Evaluation of Autophagy Process in Differentiation of Human Induced Pluripotent Stem Cells toward Insulin Producing Cells.

Background: Autophagy is an intracellular self-degradative homeostasis process which eliminates undesirable and harmful macromolecules and organelles. Autophagy is also involved in self-renewal and differentiation of induced pluripotent stem cell (iPSCs). Objective: In this study, we investigated the expression profile of autophagy marker genes in human iPSCs during their differentiation induction toward insulin producing ß-like cells. Methods: Human iPSC line, R1-hiPSC1, was used for differentiation induction toward ß-like cells. The mRNA expression of Nanog, OCT4 (pluripotency markers), SOX17, FOXA2 (endodermic markers), PTF1A, NKX6.1 (exocrine/endocrine determinants), and PDX1 were measured during differentiation stages. Autophagy was monitored by genes expression study of four autophagy markers, MAP1LC3B, BECN1, SQSTM1/P62 and ATG5, along with protein expression profile of LC3b-II during differentiation stages. Results: The mRNA expression measurement of pluripotency, endoderm and exocrine/endocrine marker genes confirmed that hiPSCs skipped pluripotency, differentiated into endoderm, passed through the pancreatic lineage commitment stage and successfully generated insulin producing ß-like cells. Expression profile of autophagy genes during differentiation stages indicated the decreased expression levels at the early stages (EB and MEI) and then increased at the definitive endoderm stages (DEI 1, DEI 2 and DE) followed by a subtractive pattern toward the end of differentiation. The results of protein expression of LC3b-II were consistent with gene expression data. Conclusion: This study demonstrated the high contribution of key autophagy genes/proteins during the differentiation of hiPSC toward ß-like cells. The enhanced autophagy levels were a prominent feature of early stages of differentiation and DE rather than the later stages.

Effects of Different Cold Preservation Solutions on the Functions of Cultured Isolated Human Hepatocytes.

BACKGROUND: Hepatocyte transplantation using isolated human hepatocytes is an alternative source that can be used for the treatment of metabolic diseases and acute liver failure as a time bridge to liver transplantation. These cells can also be used for bioartificial liver systems and in vitro study of drug toxicity. OBJECTIVE: To determine which cold preservation solution is better maintain the liver function. METHODS: We prepared 4 cold preservation solutions made of different combination of antioxidants, chelating, membrane protective, and anti-apoptotic agents as well as inhibitor of cyclophilin D. For hepatocyte isolation, we used livers obtained from unused deceased donor livers and the liver of patients with Crigler-Najjar syndrome who were candidates of partial liver transplantation. After culture and cold preservation, the level of albumin, and urea production were measured as indices of liver functionality. RESULTS: We found that albumin production significantly decreased after cold preservation in solution 1. There was no significant difference in urea production after cold preservation in solution 1 compared with control 24 h. No significant differences in albumin production were found after cold storage in solution 2 and solution 4 compared with control 24 h. Urea production significantly decreased after cold storage in solutions 2 and 4 compared with control 24 h. As a whole albumin and urea production were significantly decreased after cold preservation. Although albumin and urea production were decreased after cold preservation, but the results of albumin production of two solutions were not significantly different from that of the control group (p=0.109 and 0.951). CONCLUSION: Cold preservation of cultured human hepatocytes in solution 2 and solution 4 could maintain the function of albumin production better than other cold preservation solutions in our experiments; solution 1 was more effective on urea production of cultured human hepatocytes at 4 °C for 24 h. To determine if these hepatocytes are suitable candidates for transplantation, further studies should be performed.

Association between Interleukin-21, 23 and 27 Expression and Protein Level with Cytomegalovirus Infection in Liver Transplant Recipients.

