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Osteochondral defects are defined most typically by damages to both cartilage and subchondral bone tissue. It is challenging to develop bilayered scaffolds that regenerate both of these lineages simultaneously. In the present study, an electrospun bilayer nanofibrous scaffold was designed to repair osteochondral lesions. A nanocomposite of hydroxyapatite, strontium, and reduced graphene oxide were combined with polycaprolactone polymer to fabricate the osteogenic differentiation layer. Additionally, the chondrogenic differentiation layer was also formed using polyethersulfone polymer and benzyl hyaluronan. The physical, mechanical, and chemical properties of the scaffolds were determined, and adipose-derived mesenchymal stem cells were cultured on each layer to evaluate their biocompatibility and differentiation potential. Cell viability, mineralization, alkaline phosphatase enzyme (ALP) expression, and extracellular calcium deposition were measured using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide assay, alizarin red staining, ALP activity, and calcium deposition. Real-time polymerase chain reaction (PCR) was used to assess the expression levels of osteogenic (Collagen I, Runx II, ALP, Osteocalcin) and chondrogenic (Sox9, Collagen II (Col II), Aggrecan) genes. Finally, the osteochondral scaffold was created by electrospinning these two layers for 2 days. The scaffold was grafted into the osteochondral defect of a Wistar rat's knee. 60 days after surgery, real-time PCR, immunohistochemistry (IHC), and hematoxylin and eosin staining were performed. The expression of chondrogenic and osteogenic genes was increased compared to the control group, as confirmed by real-time PCR. Furthermore, IHC revealed a rise in Col II and Collagen X expression. Finally, in vivo and in vitro studies have shown that the electrospun bilayer scaffold is biocompatible, which facilitates osteochondral healing.

Nanotechnology plays a promising role in biomedical applications, particularly tissue engineering. Recently, the application of magnetic scaffolds and pulsed electromagnetic field (PEMF) exposure has been considered in bone tissue regeneration. In this study, 3rd generation dendrimer-modified superparamagnetic iron oxide nanoparticles (G3-SPIONs) are synthesized comprehensively characterized. Magnetic polycaprolactone (PCL) nanofibers are prepared by incorporating G3-SPIONs within the electrospinning process ,and physicochemical characteristics ,as well as cytocompatibility and cell attachment are assessed. Eventually, the osteogenic differentiation ability of adipocyte-derived mesenchymal stem cells (ADMSCs) cultured on the magnetic scaffold with and without PEMF exposure was investigated by measurement of alkaline phosphatase (ALP) activity and calcium content. The expression of specific bone markers was studied using the Real-time PCR method. According to the results, G3-SPIONs with mean size and zeta potential of 17.95 ± 3.57 nm and 22.7 mV, respectively, show a high saturation magnetization (57.75 emu/g). Adding G3-SPIONs to PCL significantly decrease nanofibers size to 495±144 nm and improves cell attachment and growth. The ADMSCs cultured on the G3-SPION-PCL scaffold in the presence of osteogenic media (OM) and exposure to PEMF expressed the highest Osteocalcin and Runx2 and showed higher calcium content as well as ALP activity. It can be concluded that the synthesized G3-SPION incorporated PCL nanofibers serve as a promising magnetic scaffold for bone regeneration. Also, utilizing the magnetic scaffold in the presence of OM and PEMF provides a synergistic effect toward osteogenic differentiation of ADMSCs. Key Words: Superparamagnetic iron oxide nanoparticles, Dendrimer, Polycaprolactone, Pulsed electromagnetic field, Bone tissue engineering

