Chitosan-coated Nanostructured Lipid Carriers Research Articles

Preparation and Optimization of MiR-375 Nano-Vector Using Two Novel Chitosan-Coated Nano-Structured Lipid Carriers as Gene Therapy for Hepatocellular Carcinoma.

Currently, there is still a lack of effective carriers with minimal side effects to deliver therapeutic miRNA. Thus, it is crucial to optimize novel drug delivery systems. MiR-375 has proven superior therapeutic potency in Hepatocellular carcinoma (HCC). The purpose of this study was to fabricate 2 novel and smart nano-carriers for the transportation efficiency of miR-375 in HCC cells and enhance its anti-tumor effects. We established the miR-375 construct through the pEGP- miR expression vector. Two nano-carriers of solid/liquid lipids and chitosan (CS) were strategically selected, prepared by high-speed homogenization, and optimized by varying nano-formulation factors. Thus, the two best nano-formulations were designated as F1 (0.5% CS) and F2 (1.5% CS) and were evaluated for miR-375 conjugation efficiency by gel electrophoresis and nanodrop assessment. Then, physio-chemical characteristics and stability tests for the miR-375 nano-plexes were all studied. Next, its efficiencies as replacement therapy in HepG2 cells have been assessed by fluorescence microscopy, flow cytometry, and cytotoxicity assay. The obtained data showed that two cationic nanostructured solid/liquid lipid carriers (NSLCs); F1 and F2 typically had the best physio-chemical parameters and long-term stability. Moreover, both F1 and F2 could form nano-plexes with the anionic miR-375 construct at weight ratios 250/1 and 50/1 via electrostatic interactions. In addition, these nano-plexes exhibited physical stability after three months and protected miR-375 from degradation in the presence of 50% fetal bovine serum (FBS). Furthermore, both nano-plexes could simultaneously deliver miR-375 into HepG2 cells and they ensure miR re-expression even in the presence of 50% FBS compared to free miR-375 (p-value < 0.001). Moreover, both F1 and F2 alone significantly exhibited minimal cytotoxicity in treated cells. In contrast, the nano-plexes significantly inhibited cell growth compared to free miR-375 or doxorubicin (DOX), respectively. More importantly, F2/miR-375 nano-plex exhibited more anti-proliferative activity in treated cells although its IC50 value was 55 times lower than DOX (p-value < 0.001). Collectively, our findings clearly emphasized the multifunctionality of the two CS-coated NSLCs in terms of their enhanced biocompatibility, biostability, conjugation, and transfection efficiency of therapeutic miR-375. Therefore, the NSLCs/miR-375 nano-plexes could serve as a novel and promising therapeutic strategy for HCC.

Preparation and Characterization of Chitosan-Coated Nanostructured Lipid Carriers (CS-NLC) Containing (6)-Gingerol and Investigating their Toxicity Against MCF-7 Breast Cancer Cell Line

Preparation and Characterization of Chitosan-Coated Nanostructured Lipid Carriers (CS-NLC) Containing (6)-Gingerol and Investigating their Toxicity Against MCF-7 Breast Cancer Cell Line

Chitosan-coated nanostructured lipid carriers for effective brain delivery of Tanshinone IIA in Parkinson’s disease: interplay between nuclear factor-kappa β and cathepsin B