BACKGROUND: Cytokines have regulatory crosstalk with CMV infection leading to manage of post-liver transplantation virus-related outcomes. OBJECTIVE: To investigate the link between IL-21, IL-23 and IL-27 mRNA and protein level with active CMV infection, which was evaluated in reactivated and non-reactivated liver transplant recipients. METHODS: Two groups of liver transplant recipients were enrolled in this study-54 without and 15 with active CMV infection. 3 EDTA-treated blood samples were taken on day 1, 4, and 7 post-liver transplantation. Plasma and buffy coats of all samples were separated. All samples were analyzed for CMV reactivation using antigenemia technique. The separated plasma of positive samples was used for viral DNA extraction and protein evaluation. For evaluating the mRNA expression level by real-time PCR, RNA extraction and cDNA synthesis were done for all samples. Also, the protein level of studied genes was estimated by ELISA. RESULTS: The expression level of IL-21, IL-23A and IL-27A cytokine genes was increased in CMV reactivated liver transplant recipients in comparison with CMV non-reactivated ones; IL-27A expression pattern was significant (p=0.001) at all sampling times. IL-21 significantly increased on the 2nd and 3rd (p=0.028 and 0.01, respectively) sampling days in CMV reactivated compared with non-reactivated patients. The expression level of IL-23A cytokine significantly increased on the 3rd (p=0.017) sampling day in CMV reactivated compared with non-reactivated liver transplant recipients. CONCLUSION: Elevation in the expression level of IL-21, IL-23A and IL-27A mRNA and protein level in CMV reactivated patients emphasized on the antiviral role of these cytokines in CMV reactivated liver transplant recipients.

Effectiveness of mesenchymal stem cell-seeded onto the 3D polylactic acid/polycaprolactone/hydroxyapatite scaffold on the radius bone defect in rat.

PURPOSE: The importance of regeneration in large bone defects forces the orthopedic surgeons to search for a proper methodology. The present experiment evaluated the capability of polylactic acid/polycaprolactone/hydroxyapatite (PLA/PCL/HA) scaffold loaded with and without mesenchymal stem cells (MSCs) on bone regeneration. METHODS: Fourier transform infrared spectrometry, X-ray diffraction, scanning electron microscopy, and rheology methodologies were used to characterize the scaffold. Forty Wistar rats were randomly divided into the four groups including the untreated defects as the control group and three other groups in which the bone defects were treated with autologous bones (autograft group), the PLA/PCL/HA scaffolds (PLA/PCL/HA group), and the MSCs-seeded scaffolds (MSCs-seeded PLA/PCL/HA group). RESULTS: Based on the qRT-PCR results, significantly higher expression levels of osteocalcin, osteopontin, and CD31 were seen in the cell-seeded scaffold group compared to the control group (P < 0.05). The CT scanning and radiographic images depicted significantly more newly formed bonny tissue in the MSCs-loaded scaffold and autograft groups than the untreated group (P < 0.001). The immunohistochemistry, biomechanical, histopathologic, and histomorphometric evaluations demonstrated significantly improved regeneration in the autograft and MSCs-loaded scaffold groups compared to the non-treated group (P < 0.05). There were significant differences between the scaffold and untreated groups in all in vivo evaluations (P < 0.05). CONCLUSION: The MSCs enhanced bone healing potential of the PLA/PCL/HA scaffold and the MSCs-seeded scaffold was comparable to the autograft as the golden treatment regimen (P > 0.05).

Photothermal/sonodynamic therapy of melanoma tumor by a gold/manganese dioxide nanocomposite: In vitro and in vivo studies.

Nanosized materials have favorable applications in nanomedicine including photothermal therapy (PTT) and sonodynamic therapy (SDT) of most malignancies. Since conventional methods of cancer treatment encounter limitations, development of other efficient routes is quite necessary. In this study, a gold/manganese dioxide nanocomposite (Au/MnO2 NC) was synthesized as a novel photo- and sono- responsive nanomaterial that was activated upon laser light of 808-nm wavelength irradiation or ultrasound exposure for cancer treatment applications. The synthesized nanocomposite comprised gold nanoparticles of about 125 ±â€¯66 nm in diameter adhered to manganese dioxide nanoroads of 77 ±â€¯30 nm in diameter and up to 2 µm length. Au/MnO2 NC represented a very good photothermal and sonodynamic conversion ability. Cytotoxicity of Au/MnO2 NC toward the C540 (B16/F10) cell line was evaluated through thermal ablation and reactive oxygen species (ROS) generation upon phototherapy and sonotherapy, respectively. Intratumoral injection of a low-dose of Au/MnO2 NC into a melanoma tumor-bearing animal model followed by laser and ultrasound radiation led to necrosis in the tumor tissue. The findings revealed that Au/MnO2 NC is aphotosensitizer/sonosensitizer for PTT and SDT of cancer.

The Effects of Cold Preservation Solutions Supplemented with UDCA and α-Lipoic Acid on the Viability and Function of Isolated Human Hepatocytes.

