Development of Iron-Based Nanoparticles
for Hyperthermia

Introduction

• Developed iron-based nanoparticles (IONPs) for targeted hyperthermia treatment.

• Used a co-precipitation method to synthesize IONPs, achieving a particle size of ~11nm.

• Functionalized IONPs with hyaluronic acid (HA) to enhance biocompatibility and prevent aggregation.

• Characterized IONPs using SEM, TEM, VSM, and XRD techniques.

• Evaluated the heating effects of IONPs and HA-coated IONPs (HA-IONPs) using a radiofrequency (RF) generator.

• Created a tissue-mimicking phantom to test the RF-induced heating in a simulated biological environment.

Situation

• Cancer is a leading cause of death worldwide, and conventional treatments often have adverse side effects.

• Hyperthermia, a minimally invasive treatment using heat to destroy cancer cells, has shown promise.

• IONPs, with their biocompatibility and superparamagnetic properties, are ideal for targeted hyperthermia.

Task

• Synthesize and characterize IONPs suitable for hyperthermia applications.

• Evaluate the RF-induced heating effects of IONPs and HA-IONPs.

• Develop a tissue-mimicking phantom to simulate the biological environment for testing.

Action

• Synthesized IONPs using a co-precipitation method.

• Functionalized IONPs with HA to improve biocompatibility and prevent aggregation.

• Characterized IONPs using SEM, TEM, VSM, and XRD techniques to analyze morphology, size, magnetic properties, and crystallinity.

• Tested the RF-induced heating effects of IONPs and HA-IONPs using a 27 MHz RF generator.

• Created a tissue-mimicking phantom using agarose, sodium chloride, and sucrose to simulate normal and abnormal tissues.

Result

• Successfully synthesized IONPs with an average size of ~11nm.

• HA-IONPs exhibited enhanced biocompatibility and stability compared to bare IONPs.

• HA-IONPs generated a temperature increase of ~10°C under RF induction, suitable for hyperthermia applications.

• The tissue-mimicking phantom showed minimal temperature changes under RF induction, indicating the safety of the technique for surrounding healthy tissues.

Conclusion

• The project successfully demonstrated the feasibility of using IONPs for RF-induced hyperthermia.

• HA-IONPs hold potential for targeted and minimally invasive cancer treatment.

• Further research and development are needed to optimize the technique and evaluate its efficacy in pre-clinical and clinical settings.

Conversion of Compound Microscope into Fluorescence Microscope for Bioimaging

Introduction

• Modified a compound microscope into a fluorescence microscope by adding different excitation sources and emission filters.

• Utilized various LEDs, including white, red, blue, green, and UV, as excitation sources.

• Added a smartphone camera interface for capturing images, enabling on-site analysis and diagnostics.

• Compared images obtained from the modified microscope to those from a high-end microscope, demonstrating comparable quality.

• Successfully imaged plant cells and endothelial cells stained with DAPI fluorophore dye.

• Developed a low-cost and portable fluorescence microscope suitable for educational and research purposes.

Situation

• Conventional fluorescence microscopes are expensive, limiting their accessibility in educational institutions and research labs.

• There is a need for a low-cost and portable fluorescence microscope that can provide comparable image quality to high-end microscopes.

Task

• Convert a compound microscope into a fluorescence microscope.

• Utilize different excitation sources and emission filters for imaging.

• Compare the image quality of the modified microscope with a high-end microscope.

Action

• Modified the eyepiece to hold a smartphone camera and emission filters.

• Used various LEDs and UV light as excitation sources.

• Prepared samples by staining them with DAPI fluorophore dye.

• Captured images using the smartphone camera and compared them to those from a high-end microscope.

Result

• Successfully converted a compound microscope into a fluorescence microscope.

• Achieved comparable image quality to a high-end microscope at a significantly lower cost.

• Demonstrated the feasibility of using a smartphone camera for on-site analysis and diagnostics.

• Successfully imaged plant cells and endothelial cells stained with DAPI.

