Transport Of Bioactive Molecules Research Articles

Immunomodulatory responses of extracellular vesicles released by gram-positive fish pathogen Streptococcus parauberis

Bacterial extracellular vesicles (BEVs) are nanosized structures that play a role in intercellular communication and transport of bioactive molecules. Streptococcus parauberis is a Gram-positive pathogenic bacterium that causes “Streptococcosis” in fish. In this study, we isolated S. parauberis-derived extracellular vesicles (SpEVs), and then physicochemical and immunomodulatory properties were determined to elucidate their biological functions. Initially, the biogenesis of SpEVs was detected using field emission scanning electron microscopy, which revealed that secretory phase SpEVs attached to the outer surface of S. parauberis. SpEVs had an average particle diameter and zeta potential of 168.3 ± 6.5 nm and −17.96 ± 2.11 mV, respectively. Field emission transmission electron microscopy analysis confirmed the presence of round or oval-shaped SpEVs with clear membrane margins. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis results showed three sharp protein bands when SpEVs were stained with Coomassie blue. In vitro toxicity of SpEVs was assayed using the murine macrophage RAW 264.7 cells and we observed no significant (p < 0.05) viability reduction up to 50 μg/mL qRT-PCR results revealed that SpEVs-treated (5 and 10 μg/mL) RAW 264.7 cells significantly (p < 0.05) induced the mRNA of proinflammatory (Il1β, Il6, and Tnfα) and anti-inflammatory (Il10) cytokines in a concentration-dependent manner. In vivo immunomodulatory effects of SpEVs were investigated by injecting SpEVs (5 and 10 μg/fish) into adult zebrafish. Transcriptional analysis based on qRT-PCR indicates significant (p < 0.05) upregulation of proinflammatory (il1β, il6, and tnfα) and anti-inflammatory (il10) genes in a concentration-dependent manner in zebrafish kidney. Further, protein expression results in zebrafish spleen tissue confirmed the immunomodulatory activity of SpEVs. In conclusion, SpEVs display the characteristics of BEVs and immunomodulatory activities, suggesting their potential application as vaccine candidate.

Plant extracellular vesicles and their potential in human health research, the practical approach.

Extracellular vesicles are small membrane particles (30-1000 nm) released by Bacteria, Eukaryotes and Archaea. They have been shown to play an important role in intracellular and intercellular communication, within and between kingdoms via transport of bioactive molecules. Thus, they can be involved in altering gene expression and regulation of physiological and pathological processes of the recipient. Their unique properties make extracellular vesicles a perfect candidate vector for targeted drug delivery or a biomarker. For a long time, animal and mainly mammal extracellular vesicles have been used in research. But for plants, there had been speculations about the existence of nanovesicles due to the presence of a cell wall. Today, awareness of plant extracellular vesicles is on the rise and their research has proved they have various functions, such as protein secretion, transport of bioactive molecules or defense against pathogens. Further potential of plant extracellular vesicles is stressed in this review.

Application of Nanomaterials to Ensure Quality and Nutritional Safety of Food

Nanomaterials (NMs) are emerging novel tools for preserving quality, enhancing shelf life, and ensuring food safety. Owing to the distinctive physicochemical characters, engineered NMs under varying sizes and dimensions have great potentials for application in the manufacturing, packaging, processing, and safety of quality agrifood. The promise of various kinds of novel NMs that are useful for food industries has opened a possibility of a new revolution in agroprocessing industries in both the emerging and advanced nations. The rapid advancement of nanoscience has provided a great impact on material science that has allowed researchers to understand every aspect of molecular complexity and its functions in life sciences. The reduced size of NMs that increase the surface area is useful in the specific target of different organs, and biodegradable nanospheres are helpful in the transport of bioactive molecules across the cellular barriers. However, nanotechnology creates a great revolution in several sections including agriculture and food industry and also reduces environmental pollution, while the toxicity of some NMs in the food industry poses a great concern to researchers for their greater application. However, most of the developed countries have regulatory control acts but developing countries do not have them yet. Therefore, for the safe use of NMs and also to minimize the health and environmental risks in both the developed and developing countries, it is indispensable to recognize the toxicity-constructed, toxicodynamic, and toxicokinetic features of NMs, which should carefully be emphasized at the home and industrial levels. The current study highlights the updates of the NMs to safeguard the quality and nutritional safety of foods at home and also at the industrial level.

The Influence of a Stressful Microenvironment on Tumor Exosomes: A Focus on the DNA Cargo.

Exosomes secreted by tumor cells, through the transport of bioactive molecules, reprogram the surroundings, building a microenvironment to support the development of the tumor. The discovery that exosomes carry genomic DNA reflecting that of the tumor cell of origin has encouraged studies to use them as non-invasive biomarkers. The exosome-mediated transfer of oncogenes suggested a new mechanism of malignant transformation that could play a role in the formation of metastases. Several studies have examined the role of tumor exosomes on the modulation of the tumor microenvironment, but relatively few have been directed to assess how stressful stimuli can influence their production and cargo. Understanding the changes in exosome loads and the production pattern of the stressed tumor cell may uncover actionable mechanisms responsible for tumor progression.

