Revolutionizing Knee Osteoarthritis Treatment: Innovative Self-Nano-Emulsifying Polyethylene Glycol Organogel of Curcumin for Effective Topical Delivery.

In the current study, self-nano-emulsifying (SNE) physically cross-linked polyethylene glycol (PEG) organogel (SNE-POG) as an innovative hybrid system was fabricated for topical delivery of water-insoluble and unstable bioactive compound curcumin (CUR). Response surface methodology (RSM) based on Optimal Design was utilized to evaluate the formulation factors. Solid fiber mechanism with homogenization was used to prepare formulations. Pharmaceutical evaluation including rheological and texture analysis, their mathematical correlations besides physical and chemical stability experiments, DSC study, in vitro release, skin permeation behavior, and clinical evaluation were carried out to characterize and optimize the SNE-OGs. PEG 4000 as the main organogelator, Poloxamer 188 (Plx188) and Ethyl Cellulose (EC) as co-gelator/nanoemulsifier agents, and PEG 400 and glycerin as solvent/co-emulsifier agents could generate SNE-POGs in PS range of 356 to 1410 nm that indicated organic base percentage and PEG 4000 were the most detrimental variables. The optimized OG maintained CUR stable in room and accelerated temperatures and could release CUR sustainably up to 72 h achieving high flux of CUR through guinea pig skin. A double-blind clinical trial confirmed that pain scores, stiffness, and difficulty with physical function were remarkably diminished at the end of 8 weeks compared to the placebo (71.68% vs. 7.03%, 62.40% vs. 21.44%, and 45.54% vs. 8.66%, respectively) indicating very high efficiency of system for treating knee osteoarthritis. SNE-POGs show great potential as a new topical drug delivery system for water-insoluble and unstable drugs like CUR that could offer a safe and effective alternative to conventional topical drug delivery system.

Exploiting Apical Sodium-Dependent Bile Acid Transporter (ASBT)-Mediated Endocytosis with Multi-Functional Deoxycholic Acid Grafted Alginate Amide Nanoparticles as an Oral Insulin Delivery System.

OBJECTIVE: Oral administration of insulin is a potential candidate for managing diabetes. However, it is obstructed by the gastrointestinal tract barriers resulting in negligible oral bioavailability. METHODS: This investigation presents a novel nanocarrier platform designed to address these challenges. In this regard, the process involved amination of sodium alginate by ethylene diamine, followed by its conjugation with deoxycholic acid. RESULTS: The resulting DCA@Alg@INS nanocarrier revealed a significantly high insulin loading content of 63.6 ± 1.03% and encapsulation efficiency of 87.6 ± 3.84%, with a particle size of 206 nm and zeta potentials of -3 mV. In vitro studies showed sustained and pH-dependent release profiles of insulin from nanoparticles. In vitro cellular studies, confocal laser scanning microscopy and flow cytometry analysis confirmed the successful attachment and internalization of DCA@Alg@INS nanoparticles in Caco-2 cells. Furthermore, the DCA@Alg@INS demonstrated a superior capacity for cellular uptake and permeability coefficient relative to the insulin solution, exhibiting sixfold and 4.94-fold enhancement, respectively. According to the uptake mechanism studies, the results indicated that DCA@Alg@INS was mostly transported through an energy-dependent active pathway since the uptake of DCA@Alg@INS by cells was significantly reduced in the presence of NaN3 by ~ 92% and at a low temperature of 4°C by ~ 94%. CONCLUSIONS: Given the significance of administering insulin through oral route, deoxycholic acid-modified alginate nanoparticles present a viable option to surmount various obstacles presented by the gastrointestinal.

The Effect of Mesenchymal Stem Cells Derived-Conditioned Media in Combination with Oral Anti-Androgenic Drugs on Male Pattern Baldness: An Animal Study.

