Structurally Flexible Triethanolamine Core PAMAM Dendrimers Are Effective Nanovectors for DNA Transfection in Vitro and in Vivo to the Mouse Thymus

2011 ◽  
Vol 22 (12) ◽  
pp. 2461-2473 ◽  
Author(s):  
Xiaoxuan Liu ◽  
Jiangyu Wu ◽  
Miriam Yammine ◽  
Jiehua Zhou ◽  
Paola Posocco ◽  
...  
Keyword(s):  
Pamam Dendrimers ◽  
Dna Transfection ◽  
Mouse Thymus

Activated protein C targets immune cells and rheumatoid synovial fibroblasts to prevent inflammatory arthritis in mice

Rheumatology ◽  
2019 ◽  
Vol 58 (10) ◽  
pp. 1850-1860 ◽  
Author(s):  
Meilang Xue ◽  
Suat Dervish ◽  
Kelly J McKelvey ◽  
Lyn March ◽  
Fang Wang ◽  
...  
Keyword(s):  
Protein C ◽  
Activated Protein C ◽  
Synovial Fibroblasts ◽  
Endothelial Protein C Receptor ◽  
Mouse Thymus ◽  
Tnf Α ◽  
Thymus Cells ◽  
Proliferation And Invasion

Abstract Objectives To investigate whether activated protein C (APC), a physiological anticoagulant can inhibit the inflammatory/invasive properties of immune cells and rheumatoid arthritis synovial fibroblasts (RASFs) in vitro and prevent inflammatory arthritis in murine antigen-induced arthritis (AIA) and CIA models. Methods RASFs isolated from synovial tissues of patients with RA, human peripheral blood mononuclear cells (PBMCs) and mouse thymus cells were treated with APC or TNF-α/IL-17 and the following assays were performed: RASF proliferation and invasion by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) and cell invasion assays, respectively; cytokines and signalling molecules using ELISA or western blot; Th1 and Th17 phenotypes in human PBMCs or mouse thymus cells by flow cytometry. The in vivo effect of APC was evaluated in AIA and CIA models. Results In vitro, APC inhibited IL-1β, IL-17 and TNF-α production, IL-17-stimulated cell proliferation and invasion and p21 and nuclear factor κB activation in RASFs. In mouse thymus cells and human PBMCs, APC suppressed Th1 and Th17 phenotypes. In vivo, APC inhibited pannus formation, cartilage destruction and arthritis incidence/severity in both CIA and AIA models. In CIA, serum levels of IL-1β, IL-6, IL-17, TNF-α and soluble endothelial protein C receptor were significantly reduced by APC treatment. Blocking endothelial protein C receptor, the specific receptor for APC, abolished the early or preventative effect of APC in AIA. Conclusion APC prevents the onset and development of arthritis in CIA and AIA models via suppressing inflammation, Th1/Th17 phenotypes and RASF invasion, which is likely mediated via endothelial protein C receptor.

scholarly journals Electroporation-Based Treatments in Urology

Cancers ◽  
2020 ◽  
Vol 12 (8) ◽  
pp. 2208 ◽  
Author(s):  
Aleksander Kiełbik ◽  
Wojciech Szlasa ◽  
Jolanta Saczko ◽  
Julita Kulbacka
Keyword(s):  
Cell Membrane ◽  
Irreversible Electroporation ◽  
Underlying Mechanism ◽  
Extracellular Proteins ◽  
Dna Transfection ◽  
Membrane Phospholipids ◽  
Cancer Antigens ◽  
Urological Cancers

The observation that an application of a pulsed electric field (PEF) resulted in an increased permeability of the cell membrane has led to the discovery of the phenomenon called electroporation (EP). Depending on the parameters of the electric current and cell features, electroporation can be either reversible or irreversible. The irreversible electroporation (IRE) found its use in urology as a non-thermal ablative method of prostate and renal cancer. As its mechanism is based on the permeabilization of cell membrane phospholipids, IRE (as well as other treatments based on EP) provides selectivity sparing extracellular proteins and matrix. Reversible EP enables the transfer of genes, drugs, and small exogenous proteins. In clinical practice, reversible EP can locally increase the uptake of cytotoxic drugs such as cisplatin and bleomycin. This approach is known as electrochemotherapy (ECT). Few in vivo and in vitro trials of ECT have been performed on urological cancers. EP provides the possibility of transmission of genes across the cell membrane. As the protocols of gene electrotransfer (GET) over the last few years have improved, EP has become a well-known technique for non-viral cell transfection. GET involves DNA transfection directly to the cancer or the host skin and muscle tissue. Among urological cancers, the GET of several plasmids encoding prostate cancer antigens has been investigated in clinical trials. This review brings into discussion the underlying mechanism of EP and an overview of the latest progress and development perspectives of EP-based treatments in urology.