Parkinson’s disease (PD) is the second most common progressive neurodegenerative disorder associated with increased oxidative stress, the underlying vital process contributing to cell death. Tanshinone IIA (TAN) is a phytomedicine with a documented activity in treating many CNS disorders, particularly PD owing to its unique anti-inflammatory and antioxidant effect. However, its clinical utility is limited by its poor aqueous solubility, short half-life, and hence low concentration reaching targeted cells. This work aimed to develop a biocompatible chitosan-coated nanostructured lipid carriers (CS-NLCs) for effective brain delivery of TAN for PD management. The proposed nanosystem was successfully prepared using a simple melt-emulsification ultra-sonication method, optimized and characterized both in vitro and in vivo in a rotenone-induced PD rat model. The developed TAN-loaded CS-NLCs (CS-TAN-NLCs) showed good colloidal properties (size ≤ 200 nm, PDI ≤ 0.2, and ζ-potential + 20 mV) and high drug entrapment efficiency (> 97%) with sustained release profile for 24 h. Following intranasal administration, CS-TAN-NLCs succeeded to achieve a remarkable antiparkinsonian and antidepressant effect in diseased animals compared to both the uncoated TAN-NLCs and free TAN suspension as evidenced by the conducted behavioral tests and improved histopathological findings. Furthermore, biochemical evaluation of oxidative stress along with inflammatory markers, nuclear factor-kabba β (NF-Kβ) and cathepsin B further confirmed the potential of the CS-TAN-NLCs in enhancing brain delivery and hence the therapeutic effect of TAN of treatment of PD. Accordingly, CS-TAN-NLCs could be addressed as a promising nano-platform for the effective management of PD.Graphical abstract

Chitosan-coated nanostructured lipid carriers for transdermal delivery of tetrahydrocurcumin for breast cancer therapy

Chitosan (Ch)-coated nanostructured lipid carriers (NLCs) have great potential for transdermal delivery with high localization of chemotherapeutics in breast cancer. This study used tetrahydrocurcumin (THC), a primary metabolite of curcumin with enhanced antioxidant and anticancer properties, as a model compound to prepare NLCs. Response surface methodology was employed to optimize THC-loaded Ch-coated NLCs (THC-Ch-NLCs) fabricated by high-shear homogenization. The optimized THC-Ch-NLCs had particle size of 244 ± 18 nm, zeta potential of −17.5 ± 0.5 mV, entrapment efficiency of 76.6 ± 0.2% and drug loading of 0.28 ± 0.01%. In vitro release study of THC-Ch-NLCs showed sustained release following the Korsmeyer-Peppas model with Fickian and non-Fickian diffusion at pH 7.4 and 5.5, respectively. THC-Ch-NLCs demonstrated significantly enhanced in vitro skin permeation, cell uptake, and remarkable cytotoxicity toward MD-MBA-231 breast cancer cells compared to the unencapsulated THC, suggesting Ch-NLCs as potential transdermal nanocarriers of THC for triple-negative breast cancer treatment.

Design of magnetic hybrid nanostructured lipid carriers containing 1,8-cineole as delivery systems for anticancer drugs: Physicochemical and cytotoxic studies

The development of versatile carriers to deliver chemotherapeutic agents to specific targets with establishing drug release kinetics and minimum undesirable side effects is becoming a promising relevant tool in the medical field. Magnetic hybrid nanostructured lipid carriers (NLC) were prepared by incorporation of 1,8-cineole (CN, a monoterpene with antiproliferative properties) and maghemite nanoparticles (MNPs) into a hybrid matrix composed of myristyl myristate coated with chitosan. Hybrid NLC characterized by DLS and TEM confirmed the presence of positively charged spherical nanoparticles of around 250 nm diameter and +10.2 mV of Z-potential. CN encapsulation into the lipid core was greater than 75 % and effectively released in 24 h. Modification of the crystalline structure of nanoparticles after incorporation of CN and MNPs was observed by XRD, DSC, and TGA analyses. Superparamagnetic NLC behavior was verified by recording the magnetization using a vibrating scanning magnetometer. NLC resulted in more cytotoxic than free CN in HepG2 and A549 cell lines. Particularly, viability inhibition of HepG2 and A549 cells was increased from 35 % to 55 % and from 38 % to 61 %, respectively, when 8 mM CN was incorporated into the lipid NPs at 24 h. Green fluorescent-labeled NLC with DIOC18 showed an enhanced cellular uptake with chitosan-coated NLC. Besides, no cytotoxicity of the formulations in normal WI-38 cells was observed, suggesting that the developed hybrid NLC system is a safe and good potential candidate for the selective delivery and potentiation of anticancer drugs.

