Cancer Nanotechnology: Methods and Protocols

Cancer Nanotechnology: Methods and Protocols
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Buy Softcover. FAQ Policy. About this book Early detection of cancer at the cellular level, even before anatomic anomalies are visible, is critical to more efficacious and cost effective diagnosis and therapeutic advances. Show all. It will be extremely useful for established investigators, students, clinical oncologists, and others interested in the field of cancer nanotechnology. Pages Grobmyer, Stephen R. Carmen et al. Show next xx. Recommended for you. Diagnostic confirmation is only possible by biopsy and histopathological analysis prior to treatment 22 — 28 with possible prior cytological evidence, 29 — 31 and lengthy and expensive diagnostic investigations that only delay the initiation of treatment should be avoided.

Methods and Protocols

Editors: Grobmyer, Stephen R., Moudgil, Brij M. (Eds.) Early detection of cancer at the cellular level, even before anatomic anomalies are visible, is critical to more efficacious and cost effective diagnosis and therapeutic advances. Molecular-Targeted Therapy for Cancer and. This detailed volume presents protocols for advancing the utility of nanotechnology in cancer research toward improving our understanding of cancer biology.

Nevertheless, the delay in the diagnosis of oral cancer has resulted in increasing the time to treatment initiation and a consequent decrease in the survival rate of patients. To increase the effectiveness of treatment and reduce side effects, the incorporation of nanotechnology-based drug delivery systems, such as polymeric nanoparticles PNPs , solid lipid nanoparticles SLNs , nanostructured lipid carriers NLCs , gold nanoparticles, hydrogels, cyclodextrin complexes, and liquid crystals LCs , represents an interesting option for drug delivery, as demonstrated in Table 1.

The genomic pathway plays a role in OSCC, whereby alterations in the genome result in the altered expression of proteins, chemical mediators, and enzymes.

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Due to oncogene activation and tumor suppressor gene inactivation, OSCC leads to the deregulation of cell proliferation and death. The genetic alterations include gene amplification, oncogene overexpression, mutations, deletions, and hypermethylation, leading to the inactivation of particular genes such as the p53 tumor suppressor gene.

Oncogenes do not play an important role in the cancer process, although they do play a role in initiation. Initiation transforms a normal cell into a premalignant cell, and it requires the inactivation of negative regulators in the cell eg, tumor suppressor genes , which is considered an important event that leads to the development of malignancy. Tumor suppressor genes are most often inactivated by point mutations, deletions, and rearrangements in both gene copies. Mutations in p53 and p16 are involved in the carcinogenesis process. The p53 gene plays a role in maintaining genomic stability, cell cycle progression, cellular differentiation, DNA repair, and apoptosis, and p16 is involved in cell cycle regulation, including cell cycle arrest and apoptosis.

Another OSCC characteristic is telomerase activity. Several oral tumors have been confirmed to have the expression of telomerase, which is strongly associated with malignancy in oral tissues. Other chemical mediators are involved in oral cancer pain, such as endothelin-1 ET-1 , proteases, and nerve growth factor. In oral cancer, ET-1 binds to the endothelin-B receptor and is expressed on nonmyelinating Schwann and dorsal root ganglion satellite cells.

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Protease-activated receptor type 2 PAR 2 is involved in oral cancer. This continual release of serine proteases in the microenvironment could produce an ongoing excitation of primary nociceptive afferents, leading to mechanical allodynia in oral cancer patients. In the microenvironment of many cancers, sensory neurons are chronically exposed to nerve growth factor NGF.

The pain mechanism in oral cancer can be established by association with perineural involvement, with invasion and proliferation of a cancer occurring within a nerve associated with pain. Angiogenesis is a crucial step in the processes of uncontrolled tumor proliferation and metastasis, and inhibiting angiogenesis is considered to be effective for treating oral cancer. Cancer cells induce the development of an exaggerated inflammatory state in the stroma, which in turn promotes cancer growth, invasion, and metastasis.

Inflammatory cells in the microenvironment, such as myeloid dendritic cells, macrophage subtypes M1 and M2 , mast cells, neutrophils, and T and B lymphocytes, secrete chemokines, prostaglandins, proteinases, and complement components that collectively adopt an exaggerated inflammatory state that promotes cancer growth, tissue invasion, and metastasis. In addition, IL-8 plays a role in the induction of the angiogenesis process.

A number of complex mechanisms are involved in the genesis and progression of oral cancer. OSSC is a multistep process in which multiple genetic events occur that alter the normal function of oncogenes and tumor suppressor genes. These events can result in the increased production of growth factors.