Aim: Polyamidoamine (PAMAM) dendrimers are attracting interest of the scientists as vehicles for nucleic acid delivery due to their suitable properties. The highly positive surface charged of PAMAM enables an adequate interaction with negatively charged microRNAs. Purpose: The purpose of this study is to investigate the anti-tumor effect of microRNA Mimic let-7b loaded in PAMAM dendrimers (G5) on Non-Small Cell Lung Cancer (NSCLC) cells. Objective: In order to increase the anti-tumor effect, chloroquine is employed to enhance the endosomal escape which is counted as a limitation in the advancement of gene delivery. Nanoparticles (NPs) were coated with natural polysaccharide "Hyaluronic Acid (HA)" to develop biodegradable carriers with targeting moiety for over-expressed CD44 receptors on NSCLC cells. The size and zeta potential measurements, gel retardation, cellular uptake, cell viability and gene expression studies were investigated for the designed delivery system. Results: DLS analysis showed monodispersed small nanoparticles, which was in agreement with TEM results. Remarkably, NPs in the cells pretreated with chloroquine exhibited the highest cytotoxicity and were capable of inducing apoptosis. In cellular uptake study, NPs labeled with Fluorescein Isothiocyanate (FITC), were successfully taken up in cancer cells. Moreover, the expression study of three genes linked with cancer initiation and development in NSCLC, including KRAS, p-21, and BCL-2 indicated a decrease in KRAS and BCL-2 (oncogenic and anti-apoptotic genes) and increase in p-21 (apoptotic gene). Conclusion: All factors considered, the results declare that application of let-7b-loaded PAMAM-HA NPs in combination with chloroquine can be a promising therapeutic option in cancer cells inhibition. This fact has frequently been highlighted by many researchers upon the potentials of micro RNA delivery in cancer cells.

Whereas several clinical trials are ongoing or have been completed examining the benefits of Ascorbic acid-based therapy of cancer patients, its efficacy at GBM treatment has not been sufficiently investigated. In the present study, the influence of L-Ascorbic acid (Vitamin C, VC) on two GBM cell lines (U87 and U251) was evaluated in terms of cytotoxicity, induction of cell cycle arrest, reactive oxygen species (ROS) production, and alteration in the level of GBM related microRNAs. The half maximal inhibitory concentration (IC50) of VC was obtained by Crystal Violet assay as 2 mM and 1.8 mM for U87 cell line following 24 and 48 h treatment, respectively. These values were obtained in a similar way for U251 cell line as 3.2 mM and 2.9 mM. Propidium iodide (PI) staining of the cells revealed that ascorbic acid caused cell cycle arrest in G2/M phase in both of studied GBM cell lines. Moreover, RT-qPCR results indicated that VC-treatment of GBM cell lines causes downregulation of Bcl-2 alongside increase in BAK-1 and BAX expressions. Flowcytometry-based DCFH assay confirmed drastic increase in reactive oxygen species (ROS) within U87 and U251 cells following VC-treatment. Eventually, study on microRNAs expression profile implied significant increase in four tumor suppressor miRNAs including miR-7, miR-34a, miR-128, and miR-182 in both of U87 and U251 cell lines after treatment with ascorbic acid. Besides, the expression levels of three onco-miRs (i.e., miR-10b in both of cell lines, miR-222 in U87 and miR-93 in U251) were significantly diminished.

Among the vaccines have been developed thus far against SARS-CoV-2, the mRNA-based ones have demonstrated more promising results regarding both safety and efficacy. Two remarkable features of the mRNA vaccines introduced by the Pfizer/BioNTech and Moderna companies are the use of (N1-methyl-pseudouridine-) modified mRNA and the microfluidics-based production of lipid nanoparticles (LNPs) as the carrier. In the present study, except Anti-Reverse Cap Analog (ARCA), no other nucleoside analogs were employed to synthesize Spike-encoding mRNA using the in vitro transcription (IVT) method. Furthermore, LNPs were prepared via the ethanol injection method commonly used for liposome formation as an alternative for microfluidics-based approaches. The produced mRNA-LNP vaccine was evaluated for nanoparticles characteristics, encapsulation and transfection efficiencies, in vitro cytotoxicity as well as stability and storability. The safety of vaccine was assessed in Balb/c mice injected with mRNA-LNPs containing 10 µg of spike-encoding mRNA. Eventually, the vaccine efficacy in inducing an immune response against SARS-CoV-2 was studied in Balb/c and C57BL/6 mice (received either 1 or 10 µg of mRNA) as well as in rhesus macaque monkeys (infused with mRNA-LNPs containing 100 µg of mRNA). The ELISA and virus neutralizing test (VNT) results showed a significant augmentation in the level of neutralizing antibodies against SARS-CoV-2. Moreover, the ELISA assay showed virus-specific IFN-γ secretion in immunized mice as a marker of TH1 cell-based immune response, whereas favorably no change in the production of IL-4 was detected.