BACKGROUND: Liver transplantation is the only treatment for end-stage and genetic liver diseases. The main burden of this treatment is the shortage of both living and cadaveric liver donors. An alternative treatment is using liver cell transplantation, which can be obtained from unused livers for transplantation. These hepatocytes should be kept ready in viable and functional situation in a frozen state to be instantly used when they would be needed. In our previous experience, we had isolated hepatocytes from unused livers. OBJECTIVE: To find a preserving solution for increasing viability and function of the isolated hepatocytes that are stored to be transplanted. METHODS: 9 cadaveric donor livers, which were not used for transplantation due to various causes such as severe steatosis, were selected to isolate hepatocytes. Various cold storage solutions were tried to find the best temperature for more viability and functionality for preservation of hepatocytes. University of Wisconsin (UW) solution and Williams E media were used as control media. 2 anti-apoptotic and anti-oxidative solutions, i.e., α-lipoic acid and ursodeoxycholic acid (UDCA), were used as cold preservatives solutions. The numbers of viable hepatocytes were estimated by trypan blue method; the functionality was assessed by the cells ability to produce urea. RESULTS: The highest number of viable and functional hepatocytes was obtained from freshly isolated cells. However, after preservation, the number of these viable hepatocytes and their functionality were not significantly different in cold storage solutions comparing to the control media used. Functionality of the isolated hepatocytes stored in UW with and without UCDA solution was similar to freshly isolated hepatocytes. CONCLUSION: Preservatives with anti-apoptotic and antioxidant activity could not increase the number of viable hepatocytes. Functionality of cold storing hepatocytes could be preserved similar to freshly isolated hepatocytes by UW solution with and without UCDA.

Evaluation of Angiotensinogen M235T and T174M Polymorphisms, Demographic and Clinical Factors in New-Onset Diabetes after Liver Transplantation in Iranian Patients.

BACKGROUND: New-onset diabetes after transplantation (NODAT) is a serious complication which runs the risk of infections, morbidity and mortality. OBJECTIVE: To evaluate M235T and T174M polymorphisms of angiotensinogen gene along with some demographic and clinical factors including age; sex; body mass index (BMI); model for end-stage liver disease (MELD) score; prednisolone, mycophenolate mofetil and tacrolimus dose; and serum level in NODAT among liver recipients. METHODS: In this study 115 patients (53 with and 62 without NODAT) who had no history of diabetes before the transplantation were investigated. Furthermore, 80 randomly selected apparently healthy people (no transplantation) were used as the control group. Two angiotensinogen polymorphisms (M235T and T174M) were studied using polymerase chain reaction restriction fragment length polymorphism (PCR-RFLP). RESULTS: Patients included 68 (59.1%) females and 47 (40.9%) males; they had a mean±SD age of 37.4±16.9 years. The M allele frequency was 55.7% (n=128) in M235T and 20.0% (n=46) in T174M polymorphisms. Binary logistic regression analysis confirmed that age (p=0.005), prednisolone dose (p<0.001) and mutated M235T polymorphism (p=0.003) were independent risk factors. CONCLUSION: Presence of M235T T allele may significantly (p<0.001) increase the NODAT risk, and increase the likelihood of developing end-stage liver disease (p=0.003). T174M T allele had a significantly (p=0.007) higher frequency in NODAT group.

Comparison of Human Mesenchymal Stem Cells Derived from Various Compartments of Human Adipose Tissue and Tunica Adventitia Layer of the Arteries Subsequent to Organ Donation.

BACKGROUND: Mesenchymal stem cells are one of the most interesting cell sources used in regenerative medicine. OBJECTIVE: In the present study, we isolated and characterized the mesenchymal stem cells from various compartments of human adipose tissue and tunica adventitia layer of the arteries. METHODS: Tissue explant culture was done from various compartments of the human adipose tissue and tunica adventitia layer of the arteries, including adipose tissue far from the vessels, perivascular tissues that are completely attached to the vessels, and tunica adventitia layer of the arteries. After the cell culture, characterization of the cells was determined at 3rd-5th passages. Flow cytometry was performed for antigen expression analysis of CD34, CD45, CD44, CD90, CD29, CD73, and CD105. For the evaluation of cell differentiation potential, adipogenic and osteogenic differentiation was conducted under appropriate protocols. RESULTS: The cells were positive for CD44, CD90, CD29, and CD73 and negative for CD34, CD45, and CD105. Adipogenic and osteogenic differentiation potentials were different among the cells from various compartments. The cells derived from perivascular tissue demonstrated better adipogenic and osteogenic differentiation. CONCLUSION: It is essential to characterize the cells from different tissues and compartments for different purposes in regenerative medicine.

