The Effect of Chitosan/Alginate/Graphene Oxide Nanocomposites on Proliferation of Mouse Spermatogonial Stem Cells.

Male survivors of childhood cancer have been known to be afflicted with azoospermia. To combat this, the isolation and purification of spermatogonial stem cells (SSCs) are crucial. Implementing scaffolds that emulate the extracellular matrix environment is vital for promoting the regeneration and proliferation of SSCs. This research aimed to evaluate the efficiency of nanocomposite scaffolds based on alginate, chitosan, and graphene oxide (GO) in facilitating SSCs proliferation. To analyze the cytotoxicity of the scaffolds, an MTT assay was conducted at 1, 3, and 7 days, and the sample containing 30 µg/mL of GO (ALGCS/GO30) exhibited the most favorable results, indicating its optimal performance. The identity of the cells was confirmed using flow cytometry with C-Kit and GFRα1 markers. The scaffolds were subjected to various analyses to characterize their properties. FTIR was employed to assess the chemical structure, XRD to examine crystallinity, and SEM to visualize the morphology of the scaffolds. To evaluate the proliferation of SSCs, qRT-PCR was used. The study's results demonstrated that the ALGCS/GO30 nanocomposite scaffold exhibited biocompatibility and facilitated the attachment and proliferation of SSCs. Notably, the scaffold displayed a significant increase in proliferation markers compared to the control group, indicating its ability to support SSC growth. The expression level of the PLZF protein was assessed using the Immunocytochemistry method. The observations confirmed the qRT-PCR results, which indicated that the nanocomposite scaffolds had higher levels of PLZF protein expression than scaffolds without GO. The biocompatible ALGCS/GO30 is a promising alternative for promoting SSC proliferation in in vitro applications.

A Drug-Eluting Injectable NanoGel for Localized Delivery of Anticancer Drugs to Solid Tumors.

Systemically administered chemotherapy reduces the efficiency of the anticancer agent at the target tumor tissue and results in distributed drug to non-target organs, inducing negative side effects commonly associated with chemotherapy and necessitating repeated administration. Injectable hydrogels present themselves as a potential platform for non-invasive local delivery vehicles that can serve as a slow-releasing drug depot that fills tumor vasculature, tissue, or resection cavities. Herein, we have systematically formulated and tested an injectable shear-thinning hydrogel (STH) with a highly manipulable release profile for delivering doxorubicin, a common chemotherapeutic. By detailed characterization of the STH physical properties and degradation and release dynamics, we selected top candidates for testing in cancer models of increasing biomimicry. Two-dimensional cell culture, tumor-on-a-chip, and small animal models were used to demonstrate the high anticancer potential and reduced systemic toxicity of the STH that exhibits long-term (up to 80 days) doxorubicin release profiles for treatment of breast cancer and glioblastoma. The drug-loaded STH injected into tumor tissue was shown to increase overall survival in breast tumor- and glioblastoma-bearing animal models by 50% for 22 days and 25% for 52 days, respectively, showing high potential for localized, less frequent treatment of oncologic disease with reduced dosage requirements.

Combined Use of Photobiomodulation and Curcumin-Loaded Iron Oxide Nanoparticles Significantly Improved Wound Healing in Diabetic Rats Compared to Either Treatment Alone.

Introduction: Here, we assess the therapeutic effects of photobiomodulation (PBM) and curcumin (CUR)-loaded superparamagnetic iron oxide nanoparticles (SPIONs), alone or together, on the maturation step of a type 1 diabetes (DM1) rat wound model. Methods: Full-thickness wounds were inflicted in 36 rats with diabetes mellitus (DM) induced by the administration of streptozotocin (STZ). The rats were randomly allocated to four groups. Group one was untreated (control); group two received CUR; group 3 received PBM (890 nm, 80 Hz, 0.2 J/cm2); group 4 received a combination of PBM plus CUR. On days 0, 4, 7, and 15, we measured microbial flora, wound closure fraction, tensile strength, and stereological analysis. Results: All treatment groups showed a substantial escalation in the wound closure rate, a substantial reduction in the count of methicillin-resistant Staphylococcus aureus (MRSA), a substantial improvement in wound strength, a substantially improvement in stereological parameters compared to the control group, however, the PBM+CUR group was superior to the other treatment groups (all, P≤0.05). Conclusion: All treatment groups showed significantly improved wound healing in the DM1 rat model. However, the PBM+CUR group was superior to the other treatment groups and the control group in terms of wound strength and stereological parameters.

Fabrication of a microfluidic device for probiotic drug's dosage screening: Precision Medicine for Breast Cancer Treatment.

Breast cancer is the most common cancer in women; it has been affecting the lives of millions each year globally and microfluidic devices seem to be a promising method for the future advancements in this field. This research uses a dynamic cell culture condition in a microfluidic concentration gradient device, helping us to assess breast anticancer activities of probiotic strains against MCF-7 cells. It has been shown that MCF-7 cells could grow and proliferate for at least 24 h; however, a specific concentration of probiotic supernatant could induce more cell death signaling population after 48 h. One of our key findings was that our evaluated optimum dose (7.8 mg/L) was less than the conventional static cell culture treatment dose (12 mg/L). To determine the most effective dose over time and the percentage of apoptosis versus necrosis, flowcytometric assessment was performed. Exposing the MCF-7 cells to probiotic supernatant after 6, 24 and 48 h, confirmed that the apoptotic and necrotic cell death signaling were concentration and time dependent. We have shown a case that these types of microfluidics platforms performing dynamic cell culture could be beneficial in personalized medicine and cancer therapy.