Background Three-dimensional (3D) cell culture methods are identified for simulating the biological microenvironment and demonstrating more similarity to in vivo circumstances. Anaplastic thyroid carcinoma (ATC) is a lethal endocrine malignancy. Despite different treatment approaches, no improvement in the survival rate of the patients has been shown. In this study, we used the 3D in vitro ATC model to investigate the cytotoxic effect of BI-847325 anticancer drug in two-dimensional (2D)- and 3D- cultured cells. Methods Human ATC cell lines, C643 and SW1736, were cultured in one percentage (w/v) sodium alginate. Spheroids were incubated in medium for one week. The reproducibility of the fabrication of alginate beads was evaluated. Encapsulation of the cells in alginate was examined by DAPI (4′,6-diamidino-2-phenylindole) staining. Survival of alginate-encapsulated cells was evaluated by CFSE (5,6-Carboxyfluorescein N-hydroxysuccinimidyl ester) staining. The population doubling times of C643 and SW1736 cell lines cultured in 2D monolayer as well as in 3D system were calculated. The cytotoxic effect of BI-847325 on 2D- and 3D- cultured cell lines was assessed for 24–72 h by MTT [3-(4,5-dimethylthiazolyl-2)-2,5-diphenyltetrazolium bromide] assay. Finally, the 3D culture results were compared with the 2D culture method. Results The half-maximal inhibitory concentration (IC50) values of BI-847325 were higher in 3D culture compared to 2D culture. The cytotoxicity data indicated that 3D in vitro models were more resistant to chemotherapy agents. Conclusions The findings of this study are beneficial for developing in vitro ATC 3D models to analyze the efficacy of different chemotherapy drugs and formulations.

Due to number of reasons such as ease of isolation and their broad differentiation capability, adult human mesenchymal stem cells (MSCs) are widely considered as of the most promising cells for regenerative medicine and tissue engineering. Nevertheless quick decrease in expression of transcription factors associated with stemness and self-renewal during ex vivo expansion of MSCs is an impediment against their therapeutic applications. Since the influence of extra cellular matrix (ECM) proteins on the fate of stem cells is well documented, the culture of MSCs on ECM-derived synthetic biomolecules is worth investigating. In the present study, a laminin-derived peptide, YIGSR was covalently immobilized on the chitosan film surface using carbodiimide chemistry and confirmed by fluorometry. The results obtained from surface characterization by atomic force microscopy (AFM) and contact angle measurement, showed no significant difference in topological features and hydrophilicity after peptide immobilization. Employment of these surfaces for culture of human adipose-derived mesenchymal stem cells demonstrated that the immobilized YIGSR peptide has a favorable effect on adhesion and maintaining viability of the MSCs as well as on the expression of stemness markers (Nanog, Oct-4, and Sox-2) in these cells.

