This conference presentation was prepared for the Optical Methods for Tumor Treatment and Detection: Mechanisms and Techniques in Photodynamic Therapy XXXI conference at SPIE BiOS, SPIE Photonics West 2023.
Quantitative phase Microscopy is an advantageous technique to retrieve 3D cell information to estimate cell dry mass and morphology. A popular method is to implement this setup in a common path configuration because it is a system robust to vibrations, and it combines phase contrast imaging with phase shifting techniques to provide high-accuracy measurements. However, this approach requires using a spatial light modulator and at least three images to calculate the phase. In our approach, we leverage the optical anisotropy of liquid crystal materials and a polarized camera to obtain four phase-shifted images simultaneously.
KEYWORDS: Breast cancer, Photodynamic therapy, Nanoparticles, In vitro testing, In vivo imaging, Mouse models, Therapeutic agents, Biomedical optics, Cancer, Safety
Multifunctional hybrid nanoparticles are being developed to carry a wide variety of therapeutic and imaging agents for multiple biomedical applications. Polysilsesquioxane (PSilQ) nanoparticles is a promising hybrid platform with numerous advantages to be used as delivery system for photodynamic therapy. In this work, we developed a redox-responsive PSilQ-based platform to transport and deliver simultaneously protoporphyrin IX (PpIX) and curcumin inside human cells. These PSilQ nanoparticles contain a high loading of PpIX (24.4 ± 2.5 %wt) and curcumin (7.6 ± 1.5 %wt), and are biodegraded inside cancer cells due to the high reducing environment. This multimodal delivery system shows a synergistic performance for the combined photo- and chemotherapy of the triple-negative breast cancer (TNBC) MDA-MB-231 cells. The safety and phototherapeutic efficacy of this PSilQ-based platform was evaluated in an orthotopic mice model of TNBC. The PSilQ nanoparticles are completely biodegraded and excreted from mice without any side effect. The efficacy data show that the PSilQ nanoparticles efficiently reduce tumor growth in the orthotopic mice model of TNBC. This work demonstrates that PSilQ nanoparticle-based platform is an excellent alternative for the combined photo- and chemotherapy of TNBC.
Photodynamic therapy (PDT) is a technique that combines light’s interaction with a photoactive substance to promote cellular death and that has been used to treat a wide range of maladies. Cancer is among the leading causes of death worldwide and has been a central issue assessed by PDT research and clinical trials over the last 35 years, but its efficiency has been hampered by photosensitizer buildup at treatment site. Nanotechnology has been addressing drug delivery problems by the development of distinct nanostructured platforms capable of increasing pharmacological properties of molecules. The association of nanotechnology’s potential to enhance photosensitizer delivery to target tissues with PDT’s oxidative damage to induce cell death has been rising as a prospect to optimize cancer treatment. In this study, we aim to verify and compare the efficiency of PDT using redox-responsive silica-based nanoparticles carrying protoporphyrin IX (PpIX) in vitro, in both tumor and healthy cells. Dose-response experiments revealed the higher susceptibility of murine melanoma cells (B16-F10 cell line) to PDT (630 nm, 50 J/cm2) when compared to human dermal fibroblasts (HDFn): after 24 h of incubation with 50 μg/mL nanoparticles solutions, approximately 80 % of B16- F10 cells were killed, while similar results were obtained in HDFn cultures when solutions over 150 μg/mL were used. Uptake and ROS generation assays suggest increased nanoparticle internalization in the tumor cell line, in comparison with the healthy cells, and greater ROS levels were observed in B16-F10 cells.
KEYWORDS: Silica, Nanoparticles, Breast cancer, Near infrared, Optical imaging, Tissue optics, In vivo imaging, Transparency, Electromagnetism, Molecules, Tumors, Nanoprobes, Luminescence
The development of novel methods for tumor detection is a burgeoning area of research. In particular, the use of silica nanoparticles for optical imaging in the near infrared (NIR) represents a valuable tool because their chemical inertness, biocompatibility, and transparency in the ultraviolet-visible and NIR regions of the electromagnetic spectrum. Moreover, silica nanoparticles can be modified with a wide variety of functional groups such as aptamers, small molecules, antibodies and polymers. Here, we report the development of a mucin 1(MUC1)-specific dye-doped NIR emitting mesoporous silica nanoparticles (MUC1-NIR-MSN) platform for the optical detection of breast cancer tissue overexpressing human tumor-associated MUC1. We have characterized the structural properties and the in vitro performance of this system. The MSN-based optical imaging probe is non-cytotoxic and targets efficiently murine mammary epithelial cancer cells overexpressing human MUC1. Finally, the ability of MUC1-NIR-MSN contrast imaging agent to selectively detect breast cancer tumors overexpressing human tumor-associated MUC1 was successfully demonstrated in a transgenic murine mouse model. The NIR imaging experiments on tumor-bearing animals showed specific accumulation of the MSN-based probe in human MUC1-positive tumors and small signal in control tumors. We envision that this MUC1-specific MSN-based optical probe has the potential to greatly aid in screening prospective patients for early breast cancer detection and in monitoring the efficacy of drug therapy.