OBJECTIVE: Androgenetic alopecia (AGA) is a prevalent form of hair loss, mainly caused by follicular sensitivity to androgens. Despite developing different anti-androgen treatment options, the success rate of these treatments has been limited. Using animal models, this study evaluated the therapeutic effects of umbilical cord (UC) stem cell conditioned media (CM) combined with oral anti-androgens for hair regeneration. MATERIALS AND METHODS: In this experimental study, Poloxamer 407 (P407) was used as a drug carrier for subcutaneous testosterone injection. AGA models were treated with oral finasteride, oral flutamide, and CM injections. Samples were thoroughly evaluated and compared using histological, stereological, and molecular analyses. RESULTS: Injecting CM-loaded hydrogel alone or combined with oral intake of anti-androgens improved hair regeneration. These treatments could promote hair growth by inducing hair follicles in the anagen stage and shortening the telogen and catagen phases. Furthermore, the combination treatment led to an upregulation of hair induction gene expression with a downregulation of inflammation genes. CONCLUSION: Through a reduction in inflammation, injection of CM-loaded hydrogel alone or combined with oral intake of anti-androgens induces the hair cell cycle with regeneration in damaged follicles. Hence, this could be a promising therapeutic method for AGA patients.

L-asparaginase immobilization in supramolecular nanogels of PEG-grafted poly HPMA and bis(α-cyclodextrin) to enhance pharmacokinetics and lower enzyme antigenicity.

L-asparaginase (ASNase) enzyme has limited therapeutic use due to its poor pharmacokinetics and immunogenicity. To overcome these obstacles, we immobilized ASNase in biocompatible poly hydroxypropyl methacrylamide (P(HPMA))-based nanogels simply formed through the host-guest inclusion complex of ASNase-conjugated random copolymer of HPMA and polyethylene glycol (PEG) acrylate (P(HPMA-MPEGA)) and α-cyclodextrin dimer (bisCD) using cystamine as a linker. The effects of bisCD and polymer concentrations on particle size, gelation time, and recovery of enzyme activity were investigated. The ASNase-conjugated bisCD nanogels were discrete, homogeneous, and spherical with a mean projected diameter of 148 ± 41 nm. ASNase immobilized in the bisCD nanogels caused cytotoxicity on HL-60 cell line with IC50 of 3 IU/ml. In-vivo rat study revealed that the immobilized ASNase reduced the enzyme antigenicity and resulted in 8.1 folds longer circulation half-life than the native enzyme. Conclusively, immobilization of ASNase in P(HPMA-MPEGA) and bisCD supramolecular nanogels could enhance the therapeutic value of ASNase in cancer chemotherapy.

Nanotechnology-based cell-mediated delivery systems for cancer therapy and diagnosis.

The global prevalence of cancer is increasing, necessitating new additions to traditional treatments and diagnoses to address shortcomings such as ineffectiveness, complications, and high cost. In this context, nano and microparticulate carriers stand out due to their unique properties such as controlled release, higher bioavailability, and lower toxicity. Despite their popularity, they face several challenges including rapid liver uptake, low chemical stability in blood circulation, immunogenicity concerns, and acute adverse effects. Cell-mediated delivery systems are important topics to research because of their biocompatibility, biodegradability, prolonged delivery, high loading capacity, and targeted drug delivery capabilities. To date, a variety of cells including blood, immune, cancer, and stem cells, sperm, and bacteria have been combined with nanoparticles to develop efficient targeted cancer delivery or diagnosis systems. The review paper aimed to provide an overview of the potential applications of cell-based delivery systems in cancer therapy and diagnosis.

Nanoencapsulation of saffron crocin into chitosan/alginate interpolyelectrolyte complexes for oral delivery: A Taguchi approach to design optimization.

Crocin, as a nutraceutical component of saffron (Crocus sativus L.), possesses numerous therapeutic effects. In the current study, a crocin-loaded chitosan/alginate (CS/ALG) nanocarrier was developed for oral delivery. The influence of preparation variables including pH and the concentrations of CS, ALG, and calcium chloride (CaCl2 ) on encapsulation efficiency (EE%) and loading efficiency (LE%) of CS/ALG nanoparticles (NPs) was evaluated by L9 Taguchi orthogonal array (OA). The results showed that at 0.25% w/v CS, 0.1% w/v ALG, pH 4.5, and absence of CaCl2 , crocin was loaded into CS/ALG NPs with EE% and LE% of 91.5% and 27.4%, respectively. Ultrasonication reduced the particle sizes (PSs) up to 100 nm, and freeze-dried NPs reproduced the particles with average size of 90 nm. Scanning electron microscopy (SEM) was successfully used to characterize the structure and morphology of freeze-dried NPs, confirming very fine NPs having sizes less than 100 nm. Crocin loaded into NPs showed higher stability in simulated gastric pH 2 compared to free crocin (2.1% and 7.5% degradation at 60 min, respectively). Furthermore, a pH-dependent sustained crocin release was observed with faster release at pH 2. Overall, the very small PS along with high encapsulation efficiency and stability can enhance crocin oral bioavailability making CS/ALG nanovehicles promising as an effective delivery system. PRACTICAL APPLICATION: Crocin as a functional component of Saffron is not sufficiently stable in gastrointestinal tract and its absorption is not complete. Chitosan/Alginate nanoparticles can encapsulate it efficiently, protect it and enhance its absorption orally. The availability and simplicity of the materials and equipments employed in current research provide the possibility to industrial scale up of the Crocin nanoparticles.