Tacrolimus-loaded chitosan-coated nanostructured lipid carriers: preparation, optimization and physicochemical characterization

Tacrolimus as an immunosuppressive drug shows poor solubility in aqueous media, accompanied by a low/variable bioavailability in peroral administration due to its high first-pass metabolism. Therefore, in this study, drug-loaded chitosan-coated nanostructured lipid carriers (CCNLCs) were prepared to overcome these limitations. Tacrolimus-loaded NLCs were prepared via solvent displacement technique and were optimized. Stearic acid (SA) and glyceryl mono stearate (GMS) were used as solid lipids, oleic acid (OA) and tween 80 were used as liquid lipid and stabilizer, respectively, in the structure of NLC. The prepared nanoparticles were characterized by size and zeta potential measurements, drug loading determinations, scanning electron microscopy (SEM), and Fourier transform infrared (FTIR) analysis. Release profile of tacrolimus from NLCs and CCNLCs was also studied. The results indicated the effect of some preparation parameters on particle size as a dependent parameter. The selected formulations had particle sizes under 100 nm. Conversion of negative zeta potential of NLCs into positive quantities in CCNLCs was attributed to the effect of chitosan coating. SEM images of nanocarriers proved nanoscale dimensions and FTIR analysis showed no unwanted chemical reactions between drug and excipients. NLCs and CCNLCs released the drug in a sustained release pattern. However, chitosan-coating attenuated the burst effect and caused more extended release. The results of this study demonstrated formation of tacrolimus-loaded NLCs and coating them with chitosan with desirable characteristics. These findings suggested that this carrier is a good candidate to overcome serious bioavailability problems of common peroral formulations of tacrolimus.

Chitosan-coated nanostructured lipid carriers of fenofibrate with enhanced oral bioavailability and efficacy

Fenofibrate is frequently used to lower cholesterol levels in cardiovascular disease. Owing to its poor solubility and high gastrointestinal permeability, it is classified as a Biopharmaceutics Classification System class II compound. The aim of this study was to improve the solubility and bioavailability of fenofibrate by formulating it as fenofibrate-loaded nanostructured lipid carriers (FFB-NLCs) and coating it with a biodegradable polymer to allow controlled drug release. Chitosan-coated nanostructured lipid carriers (CF-NLCs) were prepared via an ultrasonication method using chitosan as the biodegradable polymer, stearic acid as the solid lipid, oleic acid as the liquid lipid, and Tween 80 as the surfactant. To study encapsulation efficiency and solubility conditions, stearic acid/oleic acid ratios were varied as 80/20, 70/30, 60/40, and 50/50 (mg/mg), by adjusting chitosan ratio. Chitosan is an adhesive polymer, coating the surface of the NLC to improve its bioavailability. All NLC formulations demonstrated a particle size of approximately 200 nm and a polydispersity index below 0.3. The encapsulation efficiencies of the NLC formulations were above 85%. For CF-NLCs, the solubility and encapsulation efficiency of fenofibrate were increased when compared with those of a commercial fenofibrate formulation. The pharmacokinetic and pharmacodynamic parameters of fenofibrate in the form of CF-NLCs were improved after oral administration. CF-NLCs can be used for allowing controlled release and improving the bioavailability and stability of fenofibrate.

Preparation and characterization of chitosan-coated nanostructured lipid carriers (CH-NLC) containing cinnamon essential oil for enriching milk and anti-oxidant activity

The cinnamon essential oil (CEO) was loaded in nanostructured lipid carriers (NLC) and NLC surfaces were coated with chitosan (CH-NLC). The particle size increased significantly from 99.98 nm, in the control NLC to 107.1, 112.2, and 119.4 nm in the CH-NLC coated with 0.05%, 0.1%, and 0.15% chitosan solution, respectively. After 40-day storage the mean size of all formulations slightly increased, however, the CH-NLC indicated less growing than NLC. The PDI value of NLC was lower than the CH-NLC (0.252 in comparison to 0.325). In addition, the negative zeta potential of NLC (−17.56) was changed to +21.4 for CH-NLC. The entrapment efficiency (EE%) of CH-NLC (93.68%) was significantly (P < 0.05) higher than NLC (88.14%) which could be attributed to the higher particle size of CH-NLC system. The encapsulation stability (ES%) of CH-NLC (84.6%) was significantly higher than NLC (78.12%). Furthermore, the FTIR spectrum showed no creation of new peaks after coating the NLC however the intensity of the peaks was shifted. The endothermic peaks of NLC illustrated low melting point (38.50 °C) and less ordered crystalline structure. The stability of milk fortified with this system against oxidation during the storage time was proved by thiobarbituric acid test and peroxide value.

