Self-Assembly System Based on Cyclodextrin for Targeted Delivery of Cannabidiol

Cannabidiol (CBD) is one specific kind of the cannabinoid in Cannabis sativa L with a wide range of pharmacological activities. However, the poor water solubility and specificity of CBD limits its application in pharmaceutical field. For solving these problems, in this work, we successfully prepared a targeted carrier by grafting biotin (BIO) onto ethylenediamine-β-Cyclodextrin (EN-CD) in a single step to generate a functionalized supramolecule, named BIO-CD. Subsequently, an amantadine-conjugated cannabinoids (AD-CBD) was prepared and self-assembled with the BIO-CD. A series of methods were used to characterize the inclusion behavior and physicochemical properties of AD-CBD and BIO-CD. The results showed that AD-CBD entered the cavity of BIO-CD and formed a 1:1 host-guest inclusion complex. MTT assay and confocal laser scanning microscopy (CLSM) revealed that the targeting effect and anticancer activity of AD-CBD/BIO-CD inclusion complex against three human cancer cell lines were higher than BIO-CD, AD-CBD and free CBD. Moreover, the inclusion complex could release drugs under weakly acidic conditions. These results demonstrated that AD-CBD/BIO-CD inclusion complex possess excellent targeted and anticancer activity, which is hopeful to be applied in clinic as a new therapeutic approach.

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Targeted Liposomal Chemotherapies to Treat Triple-Negative Breast Cancer

Cancers ◽

10.3390/cancers13153749 ◽

2021 ◽

Vol 13 (15) ◽

pp. 3749

Author(s):

Yingnan Si ◽

Ya Zhang ◽

Hanh Giai Ngo ◽

Jia-Shiung Guan ◽

Kai Chen ◽

...

Keyword(s):

Targeted Delivery ◽

Laser Scanning ◽

Triple Negative ◽

Imaging System ◽

Laser Scanning Microscopy ◽

Confocal Laser ◽

Surface Binding ◽

Synthesis Inhibitor ◽

Tnbc Cell

Triple-negative breast cancers (TNBCs) are highly aggressive and recurrent. Standard cytotoxic chemotherapies are currently the main treatment options, but their clinical efficacies are limited and patients usually suffer from severe side effects. The goal of this study was to develop and evaluate targeted liposomes-delivered combined chemotherapies to treat TNBCs. Specifically, the IC50 values of the microtubule polymerization inhibitor mertansine (DM1), mitotic spindle assembly defecting taxane (paclitaxel, PTX), DNA synthesis inhibitor gemcitabine (GC), and DNA damage inducer doxorubicin (AC) were tested in both TNBC MDA-MB-231 and MDA-MB-468 cells. Then we constructed the anti-epidermal growth factor receptor (EGFR) monoclonal antibody (mAb) tagged liposomes and confirmed its TNBC cell surface binding using flow cytometry, internalization with confocal laser scanning microscopy, and TNBC xenograft targeting in NSG female mice using In Vivo Imaging System. The safe dosage of anti-EGFR liposomal chemotherapies, i.e., <20% body weight change, was identified. Finally, the in vivo anti-tumor efficacy studies in TNBC cell line-derived xenograft and patient-derived xenograft models revealed that the targeted delivery of chemotherapies (mertansine and gemcitabine) can effectively inhibit tumor growth. This study demonstrated that the targeted liposomes enable the new formulations of combined therapies that improve anti-TNBC efficacy.

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One-step preparation of antimicrobial silicone materials based on PDMS and salicylic acid: insights from spatially and temporally resolved techniques

npj Biofilms and Microbiomes ◽

10.1038/s41522-021-00223-6 ◽

2021 ◽

Vol 7 (1) ◽

Author(s):

Luca Barbieri ◽

Ioritz Sorzabal Bellido ◽

Alison J. Beckett ◽

Ian A. Prior ◽

Jo Fothergill ◽

...