DNA-mediated transfection as a model for oncogenesis

1988 ◽  
Vol 66 (6) ◽  
pp. 594-616 ◽  
Author(s):  
Mary Pat Moyer ◽  
John W. Egan ◽  
J. Bradley Aust ◽  
Rex C. Moyer
Keyword(s):  
Animal Models ◽  
Human Cancer ◽  
Neoplastic Transformation ◽  
Dna Transfer ◽  
Dna Transfection ◽  
Tumor Induction ◽  
Oncogenic Potential ◽  
Cancer Initiation

DNA transfer technology has greatly contributed to progress in understanding molecular biology and genetics. In recent years, great efforts have been expended to determine the oncogenic potential of single, defined genes or complex gene mixtures as a prelude to defining the role those genes may play in neoplastic transformation in vitro and tumor induction in vivo. This paper reviews the currently available DNA transfection techniques and their application toward understanding cancer initiation and progression, and how the in vitro and animal models may apply to human cancer.

scholarly journals Use of Polyamidoamine Dendrimers in Brain Diseases

Molecules ◽  
2018 ◽  
Vol 23 (9) ◽  
pp. 2238 ◽  
Author(s):  
Maria Florendo ◽  
Alexander Figacz ◽  
Bhairavi Srinageshwar ◽  
Ajit Sharma ◽  
Douglas Swanson ◽  
...  
Keyword(s):  
Pamam Dendrimers ◽  
Brain Diseases ◽  
Surface Functional Groups ◽  
Polyamidoamine Dendrimers ◽  
Effective Delivery ◽  
The Right ◽  
The Brain ◽  
Reproducible Manner

Polyamidoamine (PAMAM) dendrimers are one of the smallest and most precise nanomolecules available today, which have promising applications for the treatment of brain diseases. Each aspect of the dendrimer (core, size or generation, size of cavities, and surface functional groups) can be precisely modulated to yield a variety of nanocarriers for delivery of drugs and genes to brain cells in vitro or in vivo. Two of the most important criteria to consider when using PAMAM dendrimers for neuroscience applications is their safety profile and their potential to be prepared in a reproducible manner. Based on these criteria, features of PAMAM dendrimers are described to help the neuroscience researcher to judiciously choose the right type of dendrimer and the appropriate method for loading the drug to form a safe and effective delivery system to the brain.

scholarly journals Neurofilaments are obligate heteropolymers in vivo

1993 ◽  
Vol 122 (6) ◽  
pp. 1337-1350 ◽  
Author(s):  
MK Lee ◽  
Z Xu ◽  
PC Wong ◽  
DW Cleveland
Keyword(s):  
Intermediate Filaments ◽  
Intermediate Filament ◽  
Cultured Cells ◽  
Dna Transfection ◽  
Tail Domain ◽  
In Vitro Assembly ◽  
Mature Neurons ◽  
Filament Network

Neurofilaments (NFs), composed of three distinct subunits NF-L, NF-M, and NF-H, are neuron-specific intermediate filaments present in most mature neurons. Using DNA transfection and mice expressing NF transgenes, we find that despite the ability of NF-L alone to assemble into short filaments in vitro NF-L cannot form filament arrays in vivo after expression either in cultured cells or in transgenic oligodendrocytes that otherwise do not contain a cytoplasmic intermediate filament (IF) array. Instead, NF-L aggregates into punctate or sheet like structures. Similar nonfilamentous structures are also formed when NF-M or NF-H is expressed alone. The competence of NF-L to assemble into filaments is fully restored by coexpression of NF-M or NF-H to a level approximately 10% of that of NF-L. Deletion of the head or tail domain of NF-M or substitution of the NF-H tail onto an NF-L subunit reveals that restoration of in vivo NF-L assembly competence requires an interaction provided by the NF-M or NF-H head domains. We conclude that, contrary to the expectation drawn from earlier in vitro assembly studies, NF-L is not sufficient to assemble an extended filament network in an in vivo context and that neurofilaments are obligate heteropolymers requiring NF-L and NF-M or NF-H.

