Gold nanorods (GNRs) have the potential for cancer treatment as tumor-targeting photosensitizers due to their strong absorption of near-infrared light. The purpose of this project was to study the subcellular localization of GNRs in cancerous to optimize cancer treatment using irradiation at appropriate wavelength. Subcellular localization of gold nanorods has been shown to have a significant impact on the retention or exclusion of GNRs within the cells as well as cell viability and morphology. GNRs were functionalized with a fluorescent dye, incubated with cells, and then imaged using a fluorescent microscope. Our results demonstrated the localization of GNRs in cancer cell mitochondria and lysosomes, which corroborates the findings of other studies. Further studies will determine the functions of the subcellular GNRs under irradiation of laser of appropriate wavelengths. Our research could lead to a targeted, effective nanotechnology based laser immunotherapy.
We here report on the synthesis and characterization of two nanoparticles focused on cancer treatment via laser immunotherapy (LIT), which is a novel cancer treatment modality with the ability to eradicate primary tumors directly and induce an immune response to destroy distant metastases. This method makes use of a photosensitizing agent and an immunoadjuvant to aide in the efficacy and targeting of a laser treatment. In consideration of ways to bring those materials to the tumor site, we investigated the use of various nanoparticles as targeted delivery agents. Due to the tendency of cancerous cells to overexpress low-density lipoprotein receptors (LDLR) to fulfill their heightened need for cholesterol, as well as due to the previously documented ability of low-density lipoproteins (LDL) to be reconstituted with other materials, these naturally occurring nanoparticles served as the base to be combined with reduced graphene oxide (rGO) and form a therapeutic nanocompound dubbed rGO-LDL. (R)-9bMS, a small molecule inhibitor, is known to significantly hinder the proliferation of triple-negative breast cancer cells—a tumor model particularly resistant to treatment. For this reason, (R)-9bMS was selected, and the photosensitizing agent, indocyanine green (ICG) was combined with it into a nanoparticle geared towards use in LIT.
Metastases are the cause of more than 90 percent of cancer-related deaths. Current treatment methods, including chemotherapy, radiation, and surgery, fail to target the metastases effectively. One potential treatment for metastatic cancer is laser immunotherapy (LIT). LIT combines the use of a photothermal laser with an immunoadjuvant, Glycated Chitosan (GC). GC combined with single-walled carbon nanotubes (SWNTs) has proven to be a viable alternative to traditional cancer treatment methods, when under irradiation of laser with appropriate wavelength. In this study, the effects of low dose and high dose laser irradiation on metastatic pancreatic cancer cell migration were observed. It was found that low dose irradiation increased the migration rate, but the high dose irradiation significantly decreased the migration rate of the cancer cells. When using LIT, the goal is to kill tumor cells and to prompt the correct immune response. If the tumor were irradiated with a low dose, it would promote metastasis. If the dose of irradiation were too high, it would destroy the entire tumor and the immune response would not recognize the tumor. Therefore, the laser dose plays an important role in LIT, particularly when using SWNT as light absorbing agent. Our results from this study will delineate the optimal laser irradiation dose for destroying tumor cells and at the same time preserve and release tumor antigens as a precursor of antitumor immune response.
The ability to modulate myofibroblast phenotype will have an impact in wound healing, aging, cancer, and Dupuytren’s contracture development. Our project focused on investigating the effect of 808 nm laser wavelength on the presence of myofibroblast and contraction using anchored delayed release collagen lattice models. Fibroblast cells were cultured and suspended in collagen lattices in the presence or absence of glycated chitosan (GC) to mimic a wound healing environment in-vitro. The cells were allowed to generate maximum tension for 6 days followed by laser light stimulation. The total number of cells and presence of fibroblast proliferation and differentiation into myofibroblasts was determined using immunostaining followed by fluorescent microscope photography. Lattice tension generation was assessed by releasing the lattices from their anchorage and measuring their diameters. Control lattices contracted more than GC lattices. Myofibroblast differentiation was slightly higher in the control group. Repeats of the experiment are needed to confirm our results to determine if near infra-red laser radiation can be applied to treat open wounds and study tumor stromas.
Non-invasive laser immunotherapy (NLIT) is a viable alternative to traditional cancer treatment because it combines the photothermal and immunological effects of non-invasive laser irradiation and single-walled carbon nanotubes (SWNT) with an immunoadjuvant, glycated chitosan (GC). This combination forms SWNT-GC, a photosensitive immunoadjuvant, which creates a tumor-specific immunity that targets both the primary tumor and any metastasis. It is known that NLIT induces anti-tumor as well as anti-metastatic immune responses, but its immunological mechanism is not clear. The objective of this study is to clarify the role of SWNT-GC in cancer cell migration. Panc02 (non-metastatic) and Panc02-H7 (metastatic) pancreatic cancer cells were used in two-dimensional elastomer plug assays to observe the restriction of cell migration induced by SWNT, GC, and SWNT-GC individually. To replicate a three-dimensional in vivo study, a similar assay was repeated using embedded collagen lattices. Both the 2D and the 3D studies confirmed previous results indicating that GC inhibits cancer cell motility. The 2D and 3D studies also showed that SWNT-GC inhibited the migration of cancer cells, but a discrepancy was observed regarding the effect of SWNT alone. The 2D model concluded that SWNT inhibited migration while the 3D model determined that SWNT promoted migration. The results of this study will guide future work to determine the mechanism behind NLIT, including how metastases are eradicated and how the tumor specific immunity is created.
KEYWORDS: Laser irradiation, Cancer, Single walled carbon nanotubes, In vitro testing, Resistance, Optical properties, In vivo imaging, Tumors, Biomedical optics, Medical research, Biology
Laser immunotherapy (LIT) uses laser irradiation and immunological stimulation to target all types of metastases and creates a long-term tumor resistance. Glycated chitosan (GC) is the immunological stimulant used in LIT. Interestingly, GC can act as a surfactant for single-walled carbon nanotubes (SWNTs) to immunologically modify SWNTs. SWNT-GC retains the optical properties of SWNTs and the immunological functions of GC to help increase the selectivity of the laser and create a more optimal immune response. One essential aspect of understanding this immune response is knowing how laser irradiation affects cancer cells’ ability to metastasize. In this experiment, a cell migration assay was performed. A 2mm circular elastomer plugs were placed at the bottom of multi-well dishes. Pre-cancerous keratinocytes, different tumor cells, and fibroblasts were then plated separately in treated wells. Once the cells reached 100% confluence, they were irradiated by either a 980nm or 805nm wavelength laser. The goal was to determine the effects of laser irradiation and immunological stimulation on cancer cell migration in vitro, paying the way to understand the mechanism of LIT in treating metastatic tumors in cancer patients.
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