Exosomes derived from platelet-rich plasma administration in site mediate cartilage protection in subtalar osteoarthritis

Subtalar osteoarthritis (STOA) is often secondary to chronic ankle sprains, which seriously affects the quality of life of patients. Due to its etiology and pathogenesis was not studied equivocally yet, there is currently a lack of effective conservative treatments. Although they have been used for tissue repair, platelet-rich plasma-derived exosomes (PRP-Exo) have the disadvantage of low retention and short-lived therapeutic effects. This study aimed to determine whether incorporation of PRP-Exo in thermosensitive hydrogel (Gel) increased their retention in the joint and thereby playing a therapeutic role on STOA due to chronic mechanical instability established by transecting lateral ligaments (anterior talofibular ligament (ATFL)/calcaneal fibular ligament (CFL)).
PRP-Exo incorporated Gel (Exo-Gel) system, composed of <em>Poloxamer</em>-407 and <em>188</em> mixture-based thermoresponsive hydrogel matrix in an optimal ratio, was determined by its release ability of Exo and rheology of Gel response to a different temperature. The biological activity of Exo-Gel was evaluated in vitro, and the therapeutic effect of Exo-Gel on STOA was evaluated in vivo.
Exo released from Exo-Gel continuously for 28 days could promote the proliferation and migration of mouse bone mesenchymal stem cells (mBMSCs) and chondrocytes, at the same time enhance the chondrogenic differentiation of mBMSCs, and inhibit inflammation-induced chondrocyte degeneration. In vivo experiments confirmed that Exo-Gel increased the local retention of Exo, inhibited the apoptosis and hypertrophy of chondrocytes, enhanced their proliferation, and potentially played the role in stem cell recruitment to delay the development of STOA. Thus, Delivery of PRP-Exo incorporated in thermosensitive Gel provides a novel approach of cell-free therapy and has therapeutic effect on STOA.

Aquaporin 4 in Traumatic Brain Injury: From Molecular Pathways to Therapeutic Target

Traumatic brain injury (TBI) is known as an acute degenerative pathology of the central nervous system, and has been shown to increase brain aquaporin 4 (AQP4) expression. Various molecular mechanisms affect AQP4 expression, including neuronal high mobility group box 1, forkhead box O3a, vascular endothelial growth factor, hypoxia-inducible factor-1 α (HIF-1 α) sirtuin 2, NF-κB, Malat1, nerve growth factor and Angiotensin II receptor type 1. In addition, inhibition of AQP4 with FK-506, MK-801 (indirectly by targeting N-methyl-D-aspartate receptor), inactivation of adenosine A2A receptor, levetiracetam, adjudin, progesterone, estrogen, V1aR inhibitor, hypertonic saline, erythropoietin, <em>poloxamer</em> <em>188</em>, brilliant blue G, HIF-1alpha inhibitor, normobaric oxygen therapy, astaxanthin, epigallocatechin-3-gallate, sesamin, thaliporphine, magnesium, prebiotic fiber, resveratrol and omega-3, as well as AQP4 gene silencing lead to reduced edema upon TBI. This review summarizes current knowledge and evidence on the relationship between AQP4 and TBI, and the potential mechanisms involved.

Solvent-Free Fabrication of Biphasic Lipid-Based Microparticles with Tunable Structure

Lipid-based biphasic microparticles are generally produced by long and complex techniques based on double emulsions. In this study, spray congealing was used as a solvent-free fabrication method with improved processability to transform water-in-oil non-aqueous emulsions into spherical solid lipid-based particles with a biphasic structure (b-MPs).
Emulsions were prepared by melt emulsification using different compositions of lipids (Dynasan<sup>®</sup>118 and Compritol<sup>®</sup>888 ATO), surfactants (Cetylstearyl alcohol and Span<sup>®</sup>60) and hydrophilic carriers (PEGs, Gelucire<sup>®</sup>48/16 and <em>Poloxamer</em> <em>188</em>).
First, pseudo-ternary phase diagrams were constructed to identify the area corresponding to each emulsion type (coarse emulsion or microemulsion). The hydrophobicity of the lipid mostly affected the interfacial tension, and thus the microstructure of the emulsion. Emulsions were then processed by spray congealing and the obtained b-MPs were characterized in terms of thermal and chemical properties (by DSC and FT-IR), external and internal morphology (by SEM, CLSM and Raman mapping).
Solid free-flowing spherical particles (main size range 200-355 µm) with different architectures were successfully produced: microemulsions led to the formation of particles with a homogeneous internal structure, while coarse emulsions generated “multicores-shell” particles consisting of variable size hydrophilic cores evenly distributed within the crystalline lipid phase. Depending on their composition and structure, b-MPs could achieve various release profiles, representing a more versatile system than microparticles based on a single lipid phase. The formulation and technological strategy proposed, provides a feasible and cost-effective way of fabricating b-MPs with tunable internal structure and release behavior.

