Emerging methods in therapeutics using multifunctional nanoparticles

Clinical translation of nanoparticle‐based drug delivery systems is hindered by an array of challenges including poor circulation time and limited targeting. Novel approaches including designing multifunctional particles, cell‐mediated delivery systems, and fabrications of protein‐based nanoparticles have gained attention to provide new perspectives to current drug delivery obstacles in the interdisciplinary field of nanomedicine. Collectively, these nanoparticle devices are currently being investigated for applications spanning from drug delivery and cancer therapy to medical imaging and immunotherapy. Here, we review the current state of the field, highlight opportunities, identify challenges, and present the future directions of the next generation of multifunctional nanoparticle drug delivery platforms.This article is categorized under:Biology‐Inspired Nanomaterials > Protein and Virus‐Based StructuresNanotechnology Approaches to Biology > Nanoscale Systems in BiologyNovel approaches in designing nanoparticles to overcome challenges faced by traditional nanoparticle‐based drug delivery systems.Peer Reviewedhttps://deepblue.lib.umich.edu/bitstream/2027.42/155963/1/wnan1625.pdfhttps://deepblue.lib.umich.edu/bitstream/2027.42/155963/2/wnan1625_am.pd

Functional Nanomaterials in Biomedicine

The great success of nanotechnology promotes a tremendous revolution in the biomedical field. Functional nanomaterials have been widely applied for the treatment of various diseases, such as cancer, bacterial infection, diabetes, inflammation, and neurodegenerative disorders. Various therapeutic nanoplatforms have been developed with therapeutic functions and intelligent properties. However, the development of nanomedicine suffers from several challenges prior to their clinical applications. For instance, disease detection in an early stage is a critical challenge for nanomedicine. It is difficult to detect disease markers (e.g., proteins, genes, or cancer circulating cells), so nanoprobes with high sensitivity and selectivity are required. Moreover, to overcome drug resistance, it is highly desirable to develop functional nanomedicines with the combination of multiple therapeutic modalities, such as chemotherapy, photothermal therapy, photodynamic therapy, chemodynamic therapy, radiotherapy, starving therapy, and immunotherapy. Additionally, the stability and degradability of most nanomedicines in biofluids should be carefully evaluated before their administration to humans. This book provides researchers with the latest investigations and findings in this field

Biological applications of multimodal imaging involving Raman and 4Pi Raman microscopy

Raman microscopy is becoming an increasingly important label-free imaging technique. It proved to be a viable tool for life science applications allowing to analyze bacteria, cells, and tissues at the molecular level. Combining Raman microscopy with complementary imaging modalities and techniques is explored here to: (1) analyze mild traumatic brain injury (mTBI) in a combination with magnetic resonance imaging (MRI) for detecting mild, and invisible to medical imaging techniques, brain tissue damage; (2) reveal complementarity of Raman and fluorescence microscopy approaches for investigating and tracking bovine lactoferrin inside calf rectal epithelial cells in the presence of enterohemorrhagic Escherichia coli (EHEC); (3) apply Raman microscopy along-side the molecular analysis approaches (such as scanning transmission electron microscopy-energy dispersive X-ray (STEM-EDX), low energy X-ray fluorescence (LEXRF), nanoscale secondary ion mass spectrometry (Nano-SIMS)) to uncover the origin of the long-range conductance in cable bacteria; (4) develop multifunctional surface enhanced Raman scattering (SERS) platform based on calcium carbonate particles for enhancing a weak Raman scattering signal of biomolecules as well as to apply Raman microscopy for particle detection in vivo in Caenorhabditis elegans (C. elegans) worms; and (5) combine Raman microscopy and atomic force microscopy (AFM) to track Chlamydia psittaci in cells. Analysis of described above samples and phenomena is based on Raman molecular fingerprint images, where, similarly to fluorescence light microscopy, the resolution is limited by diffraction of light. Therefore, efforts are also put to enhance the resolution of Raman microscopy-based imaging by adding a 4Pi configuration to a confocal Raman microscope. As a result, a possibility to enhance the axial (also called longitudinal) resolution is investigated by constructing a 4Pi confocal Raman microscope, which is also applied to study bacteria inside cells. Results presented in this work emphasize the added value of multimodal microscopy approaches, particularly involving Raman microscopy, in a broad range of applications in bioengineering, biomedicine, and biology

Block Copolymer Based Magnetic Nanoclusters for Cancer-Theranostics: Synthesis, Characterization and In Vitro Evaluation