Recent advances in the understanding of the molecular control of these various pathways will allow for more accurate diagnosis and assessment of prognosis and might lead the way for more novel approaches for treatment and prevention. Treatment protocols for oral cavity cancers are generalized therapies based on stage, chemoradiation therapy, and induction chemotherapy for locally advanced disease. The oral administration of anticancer agents is preferred by patients for its convenience and potential for outpatient treatment.

In addition, oral administration facilitates prolonged exposure to a cytotoxic agent. Intravenous administration leads to immediate and complete bioavailability; thus, this route has the potential to be hazardous because high concentrations of drugs are delivered to normal tissues, causing greater damage to healthy tissues and increased adverse reactions. To overcome the disadvantages of current cancer treatment techniques, the scientific community has turned toward nanotechnology to develop new and more effective nanotechnology-based drug carrier systems to optimize oral, buccal, and intravenous treatment routes.

NPs can improve the stability of drugs and control their targeted delivery, allowing for a constant and uniform concentration at the site of a lesion and facilitating drug extravasation into the tumor system, thus reducing side effects. Damascelli et al evaluated the effectiveness of the intra-arterial infusion of paclitaxel incorporated in NPs based on human albumin albumin NPs for use as induction chemotherapy before definitive advanced tongue cancer treatment. In addition, paclitaxel causes severe allergic reactions with intravenous use.

Albumin NPs are attractive formulations because they can incorporate a significant amount of drugs into a particle matrix due to the different drug-binding sites present in albumin molecules. Damascelli et al reported that the intra-arterial infusion of paclitaxel in albumin nanoparticles is reproducible and effective.

In addition, NARNPs improved the biochemical status to a normal range in 7,dimethylbenz a anthracene-induced oral carcinogenesis. Yu et al also investigated the action of NPs against oral cancer. In another study, the potential antitumor activity of cisplatin-loaded nanoparticles based on PEG-poly glutamic acid block copolymers was assayed in four OSCCs.

The results showed that the growth inhibitory effects of cisplatin-loaded nanoparticles were significantly less than that for free cisplatin.

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However, the caspase-3 and -7 cascades, which are activated by a cisplatin stimulus, induced the release of cytochrome c from mitochondria and led to an irreversible commitment to apoptotic cell death in both cisplatin- and NCtreated OSC cells. Other interesting data obtained from this study revealed that nephrotoxicity, a crucial side effect of cisplatin-loaded nanoparticles, is much lower than that for free cisplatin.

Therefore, it can be interpreted that these NPs are as efficient against OSCC as free cisplatin but with much less renal toxicity. Li et al prepared NPs based on biocompatible and biodegradable hyperbranched poly ether ester polymers that possess many hydroxyl and carboxyl functional groups available for functionalization, including the covalent attachment of drug molecules.

In addition, photosensitizer-loaded nanoparticles can reach the most sensitive subcellular sites, demonstrating a capability for treating superficial oral cancer or precancerous lesions. Nevertheless, some studies revealed that some of the aforementioned polymers may lead to cytotoxicity after internalization into cells, restricting the use of NPs as a drug delivery system. In addition, the large-scale production of PNPs is also problematic and is not relevant for the pharmaceutical market. Therefore, SLNs were developed to overcome the disadvantages of PNPs because they demonstrate physical stability, protection of incorporated labile drugs from degradation, controlled release, and excellent tolerability; thus, they can be used for different routes of administration, such as parenteral, oral, dermal, ocular, pulmonary, and rectal.

SLNs are made from solid lipids at room temperature and are stabilized by surfactant. Holpuch et al tested a SLN formulation as a local oral cancer chemoprevention strategy. Furthermore, the penetration and subsequent internalization of nanoparticles within proliferating basal layer cells demonstrates the feasibility of nanoparticle formulations for local delivery and the stabilization of oral chemopreventive compounds.

However, SLNs have some limitations because the HPH process leads to drug degradation, the coexistence of different lipid modifications and colloidal species, and a low drug-loading capacity, and because of the kinetics of the distribution processes.

Cancer nanotechnology : methods and protocols / edited by Stephen R. Grobmyer and Brij M. Moudgil

To overcome these difficulties, a new generation of SLNs has emerged, ie, NLCs, which consist of solid matrix entrapping variable liquid lipid nanocompartments, as shown in Figure 1. The presence of liquid lipid nanocompartments avoids solid lipid crystallization and improves the drug payload and release because these are still controlled by a surrounding solid lipid barrier. In addition, NLCs had great effects against prostate cancer due to the enhanced intracellular uptake of NLCs by cells.