Design of ultra-fine carvedilol nanococrystals: Development of a safe and stable injectable formulation.

Carvedilol (CAR) is a strategic beta-blocker agent which its application has been limited by its very low water solubility. The present study describes a soluble form of drug based on nano-cocrystal (NCC) anti-solvent precipitation technique. The COSMOquick software was employed to select the optimum coformer (tartaric acid, TA) and organic solvent (acetone) relying on the enthalpy changes of cocrystallization and solubilization. Central Composite Design (CCD) considering the impact of CAR, TA, poloxamer 188 (stabilizer) concentrations, and anti-solvent/solvent ratio on CAR NCCs particle size (PS) could produce ultra-fine NCCs (about 1 nm). The lyophilization of NCCs investigating slow/fast freezing rates, various types and concentrations of cryprotectants and lyoprotectants indicated that PEG and trehalose (5 % w/vconcentration) under slow freezing rate could re-produce the initial PSs successfully. CAR NCCs indicated about 2000 fold increase in solubility compared with pure CAR. DSC and PXRD experiments proved that the formulations containing trehalose led to more crystalline and the ones comprising PEG led to more amorphous structures. Interestingly, the slow freezed PEG protected NCCs were physically stable for at least 18 months. In conclusion, the NCC technology could produce the first safe soluble form of CAR for treating hypertension urgencies easy for industrial scale-up.

Curcumin-incorporated crosslinked sodium alginate-g-poly (N-isopropyl acrylamide) thermo-responsive hydrogel as an in-situ forming injectable dressing for wound healing: In vitro characterization and in vivo evaluation.

Sodium alginate products have been extensively used for wound-dressing. In present study, a series of thermo-sensitive cross-linked poly(N-isopropylacrylamide) grafted sodium alginate (Alg-g-pNIPAM) copolymers were synthesized for delivery of curcumin to wound. FTIR, 1H NMR, elemental analysis and DSC showed successful polymerization and precise structure of copolymers. Thermogelation at 27-42 °C depending on the copolymer concentration, chain-length of pNIPAM and pH was observed. The optimum copolymer with proper rheological and syringeability properties showed excellent thermogelling at a wide range of pH and concentration, and could prolong the release of curcumin up to 72 h. In-vivo wound contraction and histopathological evaluations revealed that in addition to the higher efficacy in wound contraction, the curcumin formulation (Cur-F) significantly reduced the inflammation, enhanced the collagenesis and resulted in increased number of fibroblasts. Well-known anti-oxidant and anti-inflammatory properties of curcumin and in situ-forming nature of Alg-g-pNIPAM can make the system an excellent candidate for further investigations.

Microneedle Arrays Combined with Nanomedicine Approaches for Transdermal Delivery of Therapeutics.

Organic and inorganic nanoparticles (NPs) have shown promising outcomes in transdermal drug delivery. NPs can not only enhance the skin penetration of small/biomacromolecule therapeutic agents but can also impart control over drug release or target impaired tissue. Thanks to their unique optical, photothermal, and superparamagnetic features, NPs have been also utilized for the treatment of skin disorders, imaging, and biosensing applications. Despite the widespread transdermal applications of NPs, their delivery across the stratum corneum, which is the main skin barrier, has remained challenging. Microneedle array (MN) technology has recently revealed promising outcomes in the delivery of various formulations, especially NPs to deliver both hydrophilic and hydrophobic therapeutic agents. The present work reviews the advancements in the application of MNs and NPs for an effective transdermal delivery of a wide range of therapeutics in cancer chemotherapy and immunotherapy, photothermal and photodynamic therapy, peptide/protein vaccination, and the gene therapy of various diseases. In addition, this paper provides an overall insight on MNs' challenges and summarizes the recent achievements in clinical trials with future outlooks on the transdermal delivery of a wide range of nanomedicines.