Conclusion

• The project successfully developed a low-cost and portable fluorescence microscope.

• The modified microscope provides comparable image quality to high-end microscopes, making it suitable for educational and research purposes.

• The use of a smartphone camera enables on-site analysis and point-of-care diagnostics.

Neural Data Analysis for Motor Cortex Recordings

• Preprocessed and sorted neural recordings from a microelectrode array implanted in the primary motor cortex of a macaque monkey during finger movements

• Generated event-aligned raster plots, peristimulus time histograms (PETHs), and developed criteria for decoding finger movements based on neural firing rates.

• Analyzed local field potentials (LFPs) using spectrograms and created tuning curves based on spectral power in specific time-frequency windows.

Wearable for Accurate stress detection

• Develop a wearable device for accurate stress detection using multiple physiological sensors and machine learning algorithms.

• The device will measure cortisol levels, heart rate variability (HRV), skin temperature, and motion data to provide a comprehensive assessment of stress response.

• The data will be processed using advanced machine learning models to classify real-time stress levels and provide personalized feedback to the user.

• The device aims to improve stress awareness and management, potentially benefiting individuals with mental health conditions and promoting overall well-being.

Situation

• Stress is a pervasive issue with significant impacts on mental and physical health.

• Traditional methods for assessing stress are often subjective and unreliable.

• There is a need for accurate and objective stress detection tools that can provide real-time feedback and facilitate personalized stress management.

Task

• Develop a wearable device that integrates multiple physiological sensors to capture a comprehensive stress response.

• Develop a robust data processing pipeline and machine learning algorithms to accurately classify stress levels.

• Design a user-friendly interface to provide real-time feedback and facilitate personalized stress management.

Action

• Develop flexible sensors for cortisol, HRV, temperature, and motion.

• Design and fabricate a wearable device prototype.

• Collect pilot data from individuals under various stress conditions.

• Develop and train machine learning models for stress classification.

• Design and implement a user interface for real-time feedback.

Challenges and Proposed Solutions

• Accurate and reliable cortisol sensing in sweat.

• Solution: Explore novel cortisol sensing materials and optimize sensor design for sensitivity and stability.

• Effective sensor fusion and machine learning model development.

• Solution: Investigate advanced machine learning techniques and optimize feature extraction for robust stress classification.

• User-friendly interface design for personalized feedback.

• Solution: Conduct user experience (UX) research and design an intuitive interface that provides clear and actionable stress insights.

Result (Expected)

• A functional wearable device prototype capable of accurately detecting stress levels in real-time.

• A robust data processing pipeline and machine learning model for personalized stress classification.

• A user-friendly interface that provides real-time feedback and facilitates stress management.

3D-Printed Gold Nanoparticle (AuNP) for Localized Surface Plasmon Resonance (LSPR) Based Detection for Biosensing Application.

• This project utilizes gold nanoparticles (AuNPs) for biosensing, focusing on integrating them with 3D printing technology.

• Contrasts 3D printing with conventional thin-film technologies, which are labor-intensive and require high temperatures and controlled environments.

• 3D printing offers precise control over shape, geometry, and thickness, making it a user-friendly, cost-effective, and environmentally friendly alternative.

• AuNPs are chosen for their stability, biocompatibility, and ease of surface functionalization, especially in localized surface plasmon-based biosensors.

• Localized surface plasmon resonance (LSPR)-based biosensors rely on changes in the dielectric properties of the local environment, enhancing sensitivity.

• The project involves establishing 3D printing fundamentals, exploring various Au salts and polymer ligands to optimize the process.

• Characterization of 3D-printed thin films involves ellipsometry, AFM, and SEM measurements, with post-deposition annealing and O2 plasma treatment to enhance film quality and sensitivity.

• The functionalized thin film will be used for dsDNA detection using LSPR sensing to demonstrate proof of concept.

Situation

• Conventional thin-film technologies are labor-intensive, expensive, and environmentally unfriendly.

