Stability and bioactivity of chitosan as a transfection agent in primary human cell cultures: A case for chitosan-only controls

Cupino, Tanya L.; Watson, Billy A.; Cupino, Alan; Oda, Keiji; Ghamsary, Mark; Soriano, Salvador; Kirsch, Wolff M.

doi:10.1016/j.carbpol.2017.10.021

Cited by 10 publications

(6 citation statements)

References 53 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…In this experiment, two kinds of environments in vitro of DNA enzymes with different concentrations of 1 and 2 U were set, and then the compound particles with N/P ratio of 45 were placed in this condition. As shown in Figure 3b The zeta potentials of CS-5Arg and CS-5Arg/pDNA complexes 3.6 | The cytotoxicity of CS-Arg/pDNA and CS-5Arg/pDNA complexes It was known that cationic polymers possessed cytotoxicity, and it could act on cell membranes and extracellular matrix, resulting in serious side effects (Cupino et al, 2018). The cytotoxicity of CS-Arg/ pDNA complexes and CS-5Arg/pDNA complexes in HEK-293 cells were studied by CCK-8 assay.…”

Section: The Agarose Gel Electrophoresis Of the Complexesmentioning

confidence: 99%

Study on the transfection efficiency of chitosan‐based gene vectors modified with poly‐l‐arginine peptides

Zhang

Chu

Sun

et al. 2020

J Biomedical Materials Res

View full text Add to dashboard Cite

Although in a series of studies, arginine peptides had shown the ability to promote the targeting delivery efficacy, the relationship between the transfection efficiency and the length of the poly-l-arginine chain had seldom been reported. This study was aimed to explore whether the chain length of poly-l-arginine grafted on chitosan had a great significance on the transfection efficiency of entering the cells. Herein, arginine and arginine peptide modified chitosan were synthesized as gene vectors (CS-Arg and CS-5Arg) and then the chemical structures were characterized by using 1 H NMR. The CS-Arg and CS-5Arg were combined with plasmids by electrostatic interactions to form stable particles. The morphology features, Zeta potentials, and buffering capacity of the complex particles were analyzed. Afterward, the combination ability with DNA and the protection ability to DNase I were studied, and the gene transfection efficiency and cellular uptake were investigated in vitro. The results showed that the gene transfection efficiency of the chitosan was significantly enhanced by arginine-graft modification. However, there were no significant differences between the CS-Arg and the CS-5Arg. The molecular simulation results indicated that the guanidine groups of grafted arginine were shielded by chitosan molecule and the guanidine groups contributed little to the gene transfection efficiency. The results demonstrated that the increased chain length of grafted arginine had no significantly enhanced effect on the transfection efficiency, which could provide convincing evidence for the construction and application of arginine and chitosan derivatives as gene vectors, and could promote the development of gene delivery system.

show abstract

Section: The Agarose Gel Electrophoresis Of the Complexesmentioning

confidence: 99%

Study on the transfection efficiency of chitosan‐based gene vectors modified with poly‐l‐arginine peptides

Zhang

Chu

Sun

et al. 2020

J Biomedical Materials Res

View full text Add to dashboard Cite

show abstract

“…These include antitumor, antifungal, antioxidant, immunoenhancing, and wound healing properties. [ 11 ] Furthermore, chitosan is biodegradable and biocompatible, and the low costs associated with its production allow extensive usage in a multitude of applications such as pharmaceutics, [ 12 ] tissue engineering, [ 13 ] gene delivery, [ 14 ] and drug delivery systems. [ 15 ]…”

Section: Introductionmentioning

confidence: 99%

“…The deacetylation of chitin is required to overcome the practical disadvantages of chitin including high hydrophobicity, insolubility in solvents such as water (due to its intramolecular hydrogen bonds), ordinary organic solvents, and dilute acid or alkali. In contrast, chitosan biodegradable and biocompatible, and the low costs associated with its production allow extensive usage in a multitude of applications such as pharmaceutics, [12] tissue engineering, [13] gene delivery, [14] and drug delivery systems. [15] Fluorescent NCs offer several advantages and have been used extensively in the literature.…”

Section: Introductionmentioning

confidence: 99%

Biomedical and Pharmacological Uses of Fluorescein Isothiocyanate Chitosan‐Based Nanocarriers