Keyword(s):

Salicylic Acid ◽

Laser Scanning ◽

Pathogenic Bacteria ◽

Pharmaceutical Products ◽

Laser Scanning Microscopy ◽

Wound Dressings ◽

Confocal Laser ◽

Vis Spectroscopy ◽

Wide Range ◽

One Step

AbstractIn this work, we introduce a one-step strategy that is suitable for continuous flow manufacturing of antimicrobial PDMS materials. The process is based on the intrinsic capacity of PDMS to react to certain organic solvents, which enables the incorporation of antimicrobial actives such as salicylic acid (SA), which has been approved for use in humans within pharmaceutical products. By combining different spectroscopic and imaging techniques, we show that the surface properties of PDMS remain unaffected while high doses of the SA are loaded inside the PDMS matrix. The SA can be subsequently released under physiological conditions, delivering a strong antibacterial activity. Furthermore, encapsulation of SA inside the PDMS matrix ensured a diffusion-controlled release that was tracked by spatially resolved Raman spectroscopy, Attenuated Total Reflectance IR (ATR-IR), and UV-Vis spectroscopy. The biological activity of the new material was evaluated directly at the surface and in the planktonic state against model pathogenic bacteria, combining confocal laser scanning microscopy, electron microscopy, and cell viability assays. The results showed complete planktonic inhibition for clinically relevant strains of Staphylococcus aureus and Escherichia coli, and a reduction of up to 4 orders of magnitude for viable sessile cells, demonstrating the efficacy of these surfaces in preventing the initial stages of biofilm formation. Our approach adds a new option to existing strategies for the antimicrobial functionalisation of a wide range of products such as catheters, wound dressings and in-dwelling medical devices based on PDMS.

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Astrogliosis in an Experimental Model of Hypovitaminosis B12: A Cellular Basis of Neurological Disorders due to Cobalamin Deficiency

Cells ◽

10.3390/cells9102261 ◽

2020 ◽

Vol 9 (10) ◽

pp. 2261

Author(s):

Zuzanna Rzepka ◽

Jakub Rok ◽

Justyna Kowalska ◽

Klaudia Banach ◽

Justyna Magdalena Hermanowicz ◽

...

Keyword(s):

Laser Scanning ◽

Laser Scanning Microscopy ◽

Cobalamin Deficiency ◽

Confocal Laser ◽

In Vivo Model ◽

Cellular Expression ◽

Wide Range ◽

Vitro Model

Cobalamin deficiency affects human physiology with sequelae ranging from mild fatigue to severe neuropsychiatric abnormalities. The cellular and molecular aspects of the nervous system disorders associated with hypovitaminosis B12 remain largely unknown. Growing evidence indicates that astrogliosis is an underlying component of a wide range of neuropathologies. Previously, we developed an in vitro model of cobalamin deficiency in normal human astrocytes (NHA) by culturing the cells with c-lactam of hydroxycobalamin (c-lactam OH-Cbl). We revealed a non-apoptotic activation of caspases (3/7, 8, 9) in cobalamin-deficient NHA, which may suggest astrogliosis. The aim of the current study was to experimentally verify this hypothesis. We indicated an increase in the cellular expression of two astrogliosis markers: glial fibrillary acidic protein and vimentin in cobalamin-deficient NHA using Western blot analysis and immunocytochemistry with confocal laser scanning microscopy. In the next step of the study, we revealed c-lactam OH-Cbl as a potential non-toxic vitamin B12 antagonist in an in vivo model using zebrafish embryos. We believe that the presented results will contribute to a better understanding of the cellular mechanism underlying neurologic pathology due to cobalamin deficiency and will serve as a foundation for further studies.

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Magnetic poly(ε-caprolactone)/iron-doped hydroxyapatite nanocomposite substrates for advanced bone tissue engineering

Journal of The Royal Society Interface ◽

10.1098/rsif.2012.0833 ◽

2013 ◽

Vol 10 (80) ◽

pp. 20120833 ◽

Cited By ~ 115

Author(s):

A. Gloria ◽

T. Russo ◽

U. D'Amora ◽

S. Zeppetelli ◽

T. D'Alessandro ◽

...

Keyword(s):

