro Studies: Cytotoxicity assays were conducted on A549 lung cancer cell lines to evaluate the effectiveness of the FA-BSA-ethoniosomes. The sulforhodamine B (SRB) assay was used to measure cell viability after treatment with various concentrations of PTS, PTS-loaded ethoniosomes, and FA-BSA-ethoniosomes.
   

Figure 1. In vitro cytotoxicity of free PTS, PTS-loaded ENs, and FA-BSA-ENs of PTS against A-549 lung cancer cell

4. In Vivo Studies: The in vivo efficacy of the formulations was tested using a mouse model of lung cancer induced by N-nitrosodiethylamine (DENA). Mice were divided into treatment groups, receiving targeted FA-BSA-ethoniosomes, non-targeted formulations, or saline. Body weight was monitored, and tumor growth was assessed through histopathological examination of lung tissue samples after the treatment regimen.
   

Figure 2. The change in mice body weight during induction of lung cancer

Data Collection and Analysis
Data were rigorously collected through various methods. In vitro cytotoxicity was quantified using absorbance measurements from the SRB assay, while in vivo tumor reduction was evaluated through histopathological analysis and immunohistochemical staining for caspase-3 to assess apoptosis. Statistical analyses, including ANOVA, were conducted to determine the significance of the findings, ensuring robust interpretation of the results.

Novel Aspects
The study introduced several novel aspects, notably the conjugation of folic acid to bovine serum albumin (BSA) for enhanced targeting of folate receptor-overexpressing lung cancer cells. This modification improves the specificity and efficacy of the ethoniosomal delivery system compared to traditional niosomes, facilitating better cellular uptake and therapeutic impact. The use of the proniosome formulation method also presents advantages, including improved stability and solubility of PTS, which is often hindered in conventional delivery systems. Overall, the innovative methodologies and formulations developed in this research signify a promising advancement in targeted cancer therapy.

Conclusion

The successful development of the folate-bovine serum albumin-coated ethoniosomal delivery system for pterostilbene (PTS) was achieved through a series of methodical steps involving formulation optimization, in vitro characterization, and in vivo efficacy evaluation. The study utilized a 2^3 factorial design to optimize key parameters, resulting in a formulation that demonstrated high entrapment efficiency and sustained drug release, ensuring effective delivery to lung cancer cells.

The highlights of the study include the demonstrated ability of the FA-BSA-ethoniosomal formulation to selectively target folate receptor-overexpressing lung cancer cells, significantly enhancing cellular uptake and therapeutic impact. In vitro cytotoxicity assays showed that the targeted delivery system was more effective than non-targeted counterparts, leading to improved anti-tumor activity. Furthermore, in vivo studies revealed a notable reduction in tumor mass, supporting the potential of this innovative drug delivery approach as a promising strategy for lung cancer treatment. Overall, the findings indicate that folate-conjugated ethoniosomes could serve as a valuable tool in advancing targeted cancer therapies.

Reference

Hanafy, Nemany A. N., et al. “Formulation and Optimization of Folate-Bovine Serum Albumin-Coated Ethoniosomes of Pterostilbene as a Targeted Drug Delivery System for Lung Cancer: In Vitro and In Vivo Demonstrations.” Cancer Nanotechnology, vol. 14, 2023, article 49.