Sulfated hyaluronic acid gel for the treatment of rheumatoid arthritis in rats.

Rheumatoid arthritis (RA) is a chronic autoimmune disease. NSAIDS, cyclophosphamide and glucocorticoid were commonly used to treat RA in clinical application, which long-term administration of these drugs caused serious adverse reactions. Therefore, sulfated hyaluronic acid (sHA) gel (SG) was prepared to firstly treat the RA and avoid the problem of toxic side effect caused by long-term application. In vitro evaluation showed that sHA inhibited the level of reactive oxygen species and TNF-α, IL-1ß, and IL-6, and decreased the ratio of macrophage M1/M2 type, which exerted better anti-inflammatory capacity. In vivo studies showed that the injection of SG into the joint cavity of collagen-induced rheumatoid arthritis (CIA) rats could effectively treat joint swelling and reduce the level of inflammatory factors in the serum. Immunofluorescence showed that SG exerted its anti-inflammatory activity by decreasing the ratio of M1/M2 type macrophages in synovial tissue. Cartilage tissue sections showed that SG reduced bone erosion and elevated chondrocyte expression. These results confirmed that sHA is expected to be developed as a drug to treat or relieve RA, which could effectively regulate the level of macrophages in rat RA, alleviate the physiological state of inflammatory over-excitation, and improve its anti-inflammatory and antioxidant capacity.

Preparation and evaluation of lecithin/zein hybrid nanoparticles for the oral delivery of Panax notoginseng saponins.

Panax Notoginseng Saponins (PNS) has been widely used in the prevention and treatment of cardiovascular and cerebrovascular diseases such as myocardial infarction, heart failure and cerebral infarction. However, oral administration of PNS showed low bioavailability because of its instability and poor membrane permeability in the gastrointestinal tract. Here, lipoprotein-inspired hybrid nanoparticles of PNS-Lecithin-Zein (PLZ-NPs) were prepared by using a simple phase separation method, which possessed a core-shell structure, where zein was used as protein part to replace the animal origin protein to increase the resistance to acid and enzymes while lecithin was used as the lipid composition to improve the oral absorption of PNS as well as to increase the drug loading capacity of PNS into the nanocarriers. The results of stability test showed that PLZ-NPs had robust enzymolysis resistance ability for acid and digestive enzymes of gastrointestinal environments. The fluorescent resonance energy transfer (FRET) assay confirmed the ability of LZ-NPs to be intactly absorbed by Caco-2 cell monolayer. Cell transport studies demonstrated that the permeability of PLZ-NPs in Caco-2/HT29-MTX co-culture cell model was 1.5-fold that of PNS. Meanwhile, the single-pass intestinal perfusion assay proved the absorption parameter Peff of PLZ-NPs was 1.75 and 1.80 times higher than that of PNS in the ileum and jejunum, respectively. Finally, the in vivo pharmacokinetic experiment showed that the relative oral bioavailability of PLZ-NPs was 1.71-fold that of free PNS in SD rat. In summary, the employment of the Lecithin/Zein hybrid nanoparticles could be considered as a promising approach for PNS analogues.

Lecithins-Zein nanoparticles for antifungal treatment: Enhancement and prolongation of drug retention in skin with reduced toxicity.

Fungal infections are one of the major skin healthcare issues and cause significant morbidity. Ketoconazole (KC) as a broad-spectrum antifungal drug is widely used to treat skin fungal diseases. However, its therapeutic effects are limited by low concentration, short duration of drug efficacy in the skin and severe systemic toxicity. Here, the ketoconazole loaded Lecithins-Zein nanoparticles (KLZ-NPs) with core-shell structure were designed to resolve above problems. In vitro penetration test confirmed that the ketoconazole concentration of the KLZ-NPs group in the stratum corneum and deeper layers increased significantly (2.98-fold, 1.51-fold higher to free ketoconazole, respectively). Meanwhile, follicular closing technique showed the formed nanoparticles via follicle pathway into the skin had been significantly enhanced, and the results of the visual fluorescent images also confirmed it. Additionally, in the in vivo imaging experiment, the fluorescence intensity of the single applying of the DiR-LZ-NPs was higher than that of the thrice usage of the free DiR. More importantly, the results also indicated that the accumulation of nanoparticles in the liver and spleen was significantly reduced. Hence, Lecithins-Zein nanoparticles are a promising strategy to enhance the drug concentration, prolong efficacy and reduce systemic toxicity in the topical administration for antifungal treatment.

Flavonoid-rich foods (FRF): A promising nutraceutical approach against lifespan-shortening diseases.