Tissue Engineering ◽

Bone Tissue Engineering ◽

Bone Tissue ◽

Laser Scanning ◽

Magnetic Measurements ◽

Laser Scanning Microscopy ◽

Confocal Laser ◽

Hyperthermia Treatment ◽

Wide Range ◽

Iron Doped

In biomedicine, magnetic nanoparticles provide some attractive possibilities because they possess peculiar physical properties that permit their use in a wide range of applications. The concept of magnetic guidance basically spans from drug delivery and hyperthermia treatment of tumours, to tissue engineering, such as magneto-mechanical stimulation/activation of cell constructs and mechanosensitive ion channels, magnetic cell-seeding procedures, and controlled cell proliferation and differentiation. Accordingly, the aim of this study was to develop fully biodegradable and magnetic nanocomposite substrates for bone tissue engineering by embedding iron-doped hydroxyapatite (FeHA) nanoparticles in a poly(ε-caprolactone) (PCL) matrix. X-ray diffraction analyses enabled the demonstration that the phase composition and crystallinity of the magnetic FeHA were not affected by the process used to develop the nanocomposite substrates. The mechanical characterization performed through small punch tests has evidenced that inclusion of 10 per cent by weight of FeHA would represent an effective reinforcement. The inclusion of nanoparticles also improves the hydrophilicity of the substrates as evidenced by the lower values of water contact angle in comparison with those of neat PCL. The results from magnetic measurements confirmed the superparamagnetic character of the nanocomposite substrates, indicated by a very low coercive field, a saturation magnetization strictly proportional to the FeHA content and a strong history dependence in temperature sweeps. Regarding the biological performances, confocal laser scanning microscopy and AlamarBlue assay have provided qualitative and quantitative information on human mesenchymal stem cell adhesion and viability/proliferation, respectively, whereas the obtained ALP/DNA values have shown the ability of the nanocomposite substrates to support osteogenic differentiation.

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Chlorin e6-Biotin Conjugates for Tumor-Targeting Photodynamic Therapy

Molecules ◽

10.3390/molecules26237342 ◽

2021 ◽

Vol 26 (23) ◽

pp. 7342

Author(s):

Wei Liu ◽

Xingqun Ma ◽

Yingying Jin ◽

Jie Zhang ◽

Yang Li ◽

...

Keyword(s):

Photodynamic Therapy ◽

Hela Cells ◽

Tumor Targeting ◽

Laser Scanning ◽

Water Solubility ◽

Laser Scanning Microscopy ◽

Chlorin E6 ◽

Confocal Laser ◽

Oxygen Generation ◽

Targeting Ability

To improve the tumor-targeting efficacy of photodynamic therapy, biotin was conjugated with chlorin e6 to develop a new tumor-targeting photosensitizer, Ce6-biotin. The Ce6-biotin had good water solubility and low aggregation. The singlet-oxygen generation rate of Ce6-biotin was slightly increased compared to Ce6. Flow cytometry and confocal laser scanning microscopy results confirmed Ce6-biotin had higher binding affinity toward biotin-receptor-positive HeLa human cervical carcinoma cells than its precursor, Ce6. Due to the BR-targeting ability of Ce6-biotin, it exhibited stronger cytotoxicity to HeLa cells upon laser irradiation. The IC50 against HeLa cells of Ce6-biotin and Ce6 were 1.28 µM and 2.31 µM, respectively. Furthermore, both Ce6-biotin and Ce6 showed minimal dark toxicity. The selectively enhanced therapeutic efficacy and low dark toxicity suggest that Ce6-biotin is a promising PS for BR-positive-tumor-targeting photodynamic therapy.

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Quaternized Chitosan Modified Nanostructure Lipid Carriers for Topical Delivery of Coenzyme Q10

Nano LIFE ◽

10.1142/s1793984420400139 ◽

2020 ◽

Vol 10 (04) ◽

pp. 2040013 ◽

Cited By ~ 1

Author(s):

Rong Liang ◽

Yuxuan Wang ◽

Lina Wu ◽

Xinjiong Ni ◽

Cheng Yang

Keyword(s):

Delivery System ◽

Coenzyme Q10 ◽

Transdermal Delivery ◽

Laser Scanning ◽

Water Solubility ◽

Laser Scanning Microscopy ◽

Confocal Laser ◽

Nanostructured Lipid Carrier ◽

Quaternized Chitosan

Nanostructured lipid carrier (NLC) is a new colloidal delivery system which can effectively solve the problems of stability and transdermal delivery of active ingredients with poor water solubility and biocompatibility. Coenzyme Q10 (CoQ10), as a lipophilic antioxidant, has poor chemical stability due to unsaturated double bonds in its molecular structure, which limits its addition and application in cosmetics. In this study, CoQ10 NLC was prepared using the mixture of Caprylyl/Capryl Glycoside (APG) and quaternized chitosan (QCS). The particle size of the QCS–APG–NLC was around 250 nm. Compared to NLC stabilized by APG, QCS–APG–NLC has better storage stability under high temperature and light conditions. In vitro transdermal experiment analysis and confocal laser scanning microscopy (CLSM) observation found that QCS modification can effectively increase the penetration amount of CoQ10 in the skin. So, it is suggested that QCS modified APG–NLC can be used as an effective transdermal delivery system for lipophilic active components.