Anaplastic thyroid carcinoma (ATC) is the most lethal malignancy in thyroid carcinomas. Long non-coding RNAs (lncRNAs) are a member of non-coding RNAs, regulating the expression of gene. Metastasis-associated lung adenocarcinoma transcript 1 (MALAT1) is an onco-lncRNA that is overexpressed in several carcinomas including ATC. Evidence showed that MALAT1 has a crucial function in apoptosis, and cell cycle progression.In order to take advantage of 3D cell culture system in cancer investigation, we have used a 3D in vitro ATC model to determine the effect of dual MEK/Aurora kinase inhibitor BI-847325 anticancer drug on the fundamental molecular mechanisms of MALAT1-mediated gene regulation in ATC.In this study, ATC cell lines (C643 and SW1736) were grown in alginate scaffold. Encapsulated cells were treated by BI-847325. Changes in expression of MALAT1, Mcl1, miR-363-3p, and cyclinD1 were measured by qRT-PCR.MALAT1 gene expression following BI-847325 treatment was significantly downregulated in C643 and SW1736 cell lines. Reversely, miR-363-3p expression was significantly upregulated by BI-847325 in both ATC cell lines. Mcl1 expression was significantly downregulated after treatment in C643 cell lines. Moreover, the expression of this gene was not significantly reduced following BI-847325 treatment in SW1736 cell line. Additionally, cyclin D1 expression was significantly downregulated after treatment in both ATC cell lines. Altogether, the result of this study was the first report of MALAT1's molecular function in ATC and suggested that BI-847325 which inhibits both MEK and Aurora kinase family could be effective against ATC by regulating the genes involved in cell cycle and apoptosis including MALAT1and its downstream genes. Graphical abstract Schematic representation of the biological role of MALAT1 in cyclin D1, miR-363-3p and Mcl1 gene regulations. Stimulation of receptor tyrosine kinase (RTK) by growth factors (GFs) phosphorylates RAS that subsequently activates RAF. Then, RAF phosphorylates MEK. Consequently, activated MEK phosphorylates ERK downstream effector, leading to the MALAT1 gene expression. MALAT1 is a negative regulator of Mcl1 mRNA by sponging of miR-363-3p. In addition, MALAT1 leads to Axin1 and APC downregulation and Wnt/β-catenin signaling pathway activation. Stable β-catenin translocates from the cytoplasm to the nucleus and promotes cyclin D1 gene expression.

Silibinin, a polyphenolic flavonolignan, is well-known as a safe therapeutic drug without any side effects in the treatment of many malignancies especially cancerous cells. In this study, to overcome problems such as low solubility of silibinin and to enhance its delivery to cancerous cells, we encapsulated silibinin in polymersome nanoparticles. Physicochemical measurements such as dynamic light scattering, transmission electron microscopy, scanning electron microscopy, and atomic force microscopy confirmed the proper encapsulation of silibinin in nanoparticles. Furthermore, antiproliferative and apoptotic activities of silibinin encapsulated in polymersome nanoparticles (SPNs) on MDA-MB-231 breast cancer cell line were validated by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay, Annexin V/Propidium Iodide measurement, and cell cycle analysis. In addition, quantitative reverse transcription polymerase chain reaction analysis confirmed that SPNs can repress oncogenic microRNAs (miRNAs) such as miR-125b and miR-182, as well as antiapoptotic genes such as Bcl2. SPNs can also induce overexpression of proapoptotic target genes such as P53, CASP9, and BAX directly and/or indirectly (through regulation of miRNAs). Our results suggested that polymersomes can be used as stable carriers in nano-dimensions and SPNs can be considered as a promising pharmacological agent for cancer therapy.

Background New insights on cellular and molecular aspects of both oligodendrocyte (OL) differentiation and myelin synthesis pathways are potential avenues for developing a cell-based therapy for demyelinating disorders comprising multiple sclerosis. MicroRNAs (miRNA) have broad implications in all aspects of cell biology including OL differentiation. MiR-184 has been identified as one of the most highly enriched miRNAs in oligodendrocyte progenitor cells (OPCs). However, the exact molecular mechanism of miR-184 in OL differentiation is yet to be elucidated. Methods and results Based on immunochemistry assays, qRT-PCR, and western blotting findings, we hypothesized that overexpression of miR-184 in either neural progenitor cells (NPCs) or embryonic mouse cortex stimulated the differentiation of OL lineage efficiently through regulating crucial developmental genes. Luciferase assays demonstrated that miR-184 directly represses positive regulators of neural and astrocyte differentiation, i.e., SOX1 and BCL2L1, respectively, including the negative regulator of myelination, LINGO1. Moreover, blocking the function of miR-184 reduced the number of committed cells to an OL lineage. Conclusions Our data highlighted that miR-184 could promote OL differentiation even in the absence of exogenous growth factors and propose a novel strategy to improve the efficacy of OL differentiation, with potential applications in cell therapy for neurodegenerative diseases. Electronic supplementary material The online version of this article (10.1186/s13287-019-1208-y) contains supplementary material, which is available to authorized users.