It is well documented that life expectancy in developed countries at birth is going to surpass the 20th century. However, regrettably, a potential decline in life expectancy has been proposed for these nations in the 21st century due to a rapid upsurge in the prevalence of fatal degenerative diseases like cardiovascular diseases (CVD), cancer and diabetes. Collectively, these three diseases accounted for 65% of all deaths in urbanized societies and were considered as a dynamic issue for shortening the genetically determined lifespan through increased mortalities, morbidities, disabilities, immense sufferings, and premature aging. These fatal degenerative diseases and premature aging are closely associated with oxidative stress produced by the free radicals in the body. In epidemiologic studies, flavonoid-rich foods (FRF) like fruits, vegetables, and beverages have been associated as protective agents against these diseases. These also have been observed for their geroprotective effects and help in preventing premature aging and deterioration of brain function, which is related to Alzheimer's disease and dementia. In this review, we presented a comprehensive overview of the FRF for their potential role against lifespan-shortening complications, i.e., CVD, cancer, and diabetes. We also have drawn the future perspective and dietary guidelines to reduce the fatal disease burden in urban populations.

Indocyanine Green-Parthenolide Thermosensitive Liposome Combination Treatment for Triple-Negative Breast Cancer.

BACKGROUND: Certain patients with triple-negative breast cancer cannot tolerate the serious adverse effects of cytotoxic chemotherapy agents, which significantly affect the disease prognosis. PURPOSE: Research into the combined use of photosensitizers and non-cytotoxic antineoplastic drugs for the safe treatment of triple-negative breast cancer is vital. METHODS: In this study, the photosensitizer indocyanine green and the natural drug parthenolide were co-loaded into thermosensitive liposomes. Under a near-infrared irradiation, indocyanine green reached excitation levels, releasing heat, and the liposome underwent a phase transition, releasing the drug were researched. RESULTS: Thus, indocyanine green and parthenolide exert synergistic antineoplastic effects. In the nude mice xenograft MDA-MB-231 tumor model, the tumor inhibition rate of indocyanine green-parthenolide thermosensitive liposomes was approximately 2.08-fold than that of paclitaxel and demonstrated a good initial safety evaluation. CONCLUSION: Photosensitizers and non-cytotoxic antineoplastic agents in combination with nanoscale carriers should be further investigated for the treatment of tumors.

Preparation and characterization of parthenolide nanocrystals for enhancing therapeutic effects of sorafenib against advanced hepatocellular carcinoma.

A novel nanocrystals delivery system of parthenolide (PTL) was designed to combined application with sorafenib (Sora) for advanced hepatocellular carcinoma (HCC) therapy, attempting to not only improve the poor aqueous solubility of PTL, but also enhance the synergistic therapeutic effects with Sora. The PTL nanocrystals (PTL-NCs) were prepared by precipitation-high-pressure homogenization method. The formed PTL-NCs with rod morphology possessed size of 126.9 ± 2.31 nm, zeta potential of -11.18 ± 0.59 mV and drug loading of 31.11 ± 1.99%. Meanwhile, PTL in PTL-NCs exhibited excellent storage stability and sustained release behavior. The combination therapy of Sora and PTL-NCs (Sora/PTL-NCs) in vitro for HepG2 cells presented superior therapeutic effects over that of individual PTL and Sora on intracellular uptake, cell proliferation inhibition and migration inhibition. Meanwhile the strongest anti-tumor effect with 81.86% inhibition rate and minimized systemic toxicity of Sora/PTL-NCs in vivo were obtained on tumor-bearing mice compared with that of PTL (48.84%) and Sora (58.83%). Thus, these findings suggested that PTL-NCs as an effective delivery system for the synergistically used with Sora to gain an optimal response against HCC, for referenced in the industrialization of nanocrystals products for intravenous administration.

Rapamycin loaded TPGS-Lecithins-Zein nanoparticles based on core-shell structure for oral drug administration.

Rapamycin as a novel macrolide immunosuppressive agent has been commonly used in organ transplantation owing to its stronger immunosuppressive effect, non-nephrotoxicity and lower side effect. However its drawbacks of low bioavailability and big individual difference remain to be improved in clinical application. Here rapamycin loaded TPGS-Lecithins-Zein nanoparticles (RTLZ-NPs) with core-shell structure were prepared by the phase separation method. The RTLZ-NPs were approximately 190.3 nm in size, with PDI and zeta potential about 0.256 and -19.71 mV respectively. Drug entrapment and loading achieved were about 86.64 and 25.73% respectively. Meanwhile RTLZ-NPs exhibited favorable enzymolysis resistance abilities in gastrointestinal environments and enhanced uptake in Caco-2 cells. The optimum absorption sites of rapamycin in the intestine were duodenum and jejunum as single-pass intestinal perfusion assay. Upon also considering the results of Caco-2 cell assay, it could be speculated that the transport of rapamycin in vivo involved active transport as well as P-glycoprotein (P-gp) based efflux. Finally, the relative oral bioavailability of RTLZ-NPS was 4.33 fold higher than free rapamycin in SD rat. Altogether the designed nanoparticles can be an efficient oral delivery strategy for rapamycin analogues to prevent the attacks from destructive enzymes, reduce cell efflux, increase cell uptake, and then enhance the oral bioavailability.