 

Genome DNA Leakage of Adeno-Associated Virus Under Freeze-Thaw Stress

Adeno-associated virus (AAV) has become an emerging tool for human gene therapies. Currently, AAV gene therapies are subjected to multiple freeze-thaw cycles during manufacturing, storage, transportation, and administration. While studies have shown that multiple freeze-thaw cycles led to a decrease in transduction efficiency, the AAV degradation mechanism during freeze-thaw is not well understood.
Here, we have characterized the impact of freeze-thaw on AAV8 by employing a variety of assays, which revealed significant increases in the amount of free single-stranded DNA (ssDNA) in AAV8 formulations after multiple freeze-thaw cycles. Subsequent analysis using Next Generation Sequencing (NGS) revealed that the ssDNA primarily consisted of genome DNA, indicating that the increased ssDNA leaked out from AAV8.
Experiments performed using different serotypes of AAV confirmed the pervasiveness of such behavior amongst AAVs. In addition, formulation screening studies were performed to understand the impact on genome DNA leakage from AAV. The formulation screening results showed that the addition of 10% sucrose and 0.1% <em>poloxamer</em> <em>188</em> to Dulbecco’s phosphate-buffered saline (DPBS) reduced the leakage of ssDNA in AAV samples after freeze-thaw cycles compared to the base formulation of DPBS alone. These findings shed new light on the degradation mechanism of AAVs and stabilization of the AAV-based gene therapies.

Sustained Delivery of Lactoferrin Using Poloxamer Gels for Local Bone Regeneration in a Rat Calvarial Defect Model

Lactoferrin (LF) is a multifunctional milk glycoprotein that promotes bone regeneration. Local delivery of LF at the bone defect site is a promising approach for enhancement of bone regeneration, but efficient systems for sustained local delivery are still largely missing. The aim of this study was to investigate the potential of the poloxamers for sustained delivery of LF to enhance local bone regeneration. The developed LF/poloxamer formulations were liquid at room temperature (20 °C) transforming to a sustained releasing gel depot at body temperature (37 °C). In vitro release studies demonstrated an initial burst release (~50%), followed by slower release of LF for up to 72 h.

100 ML POLOXAMER 188

from CORNING
13-901-CI | 100 mL/pk: 80.00 EUR

100 G POLOXAMER 188, POWDER

from CORNING
61-161-RM | 100 g/pk: 67.00 EUR

Mouse pre-microRNA Expression Construct mir-188

from SBI
MMIR-188-PA-1 | Bacterial Streak: 684.00 EUR

ICG amine

from AAT Bioquest
188 | 1 mg: 306.00 EUR

Monkey (Cynomolgus) cDNA Normal Tissue: Pancreas

from Alpha Diagnostics
MC34-188 | 10 rxn: 415.00 EUR

Recombinant Mouse C1qtnf9 Protein

from Creative Biolabs
CTP-188 | 100ug: Ask for price

M. Tuberculosis EsxB (LHP) (CFP10) Antigen

from Creative Diagnostics
DAGA-188 | 100 µg: 724.00 EUR

miRZip-188-3p anti-miR-188-3p microRNA construct

from SBI
MZIP188-3p-PA-1 | Bacterial Streak: 684.00 EUR

miRZip-188-5p anti-miR-188-5p microRNA construct

from SBI
MZIP188-5p-PA-1 | Bacterial Streak: 684.00 EUR

JAG - 1 (188 - 204), Jagged – 1 (188 - 204), Notch Ligand

from CHI Scientific
5-01413 | 4 x 1mg: Ask for price

HIST1H1B (Ab-188) Antibody

from Cusabio
1-CSB-PA010377PA188nphHU | [ 100ul: 332.00 EUR ] [ 50ul: 335.00 EUR ]

