Runx1 Messenger RNA Delivered by Polyplex Nanomicelles Alleviate Spinal Disc Hydration Loss in a Rat Disc Degeneration Model

Vertebral disc degenerative disease (DDD) affects millions of people worldwide and is a critical factor leading to low back and neck pain and consequent disability. Currently, no strategy has addressed curing DDD from fundamental aspects, because the pathological mechanism leading to DDD is still controversial. One possible mechanism points to the homeostatic status of extracellular matrix (ECM) anabolism, and catabolism in the disc may play a vital role in the disease’s progression. If the damaged disc receives an abundant amount of cartilage, anabolic factors may stimulate the residual cells in the damaged disc to secrete the ECM and mitigate the degeneration process. To examine this hypothesis, a cartilage anabolic factor, Runx1, was expressed by mRNA through a sophisticated polyamine-based PEG-polyplex nanomicelle delivery system in the damaged disc in a rat model. The mRNA medicine and polyamine carrier have favorable safety characteristics and biocompatibility for regenerative medicine. The endocytosis of mRNA-loaded polyplex nanomicelles in vitro, mRNA delivery efficacy, hydration content, disc shrinkage, and ECM in the disc in vivo were also examined. The data revealed that the mRNA-loaded polyplex nanomicelle was promptly engulfed by cellular late endosome, then spread into the cytosol homogeneously at a rate of less than 20 min post-administration of the mRNA medicine. The mRNA expression persisted for at least 6-days post-injection in vivo. Furthermore, the Runx1 mRNA delivered by polyplex nanomicelles increased hydration content by ≈43% in the punctured disc at 4-weeks post-injection (wpi) compared with naked Runx1 mRNA administration. Meanwhile, the disc space and ECM production were also significantly ameliorated in the polyplex nanomicelle group. This study demonstrated that anabolic factor administration by polyplex nanomicelle-protected mRNA medicine, such as Runx1, plays a key role in alleviating the progress of DDD, which is an imbalance scenario of disc metabolism. This platform could be further developed as a promising strategy applied to regenerative medicine.

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The ETS1 transcription factor is required for the development and cytokine-induced expansion of ILC2

Journal of Experimental Medicine ◽

10.1084/jem.20150851 ◽

2016 ◽

Vol 213 (5) ◽

pp. 687-696 ◽

Cited By ~ 44

Author(s):

Erin C. Zook ◽

Kevin Ramirez ◽

Xiaohuan Guo ◽

Grant van der Voort ◽

Mikael Sigvardsson ◽

...

Keyword(s):

Transcription Factor ◽

Messenger Rna ◽

Critical Factor ◽

Protective Role ◽

Lymphoid Cells ◽

Allergic Lung Inflammation ◽

Factor Inhibitor ◽

Group 2

Group 2 innate lymphoid cells (ILC2s) are a subset of ILCs that play a protective role in the response to helminth infection, but they also contribute to allergic lung inflammation. Here, we report that the deletion of the ETS1 transcription factor in lymphoid cells resulted in a loss of ILC2s in the bone marrow and lymph nodes and that ETS1 promotes the fitness of the common progenitor of all ILCs. ETS1-deficient ILC2 progenitors failed to up-regulate messenger RNA for the E protein transcription factor inhibitor ID2, a critical factor for ILCs, and these cells were unable to expand in cytokine-driven in vitro cultures. In vivo, ETS1 was required for the IL-33–induced accumulation of lung ILC2s and for the production of the T helper type 2 cytokines IL-5 and IL-13. IL-25 also failed to elicit an expansion of inflammatory ILC2s when these cells lacked ETS1. Our data reveal ETS1 as a critical regulator of ILC2 expansion and cytokine production and implicate ETS1 in the regulation of Id2 at the inception of ILC2 development.

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Activation of MAT2A-RIP1 signaling axis reprograms monocytes in gastric cancer

Journal for ImmunoTherapy of Cancer ◽

10.1136/jitc-2020-001364 ◽

2021 ◽

Vol 9 (2) ◽

pp. e001364

Author(s):

Yan Zhang ◽

Hui Yang ◽

Jun Zhao ◽

Ping Wan ◽

Ye Hu ◽

...