Sonodynamic cancer therapy by a nickel ferrite/carbon nanocomposite on melanoma tumor: In vitro and in vivo studies.

Reactive oxygen species (ROS)-mediated cancer therapy using light or ultrasound (US) has been widely approached as a non-invasive and inspiring alternative treatment. Sonodynamic therapy (SDT), a non-invasive therapeutic modality of cancer, is an outcome of low-intensity US effect on cancer cells using a sonosensitizer, which results in heat and ROS production followed by cell death. The aim of this study was synthesis, characterization and cancer SDT application of a nickel ferrite/carbon nanocomposite (NiFe2O4/C), as a sonosensitizer. SDT was carried out by applying a 1.0-MHz US radiation at 1.0 W cm-2 of power density and 100% pulse ratio for 60 s. A significant C540 (B16/F10) cell killing was observed in vitro due to ROS production of 100 µg mL-1 of NiFe2O4/C upon SDT. In addition, SDT of melanoma cancer in a mouse model using intratumorally injected NiFe2O4/C of 100 µg mL-1 produced remarkable efficacious recovery in the tumor and significant necrosis (up to 60%) in histological assessments, while injection of NiFe2O4/C or US irradiation alone induced no healing effect. Therefore, SDT using NiFe2O4/C attained success in destroying melanoma cancer and can be developed and introduced as an alternative treatment strategy for melanoma cancer. In furtherance of SDT, magnetic resonance (MR) imaging (1.5 T) in an agarose phantom indicated the effectiveness of NiFe2O4/C as a negative contrast agent in transverse relaxation time-weighted imaging with a corresponding relaxation rate (r2) of 78.9 mmol L-1 s-1. The results confirmed the applicability of the nanocomposite as a theranostics agent for simultaneous SDT and MR imaging.

Capecitabine-loaded nanoniosomes and evaluation of anticancer efficacy.

Capecitabine (CAP) is an FDA-approved and frequently used chemotherapeutic agent for the treatment of various cancers. However, there are some side effects and chemoresistance limiting its use. Nanotechnological approaches can enhance the efficacy of anticancer drugs. In this study, CAP-loaded nanoniosomes were prepared. Nanoniosomes were prepared by the method of thin film hydration wherein CAP was loaded into the nanoniosomes. Nanoniosomes were then characterized by field emission scanning electron microscopy and (particle) vesicle size analysis. The cytotoxicity effect of the nanoniosomes were evaluated by the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay. CAP was loaded into the nanoniosomes and loading capacity and entrapment efficiency were determined. The vesicle size of the nanoniosomes was obtained in the nanometer scale, and CAP release profiles from the nanoniosomes were also obtained. Finally, the cytotoxicity effect of CAP and CAP-loaded nanoniosomes were evaluated toward MCF7 and PANC1 cell lines. The nanoniosomes with an amphipathic structure can penetrate into the cells with an enhanced release rate. These caused the toxicity of drug in the nanoniosomes to be higher than the free drug.

Repression of melanoma tumor in vitro and in vivo by photothermal effect of carbon xerogel nanoparticles.

Nanosized carbonaceous materials are favorable in biomedicine applications including photothermal therapy (PTT) of cancer. Since conventional strategies of cancer treatment have not responded to this serious disease, development of efficient alternative and promising strategies is highly desirable. In this study, carbon xerogel nanoparticles (CX-NPs) were synthesized as a novel photothermal nanomaterial and activated upon laser light of 808-nm wavelength for cancer phototherapy application. The synthesized CX-NPs had a spherical shape with a size of about 16 nm that showed nice photothermal conversion ability. Upon light irradiation with a power density of 1.0 W cm-2 for 15 min, a temperature increment occurred. A concentration-dependent cytotoxicity was also obtained for CX-NPs toward the C540 (B16/F10) cell line upon light irradiation, while CX-NPs presented biocompatibility in the mice model in dark. Photothermal property of CX-NPs efficiently led to reduction in the cell viability. A low-dose of CX-NPs was also applied in PTT of a melanoma tumor-bearing animal model. Based on tumor histopathological evaluations and volume change measurements in mice, a very good control of tumor situations after PTT by CX-NPs was attained. The findings revealed that CX-NPs is a good and novel photoabsorber for PTT of cancer.