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Cell-to-Cell Signaling in Xylella fastidiosa Suppresses Movement and Xylem Vessel Colonization in Grape

Molecular Plant-Microbe Interactions ◽

10.1094/mpmi-21-10-1309 ◽

2008 ◽

Vol 21 (10) ◽

pp. 1309-1315 ◽

Cited By ~ 71

Author(s):

Subhadeep Chatterjee ◽

Karyn L. Newman ◽

Steven E. Lindow

Keyword(s):

Cell Signaling ◽

Type Strain ◽

Laser Scanning ◽

Wild Type Strain ◽

Xylella Fastidiosa ◽

Laser Scanning Microscopy ◽

Confocal Laser ◽

Deficient Mutant ◽

Wild Type ◽

Wide Range

Cell-to-cell signaling mediated by a fatty acid diffusible signaling factor (DSF) is central to the regulation of the virulence of Xylella fastidiosa. DSF production by X. fastidiosa is dependent on rpfF and, although required for insect colonization, appears to reduce its virulence to grape. To understand what aspects of colonization of grape are controlled by DSF in X. fastidiosa and, thus, those factors that contribute to virulence, we assessed the colonization of grape by a green fluorescent protein–marked rpfF-deficient mutant. The rpfF-deficient mutant was detected at a greater distance from the point of inoculation than the wild-type strain at a given sampling time, and also attained a population size that was up to 100-fold larger than that of the wild-type strain at a given distance from the point of inoculation. Confocal laser-scanning microscopy revealed that approximately 10-fold more vessels in petioles of symptomatic leaves harbored at least some cells of either the wild type or rpfF mutant when compared with asymptomatic leaves and, thus, that disease symptoms were associated with the extent of vessel colonization. Importantly, the rpfF mutant colonized approximately threefold more vessels than the wild-type strain. Although a wide range of colony sizes were observed in vessels colonized by both the wild type and rpfF mutant, the proportion of colonized vessels harboring large numbers of cells was significantly higher in plants inoculated with the rpfF mutant than with the wild-type strain. These studies indicated that the hypervirulence phenotype of the rpfF mutant is due to both a more extensive spread of the pathogen to xylem vessels and unrestrained multiplication within vessels leading to blockage. These results suggest that movement and multiplication of X. fastidiosa in plants are linked, perhaps because cell wall degradation products are a major source of nutrients. Thus, DSF-mediated cell-to-cell signaling, which restricts movement and colonization of X. fastidiosa, may be an adaptation to endophytic growth of the pathogen that prevents the excessive growth of cells in vessels.

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Surface Characterization Using Wavelet Theory and Confocal Laser Scanning Microscopy

Journal of Tribology ◽

10.1115/1.1866161 ◽

2005 ◽

Vol 127 (2) ◽

pp. 394-404 ◽

Cited By ~ 34

Author(s):

Chengqing Yuan ◽

Zhongxiao Peng ◽

Xinping Yan

Keyword(s):

Surface Roughness ◽

Confocal Laser Scanning Microscopy ◽

Laser Scanning ◽

Surface Characterization ◽

Laser Scanning Microscopy ◽

Confocal Laser Scanning ◽

Scanning Microscopy ◽

Confocal Laser ◽

Wavelet Theory ◽

Wide Range

Surface characterization, particularly roughness analysis, is very important for a wide range of applications including wear assessment. This paper proposes a set of methods and techniques to acquire appropriate images using confocal laser scanning microscopy, to separate roughness, waviness, and form using wavelet theory, and to characterize surface roughness for engineering surfaces and surfaces of small particles. Two application examples on engineering surfaces and wear particles have been presented in the paper to demonstrate that the method developed in this study can be used to measure surface roughness reliably and precisely. A guide on how to determine the iris size, step size, and objective lens has been scientifically provided according to theoretical analysis and experimental results.