TPGS-modified liposomes for the delivery of ginsenoside compound K against non-small cell lung cancer: formulation design and its evaluation in vitro and in vivo.

OBJECTIVE: This work aimed at preparing ginsenoside compound K (GCK)-loaded liposomes modified with TPGS (GCKT-liposomes) to enhance solubility and targeting capability of GCK, as well as inhibit the efflux of GCK from tumour cells. METHODS: GCKT-liposomes were prepared by the thin-film hydration method and characterized by particle size, polydispersity, zeta potential and drug encapsulation efficiency. A549 cells were used as antitumour cell model to access the cellular uptake of the GCK and perform its antitumour function. The enhancement of in vivo antitumour efficacy of GCKT-liposomes was evaluated by nude mice bearing tumour model. KEY FINDINGS: The results showed that GCKT-liposomes achieved a comparatively high drug loading efficiency and reasonable particle size at the ratio of 7 : 3 (phospholipid: TPGS). The in vitro release demonstrated that the dissolution of GCK was remarkably improved by entrapping it into liposomes. In addition, GCKT-liposomes exhibited a great hypersensitizing effect on A549 cells, and the cellular uptake was enhanced. Compared with free GCK, the IC50 of GCKT-liposomes was significantly reduced (16.3 ± 0.8 vs 24.9 ± 1.0 µg/ml). In vivo antitumour assay also indicated that GCKT-liposomes achieved higher antitumour efficacy (67.5 ± 0.5 vs 40.8 ± 0.7%). CONCLUSION: The novel GCKT-liposomes significantly improved the antitumour efficacy of GCK.

[Preparation and in vitro dissolution of magnolol solid dispersion].

In this study, solid dispersion system of magnolol in croscarmellose sodium was prepared by using the solvent evaporation method, in order to increase the drug dissolution. And its dissolution behavior, stability and physical characteristics were studied. The solid dispersion was prepared with magnolol and croscarmellose sodium, with the proportion of 1∶5, the in vitro dissolution of magnolol solid dispersion was up to 80.66% at 120 min, which was 6.9 times of magnolol. The results of DSC (differential scanning calorimetry), IR (infra-red) spectrum and SEM (scanning electron microscopy) showed that magnolol existed in solid dispersion in an amorphous form. After an accelerated stability test for six months, the drug dissolution and content in magnolol solid dispersion showed no significant change. So the solid dispersion prepared with croscarmellose sodium as the carrier can remarkably improve the stability and dissolution of magnolol.

N-octyl-N-arginine-chitosan (OACS) micelles for gambogic acid oral delivery: preparation, characterization and its study on in situ intestinal perfusion.

CONTEXT: Gambogic acid (GA) can inhibit the growth of various cancer cells. However, the low bioavailability caused by insolubility, limits its clinical application. L-arginine is always used with GA to form a complex to obtain the higher solubility. Moreover, guanidyl group from arginine, which can facilitate the cellular uptake, was identified. OBJECTIVE: In this study, L-arginine and chitosan (CS) were used for the first time to prepare N-octyl-N-arginine CS (OACS), a novel amphiphilic carrier for GA with solubility- and absorption-enhancing functions; the characterization of the GA loaded OACS micelles (GA-OACS) and its absorption-enhancing effect were also investigated. MATERIALS AND METHODS: GA-OACS were prepared by the dialysis method. The formed micelles were characterized and evaluated by atomic force microscope (AFM), dynamic light scattering, differential scanning calorimeter (DSC), solubility test, in vitro release and in situ intestinal perfusion. RESULTS: The GA-OACS micelles were successfully prepared attaining a 35.3% drug loading and 82.2% entrapment efficiency. GA-OACS had a homogeneous particle size of 160.3 nm; +21.8 mv zeta potential with smooth continuous surface was observed by using AFM. DSC diagram suggested that GA was encapsulated in the micelles. Meanwhile, GA encapsulated in micelles exhibited a desirable slow release in vitro experiment. The solubility of GA in OACS micelles was increased up to 3.16 ± 0.13 mg/mL, 2320 times than that of free GA. The single pass perfusion showed that the absorption of GA-OACS micelles was enhanced 3.6-fold, 2.1-fold and 2.2-fold for jejunum, ileum and colon, respectively. DISCUSSION AND CONCLUSION: OACS provided excellent ability of drug loading, increasing solubility and enhanced absorption for GA, which indicated that OACS micelles as an oral drug delivery carrier may have potential research and application values.