Photodynamic therapy (PDT) has emerged as an alternative approach to chemotherapy and radiotherapy for cancer
treatment. The photosensitizer (PS) is perhaps the most critical component of PDT, and continues to be an area of
intense scientific research. Traditionally, PS molecules like porphyrins have dominated the field. Nevertheless, these PS
agents have several disadvantages, with low water solubility, poor light absorption, and reduced selectivity for targeted
tissues being some of the main drawbacks. Polysilsesquioxane (PSilQ) nanoparticles provide an interesting platform for
developing PS-loaded hybrid nanocarriers. Several advantages can be foreseen by using this platform such as carrying a
large payload of PS molecules; their surface and composition can be tailored to develop multifunctional systems (e.g.
target-specific); and due to their small size, nanoparticles can penetrate deep into tissues and be readily internalized by
cells. In this work, porphyrin-loaded PSilQ nanoparticles with a high payload of photosensitizers were synthesized,
characterized, and applied in vitro. The network of this nanomaterial is formed by porphyrin-based photosensitizers
chemically connected via a redox-responsive linker. Under reducing environment such as the one found in cancer cells
the nanoparticles can be degraded to efficiently release single photosensitizers in the cytoplasm. The platform was
further functionalized with polyethylene glycol (PEG) and folic acid as targeting ligand to improve its biocompatibility
and target specificity toward cancer cells overexpressing folate receptors. The effectiveness of this porphyrin-based
hybrid nanomaterial was successfully demonstrated in vitro using MDA-MB-231 breast cancer cell line.
In recent years the use of nanoparticles in medical applications has boomed. This is because the various applications that provide these materials like drug delivery, cancer cell diagnostics and therapeutics [1-5]. Biomedical applications of Quantum Dots (QDs) are focused on molecular imaging and biological sensing due to its optical properties. The size of QDs can be continuously tuned from 2 to 10 nm in diameter, which, after polymer encapsulation, generally increases to 5 – 20 nm diminishing the toxicity. The QDs prepared in our lab have a diameter between 2 to 7 nm. Particles smaller than 5 nm can interact with the cells [2]. Some of the characteristics that distinguish QDs from the commonly used fluorophores are wider range of emission, narrow and more sharply defined emission peak, brighter emission and a higher signal to noise ratio compared with organic dyes [6]. In this paper we will show our progress in the study of the interaction of quantum dots in live cells for image and Raman spectroscopy applications. We will also show the results of the interaction of quantum dots with genomic DNA for diagnostic purposes.
This work reports the synthesis, structural characterization, and optical properties of ZrO2:Yb3+-Er3+ (2–1 mol%) nanocrystals. The nanoparticles were coated with 3-aminopropyl triethoxysilane (APTES) and further modified with biomolecules, such as Biotin-Anti-rabbit (mouse IgG) and rabbit antibody-AntiKi-67, through a conjugation method. The conjugation was successfully confirmed by Fourier transform infrared, zeta potential, and dynamic light scattering. The internalization of the conjugated nanoparticles in human cervical cancer (HeLa) cells was followed by two-photon confocal microscopy. The ZrO2:Yb3+-Er3+ nanocrystals exhibited strong red emission under 970-nm excitation. Moreover, the luminescence change due to the addition of APTES molecules and biomolecules on the nanocrystals was also studied. These results demonstrate that ZrO2:Yb3+-Er3+ nanocrystals can be successfully functionalized with biomolecules to develop platforms for biolabeling and bioimaging.
Photodynamic therapy (PDT) has emerged as an alternative approach to chemotherapy and radiotherapy for cancer treatment. The photosensitizer (PS) is perhaps the most critical component of PDT, and continues to be an area of intense scientific research. Traditionally, PS molecules (e.g. porphyrins) have dominated the field. Nevertheless, these PS agents have several disadvantages, with low water solubility, poor light absorption and reduced selectivity for targeted tissues being some of the main drawbacks. Polysilsesquioxane (PSilQ) nanoparticles are crosslinked homopolymers formed by the condensation of functionalized trialkoxysilanes or bis(trialkoxysilanes). We believe that PSilQ particles provide an interesting platform for developing PS nanocarriers. Several advantages can be foreseen by using this platform such as carrying a large payload of PS molecules; their surface and composition can be tailored to develop multifunctional systems (e.g. target-specific); and due to their small size, nanoparticles can penetrate deep into tissues and be readily internalized by cells. In this work, PSilQ nanoparticles with a high payload of photosensitizers were synthesized, characterized, and applied in vitro. The network of this nanomaterial is formed by protoporphyrin IX (PpIX) molecules chemically connected via a redox-responsive linker. Under reducing environment such as the one found in cancer cells the nanoparticles can be degraded to efficiently release single photosensitizers in the cytoplasm. The phototoxicity of this porphyrin-based PSilQ nanomaterial was successfully demonstrated in vitro using human cervical (HeLa) cancer cells. We envision that this platform can be further functionalized with polyethylene glycol (PEG) and targeting ligands to improve its biocompatibility and target specificity.
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