“There is plenty of room at the bottom”. In this visionary lecture in 1959 Prof. Richard Feynman spoke of the interesting ramifications of working with matter at the atomic scale. Since then, scientists have worked relentlessly towards realizing his vision. The influence of nanobiotechnology on material science and polymer chemistry has given rise to a new field called ‘theranostics’, combining drug delivery and diagnostics within the same nanostructures, thereby enabling simultaneous diagnosis, targeted drug delivery and continued therapy monitoring. Iron oxide nanoparticles (MNPs) are one such class of MRI contrast agents that can be converted into theranostic nanomedicines for cancer therapy. However, development of a stable theranostic contrast system comprising of MNPs is complex and requires a careful balance between the therapeutic diagnostic components. We explored the potential of biodegradable hydrophilic block ionomers such as anionic poly (glutamic acid-b-ethylene glycol) and cationic poly (l-lysine-b-ethylene glycol) in formulating stable magnetic nanoclusters (MNCs). These MNCs were extensively characterized for their composition, colloidal stability and factors influencing their MRI capability. Extensive in vitro studies revealed that the anionic cisplatin-loaded MNCs showed minimal non-specific uptake, a highly preferred feature for targeted cancer therapy. Luteinizing hormone releasing hormone receptor (LHRHr) targeting significantly enhanced the uptake of these formulations in LHRHr-positive ovarian cancer cells. LHRHr targeting also helped improve the theranostic efficacy in cisplatin resistant ovarian cancer cells. One the other hand, cationic MNCs were used to demonstrate the potential of MNCs to function as stimuli-responsive theranostic systems capable of releasing the payload in the acidic milieu breast and ovarian cancer cells. These cationic MNCs also exhibited significantly enhanced T2-weighted MRI contrasts at much lower concentrations than the anionic counterparts. Finally, we successfully evaluated the feasibility of kinetically controlled flash nanoprecipitation technique using multi-inlet vortex mixer (MIVM) to formulate well-defined MNCs from non-ionic amphiphilic Pluronic tri-block copolymers. In comparison to self-assembly techniques, flash nanoprecipitation resulted in significant reduction in polydispersity. It was observed that the hydrophobic block-length of the copolymer dictates the extent of encapsulation hydrophobic therapeutic agents along with the MNPs. exhibited the potential to function as both T1 and T2 contrast agents. In summary, looking at the bigger picture, the work presented here emphasizes on the importance of product development in establishing a critical balance between the therapeutic and imaging functionalities when designing an efficient targeted theranostic nanosystems

New tools for visualising nanoparticle drug delivery

Encapsulating drugs in polymeric nanoparticles (NPs) is becoming increasingly popular for targeted and sustained drug delivery. NP drug delivery systems can increase the lifetime of therapeutics in vivo, they can improve safety by allowing a lower dose to be administered, and they are able to pass the blood brain barrier. Chapter 1 will discuss the anatomy of the brain, how it can be affected by multiple sclerosis (MS) and why NP drug delivery has an important part to play in treating this disease. Due to the nanoscale size of these drug delivery systems, it is challenging to image their uptake, distribution and fate in a biological environment. Raman scattering is a vibrational technique which can probe the chemical bonds in a sample, however, it is a very weak effect. Stimulated Raman scattering (SRS) is a dual laser technique which increases the observed Raman signal and has been used to image biological samples at video-rate. Intracellular contrast can be increased further by introducing spectroscopically bioorthogonal chemical labels to the NPs, which appear in the so called cell-silent region of the Raman spectrum. Previous work to image NPs with fluorescence and Raman microscopies will be discussed in Chapter 1. In this thesis, Raman spectroscopy was used to image bioorthogonally labelled polymeric NPs in in vitro cellular and ex vivo tissue models of the brain. The polymer poly(lactic acid-co-glycolic acid) (PLGA) was chosen as a biocompatible and biodegradable polymer widely used in drug delivery. Chapter 2 describes the synthesis of PLGA with both carbon-deuterium and alkyne modifications which both produce Raman peaks in the cell-silent region. The optimisation of NP synthesis from these labelled polymers by the emulsification-evaporation method is discussed in Chapter 3. In Chapter 4, the NPs synthesised from both the deuterium and alkyne analogues of PLGA were imaged with SRS in biological models of the brain. Tuning to the bioorthogonal peaks allowed imaging of the NPs without cellular background. Both NP analogues were imaged in primary rat microglia, the macrophages of the brain, and additionally the alkyne labelled NPs were imaged in ex vivo cortical mouse brain slices. Immunohistochemical analysis of these brain slices confirmed that the NPs were selectively taken up into microglia

Nanoparticle Design and Novel Approaches to Enhance Photothermal Cancer Therapy.