All NLCs were prepared by an emulsification and low-temperature solidification method. Nanoparticles based on noble metals, particularly gold, have an immense potential for cancer diagnosis and therapy based on their surface-plasmon resonance-enhanced light scattering and absorption.

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El-Sayed et al prepared gold nanoparticles AuNPs by the citrate reduction of chloroauric acid. The results showed that the scattering images and absorption spectra recorded from anti-EGFR antibody-conjugated AuNPs incubated with cancerous and noncancerous cells were different and provided a potential technique for oral cancer diagnostics.

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Kah et al also investigated AuNPs for the early diagnosis of oral cancer based on surface plasmon resonance. Furthermore, the use of gold nanoparticles in surface-enhanced Raman scattering enhanced Raman spectroscopy signals for the analysis of cancer-related chemical changes in saliva. Afifi et al used hamster buccal pouch carcinoma as a model for OSCC to study the effects of plasmonic photothermal therapy using AuNPs combined with visible laser irradiation. The results demonstrated an amplified decrease in proliferation rates for cancer cells upon plasmonic photothermal therapy using AuNPs in addition to maintaining no adverse effects on normal cells, which can be explained by the enhanced permeability and retention effect.

These findings indicate that AuNPs directly injected into hamster buccal pouch carcinomas can be used as a treatment for human OSCC in the future. Liposomes are unilamellar or multilamellar microscopic particles composed of membrane-like lipid layers, often phospholipids and cholesterol, surrounding aqueous compartments, , as depicted in Figure 2. Figure 2 Schematic representation of unilamellar A and multilamellar B liposomes.

Note: the arrow indicates an enlarged view of the outer layers of multilamellar liposomes. Liposomes are the most widely used drug delivery systems for the systemic administration of many drugs for decreasing drug toxicity and increasing their accumulation at target sites. Furthermore, liposome-based formulations for gene therapy, such as synthetic cationic liposomal-DNA called lipoplexes, have clear potential, particularly for oral cancer treatment.

Thus, these results showed that photodynamic therapy mediated by a liposomal formulation of aluminum phthalocyanine chloride can be effective against chemically induced oral cancer. Thus, these results are promising for oral cancer treatment. A hydrogel is a mesh of hydrophilic polymeric chains dispersed in water that is swellable and can release drugs for dissolution and disintegration through the spaces in their mesh, as shown in Figure 3. In addition to swelling, physical properties include permeability, mechanical resistance, and surface aspects that can be modulated through structural modification.

Figure 3 Hydrophilic polymeric chains network and release the drug for dissolution through the spaces of their mesh. Hydrogels are attractive for oral administration because their polymeric chains can closely interact with saliva glycoproteins, causing a mucoadhesion phenomenon. There has been a great deal of interest in the use of hydrogels as chemotherapeutic drug delivery systems for drugs including paclitaxel, doxorubicin, DTX, tamoxifen, and cisplatin.

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MH2 and CMH2 hydrogels were obtained by free radical polymerization of the methacrylate monomer N-methacryloyl-l-histidine and NIPAAm, and the authors assessed the in vitro cytotoxicity of cisplatin-loaded hydrogels. They reported that P9 hydrogels could modulate the rate of cisplatin release.

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Despite these differences, the release profiles were similar for both types of hydrogels, with an initial burst reaching a maximum concentration at approximately 2 to 3 hours. The researchers concluded that hydrogels containing both cross-linking agents can improve the chemical and mechanical properties presented when compared with hydrogels obtained with only one of the reticulating agents, making it attractive for the treatment of oral cancers because the release profile of the system occurs quickly, thus releasing the drug formulation before it is removed from the oral cavity by the salivary flow.

Emoto et al studied hydrogels obtained with cross-linkable hyaluronic acid for the intraperitoneal administration of cisplatin for extended retention and consequent action against peritoneal carcinomatosis. Afterward, ethylene glycol was added to stop the reaction, and the mixture was immediately dialyzed against water. The formation and swelling kinetics of hydrogels and the in vitro release kinetics of cisplatin from hydrogels were studied.

The tests showed that there was sustained cisplatin release within 4 days. The researchers also evaluated the antitumor effects of the intraperitoneal administration of cisplatin-loaded acid hyaluronic hydrogels using a mouse model of gastric cancer. They observed a significant reduction in the weight of the peritoneal nodules in the gel-cisplatin group, whereas no significant reduction was detected in a phosphate-buffered saline-cisplatin group.