[Study on intestinal absorption kinetics of apigenin in rats].

To establish an in situ single-way intestinal perfusion model, in order to study the intestinal absorption kinetics of AP. The concentration of AP in the perfusate was determined by HLPC. The results showed different AP concentrations in all intestinal segments, with the fastest absorption rate in duodenum, which was followed by jejunum, ileum and colon. In general, the constant absorption rate (Ka) of AP in duodenum and jejunum first increased and then decreased with the rise in drug concentration (P <0. 05); the absorption mechanism may be related to active transport and facilitated diffusion factors. The constant absorption rate (Ka) of AP in ileum and colon generally kept unchanged with the rise diffusion in drug concentration, the absorption mechanism may be related to passive.

Studies on lactoferrin nanoparticles of gambogic acid for oral delivery.

PURPOSE: Lactoferrin (Lf), a mammalian cationic iron-binding glycoprotein belonging to the transferrin (Tf) family, has been widely used in a variety of fields ranging from treating infant diarrhea and supporting newborn growth to food and pharmaceutical applications. In this study, Lf nanoparticles were firstly used as carriers of gambogic acid (GA) to enhance oral absorption and anti-cancer activity, hence reducing the related toxic effect. METHODS: Gambogic acid-lactoferrin nanoparticles (GL-NPs) were prepared by the nanoparticle albumin-bound (NAB) technology. The formed nanoparticles were characterized by DSC, TEM, etc. In situ intestinal perfusion experiment was performed to clarify the absorption mechanism of GL-NPs. Furthermore in vivo and in vitro anti-tumor activities of GL-NPs were also investigated. RESULTS: GL-NPs was successfully prepared with about 150 nm mean size, +20 mV ζ potential, 92.3 ± 7.2% encapsulation efficiency and 9.04 ± 0.7% DL; GL-NPs also exhibited a better stability and a desirable slow release in vitro experiment. The results of in situ intestinal perfusion showed a transformation of GA absorption from passive diffusion into active transport or facilitated diffusion by GL-NPs. MTT assay of GL-NPs showed almost an equal anti-proliferative effect with arginine solution of GA (Arg-GA) in HepG2 cell. The inhibitory rate against S180 tumor mice after oral administration of GL-NPs was up to 86.01% which was 1.39-folds of intravenous injection of Arg-GA. CONCLUSION: The in vitro results showed that the NAB technology was feasible for industrial production of Lf nanoparticles and the in vivo results proved that the effective GL-NPs is a promising approach for the oral delivery of GA. These obtained research works have also paved the preliminary way for the study of Lf as an oral drug delivery carrier.

[Study on intestinal absorption kinetics of gambogic acid in rats].

OBJECTIVE: To investigate the intestinal absorption kinetics of gambogic acid (GA) in rats. METHOD: In situ single-way intestinal perfusion model was established to study the intestinal absorption kinetics of GA in different absorption segments, and the concentration of GA in the perfusate was determined by HPLC. The effect of drug concentrations on intestinal absorption was also detected. RESULT: GA showed a higher absorption rate than other intestinal segments (P < 0.05) and kept unchanged in duodenum after addition in drug concentration. CONCLUSION: GA can be absorbed in all intestinal segments in rats with the higher absorption rate in duodenum. Its mechanism is passive diffusion.

Enhanced bioavailability after oral and pulmonary administration of baicalein nanocrystal.

The aim of the study was to investigate the potential of oral and pulmonary nanocrystal to enhance the bioavailability of baicalein, a bioactive flavonoid isolated from the root of Scutellaria baicalensis Georgi. So far, the nano-sized delivery system of baicalein and its pulmonary delivery have received no exploration. In the present investigation, the baicalein nanocrystal was prepared by anti-solvent recrystallization followed by high pressure homogenization. In vitro characterization was performed including particle size and distribution, Zeta potential, dissolution, scanning electron microscopy, differential scanning calorimetry and X-ray powder diffractometry. It was indicated that no crystalline change was observed after nanocrystal preparation. The baicalein nanocrystal containing only trace of stabilizer exhibited a significantly enhanced dissolution of baicalein. In vivo test was also carried out in rats and pharmacokinetic parameters of the baicalein crystal and the baicalein nanocrystal after gavage and pulmonary administration were compared, based on the simultaneous determination of baicalein and baicalin by high performance liquid chromatography. The mean relative bioavailability of oral baicalein nanocrystal was 1.67-fold that of oral baicalein crystal. The pulmonary baicalein nanocrystal had rapid and extensive absorption and had almost identical pharmacokinetic parameters to intravenous baicalein injection.