rno-miR-188 Primers

from ABM
MP-r00230 | 150 ul / 10 uM: 176.00 EUR

R-JAG (188 - 206) R - Jagged -1(188 - 206), Notch Ligand

from CHI Scientific
5-01864 | 4 x 1mg: Ask for price

CEB beta antibody (Thr235/188)

from Fitzgerald
70R-30652 | 100 ug: 327.00 EUR

rno-miR-188-3p Primers

from ABM
MPR00447 | 150 ul / 10 uM: 176.00 EUR

rno-miR-188-5p Primers

from ABM
MPR00448 | 150 ul / 10 uM: 176.00 EUR

hsa-miR-188-3p Primers

from ABM
MPH01206 | 150 ul / 10 uM: 121.00 EUR

hsa-miR-188-5p Primers

from ABM
MPH01207 | 150 ul / 10 uM: 121.00 EUR

rno??miR-188 miRNA Antagomir

from ABM
MNR01210 | 2 x 5.0 nmol: 329.00 EUR
 Poloxamer, with and without LF, increased osteoblast viability at 72 h (p < 0.05) compared to control, and the immune response from THP-1 cells was mild when compared to the suture material. In rat calvarial defects, the LF/poloxamer group had lower bone volume than the controls (p = 0.0435). No difference was observed in tissue mineral density and lower bone defect coverage scores (p = 0.0267) at 12 weeks after surgery. In conclusion, LF/poloxamer formulations support cell viability and do not induce an unfavourable immune response; however, LF delivery via the current formulation of LF200/poloxamer gel did not demonstrate enhanced bone regeneration and was not compatible with the rat calvarial defect model.

Isolation of exosome from the culture medium of Nasopharyngeal cancer (NPC) C666-1 cells using inertial based Microfluidic channel

Isolation of exosome from culture medium in an effective way is desired for a less time consuming, cost saving technology in running the diagnostic test on cancer. In this study, we aim to develop an inertial microfluidic channel to separate the nano-size exosome from C666-1 cell culture medium as a selective sample. Simulation was carried out to obtain the optimum flow rate for determining the dimension of the channels for the exosome separation from the medium.
The optimal dimension was then brought forward for the actual microfluidic channel fabrication, which consisted of the stages of mask printing, SU8 mould fabrication and ended with PDMS microchannel curing process. The prototype was then used to verify the optimum flow rate with polystyrene particles for its capabilities in the actual task on particle separation as a control outcome. Next, the microchip was employed to separate the selected samples, exosome from the culture medium and compared the outcome from the conventional exosome extraction kit to study the level of effectiveness of the prototype.
The exosome outcome from both the prototype and extraction kits were characterized through zetasizer, western blot and Transmission electron microscopy (TEM). The microfluidic chip designed in this study obtained a successful separation of exosomes from the culture medium. Besides, the extra benefit from these microfluidic channels in particle separation brought an evenly distributed exosome upon collection https://joplink.net/exosome-isolation-kits/ while the exosomes separated through the extraction kit was found clustered together. Therefore, this work has shown the microfluidic channel is suitable for continuous separation of exosomes from the culture medium for a clinical study in the future.

Characterization of surface markers on extracellular vesicles isolated from lymphatic exudate from patients with breast cancer

Background: Breast cancer is the most common cancer, and the leading cause of cancer-related deaths, among females world-wide. Recent research suggests that extracellular vesicles (EVs) play a major role in the development of breast cancer metastasis. Axillary lymph node dissection (ALND) is a procedure in patients with known lymph node metastases, and after surgery large amounts of serous fluid are produced from the axilla. The overall aim was to isolate and characterize EVs from axillary serous fluid, and more specifically to determine if potential breast cancer biomarkers could be identified.
Methods: Lymphatic drain fluid was collected from 7 patients with breast cancer the day after ALND. EVs were isolated using size exclusion chromatography, quantified and detected by nanoparticle tracking analysis, electron microscopy, nano flow cytometry and western blot. The expression of 37 EV surface proteins was evaluated by flow cytometry using the MACSPlex Exosome kit.
Results: Lymphatic drainage exudate retrieved after surgery from all 7 patients contained EVs. The isolated EVs were positive for the typical EV markers CD9, CD63, CD81 and Flotillin-1 while albumin was absent, indicating low contamination from blood proteins. In total, 24 different EV surface proteins were detected.
Eleven of those proteins were detected in all patients, including the common EV markers CD9, CD63 and CD81, cancer-related markers CD24, CD29, CD44 and CD146, platelet markers CD41b, CD42a and CD62p as well as HLA-DR/DP/DQ. Furthermore, CD29 and CD146 were enriched in Her2+ patients compared to patients with Her2- tumors.
Conclusions: Lymphatic drainage exudate retrieved from breast cancer patients after surgery contains EVs that can be isolated using SEC isolation. The EVs have several cancer-related markers including CD24, CD29, CD44 and CD146, proteins of potential interest as biomarkers as well as to increase the understanding of the mechanisms of cancer biology.