Colorectal Cancer (CRC) has the most common malignant gastrointestinal cancer which representing about13% of all malignant tumor. CRC Cancer Stem Cell is the major reasons for recurrence of disease cause of solid tumor metastasis, relapse of cancer after treatment and drug resistance. Silibinin, an herbal extract from milk thistle plant, has been identified as a potential cancer medicine that can target the signaling pathway of CSCs and change their abilities. In our study, the results of CSC confirmation test such as specific surface CD markers and ability to form colonospheres was indicated the HT-29 cells as CSC-CRC. To increase the effectiveness of Silibinin, and also, to evaluate therapeutic intentions on HT-29 cancer stem-like cells, we encapsulated Silibinin in polymersome nanoparticle and validated the anti-proliferative and apoptotic activities of this new patent by MTT assay, AnnexinV/PI method, cell cycle analysis and DAPI staining. Furthermore, the efficacy of drug on Multicellular Tumor Spheroid (MCTS) and single cell suspension was showed that SPN had succeed to decrees the expression level of CSC CD markers compared with control group. Follow by using miRNAs as a novel and minus invasive expertise for prognostic, RT-qPCR confirmed that SPNs can repress oncogenic miRNAs such as miR-221 and miR-222. Silibinin encapsulated in Polymersome Nanoparticles (SPNs) can also enhance the expression of tumor suppressor miR-34a and some of its proapoptotic target genes such as P53, BAX, CASP9, CASP3, and CASP8. Our results suggested that SPNs can be recognized as a new stimulant factor to direct the HT-29 cancer cells toward apoptosis pathways thorough modify expression of some miRNAs and their apoptotic target genes directly and/or indirectly.

Nanotechnology plays a promising role in biomedical applications, particularly tissue engineering. Recently, the application of magnetic scaffolds and pulsed electromagnetic field (PEMF) exposure has been considered in bone tissue regeneration. In this study, 3rd generation dendrimer-modified superparamagnetic iron oxide nanoparticles (G3-SPIONs) are synthesized comprehensively characterized. Magnetic polycaprolactone (PCL) nanofibers are prepared by incorporating G3-SPIONs within the electrospinning process ,and physicochemical characteristics ,as well as cytocompatibility and cell attachment are assessed. Eventually, the osteogenic differentiation ability of adipocyte-derived mesenchymal stem cells (ADMSCs) cultured on the magnetic scaffold with and without PEMF exposure was investigated by measurement of alkaline phosphatase (ALP) activity and calcium content. The expression of specific bone markers was studied using the Real-time PCR method. According to the results, G3-SPIONs with mean size and zeta potential of 17.95 ± 3.57 nm and 22.7 mV, respectively, show a high saturation magnetization (57.75 emu/g). Adding G3-SPIONs to PCL significantly decrease nanofibers size to 495±144 nm and improves cell attachment and growth. The ADMSCs cultured on the G3-SPION-PCL scaffold in the presence of osteogenic media (OM) and exposure to PEMF expressed the highest Osteocalcin and Runx2 and showed higher calcium content as well as ALP activity. It can be concluded that the synthesized G3-SPION incorporated PCL nanofibers serve as a promising magnetic scaffold for bone regeneration. Also, utilizing the magnetic scaffold in the presence of OM and PEMF provides a synergistic effect toward osteogenic differentiation of ADMSCs. Key Words: Superparamagnetic iron oxide nanoparticles, Dendrimer, Polycaprolactone, Pulsed electromagnetic field, Bone tissue engineering