Evaluation of a self-nanoemulsifying docetaxel delivery system.

A self-nanoemulsifying drug delivery system (SNEDDS) was developed as a novel route to enhance the efficacy of docetaxel lipophilic drug. SNEDDS comprised ethyl oleate, Tween 80 and poly(ethylene glycol) 600, as oil, surfactant and co-surfactant, and formed stabilized monodispersed oil nanodroplets upon dilution in water. SNEDDS represented encapsulation efficiency and loading capacity of 21.4 and 52.7%, respectively. The docetaxel release profile from the drug-loaded SNEDDS was recorded, its effectiveness against MCF-7 cell line was investigated, and an IC50 value of 0.98 ± 0.05 µg mL-1 was attained. The drug-loaded SNEDDS was administrated in rats, and the pharmacokinetic parameters of maximum concentration of 22.2 ± 0.8 µg mL-1, time to attain this maximum concentration of 230 min, and area under the curve of 1.71 ± 0.18 µg min mL-1 were obtained. The developed SNEDDS formulation can be represented as an alternative to docetaxel administration.

In vitro and in vivo tumor annihilation by near-infrared photothermal effect of a NiFe2O4/C nanocomposite.

Nanothechnology-mediated photothermal therapy (PTT) is emerging as one of the inspiring alternative modality of cancer therapy that applies near-infrared radiation. High favorability of this approach is due to its minimum invasiveness, safety of non-targeted area, quick recovery, and capable simultaneous imaging. In this approach, photoabsorbing nanomaterials convert energy of infrared light to vibrational motion and generate heat. In the present study, a nanocomposite comprised nickel ferrite and carbon (NiFe2O4/C) was synthesized, characterized and introduced as a novel photoabsorbing agent in cancer phototherapy. NiFe2O4/C was characterized by field emission scanning electron microscopy, Fourier-transform infrared spectroscopy, and X-ray diffraction patterns. A diode laser of 808 nm with a power density of 1.0 W cm-2 was selected as the light source to evaluate the photothermal property of NiFe2O4/C toward cancer repression in C540 (B16/F10) cell line and melanoma bearing tumor model in male balb/c mice. Temperature enhancement ability of NiFe2O4/C confirmed its photoabsorbing property. While NiFe2O4/C had a concentration dependent cytotoxicity on C540 (B16/F10) cell line, PTT of NiFe2O4/C activated by laser irradiation showed its destroying effect on the C540 (B16/F10) cell line. On the other hand, histological analyses and tumor volume changes were performed for the in vivo PTT of NiFe2O4/C upon intratumoral injection. The results showed that after 24 h, PTT of the nanocomposite cured the tumor properly, whereas NiFe2O4/C injection or laser exposure alone had no treatment effect. Also, 5-day post-treating the melanoma bearing tumor model indicated that the level of necrosis significantly increased during this time in the PTT treated mouse. Therefore, PTT using NiFe2O4/C is proposed as a promising procedure for the melanoma cancer therapy.

Evaluation of a nanocomposite of PEG-curcumin-gold nanoparticles as a near-infrared photothermal agent: an in vitro and animal model investigation.

Hyperthermia is a promising alternative modality for the conventional cancer treatments. Nanoparticle-mediated photothermal therapy (PTT) has been widely applied for hyperthermia cancer therapy by a near-infrared light irradiation. Some special nanoparticles can convert light energy into heat and destroy the tumor cells. Inspired from the photothermal efficacy of the gold nanoparticles, here we synthesized, characterized, and applied novel photothermal polyethylene glycol-curcumin-gold nanoparticles (PEG-Cur-Au NPs) in cancer PTT. The effect of PEG-Cur-Au NPs upon irradiation by an 808-nm laser on C540 (B16/F10) cell line as well as implanted (bearing) melanoma tumor in inbred C57 mice was investigated. In vitro temperature increment, cell viability evaluation, and histological analyses were performed. The results showed a dose-dependent cytotoxicity of PEG-Cur-Au NPs toward C540 (B16/F10) cell line at concentrations ≥ 25 µg mL-1 with an IC50 value of 42.7 µg mL-1 in dark (and with no toxicity for 10 µg mL-1). On the other hand, 808-nm laser irradiation alone (without using PEG-Cur-Au NPs) for 10 min induced killing effect on the C540 (B16/F10) cell line in a laser power-dependent manner at power density > 0.5 W cm-2 (no toxicity for 0.5 W cm-2). However, PPT using PEG-Cur-Au NPs was tremendously observed after laser illumination. Even under laser irradiation at a power density of 0.5 W cm-2 of PEG-Cur-Au NPs of concentrations < 10 µg mL-1, PTT of the cells was substantial. Histological analyses and volume measurements of the induced tumors in the mice revealed an appropriate control of the tumors upon PTT by PEG-Cur-Au NPs. Combination of PEG-Cur-Au NP administration and 808-nm diode laser irradiation destroyed the melanoma cancer cells in the animal model.