Paclitaxel-loaded polypeptide-polyacrylamide nanomicelles overcome drug-resistance by enhancing lysosomal membrane permeability and inducing apoptosis.

The aim of the current project was to investigate the in vitro properties of Paclitaxel (PTX)-loaded pHPMA5kD -pHis5kD -pLeu3kD nanomicelles (NMs) on multidrug resistance cell line. Circular dichroism analysis was done to investigate the effect of pH on the secondary structure of the copolymer. Cytotoxicity assay together with fluorescence imaging and flow cytometry were performed to get an insight about toxicity and cellular uptake mechanism of NMs. Acridine orange assay, rhodamine 123 (Rh123) accumulation assay, and apoptosis analysis were conducted for further investigation. It was found that the secondary structure of the copolymer changed in response to pH, PTX-loaded NMs had higher cytotoxicity on both drug-sensitive (MES-SA and MCF-7) and multidrug resistant cells (MES-SA/DX5) compared to free PTX, and interestinly free copolymer inhibited the growth of MES-SA/DX5 cells while it was nontoxic on drug-sensitive cells. Moreover, the copolymer was able to induce lysosome membrane permeation and increase Rh123 accumulation inside cells indicating inhibition of the P-gp efflux pumps. Finally, apoptosis was strongly induced in MES-SA/DX5 cells upon treatment with PTX-loaded NMs. It can be concluded that the designed hybrid copolymer is a good candidate for in vivo assay and developing a powerful system against multidrug resistance tumors.

Freeze-Drying of Pharmaceutical and Nutraceutical Nanoparticles: The Effects of Formulation and Technique Parameters on Nanoparticles Characteristics.

Nanoparticles (NPs) are of the most interesting novel vehicles for effective drug delivery to humans. Freeze drying is known as an engaging process to improve the long lasting stability of NPs formulations. This study aims to elucidate the importance of various parameters involving in freeze-drying of the most common pharmaceutical/nutraceutical NPs including nanosuspensions, nanocrystals (NCs), cocrystals/nanococrystals, nanoemulsions (NEs), nanocapsules (NCPs) and nanospheres (NSPs). Regarding this, the therapeutic goals of NPs and specifications of drug must be considered. According to our survey, the most influential factors for achieving optimum results include type and concentration of cryoprotectant/lyoprotectant, stabilizer structure and concentration, the NPs concentration in solution, freezing, annealing, and drying rate, the interaction between protectants and stabilizer, solvent type and antisolvent to solvent ratio. The study shows that for each class of NPs, specific variables are of highest significance and should be optimized. For instance, about NCs, freezing rate and antisolvent/solvent ratio should be particularly considered and for emulsified NPs, the best results have been obtained by 5-20% of saccharides as cryoprotectants. These findings suggest that to obtain a product with the lowest aggregation and particle size (PS), optimization of the effective factors in formulation and lyophilization process are essential.

Author Correction: Biosurfactant Production by Lactic Acid Bacterium Pediococcus dextrinicus SHU1593 Grown on Different Carbon Sources: Strain Screening Followed by Product Characterization.

An amendment to this paper has been published and can be accessed via a link at the top of the paper.

Effect of different alcoholic-alkaline treatments on physical and mucoadhesive properties of tapioca starch.