Keyword(s):

Gastric Cancer ◽

Cell Metabolism ◽

Vital Role ◽

Methionine Metabolism ◽

Promoter Regions ◽

Normal Tissues ◽

Methionine Cycle

BackgroundThe activation of tumor-associated macrophages (TAMs) facilitates the progression of gastric cancer (GC). Cell metabolism reprogramming has been shown to play a vital role in the polarization of TAMs. However, the role of methionine metabolism in function of TAMs remains to be explored.MethodsMonocytes/macrophages were isolated from peripheral blood, tumor tissues or normal tissues from healthy donors or patients with GC. The role of methionine metabolism in the activation of TAMs was evaluated with both in vivo analyses and in vitro experiments. Pharmacological inhibition of the methionine cycle and modulation of key metabolic genes was employed, where molecular and biological analyses were performed.ResultsTAMs have increased methionine cycle activity that are mainly attributed to elevated methionine adenosyltransferase II alpha (MAT2A) levels. MAT2A modulates the activation and maintenance of the phenotype of TAMs and mediates the upregulation of RIP1 by increasing the histone H3K4 methylation (H3K4me3) at its promoter regions.ConclusionsOur data cast light on a novel mechanism by which methionine metabolism regulates the anti-inflammatory functions of monocytes in GC. MAT2A might be a potential therapeutic target for cancer cells as well as TAMs in GC.

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Reversal of senescence-associated beta-galactosidase expression during in vitro three-dimensional tissue-engineering of human chondrocytes in a polymer scaffold

Scientific Reports ◽

10.1038/s41598-021-93607-9 ◽

2021 ◽

Vol 11 (1) ◽

Author(s):

Shojiro Katoh ◽

Atsuki Fujimaru ◽

Masaru Iwasaki ◽

Hiroshi Yoshioka ◽

Rajappa Senthilkumar ◽

...

Keyword(s):

Regenerative Medicine ◽

Replicative Senescence ◽

Cultured Cells ◽

Three Dimensional ◽

Human Chondrocytes ◽

Cartilage Tissue ◽

Optimal Method ◽

Beta Galactosidase

AbstractRegenerative medicine applications require cells that are not inflicted with senescence after in vitro culture for an optimal in vivo outcome. Methods to overcome replicative senescence include genomic modifications which have their own disadvantages. We have evaluated a three-dimensional (3D) thermo-reversible gelation polymer (TGP) matrix environment for its capabilities to reverse cellular senescence. The expression of senescence-associated beta-galactosidase (SA-βgal) by human chondrocytes from osteoarthritis-affected cartilage tissue, grown in a conventional two-dimensional (2D) monolayer culture versus in 3D-TGP were compared. In 2D, the cells de-differentiated into fibroblasts, expressed higher SA-βgal and started degenerating at 25 days. SA-βgal levels decreased when the chondrocytes were transferred from the 2D to the 3D-TGP culture, with cells exhibiting a tissue-like growth until 42–45 days. Other senescence associated markers such as p16INK4a and p21 were also expressed only in 2D cultured cells but not in 3D-TGP tissue engineered cartilage. This is a first-of-its-kind report of a chemically synthesized and reproducible in vitro environment yielding an advantageous reversal of aging of human chondrocytes without any genomic modifications. The method is worth consideration as an optimal method for growing cells for regenerative medicine applications.

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Induction of Winter Flounder Antifreeze Protein Messenger RNA at 4 C in vivo and in vitro

Physiological Zoology ◽

10.1086/physzool.59.6.30158614 ◽

1986 ◽

Vol 59 (6) ◽

pp. 679-695 ◽

Cited By ~ 5

Author(s):

Jeffrey L. Price ◽

Brian B. Gourlie ◽

Yuan Lin ◽

Ru Chih C. Huang

Keyword(s):

Messenger Rna ◽

Antifreeze Protein ◽

Winter Flounder

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The Role of microRNAs in Osteoporosis Diagnostics

Osteologie/Osteology ◽

10.1055/a-1514-1800 ◽

2021 ◽

Vol 30 (03) ◽

pp. 222-229

Author(s):

Matthias Hackl ◽

Elisabeth Semmelrock ◽

Johannes Grillari

Keyword(s):