Electrofluidic control of bioactive molecule delivery into soft tissue models based on gelatin methacryloyl hydrogels using threads and surgical sutures

The delivery of bioactive molecules (drugs) with control over spatial distribution remains a challenge. Herein, we demonstrate for the first time an electrofluidic approach to controlled delivery into soft tissue models based on gelatin methacryloyl (GelMA) hydrogels. This was achieved using a surgical suture, whereby transport of bioactive molecules, including drugs and proteins, was controlled by imposition of an electric field. Commonly employed surgical sutures or acrylic threads were integrated through the hydrogels to facilitate the directed introduction of bioactive species. The platform consisted of two reservoirs into which the ends of the thread were immersed. The anode and cathode were placed separately into each reservoir. The thread was taken from one reservoir to the other through the gel. When current was applied, biomolecules loaded onto the thread were directed into the gel. Under the same conditions, the rate of movement of the biomolecules along GelMA was dependent on the magnitude of the current. Using 5% GelMA and a current of 100 µA, 2 uL of fluorescein travelled through the hydrogel at a constant velocity of 7.17 ± 0.50 um/s and took less than 8 minutes to exit on the thread. Small molecules such as riboflavin migrated faster (5.99 ± 0.40 μm/s) than larger molecules such as dextran (2.26 ± 0.55 μm/s with 4 kDa) or BSA (0.33 ± 0.07 μm/s with 66.5 kDa). A number of commercial surgical sutures were tested and found to accommodate the controlled movement of biomolecules. Polyester, polyglactin 910, glycolide/lactide copolymer and polyglycolic acid braided sutures created adequate fluid connection between the electrodes and the hydrogel. With a view to application in skin inflammatory diseases and wound treatment, wound healing, slow and controlled delivery of dexamethasone 21-phosphate disodium salt (DSP), an anti-inflammatory prodrug, was achieved using medical surgicryl PGA absorbable suture. After 2 hours of electrical stimulation, still 81.1% of the drug loaded was encapsulated within the hydrogel.

Role of microsphere as drug carrier for osteogenic differentiation

Drug delivery system in sustainable manner has a great potential in biomedical applications. Along with its biocompatibility, microspheres ability to encapsulate and promote sustained release of drugs or growth factor makes them an ideal carrier for the transport of bioactive molecules for tissue regeneration and controlled drug delivery applications. Additionally, the injectable form of small spherical microsphere facilitates the accurate drug delivery. These drug carrier microspheres have significant role in osteogenic differentiation. The invasive delivery of bioactive molecules promotes bone regeneration. The drug stimulates the osteogenic differentiation of stem cell and promotes the damaged tissue for self-repair. This review elaborates the role of polymeric microspheres and the composite microspheres as a drug carrier for osteogenic differentiation.

A Role of Vesicular Transduction of Intercellular Signals in Cancer Development

Export of biologically active compounds is essential for any living cell. Transport of bioactive molecules through a cellular membrane can be active, or passive, or vesicular. In the past decade, vesicular transduction of intercellular signals has attracted great interest in the scientific community. An extremely important role of the vesicle transduction has been established for almost all processes in a living body. Not only profiles of protein and RNA expression in a cell, but also its secretome change during various pathologies, including cancer development. The enhanced secretion of vesicles by transformed cells is one important factor in creating a special microenvironment that favors tumor progression. At present, a role of exosomes has been demonstrated for such important processes as an epithelial-mesenchymal transition, angiogenesis, metastatic niche formation, chemotherapeutic resistance, and interaction with the immune system. The special biological role of the extracellular vesicles and their basic differences depend on their molecular composition. Therefore, special protein and lipid markers are responsible for a vesicular targeted delivery with information due to the preferable interaction with cells of a definite type. The exosomes of cancer cells can facilitate apoptosis or growth of neighboring malignant cells depending on the exosome composition. These and other special features of the extracellular vesicles make studies of their composition and role especially interesting and attract significant attention from researchers. Despite the rapid progress in this field, there are still many unresolved problems, such as a search for specific markers which allow identification of different types of vesicles or vesicles secreted by distinct cells, as well as screening of vesicular markers of cancers and other diseases that are associated with disorders in a functioning immune system. This review is mainly focused on the role of intercellular vesicular transport of bioorganic molecules in cancer progression. We believe that a successful treatment of oncological diseases is impossible without an understanding of the intercellular communication of both cancer cells between each other and with other systems of an organism and with a concept of an active participation of the cell-secreted vesicles in this process.