Hybrid Nanostructured Films for Topical Administration of Simvastatin as Coadjuvant Treatment of Melanoma

This work aims at (1) assessing the potential of repurposing simvastatin (SV) to support the most common therapies against melanoma and (2) developing an innovative topical adhesive film, composed by chitosan-coated nanostructured lipid carriers (Ch-NLC) used as drug vehicle. A factorial design approach was employed as the basis for the formulation development. Optimized Ch-NLC displayed a particle size of 108 ± 1 nm, a polydispersity index of 0.226, a zeta potential of 17.0 ± 0.6 mV, as well as an entrapment efficiency of 99.86 ± 0.08%, and SV loading of 14.99 ± 0.01%. The performance of SV-Ch-NLC films was assessed in terms of release, permeation, and adhesion, as critical quality attributes. Cutaneous tolerability and in vitro cytotoxicity studies were performed to warrant film safety and drug effectiveness, respectively. The topical films provided a sustained release kinetic profile of SV and were classified as nonirritant systems. The encapsulation of SV increased cytotoxicity in melanoma cells. The key role of squalene as nanostructuring agent of the lipid nanoparticle matrix and as permeation enhancer was highlighted, suggesting its key action for potentiating skin permeation and uptake into melanoma cells. Topical SV-Ch-NLC films are thus able to provide an in situ extended drug delivery and useful as coadjuvant treatment of melanoma skin lesions.

Mucoadhesive chitosan-coated nanostructured lipid carriers for oral delivery of amphotericin B

This study describes the properties of an amphotericin B-containing mucoadhesive nanostructured lipid carrier (NLC), with the intent to maximize uptake within the gastrointestinal tract. We have reported previously that lipid nanoparticles can significantly improve the oral bioavailability of amphotericin B (AmpB). On the other hand, the aggregation state of AmpB within the NLC has been ascribed to some of the side effects resulting from IV administration. In the undissolved state, AmpB (UAmpB) exhibited the safer monomeric conformation in contrast to AmpB in the dissolved state (DAmpB), which was aggregated. Chitosan-coated NLC (ChiAmpB NLC) presented a slightly slower AmpB release profile as compared to the uncoated formulation, achieving 26.1% release in 5 hours. Furthermore, the ChiAmpB NLC formulation appeared to prevent the expulsion of AmpB upon exposure to simulated gastrointestinal pH media, whereby up to 63.9% of AmpB was retained in the NLC compared to 56.1% in the uncoated formulation. The ChiAmpB NLC demonstrated mucoadhesive properties in pH 5.8 and 6.8. Thus, the ChiAmpB NLC formulation is well-primed for pharmacokinetic studies to investigate whether delayed gastrointestinal transit may be exploited to improve the systemic bioavailability of AmpB, whilst simultaneously addressing the side-effect concerns of AmpB.

Preparation of Chitosan-Coated Nanostructured Lipid Carriers (CH-NLCs) to Control Iron Delivery and Their Potential Application to Food Beverage System.

Nanostructured lipid carriers (NLCs) and chitosan-coated NLCs (CH-NLCs) were prepared by a combined double emulsion and melt dispersion method. The physicochemical properties of them were investigated to determine the optimum conditions. At 3% emulsifier concentration, the NLCs showed about 191.0 ± 4.1 nm mean particle size and low polydispersity index value compared to those observed under other conditions. The encapsulation efficiency of CH-NLCs was 83.9% which was about 10% higher than that of NLCs. In vitro release test showed that low release amount of iron from the NLCs (11%) and CH-NLCs (5%) in simulated gastric condition within 3 h, whereas the iron release amounts of the NLCs and CH-NLCs were high (over 77%) in simulated intestinal condition because both NLCs and CH-NLCs were totally destructed in the intestinal condition after 3 h. In the thiobarbituric acid test, the absorbance of milk-fortified NLC and CH-NLCs was lower than that of original milk. Based on these results, CH-NLCs showed great potential for iron fortification in milk.

