The use of biotinylated-EGF-modified gelatin nanoparticle carrier to enhance cisplatin accumulation in cancerous lungs via inhalation
Introduction
Lung cancer is one of the most harmful forms of cancer. The long-term survival rate of lung cancer patients treated by conventional modalities, such as surgical resection, radiation, and chemotherapy remains far from satisfactory. Systemic drug delivery is rarely successful because only a limited dosage of the chemotherapeutic drugs target lung tumor sites, even when administered at a high dose [1]. Most chemotherapeutic drugs act on normal cells, inhibiting their growth; this makes the patient extremely weak and can even result in death.
Cisplatin (cis-dichlorodiammineplatinum (II); CDDP) is a key drug used in the chemotherapy of cancers, including gastrointestinal, genitourinary and lung cancer; cisplatinum-based chemotherapy is accepted as a standard first-line treatment for advanced non-small cell lung carcinoma (NSCLC) [2]. Chemotherapy with cisplatin is associated with various secondary effects, such as anemia, nausea, vomiting, neurotoxicity and nephrotoxicity [3], which impair the patient's quality of life and can even be life-threatening in preexisting conditions. Due to these side effects, alternative methods of administering toxic cisplatin are needed.
For the reduction of side effects, specific drug delivery systems (DDSs) have been investigated. Several polymers, of natural as well as synthetic origin, have been used as reservoirs for delivery of cisplatin, such as polymeric micelles [3], poly(γ,l-glutamic acid)(γ-PGA) [4], polylactic acid (PLA) [5], etc. In this study, gelatin was selected as the raw material to prepare the drug carrier because of its biocompatibility and biodegradability [6], [7]. Its degradation rate can be regulated by the degree of crosslinking to control drug release rate. Konishi et al. showed that a gelatin hydrogel containing CDDP could release the drug over a long period, resulting in a superior anticancer effect [8]. However, there have been few published studies on the formulation of gelatin nanoparticles (GPs) with cisplatin. In this study, we have successfully prepared gelatin nanoparticles complexed with CDDP (GP–Pt).
Another way to reduce the side effects of drugs is active targeting. A tumor-specific ligand can be employed for more specific recognition of and interaction with cancer cells, reducing the effect on normal cells. The overexpression of epidermal growth factor receptor (EGFR) on human tumors, especially on NSCLC, has been reported [9]. In the present study, epidermal growth factor (EGF) was conjugated with the gelatin–cisplatin nanocomplex as a targeting vehicle for lung cancer treatment.
The objective of the present work was to evaluate the anticancer efficiency and specific targeting ability of a cisplatin–gelatin nanomedicine with affinity for EGFR. We developed a gelatin nanoparticle (GP) complex with CDDP (GP–Pt) surface-modified with NeutrAvidinFITC-biotinylated epidermal growth factor (bEGF), abbreviated as GP–Pt–bEGF, to target the tumor site and be taken up by EGFR-mediated endocytosis in tumor cells overexpressing EGFR. In vitro anticancer efficiency was examined by the MTT assay. The in vivo anticancer effect was evaluated by intratumor injection in a mouse subcutaneous model. Aerosol delivery was adopted to allow inhaled chemotherapy for metastatic lung cancer.
Section snippets
Reagent and chemicals
Gelatin type A (derived from porcine skin, bloom 175), cisplatin, Ham's F12k medium, Bicinchoninic Acid (BCA) protein assay kit, bovine serum albumin (BSA), Ethylenediaminetetraacetic acid (EDTA), 1-(4,5-Dimethylthiazol-2-yl)-3,5-diphenyl- formazan (MTT), hematoxylin and eosin were purchased from Sigma–Aldrich (Saint Louis, MO, USA). Fetal calf serum (FCS) was from Biological industries (Kibbutz Beit Haemek, Israel). Trypsin–EDTA, penicillin/streptomycin, and phosphate-buffered saline (PBS)
Results
A schematic representation of the cisplatin with different formulations is represented in Fig. 1. Gelatin nanoparticles (GPs) were prepared by the two desolvation method through acetone addition (Fig. 1(a)). Cisplatin loaded GPs (GP–Pt) were formed via a ligand exchange reaction of Pt(II) from the chloride to the carboxyl group in the GPs (Fig. 1(b)). Finally, surface modification of GP–Pt was performed by NeutrAvidinFITC-biotinylated-EGF conjugation. The EGFR-targeted formulation of CDDP is
Discussion
Many research reports about CDDP delivery systems with polymer materials (such as PGA [4], and PEG-P(Glu) block copolymers [19]) have indicated possible interactions between the carboxyl groups of the polymer and CDDP. The functional groups of the natural polymer gelatin, such as carboxyl, hydroxyl, and amino groups, are available for chemical reaction. Based on their nature, it is conceivable for gelatin to interact with CDDP because it contains many carboxyl groups. The FT-IR spectrum (Fig. 2
Conclusion
A polymer-anticancer drug conjugate, gelatin nanoparticle (GPs), was developed as a carrier of CDDP to enhance its therapeutic effects and reduce its side effects. GP–Pt–bEGF with EGFR affinity produced Pt concentration in cells highly expressing EGFR. The in vitro anticancer study showed that GP–Pt–bEGF was more potent than free CDDP or GP–Pt, due to a rapid onset of action on the cell cycle and a lower IC50 for the inhibition of A549 cell growth. The in vivo anticancer experiment showed that
References (35)
Twenty-five years of treating advanced NSCLC: what have we achieved?
