Gold-Polystyrene Core-Shell Hybrid Nanoparticles Mediated Highly Efficient Intracellular Delivery Using Light Pulses
25 Jan 2021-
TL;DR: In this article, hybrid nanoparticles composed of gold and polystyrene are used to mediate the photoporation for the successful delivery of propidium iodide dye, quantum dots, and plasmid into CL1-0, AGS, and P-19 cells.
Abstract: Cellular transfection is a method by which exogenous biomolecules can be introduced into the cells. In the fields of molecular and cellular biology, cellular transfection is considered an essential tool, specifically for applications such as drug delivery, cellular therapy, and biomedical imaging. Photoporation based cellular transfection approach uses high-intensity light energy to create membrane pores, sometimes nanoparticles are incorporated to achieve the result at low intensity. In this work, hybrid nanoparticles, composed of gold and polystyrene are used to mediate the photoporation. Laser fluence, exposure time, wavelength, the concentration of nanoparticles, and concentration of exogenous molecules are optimized for the successful delivery of propidium iodide dye, quantum dots, and plasmid into CL1-0, AGS, and P-19 cells. The best results (delivery efficiency of 92% and cell viability of 98%) are achieved in delivering propidium iodide dye into CL1-0 cells.
TL;DR: In this article , a one-pot surface-enhanced Raman scattering (SERS) based immunoassay was used to detect SARS-CoV-2, without any washing process using a portable Raman spectrometer.
Abstract: Rapid and sensitive diagnostics of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is of utmost importance to control the widespread coronavirus disease 2019 (COVID-19) upsurge. This study demonstrated a novel one-pot surface-enhanced Raman scattering (SERS) based immunoassay to detect SARS-CoV-2, without any washing process using a portable Raman spectrometer. The SERS-immune assay was designed using a regular digital versatile disk (DVD) substrate integrated with Raman reporter labeled silver nanoparticles for double clamping effects. The disks were molded to form nanopillar arrays and coated with silver film to enhance the sensitivity of immunoassay. The SERS platform demonstrated a limit of detection (LoD) up to 50 pg mL-1 for SARS-CoV-2 spike protein and virus-like-particle (VLP) protein in phosphate buffer saline within a turnaround time of 20 minutes. Moreover, VLP protein spiked in untreated saliva achieved an LoD of 400 pg mL-1, providing a cycle threshold (Ct) value range of 30-32, closer to reverse transcription-polymerase chain reaction (RT-PCR) results (35-40) and higher than the commercial rapid antigen tests, ranging from 25-28. Therefore, the developed one-pot SERS based biosensor exhibited highly sensitive and rapid detection of SARS-CoV-2, which could be a potential point-of-care platform for early and cost-effective diagnosis of the COVID-19 virus.
TL;DR: The present challenges that need urgent attention are outlined to translate the progress in between in vitro nanomedicine to clinical research, which may eventually lead to advance clinical therapeutics for chronic diseases.
Abstract: Gene delivery for molecular-level therapeutics is regarded as a prospective remedial route to cure human diseases by the medical community. The major challenge for delivering genes in vivo is the lack of suitable delivery vehicles possessing high transfection efficiencies and low cytotoxicity. Currently, viral vectors such as retroviruses, lentiviruses, adenoviruses, adeno-associated viruses (AAV), and herpes simplex viruses (HSV) are being used as successful vectors at clinical trial levels. However, their use has raised major concerns related to insertion mutagenesis and immunogenicity in the medical research community. To address these issues, several non-viral gene delivery vectors are being explored. Among these non-viral vectors, multifunctional nanoparticles have shown superior performance in terms of enhanced gene stability, shielding of cargo from nuclease degradation, and improved passive/active targeting. This review focuses on the explicit role of various non-viral, multifunctional nanoparticles such as lipid-based nanoparticles, quantum dots (QDs), carbon nanotubes, magnetic nanoparticles, silica nanoparticles, and polymer-based nanoparticles, in distinct gene delivery strategies namely, image-guided gene delivery, optically-trackable and optically-activated gene therapy, combinational gene therapy, and present their proficiency in crossing the biological barriers. Furthermore, we highlight the applications of multifunctional nanoparticles as efficient nanovehicles in gene therapy of infectious diseases, cancers, and brain dysfunctional diseases. More importantly, we discuss the in vitro and in vivo toxicity assessments of these multifunctional nanoparticles. Summarily, we outline the present challenges that need urgent attention to translate the progress in between in vitro nanomedicine to clinical research, which may eventually lead us to advance clinical therapeutics for chronic diseases.