Bone Mass ◽

Bone Metabolism ◽

Messenger Rna ◽

Biological Fluids ◽

Clinical Diagnostics ◽

Bone Architecture ◽

Estrogen Metabolism ◽

Age Related

AbstractMicroRNAs (miRNAs) are short (18–24 nucleotides) non-coding RNA sequences that regulate gene expression via binding of messenger RNA. It is estimated that miRNAs co-regulate the expression of more than 70% of all human genes, many of which fulfil important roles in bone metabolism and muscle function. In-vitro and in-vivo experiments have shown that the targeted loss of miRNAs in distinct bone cell types (osteoblasts and osteoclasts) results in altered bone mass and bone architecture. These results emphasize the biological relevance of miRNAs for bone health.MiRNAs are not only considered as novel bone biomarkers because of their biological importance to bone metabolism, but also on the basis of other favorable properties: 1) Secretion of miRNAs from cells enables “minimally invasive” detection in biological fluids such as serum. 2) High stability of miRNAs in serum enables the retrospective analysis of frozen blood specimens. 3) Quantification of miRNAs in the serum is based on the RT-PCR - a robust method that is considered as the gold standard for the analysis of nucleic acids in clinical diagnostics.With regard to osteoporosis, it has been shown that many of the known risk factors are characterized by distinct miRNA profiles in the affected tissues: i) age-related loss of bone mass, ii) sarcopenia, iii) changes in estrogen metabolism and related changes Loss of bone mass, and iv) diabetes. Therefore, numerous studies in recent years have dealt with the characterization of miRNAs in the serum of osteoporosis patients and healthy controls, and were able to identify recurring miRNA patterns that are characteristic of osteoporosis. These novel biomarkers have great potential for the diagnosis and prognosis of osteoporosis and its clinical outcomes.The aim of this article is to give a summary of the current state of knowledge on the research and application of miRNA biomarkers in osteoporosis.

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Factors affecting the time of formation of the mouse blastocoele

Development ◽

10.1242/dev.41.1.79 ◽

1977 ◽

Vol 41 (1) ◽

pp. 79-92

Author(s):

Rosita Smith ◽

Anne McLaren

Keyword(s):

Cytochalasin B ◽

Critical Factor ◽

Cell Number ◽

Experimental Manipulation ◽

Factors Affecting ◽

Elapsed Time ◽

Developmental Age ◽

Culture Formation

In normal mouse embryos developing in vivo, the first appearance of the blastocyst cavity was found to be associated more closely with developmental age, judged by cell number, than with chronological age, i.e. elapsed time since ovulation. When development was slowed by in vitro culture, formation of the blastocoele was delayed. However, cell number itself was not a critical factor, since the number of cells per embryo could be doubled or tripled or halved by experimental manipulation without substantially affecting the timing of blastocoele formation. Experiments in which one cell division was suppressed with cytochalasin-B, leading to tetraploidy, showed that the number of cell divisions since fertilization was also not critical. A possible role is suggested either for nucleocytoplasmic ratio, or for the number of nuclear or chromosomal divisions or DNA replications since fertilization, all of which increase during cleavage.

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Differential Effects of Halothane and Thiopental on Surfactant Protein C Messenger RNA In Vivo and In Vitro in Rats

Anesthesiology ◽

10.1097/00000542-200009000-00030 ◽

2000 ◽

Vol 93 (3) ◽

pp. 805-810 ◽

Cited By ~ 17

Author(s):

Catherine Paugam-Burtz ◽

Serge Molliex ◽

Bernard Lardeux ◽

Corinne Rolland ◽

Michel Aubier ◽

...

Keyword(s):

Messenger Rna ◽

Protein C ◽

Alveolar Type ◽

Type Ii Cells ◽

Type Ii ◽

Alveolar Type Ii Cells ◽

Mechanically Ventilated ◽

Mrna Content

Background Pulmonary surfactant is a complex mixture of proteins and phospholipids synthetized by alveolar type II cells. Volatile anesthetics have been shown to reduce surfactant phospholipid biosynthesis by rat alveolar type II cells. Surfactant-associated protein C (SP-C) is critical for the alveolar surfactant functions. Our goal was to evaluate the effects of halothane and thiopental on SP-C messenger RNA (mRNA) expression in vitro in rat alveolar type II cells and in vivo in mechanically ventilated rats. Methods In vitro, freshly isolated alveolar type II cells were exposed to halothane during 4 h (1, 2, 4%) and 8 h (1%), and to thiopental during 4 h (10, 100 micrometer) and 8 h (100 micrometer). In vivo, rats were anesthetized with intraperitoneal thiopental or inhaled 1% halothane and mechanically ventilated for 4 or 8 h. SP-C mRNA expression was evaluated by ribonuclease protection assay. Results In vitro, 4-h exposure of alveolar type II cells to thiopental 10 and 100 micrometer increased their SP-C mRNA content to 145 and 197%, respectively, of the control values. In alveolar type II cells exposed for 4 h to halothane 1, 2, and 4%, the SP-C mRNA content increased dose-dependently to 160, 235, and 275%, respectively, of the control values. In vivo, in mechanically ventilated rats, 4 h of halothane anesthesia decreased the lung SP-C mRNA content to 53% of the value obtained in control (nonanesthetized, nonventilated) animals; thiopental anesthesia increased to 150% the lung SP-C mRNA content. Conclusions These findings indicate that halothane and thiopental used at clinically relevant concentrations modulate the pulmonary SP-C mRNA content in rats. In vivo, the additive role of mechanical ventilation is suggested.