• There is a need for a low-cost, environmentally friendly, and user-friendly alternative for producing high-quality thin films.

• AuNPs have exceptional properties for biosensing applications, including stability, biocompatibility, and ease of surface functionalization.

Task

• Develop a novel 3D printing setup and method for high-throughput, low-cost fabrication of complex structures and thin films.

• Develop a reliable method for the mass-scale 3D printing of AuNP-based thin films.

• Develop an effective method for the functionalization of 3D-printed AuNP-based thin films.

• Establish the optical setup for LSPR sensing and demonstrate its reliability.

• Demonstrate the LSPR sensing reliability by detecting dsDNA using functionalized 3D-printed AuNP-based thin films.

Action

• Establish the fundamentals of 3D printing, focusing on metal precursors, neutral polymer ligands, and reducing agents.

• Explore various Au(I/II/III) salts and polymer ligands to optimize the 3D printing process.

• Characterize the 3D-printed thin films using ellipsometry, AFM, and SEM measurements.

• Perform post-deposition annealing and O2 plasma treatment to enhance film quality and sensitivity.

• Functionalize the 3D-printed AuNP-based thin films using a biotin-streptavidin conjugate.

• Establish the optical setup for LSPR sensing and demonstrate its reliability for dsDNA detection.

Result

• Successfully developed a novel 3D printing setup and method for fabricating AuNP-based thin films.

• Achieved mass-scale 3D printing of AuNP-based thin films with high reliability and consistency.

• Developed an effective method for functionalizing 3D-printed AuNP-based thin films using a biotin-streptavidin conjugate.

• Successfully established the optical setup for LSPR sensing and demonstrated its reliability for dsDNA detection.

Conclusion

• The project successfully developed a new, low-cost, high-throughput platform for LSPR-based biosensing applications.

• This platform has the potential to serve as a point-of-care diagnostic tool for various diseases, including cancer, Alzheimer's, and infectious diseases.

• The project's success promises to revolutionize biosensing through innovative 3D printing technology.

Natural Killer Cells Incubated With Hyaluronic Acid-Functionalized Superparamagnetic Iron Oxide Nanoparticles for Prostate Cancer

• Prostate cancer remains a significant global health burden, necessitating novel targeted therapies.

• This project proposes combining natural killer (NK) cell immunotherapy with hyaluronic acid (HA)-functionalized superparamagnetic iron oxide nanoparticles (SPIONS) for targeted prostate cancer treatment.

• NK cells possess inherent tumor-homing and cytotoxic properties.

• HA offers tumor-targeting capabilities by binding to CD44, overexpressed on prostate cancer cells.

• SPIONS provide magnetic targeting and hyperthermia potential.

• This strategy aims to create a synergistic and effective therapeutic approach by combining these elements.

Situation

• Prostate cancer is a leading cause of cancer-related deaths globally, with a significant impact on patients' quality of life.

• Current treatments have limitations, including side effects and the development of resistance.

• NK cell immunotherapy holds promise but faces challenges in tumor infiltration and the immunosuppressive tumor microenvironment.

Task

• Synthesize and characterize HA-SPIONs.

• Evaluate their targeting of NK cells in vitro.

• Assess the in vivo tumor-targeting and therapeutic efficacy of HA-SPION-labeled NK cells in prostate cancer models.

Action

• Synthesize HA-SPIONs using co-precipitation of iron salts and coat with HA using EDC/NHS chemistry.

• Optimize HA density on SPIONs for maximum CD44 binding.

• Characterize HA-SPIONs using DLS, TEM, ICP-MS, and VSM.

• Isolate NK cells from healthy donors and prostate cancer patients.

• Assess HA-SPION binding, uptake, and effects on NK cell viability, phenotype, and function.

• Establish subcutaneous and orthotopic prostate tumor models in mice.

• Inject HA-SPION-labeled or unlabeled NK cells and apply an external magnetic field.

• Monitor NK cell biodistribution and tumor infiltration using MRI and flow cytometry.

• Assess tumor growth and metastasis using bioluminescence imaging and histology.

