Incorporation of Synthetic mRNA in Injectable Chitosan-Alginate Hybrid Hydrogels for Local and Sustained Expression of Exogenous Proteins in Cells

Steinle, Heidrun; Ionescu, Tudor-Mihai; Schenk, Selina; Golombek, Sonia; Kunnakattu, Silju-John; Özbek, Melek Tutku; Schlensak, Christian; Wendel, Hans Peter; Avci‐Adali, Meltem

doi:10.3390/ijms19051313

Cited by 26 publications

(37 citation statements)

References 59 publications

(65 reference statements)

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“…Injection of an MSR releasing OVA as antigen enhances systemic helper T cells TH1 and TH2 serum antibody and cytotoxic T‐cell levels compared to bolus injections. Steinle et al . evaluated similar degradable, injectable chitosan‐alginate hybrid hydrogels for the delivery of mRNA to HEK293 cells that were both incorporated into the hydrogels; a simplistic in vitro assay showed enhanced expression of their reporter gene over 3 weeks; no in vivo studies were carried out.…”

Section: Introductionmentioning

confidence: 99%

Injectable Biodegradable Chitosan‐Alginate 3D Porous Gel Scaffold for mRNA Vaccine Delivery

Yan

Chen

Zhang

et al. 2018

Macromolecular Bioscience

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mRNA vaccines have proven to be more stable, effective, and specific than protein/peptide-based vaccines in stimulating both humoral and cellular immune response. However, mRNA’s fast degradation rate and low-transfection efficiency in vivo impede its potential in vaccination. Recent research in gene delivery has focused on nonviral vaccine carriers and either implantable or injectable delivery systems to improve transgene expression in vivo. Here, an injectable chitosan-alginate gel scaffold for the local delivery of mRNA vaccines is reported. Gel scaffold biodegradation rates and biocompatibility are quantified. Scaffold-mediated mRNA in vivo transgene expression as well as ovalbumin antigen specific cellular and humoral immune responses are evaluated in vivo. Luciferase reporter protein expression resulting from mRNA lipoplex-loaded gel scaffolds is five times higher than systemic injection. Compared to systemic injections of naked mRNA or mRNA:lipoplexes, elevated levels of T cell proliferation and IFN-γ secretion are seen with in vivo scaffold-mediated mRNA lipoplex delivery. Furthermore, a humoral response (ovalbumin antigen specific IgG levels) is observed as early as week 1 for scaffold-mediated mRNA lipoplex delivery, while protein-based immunization did not elicit IgG production until 2 weeks post-injection. Results suggest that injectable scaffold mRNA vaccine delivery maybe a viable alternative to traditional nucleic acid immunization methods.

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Section: Introductionmentioning

confidence: 99%

Injectable Biodegradable Chitosan‐Alginate 3D Porous Gel Scaffold for mRNA Vaccine Delivery

Yan

Chen

Zhang

et al. 2018

Macromolecular Bioscience

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“…Especially, if a conversion of cell fate for example differentiation of MSCs into chondrocytes [39] or reprogramming of cells to pluripotency [38] is required. Thus, for sustained mRNA delivery and prolonged protein expression, various strategies using nanoparticles [40], hydrogels [41], and biomaterials such as collagen sponges [42] have been investigated. Thereby, the regenerative potential can be further improved.…”

Section: Discussionmentioning

confidence: 99%

Exogenous Delivery of Link N mRNA into Chondrocytes and MSCs—The Potential Role in Increasing Anabolic Response

Tendulkar

Ehnert

Sreekumar

et al. 2019

IJMS

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Musculoskeletal disorders, such as osteoarthritis and intervertebral disc degeneration are causes of morbidity, which concomitantly burdens the health and social care systems worldwide, with massive costs. Link N peptide has recently been described as a novel anabolic stimulator for intervertebral disc repair. In this study, we analyzed the influence on anabolic response, by delivering synthetic Link N encoding mRNA into primary human chondrocytes and mesenchymal stromal cells (SCP1 cells), Furthermore, both cell types were seeded on knitted titanium scaffolds, and the influence of Link N peptide mRNA for possible tissue engineering applications was investigated. Synthetic modified Link N mRNA was efficiently delivered into both cell types and cell transfection resulted in an enhanced expression of aggrecan, Sox 9, and type II collagen with a decreased expression of type X collagen. Interestingly, despite increased expression of BMP2 and BMP7, BMP signaling was repressed and TGFβ signaling was boosted by Link N transfection in mesenchymal stromal cells, suggesting possible regulatory mechanisms. Thus, the exogenous delivery of Link N peptide mRNA into cells augmented an anabolic response and thereby increased extracellular matrix synthesis. Considering these findings, we suppose that the cultivation of cells on knitted titanium scaffolds and the exogenous delivery of Link N peptide mRNA into cells could mechanically support the stability of tissue-engineered constructs and improve the synthesis of extracellular matrix by seeded cells. This method can provide a potent strategy for articular cartilage and intervertebral disc regeneration.

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“…When alginate was used on its own, a fast release was shown, and this was attributed to electrostatic repulsive forces between the negatively charged alginate and the negatively charged synthetic mRNA. Thus, combining the negatively charged alginate with the positively charged chitosanglycerophosphate resulted in a hybrid thermogel capable of sustained release of synthetic mRNA, at an optimal rate, faster than that of chitosan alone, but slower than that of alginate alone (Steinle et al, 2018).…”

Section: Chitosan Based Thermogelsmentioning

confidence: 99%

Hybrid Thermo-Responsive Polymer Systems and Their Biomedical Applications

et al. 2020

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Targeted and controlled drug delivery employing "smart materials" is a widely investigated field, within which stimuli-responsive polymers, particularly those which are thermo-responsive, have received considerable attention. Thermo-responsive polymers have facilitated the formulation of in situ gel forming systems which undergo a sol-gel transition at physiological body temperature, and have revolutionized the fields of tissue engineering, cell encapsulation, and controlled, sustained delivery of both drugs and genes. However, the use of single thermo-responsive polymers in the creation of these systems has posed numerous problems in terms of physico-mechanical properties, such as poor mechanical strength, high critical gelation concentrations (CGC) resulting in increased production costs and solutions that are too viscous, toxicity, as well as gelation temperatures that are incompatible with physiological body temperatures. Hybridization of these thermo-responsive polymers with other polymers has therefore been employed, resulting in the creation of tailor-made drug delivery systems that have optimal gelation temperatures and concentrations, ideal viscosities and improved gel strengths. This article reviews various thermo-responsive polymers that have been employed in the formulation of thermo-gelling systems. Special attention has been given to the hybridization of each of these polymers, the resulting systems that have been created, and their biomedical applications.

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Incorporation of Synthetic mRNA in Injectable Chitosan-Alginate Hybrid Hydrogels for Local and Sustained Expression of Exogenous Proteins in Cells

Cited by 26 publications

References 59 publications

Injectable Biodegradable Chitosan‐Alginate 3D Porous Gel Scaffold for mRNA Vaccine Delivery

Injectable Biodegradable Chitosan‐Alginate 3D Porous Gel Scaffold for mRNA Vaccine Delivery

Exogenous Delivery of Link N mRNA into Chondrocytes and MSCs—The Potential Role in Increasing Anabolic Response

Hybrid Thermo-Responsive Polymer Systems and Their Biomedical Applications

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