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Relative messenger RNA expression profiling of collagenases and aggrecanases in human articular chondrocytes in vivo and in vitro

Arthritis & Rheumatism ◽

10.1002/art.10531 ◽

2002 ◽

Vol 46 (10) ◽

pp. 2648-2657 ◽

Cited By ~ 262

Author(s):

Brigitte Bau ◽

Pia M. Gebhard ◽

Jochen Haag ◽

Thomas Knorr ◽

Eckart Bartnik ◽

...

Keyword(s):

Expression Profiling ◽

Messenger Rna ◽

Articular Chondrocytes ◽

Human Articular Chondrocytes ◽

Rna Expression ◽

Rna Expression Profiling

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Regenerative Medicine and Angiogenesis; Challenges and Opportunities

Advanced Pharmaceutical Bulletin ◽

10.34172/apb.2020.061 ◽

2020 ◽

Vol 10 (4) ◽

pp. 490-501

Author(s):

Mozhgan Jahani ◽

Davood Rezazadeh ◽

Parisa Mohammadi ◽

Amir Abdolmaleki ◽

Amir Norooznezhad ◽

...

Keyword(s):

Regenerative Medicine ◽

Large Scale ◽

Diffusion Limit ◽

Blood Vessel Development ◽

Engineering Technique ◽

Challenges And Opportunities ◽

Vessel Development ◽

Tissue Engineering Technique

Blood vessel development is one of the most prominent steps in regenerative medicine due tothe restoration of blood flow to the ischemic tissues and providing the rapid vascularizationin clinical-sized tissue-engineered grafts. However, currently tissue engineering technique isrestricted because of the inadequate in vitro/in vivo tissue vascularization. Some challenges likeas transportation in large scale, distribution of the nutrients and poor oxygen diffusion limit theprogression of vessels in smaller than clinically relevant dimensions as well in vivo integration.In this regard, the scholars attempted to promote the vascularization process relied on the stemcells (SCs), growth factors as well as exosomes and interactions of biomaterials with all of themto enable the emergence of ideal microenvironment which is needed for treatment of unhealthyorgans or tissue regeneration and formation of new blood vessels. Thus, in the present reviewwe aim to describe these approaches, advances, obstacles and opportunities as well as theirapplication in regeneration of heart as a prominent angiogenesis-dependent organ.

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Importance of color stability of the esthetic restoration materials

International Journal of Community Medicine and Public Health ◽

10.18203/2394-6040.ijcmph20214618 ◽

2021 ◽

Vol 8 (12) ◽

pp. 6096

Author(s):

Thuraya Abdulrahim Basudan ◽

Ibrahim Alhussain Bahshan ◽

Mohammed Khashman Almutairi ◽

Nwaf Ahmed Alkadi ◽

Jehad Aymen Al Qiriaqri ◽

...

Keyword(s):

Dental Care ◽

Dental Health ◽

Health Sector ◽

Color Stability ◽

Vital Role ◽

Medical Treatments ◽

The Public ◽

Human Use

Color stability has a vital role in several factors. Including the cosmetic appearance, confidence while smiling, and facial emotions, and in some patients, it denoted healthy and vital dental health. Dental discoloration denotes the lack of dental care. This varies from excessive use of external artificial coloration materials such as coffee, tea, and fizzy drinks. Also, in some cases, the dental discoloration is due to internal pathological conditions or due to medical treatments usage. Radiation has been remarked as one of the causes that cause dental discoloration. In addition to that, chemotherapy was also associated with dental discoloration. Some medications, such as tetracycline and antihistamines, were linked to dental discoloration. The literature discussed in-vitro experiments for the substances affecting the teeth' color status. Coffee was the most significant cause for dental discoloration, either human use (in-vivo) or laboratory experiments (in-vitro). The sociodemographic status was linked to the variance of dental discoloration. Future recommendations are concerned about the public health sector. The WHO should offer comprehensive dental care for all people all over the world, and not exclusive for specific socioeconomic areas. Practical plans for screening dental pathologies should be investigated and for systematic pathologies that might be associated with dental issues.

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