TL;DR: This review article will emphasize the basic concept and working mechanism associated with electroporation, single cell Electroporation and biomolecular delivery using micro/nanofluidic devices, their fabrication, working principles and cellular analysis with their advantages, limitations, potential applications and future prospects.
Abstract: © 2018 IOP Publishing Ltd. The ability to deliver foreign molecules into a single living cell with high transfection efficiency and high cell viability is of great interest in cell biology for applications in therapeutic development, diagnostics and drug delivery towards personalized medicine. Many chemical and physical methods have been developed for cellular delivery, however most of these techniques are bulk approach, which are cell-specific and have low throughput delivery. On the other hand, electroporation is an efficient and fast method to deliver exogenous biomolecules such as DNA, RNA and oligonucleotides into target living cells with the advantages of easy operation, controllable electrical parameters and avoidance of toxicity. The rapid development of micro/nanofluidic technologies in the last two decades, enables us to focus an intense electric field on the targeted cell membrane to perform single cell micro-nano-electroporation with high throughput intracellular delivery, high transfection efficiency and cell viability. This review article will emphasize the basic concept and working mechanism associated with electroporation, single cell electroporation and biomolecular delivery using micro/nanoscale electroporation devices, their fabrication, working principles and cellular analysis with their advantages, limitations, potential applications and future prospects.
TL;DR: The author demonstrates invasive and noninvasive with time and non-time resolved SCA and suggests that single cell analysis is possible with capillary electrophoresis (CE) combined with a detection method such as electrochemical detection (ED), laser induced fluorescence (LIF) detection and mass spectrometry (MS).
Abstract: The Special Issue of Micromachines entitled “Micro/Nanofluidic Devices for Single Cell Analysis” covers recent advancements regarding the analysis of single cells by different microfluidic approaches. To understand cell to cell behavior with their organelles and their intracellular biochemical effect, single cell analysis (SCA) can provide much more detailed information from small groups of cells or even single cells, compared to conventional approaches, which only provide ensemble-average information of millions of cells together. Earlier reviews provided single cell analysis using different approaches [1–3]. The author demonstrates invasive and noninvasive with time and non-time resolved SCA ; whereas some other literature provided destructive (with dyes, DNA, RNA, proteins and amino acids) and nondestructive (electroporation, impedance measurement and fluorescence based methods) cellular content analysis using microfluidic devices . Further literature also suggest that single cell analysis is possible with capillary electrophoresis (CE) combined with a detection method such as electrochemical detection (ED), laser induced fluorescence (LIF) detection and mass spectrometry (MS) [4,5]. [...]
01 Jan 2020
TL;DR: This chapter mainly focuses on different physical drug-delivery techniques such as electroporation, optoporation, mechanopsoration, magnetoporation and hybrid techniques along with their working mechanisms, advantages, disadvantages, and limitations.
Abstract: Delivery of exogenous materials or cargo such as drugs, proteins, peptides, and nucleic acids into cells is a vital segment in molecular and cellular biology for potential cellular therapy and drug-discovery applications contributing toward personalization of medicine. Over the years, drug-delivery techniques have been developed in order to gain more control over the drug dosage, targeted delivery, and to minimize side effects. The major drug-delivery techniques can be classified as viral, chemical, and physical methods. Viral vectors are prominently used for gene therapy; however, they are cell-specific and have an immune response with high toxicity. Chemical methods are often limited by the low efficiency of plasmid delivery into different cell types due to plasmid degradation and toxicity. Considering these limitations, different physical methods such as photoporation, gene gun, hydrodynamic injection, electroporation, and mechanoporation, etc., are being widely developed for highly efficient cargo delivery with low toxicity. These methods are able to create transient hydrophilic membrane pores to deliver cargos into cells using different physical energies. Currently, ex vivo cargo delivery is widely studied while few in vivo applications have been developed. Concerning several obstacles to cargo delivery into cells, this chapter mainly focuses on different physical drug-delivery techniques such as electroporation, optoporation, mechanoporation, magnetoporation, and hybrid techniques along with their working mechanisms, advantages, disadvantages, and limitations. An insight into the future prospects and real-time applications of these techniques is also discussed.