Understanding the Role and Clinical Applications of Exosomes in Gynecologic Malignancies: A Review of the Current Literature

Background: Gynecologic malignancies are those which arise in the female reproductive organs of the ovaries, cervix, and uterus. They carry a great deal of morbidity and mortality for patients, largely due to challenges in diagnosis and treatment of these cancers. Although advances in technology and understanding of these diseases have greatly improved diagnosis, treatment, and ultimately survival for patients with gynecologic malignancies over the last few decades, there is still room for improvements in diagnosis and treatment, for which exosomes may be the key. This paper reviews the current knowledge regarding gynecologic tumor derived-exosomal genetic material and proteins, their role in cancer progression, and their potential for advancing the clinical care of patients with gynecologic cancers through novel diagnostics and therapeutics.
Literature review: Ovarian tumor derived exosome specific proteins are reviewed in detail, discussing their role in ovarian cancer metastasis. The key microRNAs in cervical cancer and their implications in future clinical use are discussed. Additionally, uterine cancer-associated fibroblast (CAF)-derived exosomes which may promote endometrial cancer cell migration and invasion through a specific miR-148b are reviewed. The various laboratory techniques and commercial kits for the isolation of exosomes to allow for their clinical utilization are described as well.
Conclusion: Exosomes may be the key to solving many unanswered questions, and closing the gaps so as to improve the outcomes of patients with gynecologic cancers around the world. The potential utilization of the current knowledge of exosomes, as they relate to gynecologic cancers, to advance the field and bridge the gaps in diagnostics and therapeutics highlight the promising future of exosomes in gynecologic malignancies.

Pathogenic Mechanisms of Preeclampsia with Severe Features Implied by the Plasma Exosomal miRNA Profile

Preeclampsia is a complication of pregnancy characterised by high blood pressure and organ damage after 20 gestational weeks. It is associated with high maternal and fetal morbidity and mortality; however, at present, there is no effective prevention or treatment for this condition. Previous studies have revealed that plasma exosomal miRNAs from pregnant women with preeclampsia could serve as biomarkers of pathogenic factors. However, the roles of plasma exosomal miRNAs in preeclampsia with severe features (sPE), which is associated with poorer pregnancy outcomes, remain unknown.
Thus, the aims of this study were to characterise plasma exosomal miRNAs in sPE and explore the related pathogenic mechanisms using bioinformatic analysis. Plasma exosomes were isolated using a mirVana RNA isolation kit.
The exosomal miRNAs were detected using high-throughput sequencing and the miRNAs related to Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways and gene ontology (GO) terms were analysed using the clusterProfiler package of R. Fifteen miRNAs exhibited increased expression and fourteen miRNAs exhibited reduced expression in plasma exosomes from women with sPE as compared to normal pregnant women.
Further, gene set enrichment analysis revealed that the differentially expressed plasma exosomal miRNAs were related to the stress response and cell junction regulation, among others. In summary, this study is the first to identify the differentially expressed plasma exosomal miRNAs in sPE. These findings highlight promising pathogenesis mechanisms underlying preeclampsia.

Exosome Isolation kit (for cell culture media)

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Exosome Isolation kit (for cell culture media)

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Exosome Isolation kit (for cell culture media)

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Exosome Isolation kit (for stem cell culture media)

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Exosome Isolation kit (for stem cell culture media)

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Reagent for Total Exosome Isolation (Culture Media Supplement)

50 ml 2071 EUR

VEX Exosome Isolation Reagent (from cell culture media)

50 ml 589 EUR

Exosome Isolation kit (for urine)

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Exosome Isolation kit (for urine)

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Exosome Isolation kit (for urine)

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