Investigation of amyloid formation inhibition of chemically and biogenically from Citrus aurantium L. blossoms and Rose damascena oils of gold nanoparticles: Toxicity evaluation in rat pheochromocytoma PC12 cells.

Fibrillation inhibition effects of chemically and biogenically gold nanoparticles (GNPs) were investigated in vitro using human insulin as a model for fibrillation of protein. This inspection was followed using the Congo red assay, thioflavin T fluorescence measurements, transmission electron microscopy, and evaluation of cytotoxicity effects on rat pheochromocytoma PC12 cells. Biogenic GNPs were synthesized using oil extracts of Citrus aurantium L. blossoms and Rose damascena blossoms as reducing and concomitant agents. Congo red assay showed development of fibril formation of insulin at acidic media at 60°C over a period of 48h. In these circumstances, transmission electron micrographs confirmed the progress of fibril state from globular chains to amyloid. However, the results of ThT fluorescence measurements indicated a concentration-dependent inhibiting effect of chemically synthesized GNPs on insulin fibrillation in vitro, simultaneously by conversion of the formed fibrils into amorphous aggregates. Furthermore, biogenic GNPs were found to more effectively inhibit the fibril formation, compared to chemically synthesized GNPs. Accordingly, just 0.05nmolL-1 of the biogenic GNPs showed similar inhibition property of chemically synthesized GNPs with a concentration of 10nmolL-1. Both types of GNPs diminished toxicity of insulin fibrils in rat pheochromocytoma PC12 cells viability.

Stem Cell-Based Therapies and Tissue Engineering of Trachea as Promising Therapeutic Methods in Mustard Gas Exposed Patients.

Tissue engineering and cell-based therapies are promising therapeutic approaches in structural and functional defects of the trachea. Researchers have focused on these approaches to overcome the complications related to such diseases. Patients exposed to mustard gas suffer from massive damage to the respiratory system. Current treatment plans are only palliative and include anti-inflammatory drugs, broncholytics, long-acting ß2-agonists, and inhaled corticosteroids. As mustard gas exposure leads to chronic airway inflammation, it seems that tracheobronchomalacia, because of chronic inflammation and weakness of the supporting cartilage, is an important factor in the development of chronic and refractory respiratory symptoms. The previous studies show that regenerative medicine approaches have promising potential to improve the life quality of patients suffering from tracheal defects. It seems that the engineered tracheal graft may improve the respiratory function and decrease symptoms in patients who suffer from asthma-like attacks due to mustard gas exposure. There are several successful case reports on the transplantation of stem cell-based bioartificial grafts in structural airway diseases. Therefore, we hope that the reconstruction of tracheobronchial structure can lead to a decrease in respiratory difficulties in mustard gas-exposed patients who suffer from tracheomalacia. In the present review, we summarize the main aspects of tracheal tissue engineering and cell-based therapies and the possibilities of the application of these approaches in mustard gas-exposed patients.

A novel self-nanoemulsifying formulation for sunitinib: Evaluation of anticancer efficacy.

Breast cancer is the top cancer and a main cause of death among women. The incidence of this cancer is increasing in the world. Sunitinib maleate is an oral, small-molecule, multi-targeted receptor tyrosine kinase inhibitor that inhibits tumor cell proliferation and angiogenesis, and has been administrated as an anticancer drug. Self-nanoemulsifying drug delivery system (SNEDDS) is an isotopic mixture of an oil, a surfactant and usually a co-surfactant, which can spontaneously form fine oil-in-water nanoemulsion in aqueous media. Here, a SNEDDS composed of 15% ethyl oleate (as an oil phase), 30% tween 80 (as a surfactant), and 55% PEG 600 (as a co-surfactant) was prepared and developed as a carrier for sunitinib. The average droplet size of sunitinib-loaded SNEDDS was 29.5±6.3nm with a stability of more than one month. Sunitinib release from SNEDDS was enhanced accompanied by a controlled dissolution of the drug. Cytotoxicity studies on 4T1 and MCF-7 cell lines indicated a toxicity enhancement in sunitinib by SNEDDS. To inspect the bioavailability of the drug-loaded SNEDDS after oral administration with a dose of 50mgkg-1, the maximum plasma concentration and the mean area under the plasma concentration-time curve were measured. It was found that these parameters were increased 1.45- and 1.24-times respectively, compared to a drug suspension.

