Dataset Information

Development and Characterization of Bioinspired Lipid Raft Nanovesicles for Therapeutic Applications.

ABSTRACT: Lipid rafts are highly ordered regions of the plasma membrane enriched in signaling proteins and lipids. Their biological potential is realized in exosomes, a subclass of extracellular vesicles (EVs) that originate from the lipid raft domains. Previous studies have shown that EVs derived from human placental mesenchymal stromal cells (PMSCs) possess strong neuroprotective and angiogenic properties. However, clinical translation of EVs is challenged by very low, impure, and heterogeneous yields. Therefore, in this study, lipid rafts are validated as a functional biomaterial that can recapitulate the exosomal membrane and then be synthesized into biomimetic nanovesicles. Lipidomic and proteomic analyses show that lipid raft isolates retain functional lipids and proteins comparable to PMSC-EV membranes. PMSC-derived lipid raft nanovesicles (LRNVs) are then synthesized at high yields using a facile, extrusion-based methodology. Evaluation of biological properties reveals that LRNVs can promote neurogenesis and angiogenesis through modulation of lipid raft-dependent signaling pathways. A proof-of-concept methodology further shows that LRNVs could be loaded with proteins or other bioactive cargo for greater disease-specific functionalities, thus presenting a novel type of biomimetic nanovesicles that can be leveraged as targeted therapeutics for regenerative medicine.

SUBMITTER: Ramasubramanian L

PROVIDER: S-EPMC9756296 | biostudies-literature | 2022 Dec

REPOSITORIES: biostudies-literature

ACCESS DATA

Publications

Development and Characterization of Bioinspired Lipid Raft Nanovesicles for Therapeutic Applications.

Ramasubramanian Lalithasri L Jyothi Harsha H Goldbloom-Helzner Leora L Light Brandon M BM Kumar Priyadarsini P Carney Randy P RP Farmer Diana L DL Wang Aijun A

ACS applied materials & interfaces 20221130 49

Lipid rafts are highly ordered regions of the plasma membrane enriched in signaling proteins and lipids. Their biological potential is realized in exosomes, a subclass of extracellular vesicles (EVs) that originate from the lipid raft domains. Previous studies have shown that EVs derived from human placental mesenchymal stromal cells (PMSCs) possess strong neuroprotective and angiogenic properties. However, clinical translation of EVs is challenged by very low, impure, and heterogeneous yields. ...[more]

PMID: 36448709

Similar Datasets

Project description:Extracellular vesicles (EVs) have shown great promise as drug delivery system (DDS). However, their complex and costly production limit their development for clinical use. Interestingly, the plant kingdom can also produce EV-like nanovesicles that can easily be isolated and purified from a large quantity of raw material at a high yield. In this study, olive-derived nanovesicles (ODNVs) were isolated from raw fruits using serial centrifugations and their physical and biological features characterized to demonstrate their promising potential to be used as a DDS. Nanotracking particle analysis indicated an average size of 109.5 ± 3.0 nm and yield of 1012 ODNVs/mL for the purest fraction. Microscopy imaging, membrane fluidity assay and lipidomics analysis showed the presence of a rich lipid bilayer that significantly varied between different sources of ODNVs but showed a distinct signature compared to human EVs. Moreover, ODNVs were enriched in PEN1 and TET8 compared to raw fruits, suggesting an extracellular origin. Interestingly, ODNVs size and yield stayed unchanged after exposure to high temperature (70 °C for 1 h), wide pH range (5-10), and 50-100 nm extrusion, demonstrating high resistance to physical and chemical stresses. This high resistance allowed ODNVs to stay stable in water at 4 °C for a month, or with the addition of 25 mM trehalose for long-term freezing storage. Finally, ODNVs were internalized by both 2D and 3D cell culture without triggering significant cytotoxicity and immunogenicity. Importantly, the anticancer drug doxorubicin (dox) could be loaded by passive incubation within ODNVs and dox-loaded ODNVs decreased cell viability by 90% compared to only 70% for free dox at the same concentration, indicating a higher efficiency of drug delivery by ODNVs. In addition, this high cytotoxicity effect of dox-loaded ODNVs was shown to be stable after a 2-week storage at 4 °C. Together, these findings suggested that ODNVs represent a promising candidate as drug nanocarrier for various DDS clinical applications, as demonstrated by their biocompatibility, high resistance to stress, good stability in harsh environment, and improvement of anticancer drug efficacy.