Alcoholic-alkaline treatment (AAT) is a physical modification method for the production of granular cold water swelling starch and should be optimized for each starch source. The main objective of this research was to investigate physical and mucoadhesive properties of tapioca starch (TS) modified by different AATs. Improvement of cold-water absorption, solubility, rheological properties at low temperatures, clarity, freeze-thaw stability, and mucoadhesion was positively correlated with the alkali amount and reaction temperature and negatively correlated with ethanol content. Morphological studies demonstrated different degrees of swelling, birefringence loss, and surface wrinkling of granules depending on modification degree. Starch pastes, modified in a higher degree, showed a change from rheopexy to thixotropy and from translucency to turbidity over time. The highest quality along with maintaining granular integrity was obtained by treating starch (10 g) with 30 g alkaline solution (2.5 M) and 110 g aqueous ethanol (40%) at 25 °C. The characteristics of this sample were higher than those of corn counterpart except for viscosity, consistency, and freeze-thaw stability and were almost similar to those of thermally gelatinized TS (TGTS). Therefore, this AA-modified TS can be an alternative for TGTS in instant and heat-sensitive foods and delivery of bioactives as a mucoadhesive polymer.

Efficacy of oral administration of licorice as an adjunct therapy on improving the symptoms of patients with Parkinson's disease, A randomized double blinded clinical trial.

ETHNOPHARMACOLOGICAL RELEVANCE: Licorice preparations are used as neuroprotective remedies in Persian ethnomedicine, in order to prevent from disabilities in neurodegenerative conditions like Parkinson's disease (PD). AIM OF THE STUDY: This study was designed to determine the licorice (root of Glycyrrhiza glabra L.) effectiveness as an adjunct treatment in the PD management. MATERIAL AND METHODS: In this double-blinded trial, 128 patients were assessed for eligibility criteria. Seventy-eight patients were ineligible and 11 of them refused from participating. Thirty-nine PD patients (YAHR staging ≤ 3) were divided into two groups by random. The patients received oral licorice or placebo syrups with a dose of 5 cc, twice a day for 6 months. High-performance liquid chromatography and spectrophotometric instruments determined licorice syrup constituents. The patients' situation for Unified Parkinson's rating scale (UPDRS) was assessed every 6 weeks for the duration of six months. In addition, patients' blood pressure, blood glucose, sodium and potassium levels, quality of life and dizziness were determined. RESULTS: Six weeks after intervention, total UPDRS, daily activities and tremor were significantly improved with a considerable effect size. A significant better motor test and rigidity scores were observed 4 months after licorice intake (p > 0.05). No electrolyte abnormality, significant changes in blood pressure or blood glucose levels were observed during the study. Each 5cc of syrup contained 136 mg of licorice extract with 12.14 mg glycyrrhizic acid, and also 136 µg of polyphenols. CONCLUSION: The licorice intake could improve the symptoms in PD patients without serious adverse events.

Nanoparticulate delivery of irinotecan active metabolite (SN38) in murine colorectal carcinoma through conjugation to poly (2-ethyl 2-oxazoline)-b-poly (L-glutamic acid) double hydrophilic copolymer.

SN-38 is the active metabolite of irinotecan, an FDA-approved chemotherapeutic agent indicated for colorectal carcinoma, which would not be clinically applicable due to its very poorly soluble and hydrolytic degradation properties. To overcome these limitations, it was proposed to conjugate SN38 to residing carboxylic acid residues in poly (2-ethyl 2-oxazoline) block poly (L-glutamic acid), inducing nano-assembly in aqueous medium. Following a series of reactions including poly (2-ethyl oxazoline) macro-initiated ring opening polymerization of N-carboxyanhydride, deprotection of benzyl group and chemical conjugation of SN38 via biodegradable ester linkage, the as-synthesized product was characterized by dynamic light scattering, ζ potential and transmission electron microscopy. The resulting particles presented about 90% loading efficiency with a mean size of 90 nm. Upon incubation with colorectal carcinoma CT26 cell line, higher association of SN-38 fluorescence and significantly more specific cytotoxicity was noticed for the SN38 conjugated particles than free drug. Therapeutic applicability of the as-synthesized product was evaluated in CT26 allograft tumor model in BALB/c mice, showing superior efficiency of the SN38 conjugated particles particularly in tumors with sizes larger than 200 mm3 than parent irinotecan and reduced mortality rate by 2.5 times. Conclusively, the poly (2-ethyl 2-oxazoline) decorated nano-conjugates of poly (L-glutamic acid) and SN38 can be regarded as a novel and potentially efficient drug delivery system for advanced colorectal carcinoma.

A randomized controlled double blinded trial to evaluate efficacy of oral administration of black strap molasses (sugarcane extract) in comparison with polyethylene glycol on pediatric functional constipation.