• Evaluate survival and toxicity in treated mice.

Result

• Successfully synthesized HA-SPIONs with optimal size, HA density, and magnetic properties.

• HA-SPION-labeled NK cells exhibited enhanced cytotoxicity against prostate cancer cells in vitro.

• In vivo studies demonstrated enhanced tumor homing and retention of HA-SPION-labeled NK cells.

• Targeted NK cell therapy resulted in improved tumor suppression, reduced metastasis, and prolonged survival.

Conclusion

• The project successfully developed a novel approach combining NK cell immunotherapy with HA-functionalized SPIONS for targeted prostate cancer therapy.

• This strategy has the potential to overcome limitations of current treatments and improve patient outcomes.

• The findings may have broader implications for treating other cancers where CD44 is overexpressed.

Development of Microfluidic Devices for Studying Organ Transplant Rejection

• Organ transplantation is a life-saving treatment for end-stage organ failure, but long-term graft survival remains a challenge due to complex immune-mediated rejection processes.

• Understanding the intricate interactions between recipient immune cells and donor cells is crucial for developing effective immunosuppressive therapies.

• Current in vitro methods often fail to capture the dynamic nature of these interactions and the influence of the microenvironment.

• This project proposes the development of a novel microfluidic device that mimics the capillary-like microenvironment, allowing real-time monitoring of immune cell-donor cell interactions in a physiologically relevant setting.

• The device will feature an endothelial cell-lined lumen surrounded by a layer of donor cells, enabling the infusion of recipient immune cells and the introduction of immunosuppressive drugs to study their effects on cellular interactions.

Situation

• Organ transplantation is a life-saving intervention for patients with end-stage organ failure, but long-term success is hindered by immune-mediated rejection.

• There is a pressing need for targeted strategies to prevent rejection and enhance graft survival.

• Conventional in vitro methods fail to capture the dynamic nature of immune cell interactions and the impact of the microenvironment.

• Animal models, while offering valuable insights, do not always accurately mimic human physiology and immunology.

• Microfluidics has emerged as a powerful tool for investigating complex biological processes, providing precise control over the cellular microenvironment and enabling real-time visualization of cell-cell interactions.

Task

• Develop a novel microfluidic device that replicates the vascularized microenvironment of a transplanted organ.

• Study immune cell-donor cell interactions in a physiologically relevant context.

• Allow real-time monitoring of dynamic cellular interactions and assessing immunosuppressive drug effects.

• Gain insights into the mechanisms underlying transplant rejection and contribute to developing targeted immunomodulatory strategies to reduce rejection and improve long-term graft survival.

Action

• Design and fabricate a microfluidic device with a central lumen lined with endothelial cells and a surrounding layer of donor cells.

• Seed primary human endothelial cells and donor cells in the device to create a physiologically relevant microenvironment.

• Infuse recipient immune cells (peripheral blood mononuclear cells) through the lumen to study their interactions with donor cells.

• Introduce immunosuppressive drugs into the device to investigate their effects on cellular interactions and graft survival.

• Monitor cellular interactions in real-time using live-cell imaging techniques.

• Analyze the effects of immunosuppressive drugs on cell-cell interactions and the overall cellular microenvironment.

Result

• Successfully developed a microfluidic device that mimics the vascularized microenvironment of a transplanted organ.

• Achieved real-time monitoring of immune cell-donor cell interactions in a physiologically relevant setting.

• Gained insights into the dynamic cellular processes involved in transplant rejection.

• Evaluated the effects of immunosuppressive drugs on cellular interactions and the microenvironment.

• Identified potential therapeutic targets and approaches for controlling organ rejection.

Conclusion

• The project successfully developed a novel microfluidic platform for studying immune cell interactions in organ transplantation.

• The device provides a valuable tool for investigating transplant rejection mechanisms and facilitating the development of targeted immunomodulatory approaches.

• The insights gained from this study contribute to improving long-term graft survival and benefiting patients in need of organ transplants.