Caprifico

Polycarpou

Foot

et al. 2020

Macromolecular Bioscience

View full text Add to dashboard Cite

with a DDA ranging between 70% and 85% becomes soluble in dilute acidic solutions such as acetic acid or formic acid. [1] Indeed, the primary amino groups of chitosan have a pK a of ≈6.5 so that, following protonation, they confer upon chitosan the feature of solubility in weakly acidic aqueous solution, allowing the polymer to be easily manipulated. [3] The DDA of chitosan plays a key role in determining its physicochemical properties, which are affected by the proportion of free amino groups (-NH 2) remaining on the polymeric chain upon deacetylation. Both the primary amino and secondary hydroxyl groups behave as reactive functional groups on chitosan. Indeed, these groups allow chitosan to be susceptible to chemical modifications such as acylation, tosylation, quaternization, alkylation, and O-carboxymethylation. [4] As a result, the chitosan structure can be functionalized and optimized to improve drug loading or release. [5] Chitosan-based nanocarriers (ChNCs), including nanoparticles (NPs), micelles, or polyplexes, have received significant attention for their numerous advantageous features such as natural sourcing, biodegradability, easy functionalizations, and low toxicity. [6] The mucoadhesive property of chitosan is key for drug delivery purposes. [7] Indeed, chitosan is a positively charged polymer that can form electrostatic bonds with the negatively charged mucous layer, made of mucin glycoproteins that cover the epithelial cells of the mucosa. The establishment of these bonds allows drug-loaded ChNCs to exhibit higher absorption and retention times at the target, while reducing dosing frequency. [8] Furthermore, chitosan is used as permeation enhancer since increases the uptake of the drugs through a transient and reversible opening of the tight junctions (TJs) protecting the paracellular pathway between endothelial cells in the so-called blood-brain barrier (BBB). [9] This is due to F-actin depolymerization and leakage of the TJ protein zonula occludens-1. [7] The small size and large surface area of NCs allow their passage through biological membranes, such the BBB, and accumulation at the intracellular (such as lysosomes) or intranuclear (DNA or RNA) target site. [10] ChNCs are distinctive for their ability to protect the encapsulated therapeutic agent and improve its bioavailability by altering the pharmacokinetics. [10] The endocytic mechanisms responsible for the internalization of ChNCs as a drug delivery system may differ according to the cell type and the drug to be delivered. Numerous biological and pharmacological properties characterize chitosan. These include antitumor, antifungal, antioxidant, immunoenhancing, and wound healing properties. [11] Furthermore, chitosan is Chitosan-based nanocarriers (ChNCs) are considered suitable drug carriers due to their ability to encapsulate a variety of drugs and cross biological barriers to deliver the cargo to their target site. Fluorescein isothiocyanatelabeled chitosan-based NCs (FITC@ChNCs) are used extensively in biomedical and pharmacological...

show abstract

“…As a natural degradable cationic polymer, chitosan (CS), with good stability and cytocompatibility, could enhance drug internalization [23,24] and has already been used for the delivery of DNA and RNA [25][26][27][28]. Via electrostatic interactions, CS can bind miRNAs to form relatively stable CS-miRNA nanoplexes (NPs) with a positive charge.…”

Section: Introductionmentioning

confidence: 99%

Chitosan-miRNA functionalized microporous titanium oxide surfaces via a layer-by-layer approach with a sustained release profile for enhanced osteogenic activity

Liu

et al. 2020

J Nanobiotechnol

View full text Add to dashboard Cite

Background The biofunctionalization of titanium implants for high osteogenic ability is a promising approach for the development of advanced implants to promote osseointegration, especially in compromised bone conditions. In this study, polyelectrolyte multilayers (PEMs) were fabricated using the layer-by-layer approach with a chitosan-miRNA (CS-miRNA) complex and sodium hyaluronate (HA) as the positively and negatively charged polyelectrolytes on microarc-oxidized (MAO) Ti surfaces via silane-glutaraldehyde coupling. Methods Dynamic contact angle and scanning electron microscopy measurements were conducted to monitor the layer accumulation. RiboGreen was used to quantify the miRNA loading and release profile in phosphate-buffered saline. The in vitro transfection efficiency and the cytotoxicity were investigated after seeding mesenchymal stem cells (MSCs) on the CS-antimiR-138/HA PEM-functionalized microporous Ti surface. The in vitro osteogenic differentiation of the MSCs and the in vivo osseointegration were also evaluated. Results The surface wettability alternately changed during the formation of PEMs. The CS-miRNA nanoparticles were distributed evenly across the MAO surface. The miRNA loading increased with increasing bilayer number. More importantly, a sustained miRNA release was obtained over a timeframe of approximately 2 weeks. In vitro transfection revealed that the CS-antimiR-138 nanoparticles were taken up efficiently by the cells and caused significant knockdown of miR-138 without showing significant cytotoxicity. The CS-antimiR-138/HA PEM surface enhanced the osteogenic differentiation of MSCs in terms of enhanced alkaline phosphatase, collagen production and extracellular matrix mineralization. Substantially enhanced in vivo osseointegration was observed in the rat model. Conclusions The findings demonstrated that the novel CS-antimiR-138/HA PEM-functionalized microporous Ti implant exhibited sustained release of CS-antimiR-138, and notably enhanced the in vitro osteogenic differentiation of MSCs and in vivo osseointegration. This novel miRNA-functionalized Ti implant may be used in the clinical setting to allow for more effective and robust osseointegration.

show abstract

Stability and bioactivity of chitosan as a transfection agent in primary human cell cultures: A case for chitosan-only controls

Cited by 10 publications

References 53 publications

Study on the transfection efficiency of chitosan‐based gene vectors modified with poly‐l‐arginine peptides

Study on the transfection efficiency of chitosan‐based gene vectors modified with poly‐l‐arginine peptides

Biomedical and Pharmacological Uses of Fluorescein Isothiocyanate Chitosan‐Based Nanocarriers

Chitosan-miRNA functionalized microporous titanium oxide surfaces via a layer-by-layer approach with a sustained release profile for enhanced osteogenic activity

Contact Info

Product

Resources

About