Erlotinib-loaded albumin nanoparticles: A novel injectable form of erlotinib and its in vivo efficacy against pancreatic adenocarcinoma ASPC-1 and PANC-1 cell lines.

Erlotinib was loaded on albumin nanoparticles for the first time and the cytotoxic effect of the resulting nanoparticles against ASPC-1 and PANC-1 pancreatic adenocarcinoma cell lines was evaluated. The carrier (albumin nanoparticles, ANPs) was synthesized by desolvation method using a mixed solvent followed by thermal crosslinking for stabilization. ANPs and the drug-loaded ANPs were characterized by field emission scanning and transmission electron microscopies, particle size analysis and Fourier transform infrared spectroscopy. The nanoformulation had a size of <14nm with a good monodispersity. Drug loading and encapsulation efficiencies were evaluated as 27 and 44%. Cytotoxicity assays after 72h revealed the potential of ANPs to improve erlotinib toxicity (54% against 34% of free drug toward ASPC-1 cell line, and 52% against 30% toward PANC-1 cell line). Values of IC50 were obtained for both cell lines and indicated significant reduction in the erlotinib dose necessary for killing the cells, while, ANPs were completely safe. The results demonstrated that erlotinib-loaded ANPs had a remarkable potential for pancreatic cancer drug delivery.

The Immunomodulatory Effect of Bone-Marrow Mesenchymal Stem Cells on Expression of TLR3 and TLR9 in Mice Dendritic Cells.

BACKGROUND: Mesenchymal stem cells (MSCs) are multipotent cells with immunomodulatory effect on immune cells including dendritic cells (DCs). DCs are the most potent antigen-presenting cells (APC). MSCs have been found to modulate both differentiation and function of DCs. DCs express a broad range of Toll-like receptors (TLR), which play a critical role in DCs maturation and function. OBJECTIVE: To evaluate expression level of TLR3 and TLR9 transcripts in DCs following treatment with MSCs supernatant. METHODS: MSCs and DCs were derived from adult BALB/c mice bone marrow and spleen, respectively. MSCs supernatant was harvested following 24, 48, and 72 hours. Isolated DCs were treated with MSCs supernatant and incubated for 24 and 48 hours. TLR3 and TLR9 transcript levels were evaluated using real-time PCR. RESULTS: The results showed that 48 and 72 hours MSCs supernatant significantly decreased the expression of TLR3 in DCs following 24 and 48 hours incubation in comparison with untreated cells (p<0.01). Moreover, 48 hours of treatment with 24, 48 and 72 hours MSCs supernatant significantly decreased TLR9 transcript level (p<0.05). CONCLUSION: TLR3 and TLR9 mRNA expression decreases in DCs after incubation with MSCs culture supernatant. This confirms the immunomodulatory role of MSCs in cell-base therapy.

Interleukin-28B rs12979860 C/T Polymorphism and Acute Cellular Rejection after Liver Transplantation.

BACKGROUND: Interleukin-28 (IL-28B) rs12979860 C/T polymorphism is a known predictor of sustained virological response after antiviral treatment in hepatitis C. IL-28B affects the innate immune system as well as intrahepatic expression level of interferon-stimulated genes. OBJECTIVE: To investigate the effect of recipient IL-28B polymorphism on occurrence of acute rejection after liver transplantation. METHODS: 140 liver allograft recipients were selected. Acute rejection episodes were recorded in 39 patients (AR group); the remaining had normal graft function (non-AR group). 70 normal subjects were also studied as the control group. The IL-28B rs12979860 was genotyped through PCR-RFLP method. RESULTS: No significant difference was found between AR and non-AR groups in terms of genotype and allele frequency. However, the CC genotype was significantly (p<0.001) more frequent in patients than in the control group; the C allele variants increased the risk of end-stage liver disease (OR: 2.60). CONCLUSION: Liver damage in association with the carriage of IL-28B C allele is associated with a higher likelihood of developing cirrhosis.