ETHNOPHARMACOLOGICAL RELEVANCE: As a significant health issue, Functional Constipation (FC) has a high prevalence of 0.7%-29% worldwide. In Persian ethnomedicine, several formulations of sugarcane extracts including faniz, shekar-e- sorkh and black strap molasses have been used to treat pediatric constipation. AIM OF THE STUDY: To evaluate efficacy and safety of oral intake of black strap molasses syrup (BSM) on FC in children. MATERIALS AND METHODS: A randomized controlled double blinded trial was performed on children aged 4-12 years diagnosed with FC according to ROMIII Constipation Measurement Questionnaire. Ninety-two patients were randomly divided in two groups of BSM or PEG syrups (40%) with a dose of 1 mL/kg body weight/day for 1 month. Symptoms were evaluated every two weeks. Possible serological side-effects and changes in children's body weight were investigated. BSM syrup constituents were determined by pharmacognostic methods. RESULTS: Treatment was successful with both interventions with no significant difference between the groups. Defecation per week was significantly improved in both groups. Symptoms including volitional stool retention, large diameter stool, painful or hard stool and large fecal mass in the rectum decreased significantly two and four weeks after intervention (P < 0.05). There was no significant difference between groups except for the rate of large diameter stool. No side -effects were observed. The BSM naturally contained polyphenols (960 µg/mL), potassium (12430 µg/mL), iron (80 µg/mL) and calcium (3320 µg/mL). CONCLUSION: BSM and PEG syrups had similar efficacy on FC. Compared with PEG, BSM syrup contained different natural micronutrients.

Biosurfactant Production by Lactic Acid Bacterium Pediococcus dextrinicus SHU1593 Grown on Different Carbon Sources: Strain Screening Followed by Product Characterization.

The present study focused on producing and characterizing a type of biosurfactant (BS) derived from lactic acid bacteria (LAB) and its potential applications in pharmaceutical and food industries due to the preference of employing nonpathogenic organisms in bioprocesses. To this aim, several screening approaches were applied to identify an efficient BS-producing strain from a set of LAB, and Pediococcus dextrinicus SHU1593 was selected as the most operative one. The BS produced by P. dextrinicus was isolated and structurally characterized as a lipoprotein with an approximately equal ratio of lipids (~52% (w/w)) and proteins (47% (w/w)). It reduced the surface tension (ST) of phosphate-buffered saline (PBS) from 72.80 ± 0.10 to 39.01 ± 0.32 mN/m. The results also indicated the potential of developing low-cost strategies aimed at the production of efficient LAB-derived BSs which are structurally and quantitatively similar to the ones obtained from conventional media. Finally, given the physical and functional characterization (i.e. critical micelle concentration (CMC), emulsification index (%E24), stability, as well as antimicrobial and anti-adhesive activities) of the BS produced in the present study, it can be introduced as a promising candidate to be employed in plenty of areas in pharmaceutical and food industries.

Reversing multi-drug tumor resistance to Paclitaxel by well-defined pH-sensitive amphiphilic polypeptide block copolymers via induction of lysosomal membrane permeabilization.

A series of hybrid di-block copolymers of poly(l- glutamic acid-b-l- leucine) (PGA-PLeu), methoxy poly (ethylene glycol)-b-poly(l-leucine) (PEG-PLeu), methoxy poly(ethylene glycol)-b-poly(γ-benzyl-l-glutamic acid) (PEG-PBLG) and tri-block copolymers of poly(ethylene glycol)-b-poly(l-glutamic acid-co-null-leucine) (PEG-PGA-PLeu) were synthesized through sequential HMDS-mediated ring-opening polymerization (ROP). Chemical structure of copolymers was studied by FTIR and 1H-NMR and their molecular weight was determined by 1H-NMR and gel permeation chromatography (GPC). Copolymers self-assembled into nanomicelles with particle size (PS) of 65 to 139 nm. Higher fraction of polyleucine (% fPLeu) led to significantly larger PS, lower critical aggregation concentration (CAC) and higher drug loading content (DLC%). In addition, introducing PEG segment led to significant decrease in PS, increase of CAC and DLC%. Apart from copolymer composition, DLC% changed by the method with significantly higher loading for solid dispersion. Remarkably, the release of PTX from PEG-PGA-PLeu tri-block copolymers was highly dependent on pH, revealing a relatively two-fold faster release at pH 5 than pH 7.4. CD spectroscopy showed transition to α-helix secondary structure at acidic pH. Hemocompatibility assay confirmed that copolymers were absolutely hemocompatible at physiological pH. MTT assays demonstrated that unlike MCF7 and 4T1 cells that PTX-loaded nanoparticles (PTX-NPs) exhibited similar antitumor activity, ten-fold higher toxicity was recognized in multidrug-resistant uterine sarcoma cells (MES-SA/DX5). Fluorescent imaging and flow cytometric analysis of cellular uptake showed that nanoparticles' uptake was time-dependent. It was also revealed that higher toxicity of the PTX-NPs could be due to ability of copolymer to inhibit P-gp pumps and induce lysosomal membrane permeabilization (LMP).