scholarly journals Are PAMAM dendrimers safe for ocular applications? -- Toxicological evaluation in ocular cells and intraocular tissues

2020 ◽  
Author(s):  
Chen Qin ◽  
Junmei Tang ◽  
Shimin Wen ◽  
SiQing Zhu ◽  
Dong Liu ◽  
...  
Keyword(s):  
Cell Metabolism ◽  
Pamam Dendrimers ◽  
Toxicological Evaluation ◽  
Oxygen Generation ◽  
Ocular Irritation ◽  
In Vivo Experiments ◽  
Polymeric Delivery ◽  
Good Biocompatibility

Abstract Backgrounds: The human eye is a sophisticated and sensitive sensory organ. Due to its dynamic and static barriers, efficient drug therapy for eye diseases is problematic. Polymeric delivery systems have been rapidly developed to overcome this problem, where polymeric dendrimers have received much attention for their unique structures. However, there is insufficient research on whether dendrimer nanomaterials could be used in ophthalmology. Poly (amidoamine) (PAMAM) is a kind of commercialized and extensively used dendrimer. Herein the ocular cytotoxicity and biosafety of PAMAM dendrimers were deeply evaluated by conducting in vitro and in vivo experiments on ocular systems. The effects of generation (G4.0, G5.0, and G6.0) and concentration on cell metabolism, apoptosis, and oxidative damage were carried out. Ocular irritation and intravitreal injection effects were also measured. Results: The results showed that the cytotoxicity of PAMAM increased with increasing generation number. PAMAM at a concentration that below 50 μg/mL was less harmful to the ocular tissues, whereas it caused apparent damage when above 50 μg/mL in the investigated situation. Moreover, singlet oxygen generation was detected in the cells after 50 μg/mL (and above) PAMAM treatment. The in vivo results show that there is no macroscopic structural change observed from fundus and histopathological section images, whereas it shows more functional impairment according to optical coherence tomography (OCT) and electroretinogram (ERG) when treated by 100 μg/mL PAMAM. Conclusion: Overall, a higher concentration of PAMAM, such as above 50 μg/mL, may cause ocular functional damage. The concentrations that are (or lower than) 50 μg/mL showed good biocompatibility and biosafety in human ocular cells and tissues.

Reciprocal Enhancement of Gene Transfer by Combinatorial Adenovirus Transduction and Plasmid DNA Transfection in Vitro and in Vivo

1999 ◽  
Vol 10 (14) ◽  
pp. 2407-2417 ◽  
Author(s):  
Edward J. Dunphy ◽  
Rebecca A. Redman ◽  
Hans Herweijer ◽  
Timothy P. Cripe
Keyword(s):  
Gene Transfer ◽  
Plasmid Dna ◽  
Dna Transfection

scholarly journals Early T-cell progenitors are the major granulocyte precursors in the adult mouse thymus

Blood ◽  
2013 ◽  
Vol 121 (1) ◽  
pp. 64-71 ◽  
Author(s):  
Maria Elena De Obaldia ◽  
J. Jeremiah Bell ◽  
Avinash Bhandoola
Keyword(s):  
T Cell ◽  
Cell Lineage ◽  
Cell Repertoire ◽  
Mouse Thymus ◽  
Thymic Development ◽  
Genetic Ablation ◽  
Cell Lineages ◽  
Repertoire Selection

Abstract The mouse thymus supports T-cell development, but also contains non–T-cell lineages such as dendritic cells, macrophages, and granulocytes that are necessary for T-cell repertoire selection and apoptotic thymocyte clearance. Early thymic progenitors (ETPs) are not committed to the T-cell lineage, as demonstrated by both in vitro and in vivo assays. Whether ETPs realize non–T-cell lineage potentials in vivo is not well understood and indeed is controversial. In the present study, we investigated whether ETPs are the major precursors of any non–T-lineage cells in the thymus. We analyzed the development of these populations under experimental circumstances in which ETPs are nearly absent due to either abrogated thymic settling or inhibition of early thymic development by genetic ablation of IL-7 receptorα or Hes1. Results obtained using multiple in vivo approaches indicate that the majority of thymic granulocytes derive from ETPs. These data indicate that myelolymphoid progenitors settle the thymus and thus clarify the pathways by which stem cells give rise to downstream blood cell lineages.

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