Influence of different surfactants on the physicochemical properties of elastic liposomes

Elastic liposomes are capable to improve drug transport through the skin by acting as penetration enhancers due to the high fluidity and elasticity of the liposome membranes. Therefore, elastic liposomes were prepared and characterized to facilitate the transdermal transport of bioactive molecules. Liposomes consisted of dimyristoylphosphatidylcholine (DMPC) as the structural component, with different surfactants derived from lauric acid as elastic components: C12E5 (polyoxyethylene-5-lauryl ether), PEG4L (polyethyleneglycol-4-lauryl ester), PEG4DL (polyethylene glycol-4-dilauryl ester), PEG8L (polyethylene glycol-8-lauryl ester) and PEG8DL (polyethylene glycol-8-dilauryl ester). The elastic liposomes were characterized in terms of their phospholipid content, mean diameter, size distribution, elasticity and stability during storage, as well as their ability to incorporate surfactant and permeate through 50 nm pore size membranes. The results showed that the phospholipid phase transition temperature, the fluidity of the lipid bilayer resulting from incorporation of the surfactant and the preservation of particle integrity were factors determining the performance of the elastic liposomes in permeating through nanoporous membranes. The best results were obtained using DMPC combined with the surfactants PEG8L or PEG8DL. The findings demonstrate the potential of using elastic liposomes for transdermal administration of drugs.

Exosomes: A Promising Factor Involved in Cancer Hypoxic Microenvironments

As a significant tumor feature, hypoxia can trigger cancer adaptive processes, induce malignant phenotype development, and promote drug resistance. Previous studies demonstrated that exosomes are critical during these procedures. Exosomes are small vesicles formed in vesicular bodies in the endosomal network. These small vesicles are mainly involved in the transport of bioactive molecules between cells. Exosomes are also involved in the mediation of some cellular communications depending on derived donor cells; thus, recipient cells undergo phenotypic changes. Furthermore, hypoxia can remarkably stimulate exosomal secretion; for instance, nucleic acids and proteins as transmission signals in exosomes in a tumor microenvironment are involved in various functions, such as inducing intratumoral heterogeneity, altering immunological responses, producing cancer-associated fibroblasts, and promoting angiogenesis and metastasis. Moreover, exosome contents resemble those of a donor cell; this finding indicates that exosomes may also be regarded as suitable biomarkers of hypoxia status. Therefore, exosomes can be used to facilitate diagnosis and prognosis with minimal invasive procedures. Further studies on exosomes in cancer may provide new therapeutic strategies.

Exosome in Tumour Microenvironment: Overview of the Crosstalk between Normal and Cancer Cells

Cancer development is a multistep process in which exosomes play important roles. Exosomes are small vesicles formed in vesicular bodies in the endosomal network. The major role of exosomes seems to be the transport of bioactive molecules between cells. Depending on the cell of origin, exosomes are implicated in the regulation of several cellular events, with phenotypic consequences in recipient cells. Cancer derived exosomes (CCEs) are important players in the formation of the tumour microenvironment by (i) enabling the escape of tumour cells to immunological system and help initiating the inflammatory response; (ii) acting in the differentiation of fibroblasts and mesenchymal cells into myofibroblasts; (iii) triggering the angiogenic process; and (iv) enhancing the metastatic evolution of the tumour by promoting epithelial to mesenchymal transformation of tumour cells and by preparing the tumour niche in the new anatomical location. Since the finding that exosomes content resembles that of the cell of origin, they may be regarded as suitable biomarkers for cancer diagnosis, allowing for diagnosis and prognosis via a minimal invasive procedure. Exosome involvement in cancer may open new avenues regarding therapeutics, such as vectors for targeted drug delivery.

Serial subculture and relative transport of human endothelial cells in serum-free, defined conditions

Endothelial cells bind, process, and transport bioactive molecules and thus provide an interactive interface between the plasma and adjacent tissues. Various hormones and factors induce endothelial cells to synthesize and secrete interactive factors. However, study of endothelial cell synthesis, processing, and transport of these bioactive molecules has been impeded because of the serum requirement for cell growth. Many of these bioactive molecules are derived from or are modified by serum components. We have developed a short-term culture system that supports sequential subculturing of endothelial cells in a serum-free culture medium on a defined extracellular matrix. The cells have a doubling rate of 33 h and the total cell number can be expanded more than 800-fold. Expression of specific markers; factor VIII related antigen-von Willebrand factor, Wiebel-Palade bodies, a cobblestone appearance of confluent cell monolayers, and angiotensin-converting enzyme activity confirm the normal morphologic integrity and biochemical function of these cultures. Using this defined serum-free medium, we have grown human endothelial cell monolayers on porous polycarbonate membrane inserts, thereby creating an upper and a lower chamber that models the vascular architecture and demonstrates an inverse relationship between transport and molecular weight. By eliminating serum components, this model system should facilitate the study of endothelial cell binding, metabolism, and transport of bioactive molecules and may contribute to a better understanding of the blood-tissue interface.