Rapid advances in bioinformatics and nanotechnology have sparked pre-clinical development of innovative therapies with potential to transform approaches to non-specific clinical practices such as chemotherapy and radiation. One of few nanoparticle-based treatments in clinical trials is photothermal therapy (PTT), which is localized by near infrared light activation of heat-producing gold nanoshells. Here we demonstrate nanoparticle-mediated PTT as a multifunctional platform to address key challenges of cancer medicine, to improve patient tolerance and long-term survival. We present our work in two sections: enhancing efficacy in metastatic settings, and increasing specificity to reduce associated toxicity. In the first section, we focus on the efficacy of PTT against breast cancer stem cells (BCSCs) and tumor-mediated immunosuppressive signaling – vital drivers of cancer growth and metastasis. First we study PTT via highly crystallized iron oxide nanoparticles (HCIONPs) in human breast cancer cells in immune-compromised mice. PTT inhibits both epithelial-like (ALDH+) and mesenchymal-like (CD44+/CD24-) BCSCs and BCSC-driven secondary tumor formation. PTT prior to surgery prevents lymph node metastasis. Next we evaluate HCIONP-mediated PTT and cancer immunotherapy (PD-L1 antibody) in immune-competent mice. PTT significantly reduces mouse ALDH+ BCSCs when given alone and in combination with PD-L1 antibody. Combination treatment reveals promising reductions in tumor growth and formation of lung macrometastases. Furthermore, increases of key inflammatory cytokines and immune cell-attracting chemokines suggest the potential to enhance T-cell tumor infiltration to trigger a systemic, cancer (stem) cell-specific immune response. In the second section, we focus on development of optimized targeted nanoparticle formulations, applicable for PTT, to improve specificity and efficiency of cancer therapy. First we report a new technique – ‘living’ PEGylation – to control the density and composition of heterobifunctional poly(ethylene glycol) (HS-PEG-R) on gold nanoparticles. Applications we demonstrate include control of targeting ligand (HS-PEG-RGD) density to maximize nanoparticle targeting efficiency, and development of double-charged, stealthy nanoparticles (optimal HS-PEG-NH2:HS-PEG-COOH ratio) to minimize immune cell uptake. Lastly, we describe targeted, theranostic nanocomposites with a core-satellite structure for PTT and magnetic resonance imaging. A facilely produced “clickable” targeting peptide enables precise control over attachment to the nanoparticles to prevent steric hindrance and optimize binding to the target receptor.PhDPharmaceutical SciencesUniversity of Michigan, Horace H. Rackham School of Graduate Studieshttp://deepblue.lib.umich.edu/bitstream/2027.42/116764/1/hpaholak_1.pd

Clinical translation of nanomaterials for early detection of genetic abnormalities in fetus and retinopathy in neonates and adults

Nuclear medicine covers a wide variety of radionuclides to meet demands of disease. In the current study, first we have looked at the application of mono-amine mono-amide ligands for Re(V), 99mTc(V), and 186Re(V) with respect to bombesin for receptor targeting in the pancreas. While procedures for synthesizing 99mTc complexes is similar to other reported procedures, rhenium complexes were synthesized using [ReO(citrate)2] - as the starting material, simplifying purification and isolation. Further studies for the 222-MAMA-BBN complex set included biodistribution studies, which determined that the 99mTc-BBN complex binds to GRP receptors in the pancreas, [about]3% ID/g. The 323-MAMA complex and derivatives were investigated to determine if the 222- or the 323-MAMA backbone provide: an easier preparation, a better framework for chelating given metals, and better transport as a targeting receptor. It is found that, in comparative studies, the 222-MAMA derivates are more preferred in chelation. However, in either case, once the metal is chelated, there is no conversion of products upon the addition of a more preferred ligand system. Another avenue of target therapy being pursued is the study of 105Rh. We are specifically looking at the study of chelation with tetrathioether complexes, to rhodium(III) to translate to the radiotracer scale. Three product isomers are formed in the reaction of rhodium, using SnCl2, with 222-S4-diAcOH. The carboxylate arm can either be free dangling, one bound to the metal, or both (removing bound chlorides respectively); all of these isomers can be easily separated using HPLC. These three species will be avoided when translated to the ligand bombesin analog. Future research in this area will be done with the 105Rh radiotracer for biological applications.Includes biblographical reference

Micro/Nano Manufacturing

Micro manufacturing involves dealing with the fabrication of structures in the size range of 0.1 to 1000 µm. The scope of nano manufacturing extends the size range of manufactured features to even smaller length scales—below 100 nm. A strict borderline between micro and nano manufacturing can hardly be drawn, such that both domains are treated as complementary and mutually beneficial within a closely interconnected scientific community. Both micro and nano manufacturing can be considered as important enablers for high-end products. This Special Issue of Applied Sciences is dedicated to recent advances in research and development within the field of micro and nano manufacturing. The included papers report recent findings and advances in manufacturing technologies for producing products with micro and nano scale features and structures as well as applications underpinned by the advances in these technologies

Cancer Nanomedicine

This special issue brings together cutting edge research and insightful commentary on the currentl state of the Cancer Nanomedicine field