Project description:Protein S-acylation (palmitoylation), a reversible post-translational modification, is critically involved in regulating protein subcellular localization, activity, stability, and multimeric complex assembly. However, proteome scale characterization of S-acylation has lagged far behind that of phosphorylation, and global analysis of the localization of S-acylated proteins within different membrane domains has not been reported. Here we describe a novel proteomics approach, designated palmitoyl protein identification and site characterization (PalmPISC), for proteome scale enrichment and characterization of S-acylated proteins extracted from lipid raft-enriched and non-raft membranes. In combination with label-free spectral counting quantitation, PalmPISC led to the identification of 67 known and 331 novel candidate S-acylated proteins as well as the localization of 25 known and 143 novel candidate S-acylation sites. Palmitoyl acyltransferases DHHC5, DHHC6, and DHHC8 appear to be S-acylated on three cysteine residues within a novel CCX(7-13)C(S/T) motif downstream of a conserved Asp-His-His-Cys cysteine-rich domain, which may be a potential mechanism for regulating acyltransferase specificity and/or activity. S-Acylation may tether cytoplasmic acyl-protein thioesterase-1 to membranes, thus facilitating its interaction with and deacylation of membrane-associated S-acylated proteins. Our findings also suggest that certain ribosomal proteins may be targeted to lipid rafts via S-acylation, possibly to facilitate regulation of ribosomal protein activity and/or dynamic synthesis of lipid raft proteins in situ. In addition, bioinformatics analysis suggested that S-acylated proteins are highly enriched within core complexes of caveolae and tetraspanin-enriched microdomains, both cholesterol-rich membrane structures. The PalmPISC approach and the large scale human S-acylated protein data set are expected to provide powerful tools to facilitate our understanding of the functions and mechanisms of protein S-acylation.

Project description:Vanillin is a natural compound easily extracted from plants. It has neuroprotective, anti-carcinogenic, antioxidant, antimicrobial, and anti-biofilm properties. It also presents high volatility, high hydrophilicity, and low bioavailability. Nanomaterials can be used to improve pharmacodynamics, solubility, and stability and to enhance pharmacokinetics. In this work, non-ionic surfactant vesicles were synthesized as vanillin carriers: neutral niosomes formed by Span60 and cholesterol, positive charged niosomes formulated with cetyltrimethylammonium bromide (CTAB), and negatively charged niosomes formulated with sodium dodecyl sulfate (SDS). Niosomes synthesis was carried out with two commonly used methods: thin film hydration (TFH) and ethanol injection method (EIM). The niosomes synthesized were used to prepare two different materials: (i) a powder containing the lyophilized noisome with vanillin systems and (ii) a gelatin matrix film containing niosomes with vanillin. Lyophilization was carried out using maltodextrin as a cryoprotectant. The lyophilization of colloidal structures allows for storage at room temperature for long periods of time, keeping their organoleptic characteristics invariable. Niosomes were characterized before and after the lyophilization process in terms of morphological characterization, size, polydispersity index (PDI), and zeta potential. Moreover, niosomes cargo was evaluated by calculating the encapsulation efficiency (EE) and loading capacity (LC). Results showed that the use of the TFH method allowed us to obtain niosomes of 255 nm with high EE (up to 40%) and LC values higher than EIM. The lyophilization process decreased the LC of the vesicles prepared, but this decrease was mitigated by up to 20% when ionic surfactants were used on the membrane bilayer. Gelatin films are biodegradable materials suitable for food packing applications. The incorporation of a natural compound with antimicrobial activity would be a clear advantage for such an application. The films prepared were characterized in terms of morphology, water solubility, color, and transparency. Niosomes synthesized by thin film hydration had better chemical and physical properties to load vanillin. Especially in the case of application in films, niosomes with a negative charge, formed by SDS, and vanillin loaded gave better mechanical and chemical characteristics to the film.

Dataset Information

Development and Characterization of Bioinspired Lipid Raft Nanovesicles for Therapeutic Applications.

Publications

Development and Characterization of Bioinspired Lipid Raft Nanovesicles for Therapeutic Applications.

Similar Datasets

OmicsDI is part of the ELIXIR infrastructure

Tweets