Ann Oncol
(2004)- et al.
Poly(γ,L-glutamic acid)–cisplatin conjugate effectively inhibits human breast tumor xenografted in nude mice
Biomaterials
(2006) - et al.
Preparation and evaluation of drug-loaded gelatin nanoparticles for topical ophthalmic use
Eur J Pharm Biopharm
(2004) - et al.
In vivo anti-tumor effect through the controlled release of cisplatin from biodegradable gelatin hydrogel
J Control Release
(2003) - et al.
Targeting efficiency and biodistribution of biotinylated-EGF-conjugated gelatin nanoparticles administered via aerosol delivery in nude mice with lung cancer
Biomaterials
(2008) - et al.
Preparation of surface modified protein nanoparticles by introduction of sulfhydryl groups
Int J Pharm
(2000) - et al.
Paclitaxel-loaded PLGA nanoparticles: potentiation of anticancer activity by surface conjugation with wheat germ agglutinin
J Control Release
(2005) - et al.
Cisplatin delivery from poly(acrylic acid-co-methyl methacrylate) microparticles
J Control Release
(2005) - et al.
In vivo anti-tumor effect of dual release of cisplatin and adriamycin from biodegradable gelatin hydrogel
J Control Release
(2005) - et al.
Therapeutic efficacy of anti-ErbB2 immunoliposomes targeted by a phage antibody selected for cellular endocytosis
Biochim Biophys Acta
(2002)
Radiosensitization by intratumoral administration of cisplatin in a sustained-release drug delivery system
Radiother Oncol
Paclitaxel liposome aerosol treatment induces inhibition of pulmonary metastases in murine renal carcinoma model
Clin Cancer Res
Cisplatin-incorporating polymeric micelles (NC-6004) can reduce nephrotoxicity and neurotoxicity of cisplatin in rats
Br J Cancer
Anti-tumor effect of cisplatin incorporated into polylactic acid microcapsules
Artif Organs
Gelatin-tricalcium phosphate membrane modified with NGF and cultured schwann cells for peripheral nerve repair: a tissue engineering approch
Biomedical Engineering: Applications, Basis and Communications
Overexpression of the epidermal growth factor receptor and its ligand transforming growth factor alpha is frequent in resectable non-small cell lung cancer but does not predict tumor progression
Clin Cancer Res
Gelatin nanoparticles by two step desolvation – a new preparation method, surface modifications and cell uptake
J Microencapsul
Cited by (241)
Nano-based carriers for pulmonary drug delivery: A review on the available drug delivery applications and toxicity issues
2024, Journal of Drug Delivery Science and TechnologyBiomedical,clinical and environmental applications of platinum-based nanohybrids: An updated review
2023, Environmental ResearchExtracellular matrix component-derived nanoparticles for drug delivery and tissue engineering
2023, Journal of Controlled ReleaseSynthetic beidellite clay as nanocarrier for delivery of antitumor oxindolimine-metal complexes
2023, Journal of Inorganic BiochemistryNanomedicine for targeting the lung cancer cells by interpreting the signaling pathways
2022, Journal of Drug Delivery Science and TechnologyPulmonary delivery of size-transformable nanoparticles improves tumor accumulation and penetration for chemo-sonodynamic combination therapy
2022, Journal of Controlled ReleaseCitation Excerpt :Taking into account that the translocation of nanoparticles depends on particle size with nanocarriers of hydrodynamic size <34 nm being capable of traveling from the lung to the bloodstream and lymph nodes [18], those with sustained release characteristics and particle size >50 nm were used to extend lung retention, achieving enhanced drug concentrations in the lungs [19–22]. Third, due to the fact that inhaled nanomedicines favor direct access of nanoparticles to the lung tumor, surface-engineered nanoparticles have been applied to penetrate and distribute into tumors with or without active tumor site targeting [16,23,24]. It is well documented that nanoparticles with particle size of <30 nm were conductive to tumor penetration [25–27].