Ex vivo and In vivo Evaluation of Chitosan Coated Nanostructured Lipid Carriers for Ocular Delivery of Acyclovir.

Herpes keratitis is the most common infectious cause of blindness in the developed world. It may be treated by acyclovir (ACV), however this antiviral drug is poorly soluble with low ocular bioavailability requiring high and frequent dosing. Nanostructured lipid carriers (NLCs) were investigated to improve the ocular bioavailability of ACV by enhancing corneal penetration as well as prolonging the exposure of infected cells to the antiviral agent. Cell uptake studies, ex vivo tolerance and cell uptake efficacy as well as in vivo corneal permeation of the developed lipid based formulations were investigated. NLCs were fabricated by the hot microemulsion technique and coated with 0.5% w/v chitosan. NLCs were capable of increasing the cell uptake of encapsulated fluorescein and ACV as examined by fluorescence microscopy and high performance liquid chromatography (HPLC) respectively. When entrapped in NLCs, the antiviral efficacy of ACV was increased by 3.5 fold after 24 hrs of exposure. The in vivo corneal permeation of the formulation was studied on Albino rabbits with NLCs capable of increasing the corneal bioavailability by 4.5 fold when compared to a commercially available ACV ophthalmic ointment. NLCs enhanced the ocular bioavailability and antiviral properties of ACV through cell internalisation, sustained release, and increased corneal permeation.

Chitosan coated nanostructured lipid carriers for brain delivery of proteins by intranasal administration

The remarkable increase in the prevalence of neurodegenerative diseases has become a serious public health problem. Considering the lack of effective treatments to address these diseases and the difficulties in accessing the brain due to the blood–brain barrier (BBB), to attain a successful strategy to improve drug delivery to the brain, the administration route becomes a point of interest. The intranasal route provides a non-invasive method to bypass the BBB. Moreover, the development of new technologies for the protection and delivery of peptides is an interesting approach to consider. Thus, in this work, a suitable chitosan coated nanostructured lipid carrier (CS-NLC) formulation with the capacity to reach the brain after being intranasally administered was successfully developed and optimized. The optimal formulation displayed a particle size of 114nm with a positive surface charge of +28mV. The in vitro assays demonstrated the biocompatibility of the nanocarrier and its cellular uptake by 16HBE14o- cells. Furthermore, no haemagglutination or haemolysis processes were observed when the particles were incubated with erythrocytes, and no toxicity signals appeared in the nasal mucosa of mice after the administration of CS-NLCs. Finally, the biodistribution study of CS-NLC-DiR demonstrated an efficient brain delivery of the particles after intranasal administration. In conclusion, CS-NLC can be considered to be a safe and effective nanocarrier for nose-to-brain drug delivery; however, to obtain a higher concentration of the drug in the brain following intranasal administration, further modifications are warranted in the CS-NLC formulation.

Nanoencapsulation Enhances Epigallocatechin-3-gallate Stability and Its Antiatherogenic Bioactivities in Macrophages

We have successfully synthesized (-)-epigallocatechin-3-gallate (EGCG) encapsulated nanostructured lipid carriers (NLCE) and chitosan-coated NLCE (CSNLCE) using natural lipids, surfactant, chitosan, and EGCG. Nanoencapsulation dramatically improved EGCG stability. CSNLCE significantly increased EGCG content in THP-1-derived macrophages compared with nonencapsulated EGCG. As compared to 10 μM nonencapsulated EGCG, both NLCE and CSNLCE at the same concentration significantly decreased macrophage cholesteryl ester content. NLCE and CSNLCE significantly decreased mRNA levels and protein secretion of monocyte chemoattractant protein-1 (MCP-1) levels in macrophages, respectively. These data suggest that nanoencapsulated EGCG may have a potential to inhibit atherosclerotic lesion development through decreasing macrophage cholesterol content and MCP-1 expression.