Over the last decades, magnificent progress in the field of nanopharmaceuticals mostly with sizes smaller than 100 nm has led to the development of novel delivery systems and brightened the hope of finding new approaches to combat threatening diseases including cancer. So far, numerous efforts have been made to develop appropriate delivery systems with favorable features such as acceptable toxicity profile, high cellular uptake, low immunogenicity, and stable physicochemical properties along with distribution of the therapeutic molecule specifically to the site of action, without affecting healthy organs and tissues. Non-viral delivery systems have always been suitable options for delivery purposes. Polymers, liposomes, and inorganic delivery systems are all of the available choices in non-viral delivery systems, with each possessing their own advantages and pitfalls. This current review presents the recent advances about the application of various non-viral nanocarriers in the delivery of diverse therapeutic agents especially in cancer treatment. Targeting ligands as an important part of designing targeted nanocarriers to the site of interest or intra-cellular environment and opportunities and challenges of nano-based systems for drug and gene delivery are also discussed.

Many biological mechanisms including cellular metabolism and cell death are regulated by mitochondria known as powerhouse of the cell. Recently, let-7b, a tumour-suppressor microRNA has been detected in mitochondria of human cells targeting several mitochondrial-encoded respiratory chain genes. Triphenylphosphonium cation (TPP) is one of the major classes of mitochondriotropics that possess the ability of specifically targeting the mitochondria. PAMAM dendrimers are one of the most available agents in gene delivery due to their well-defined and beneficial features such as large density of surface functional groups. Hyaluronic acid (HA), a natural polysaccharide has been demonstrated to have the abilities such as good biocompatibility and targeting CD44 overexpressed receptors on non-small cell lung cancer (NSCLC) cells. In this research, let-7b-PAMAM (G5)-TPP and let-7b-PAMAM (G5)-TPP-HA nano-carriers were designed to deliver let-7b miRNA mimic to NSCLC cells' mitochondria as a novel way of cancer cells inhibition. Nano-carriers were capable of being successfully taken up by A549 cells and localised in mitochondria environment. Let-7b loaded nanoparticles reduced cell viability and induced apoptosis significantly. Expression of genes involved in mitochondrial oxidative function was decreased resulting in nanoparticles effect on mitochondria. Application of mitochondria targeted-miRNA delivery systems could regulate cellular functions to inhibit lung cancer.

Magnetic iron oxide nanoparticles are a well-explored class of nanomaterials known for their high magnetization and biocompatibility. They have been used in various biomedical applications such as drug delivery, biosensors, hyperthermia, and magnetic resonance imaging (MRI) contrast agent. It is necessary to surface modify the nanoparticles with a biocompatible moiety to prevent their agglomeration and enable them to target to the defined area. Dendrimers have attracted considerable attention due to their small size, monodispersed, well-defined globular shape, and a relative ease incorporation of targeting ligands. In this study, superparamagnetic iron oxide nanoparticles were synthesized via a coprecipitation method. The magnetic nanoparticles (MNPs) had been modified with (3-aminopropyl) triethoxysilane, and then polyamidoamine functionalized MNPs had been synthesized cycling. Various characterization techniques had been used to reveal the morphology, size, and structure of the nanoparticles such as scanning electron microscopy, transmission electron microscope, X-ray diffraction analysis, and vibrating sample magnetometer, Fourier-transform infrared spectroscopy and zeta potential measurements. In addition, the cytotoxicity property of G3-dendrimer functionalized MNPs were evaluated using 3-[4,5-dimethylthiazol-2-yl]-2, 5-diphenyl tetrazolium bromide assay which confirmed the biocompatibility of the nanocomposites. Dendrimer functionalized MNPs are able to act as contrast agents for MRI and magnetic fluid hyperthermia mediators. A superior heat generation was achieved for the given concentration according to the hyperthermia results. MRI results show that the synthesized nanocomposites are a favorable option for MRI contrast agent. We believe that these dendrimer functionalized MNPs have the potential of integrating therapeutic and diagnostic functions in a single carrier.