Effect of PEGylation on assembly morphology and cellular uptake of poly ethyleneimine-cholesterol conjugates for delivery of sorafenib tosylate in hepatocellular carcinoma.

Introduction: Sorafenib (SFB) is an FDA-approved chemotherapeutic agent with a high partition coefficient (log P = 4.34) for monotherapy of hepatocellular carcinoma (HCC). The oral bioavailability is low and variable, so it was aimed to study the application of the polymeric nanoassembly of cholesterol conjugates of branched polyethyleneimine (PEI) for micellar solubilization of SFB and to investigate the impact of the polymer PEGylation on the physicochemical and cellular characteristics of the lipopolymeric dispersions. Methods: Successful synthesis of cholesterol-PEI lipopolymers, either native or PEGylated, was confirmed by FTIR, 1H-NMR, pyrene assay methods. The nanoassemblies were also characterized in terms of morphology, particle size distribution and zeta-potential by TEM and dynamic light scattering (DLS). The SFB loading was optimized using general factorial design. Finally, the effect of particle characteristics on cellular uptake and specific cytotoxicity was investigated by flow cytometry and MTT assay in HepG2 cells. Results: Transmission electron microscopy (TEM) showed that PEGylation of the lipopolymers reduces the size and changes the morphology of the nanoassembly from rod-like to spherical shape. However, PEGylation of the lipopolymer increased critical micelle concentration (CMC) and reduced the drug loading. Moreover, the particle shape changes from large rods to small spheres promoted the cellular uptake and SFB-related cytotoxicity. Conclusion: The combinatory effects of enhanced cellular uptake and reduced general cytotoxicity can present PEGylated PEI-cholesterol conjugates as a potential carrier for delivery of poorly soluble chemotherapeutic agents such as SFB in HCC that certainly requires further investigations in vitro and in vivo.

Pharmacokinetics and Biodistribution of Pegylated Methotrexate after IV Administration to Mice.

The efficacy of methotrexate (MTX) as an antimetabolite chemotherapeutic agent highly depends on its blood circulation half-life. In our previous study, different conjugates of MTX (MTX-PEG) were synthesized, their physicochemical properties were investigated and MTX-PEG5000 was finally selected as optimum drug-conjugate for further investigations. In the current work, first the stability of MTX-PEG5000 was studied at 37 °C and the results indicated its high stability in plasma (T1/2 = 144 h) and a relatively rapid degradation in tissue homogenate (T1/2 = 24 h). The study of protein binding pointed out that the conjugate was highly protein-bound (95%). The results of pharmacokinetic studies in mice indicated that MTX-PEG5000 had longer plasma distribution and elimination half-lives compared to free MTX (T1/2 α 9.16 min for MTX-PEG5000 versus 2.45 min for MTX and T1/2 ß 88.44 for MTX-PEG5000 versus 24.33 min for MTX). Pharmacokinetic parameters also showed higher area under the curve (AUC) of conjugate compared to parent drug (12.33 mg.mL-1.min for MTX-PEG5000 versus 2.64 mg.mL-1.min for MTX). The biodistribution studies demonstrated that MTX-PEG5000 did not highly accumulate in liver and intestine and had a mild and balanced distribution to other organs. Also, the conjugate was measurable in tissues up to 48 h after injection and was detected in the brain, suggesting the possibility of delivering drug to brain tumors.