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Morphology of powerful suction organs from blepharicerid larvae living in raging torrents

BMC Zoology ◽

10.1186/s40850-019-0049-6 ◽

2019 ◽

Vol 4 (1) ◽

Cited By ~ 4

Author(s):

Victor Kang ◽

Richard Johnston ◽

Thomas van de Kamp ◽

Tomáš Faragó ◽

Walter Federle

Keyword(s):

Laser Scanning ◽

Close Contact ◽

Laser Scanning Microscopy ◽

Confocal Laser ◽

Wide Range ◽

Flow Speeds ◽

Micro Tomography ◽

Structural Adaptations ◽

Attachment Devices

Abstract Background Suction organs provide powerful yet dynamic attachments for many aquatic animals, including octopus, squid, remora, and clingfish. While the functional morphology of suction organs from some cephalopods and fishes has been investigated in detail, there are only few studies on such attachment devices in insects. Here we characterise the morphology and ultrastructure of the suction attachment organs of net-winged midge larvae (genus Liponeura; Diptera: Blephariceridae) – aquatic insects that live on rocks in rapid alpine waterways where flow speeds can reach 3 m s− 1 – using scanning electron microscopy, confocal laser scanning microscopy, and X-ray computed micro-tomography (micro-CT). Furthermore, we study the function of these organs in vivo using interference reflection microscopy. Results We identified structural adaptations important for the function of the suction attachment organs in L. cinerascens and L. cordata. First, a dense array of spine-like microtrichia covering each suction disc comes into contact with the substrate upon attachment, analogous to hairy structures on suction organs from octopus, clingfish, and remora fish. These spine-like microtrichia may contribute to the seal and provide increased shear force resistance in high-drag environments. Second, specialised rim microtrichia at the suction disc periphery were found to form a continuous ring in close contact and may serve as a seal on a variety of surfaces. Third, a V-shaped cut on the suction disc (“V-notch“) is actively opened via two cuticular apodemes inserting on its flanks. The apodemes are attached to dedicated V-notch opening muscles, thereby providing a unique detachment mechanism. The complex cuticular design of the suction organs, along with specialised muscles that attach to them, allows blepharicerid larvae to generate powerful attachments which can withstand strong hydrodynamic forces and quickly detach for locomotion. Conclusion The suction organs from Liponeura are underwater attachment devices specialised for resisting extremely fast flows. Structural adaptations from these suction organs could translate into future bioinspired attachment systems that perform well on a wide range of surfaces.

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Design of new acid-activated cell-penetrating peptides for tumor drug delivery

PeerJ ◽

10.7717/peerj.3429 ◽

2017 ◽

Vol 5 ◽

pp. e3429 ◽

Cited By ~ 12

Author(s):

Jia Yao ◽

Yinyun Ma ◽

Wei Zhang ◽

Li Li ◽

Yun Zhang ◽

...

Keyword(s):

Drug Delivery ◽

Targeted Delivery ◽

Laser Scanning ◽

Critical Role ◽

Cell Penetrating Peptides ◽

Laser Scanning Microscopy ◽

Confocal Laser ◽

Ph Response ◽

Cell Penetrating ◽

Ph Dependent

TH(AGYLLGHINLHHLAHL(Aib)HHIL-NH2), a histidine-rich, cell-penetrating peptide with acid-activated pH response, designed and synthesized by our group, can effectively target tumor tissues with an acidic extracellular environment. Since the protonating effect of histidine plays a critical role in the acid-activated, cell-penetrating ability of TH, we designed a series of new histidine substituents by introducing electron donating groups (Ethyl, Isopropyl, Butyl) to the C-2 position of histidine. This resulted in an enhanced pH-response and improved the application of TH in tumor-targeted delivery systems. The substituents were further utilized to form the corresponding TH analogs (Ethyl-TH, Isopropyl-TH and Butyl-TH), making them easier to protonate for positive charge in acidic tumor microenvironments. The pH-dependent cellular uptake efficiencies of new TH analogs were further evaluated using flow cytometry and confocal laser scanning microscopy, demonstrating that ethyl-TH and butyl-TH had an optimal pH-response in an acidic environment. Importantly, the new TH analogs exhibited relatively lower toxicity than TH. In addition, these new TH analogs were linked to the antitumor drug camptothecin (CPT), while butyl-TH modified conjugate presented a remarkably stronger pH-dependent cytotoxicity to cancer cells than TH and the other conjugates. In short, our work opens a new avenue for the development of improved acid-activated, cell-penetrating peptides as efficient anticancer drug delivery vectors.

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