Fabrication of electrospun chitosan nanofibers is still a controversial issue in publications. Although regarding the lots of reports, mixtures of chitosan with a hydrophilic synthetic polymer such as polyethylene oxide (PEO) have been electrospun successfully, abundance of partly contradictory protocols in which one variable has been surveyed in each study is unfortunately baffling. In the present study, influence of three considerable parameters including the average molecular weight of chitosan, chitosan solution concentration and the mass ratio of polyethylene oxide to chitosan at the mixtures on electrospinning possibility as well as the quality of as-spun fibers is investigated. Eventually, the necessities for obtaining the best results are introduced followed by further analysis of optimized nanofibers using atomic force microscopy. According to our results, the blend solutions prepared from the low molecular weight (LMW) chitosan and PEO are efficient for reproducible production of bead-free electrospun nanofibers even in low proportion of polyethylene oxide.

Template-mediated self-assembly synthesis has produced a diverse range of biomimetic materials with unique physicochemical properties. Here, we fabricated novel fluorescent three-dimensional (3-D) hydroxyapatite (HAP) nanorod-assembled microspheres using iron quantum cluster (FeQC) as a hybrid template, containing three organic components: hemoglobin chains, piperidine, and iron clusters. The material characterization indicated that the synthesized HAP possessed a uniform rod-like morphology, ordered 3-D architecture, high crystallinity, self-activated fluorescence, and remarkable photostability. Our study proposed that this FeQC template is a promising regulating agent to fabricate fluorescent self-assembled HAP microspheres with a controlled morphology. The effect of HAP on stem cell fate and their osteogenic differentiation was investigated by culturing human bone marrow-derived mesenchymal stromal/stem cells (BMSCs) with HAP microspheres. Significant increases in collagen matrix production and gene expression of osteogenic markers, including osteocalcin (OCN), Runt-related transcription factor 2 (Runx2), bone sialoprotein (BSP) and alkaline phosphatase (ALP), were observed compared to the controls after 21 days of culture. Taken together, our data suggest that synthetic HAP nanorod-assembled microspheres represent a promising new biomaterial which exhibits enhanced fluorescent properties and osteoinductive effects on human BMSCs.

Table S2. Primers for qRT-PCR of genes and miRNAs. (DOCX 13 kb)

Table S1. Primers for 3â ˛UTR cloning. (DOCX 12 kb)

Introduction: Camphor is a natural antioxidant with anti-inflammatory and tissue repair properties. Nephrotoxicity is the most important side effect of gentamicin (GEM) administration. Therefore, investigating the effect of natural antioxidants can resolve this complication. Objectives: We aimed to assay the effect of camphor on biochemical factors and gene expression of antioxidant enzymes (catalase [CAT], glutathione peroxidase [GPX]) and inflammatory markers (tumor necrosis factor-alpha [TNF-α], nuclear factor kappa-B [NF-κB], interleukine-6 [IL-6]), and apoptotic indices (BCL2-associated X protein [Bax], B-cell lymphoma 2 [Bcl-2], caspase-3)], against GEM-induced nephrotoxicity in rats. Materials and Methods: Thirty adult male Wistar rats were allocated to five groups. Positive control and treatment groups were given GEM to induce nephrotoxicity. Animal treatment groups were treated with camphor in olive oil for 12 days. Renal biopsies, serum, extraction of renal tissue and urine of rats were taken after the twelfth day. Biopsies were examined for structural changes using a light microscope, moreover, apoptosis, desired biochemical and inflammatory factors, were investigated by suitable methods. Results: Camphor had no effect on biochemical factors, including malondialdehyde (MDA), glutathione (GSH), nitric oxide (NO), urea, creatinine and urine protein. However, it reduced the gene expression of TNF-α, NF-κB, IL-6, Bax, and caspase-3 and increased the gene expression of GPX and CAT and Bcl-2. Moreover, camphor improved kidney histopathological changes in the camphor groups in comparison with the GEM group. Conclusion: Camphor can be useful in the attenuation of GEM-induced nephrotoxicity based on expression levels of examined enzymes and factors and improving kidney histopathological changes.

Construction of scaffolds which are similar to natural niches regarding both biochemical composition and mechanical characteristics has gained great attention in the field of tissue engineering. However, application of natural polymers, such as hyaluronic acid, is challenging in construction of scaffolds due to physicochemical properties, difficult to use in electrospinning and low cell adhesion rate. In this study, HA was acetylated to make it soluble in high polarity solvent and blended with PCL for construction of nanofibrous composite (ac-HA/PCL) scaffolds. Chondroinductivity of the constructed scaffolds was investigated using human mesenchymal stem cells (hADSCs). The presence of acetyl groups, as well as morphology and biocompatibility of the composite scaffolds were characterized by HNMR, FTIR, SEM and MTT assay respectively. Expression of cartilage-specific genes (SOX9, Col II and Aggrecan) was monitored by Real-time PCR. Significant increase in expression of Sox9 and Col II as the markers of chondrogenic differentiation as well as the results of Alcian blue staining, indicated the chondro-inductive potential of HA/PCL nanofibrous scaffolds. Acetylated HA was biocompatible with chondroinductivity features, therefore it not only had the positive characteristics of natural HA, but also enhanced the cellular attachment and application potential.

Expansion of human induced pluripotent stem cells (h-iPSCs) on mouse derived feeder layers or murine cells secretions such as Matrigel hamper their clinical applications. Alternative methods have introduced novel substrates as stem cell niches or/and optimized combinations of humanized soluble factors as fully defined mediums. Accordingly vitronectin as a main part of ECM have been commercialized significantly as a stem cell niche-forming substrate. In this work, we used a functional peptide derived from vitronectin (VTN) and co-immobilized it with FGF-2 (as an indisputable ingredient of defined culture mediums) on chitosan film surface. After chemical and physical characterization of the pristine chitosan surface as well as ones modified by VTN or/and FGF-2, h-iPS cells were cultured on them at the xeno/feeder-free conditions. Our results demonstrated that co-immobilization of these two biomolecules has a synergistic effect on adhesion and clonal growth of h-iPS cells with maintained expression of pluripotency markers in a FGF-2 density-dependent manner. This is the first report of co-immobilization of an ECM derived molecule and a growth factor for stem cell culture.

Patients with poor prognosis in thyroid cancers resistant to currently available therapeutic modalities are often in search of a new treatment. The epidermal growth factor receptor (EGFR) vIII, a ligand-independent, constitutively active, mutated form of EGFR has been shown to play a role in the pathogenesis of some cancers. Consequently, the immunohistochemical detection of EGFRvIII with novel camel antibodies, which are valuable for their ability to interact with less antigenic epitopes in contrast to the conventional antibodies, might be worthy in diagnostic techniques of thyroid neoplasms. EGFRvIII was evaluated on paraffin-embedded tissue specimens of 40 samples of follicular carcinomas, papillary carcinomas, medullary carcinomas, follicular adenomas, and goiter of the thyroid gland by immunohistochemistry. Positive immunostaining of neoplastic tissues with camel and rabbit polyclonal, as a control, were 81.3% and 39.1%, respectively. No goiter tissue was stained with either antibody preparation. Also, the results showed that the sensitivity of camel heavy chain antibodies (65%) is higher in contrast to conventional rabbit under the same conditions (39.1%). Considering the results of this study, exploiting the smaller heavy chain antibodies of camels against EGFRvIII seems promising in the diagnosis procedures of thyroid neoplasms.