Structure, Distribution, Regulation, and Function of Splice Variant Isoforms of Nitric Oxide Synthase Family in the Nervous System.

Nitric oxide (NO) is a small molecule produced by nitric oxide synthase (NOS) with various physio-pathological functions in the body. There are three main NOS isoforms, including the endothelial (eNOS), inducible (iNOS), and neuronal NOS (nNOS), that exist in the peripheral organs and nervous systems of humans and rodents. Moreover, NOS includes other identified NOS isoforms, such as retinal Muller glial cells (mNOS), mitochondrial (mtNOS), penile (PnNOS), testis-specific (TnNOS), and invertebrate Drosophila NOS (dNOS), which are the lesser-known types. It is proposed that the versatile functions of NOS isoforms depend on various NOS splice variant subtypes and their expression in the neural (e.g., brain, and spinal cord) and non-neuronal tissues (e.g., lung, kidney, liver, and GI tract). Therefore, this review summarizes the NOS subtypes, splice variants, targeted splicing expression in the body, and their proposed physio-pathological functions. At last, alternative NOS subtypes and isoforms, which have previously received scant attention, will be addressed in this article.

Stem Cell Differentiation into Cardiomyocytes: Current Methods and Emerging Approaches.

Cardiovascular diseases (CVDs) are globally known to be important causes of mortality and disabilities. Common treatment strategies for CVDs, such as pharmacological therapeutics impose serious challenges due to the failure of treatments for myocardial necrosis. By contrast, stem cells (SCs) based therapies are seen to be promising approaches to CVDs treatment. In such approaches, cardiomyocytes are differentiated from SCs. To fulfill SCs complete potential, the method should be appointed to generate cardiomyocytes with more mature structure and well-functioning operations. For heart repairing applications, a greatly scalable and medical-grade cardiomyocyte generation must be used. Nonetheless, there are some challenges such as immune rejection, arrhythmogenesis, tumorigenesis, and graft cell death potential. Herein, we discuss the types of potential SCs, and commonly used methods including embryoid bodies related techniques, co-culture, mechanical stimulation, and electrical stimulation and their applications, advantages and limitations in this field. An estimated 17.9 million people died from CVDs in 2019, representing 32 % of all global deaths. Of these deaths, 85 % were due to heart attack and stroke.

Investigation of toxicity effect of TiCN coated on 304 SS and 410 SS substrates in rat fibroblasts and B-lymphocytes.

In the present study, TiCN thin films were coated on AISI 304 and AISI 410 stainless steel (SS) substrates by Cathodic Arc Physical Vapor Deposition method. TiCN-coated substrates were confirmed by the XRD analysis results. Dense morphology and fine-grained surface of TiCN film were established by SEM images. Cellular toxicity of the coated 304 SS and 410 SS substrates was investigated in the fibroblasts and B-lymphocyte. In respect to that, we have shown coated substrates cytotoxicity, oxidative stress as well as cell viability, reactive oxygen species (ROS), lipid peroxidation (MDA), protein carbonyl, glutathione oxidase (GSSG), and glutathione reductase (GSH) assessment, releasing cytochrome c (Cytc), lysosomal membrane destabilization (AO) may lead to cell death signaling. Our results showed that the coated 304 SS and 410 SS substrates induced cells dysfunction via a significant increase in ROS production, MDA (P < 0.01 and P < 0.001), protein carbonyl (P < 0.05), and GSSG (P < 0.05 and P < 0.01) that correlated to cytochrome c release (P < 0.01). In addition, increased disturbance in oxidative phosphorylation was also shown by the decrease in cell viability (P < 0.001) and GSH (P < 0.01 and P < 0.001) in the coated 304 SS and 410 SS substrates-treated fibroblast and B-lymphocytes. The coated 304 SS and 410 SS substrates contacted cells and trafficked to the lysosomes and this is followed by lysosomal damage, leading to apoptosis/Necrosis. Our results indicated that these materials cause cellular dysfunction and subsequent oxidative stress leading to cognitive impairment in the rat fibroblasts and B-lymphocytes cells.

Gold nanorods decorated polycaprolactone/cellulose acetate hybrid scaffold for PC12 cells proliferation.

Synthetic and natural polymers have recently received considerable attention due to the exclusive potential for supporting the regenerative cellular processes in peripheral nerve injuries (PNIs). Gold nanorods (GNRs) decorated polycaprolactone (PCL)/cellulose acetate (CA) nanocomposite (PCL/CA/GNR) were fabricated via electrospinning to improve PC12 cells attachment and growth or scaffold cues. Transmission electron microscopy (TEM) corroborated the GNR distribution (23 ± 2 nm length and 3/1 Aspect ratio) and suitable average dimension of 800 nm for the fibers; also, scanning electron microscopy (SEM) represented block-free and smooth fibers without perturbation. Because of gold nanorods incorporation, electrical conductivity of PCL/CA/GNR increased ~21%. Water contact angle data emphasized PCL/CA/GNR surface is more wettable that PCL/CA (<90° at 62 s). Real-time PCR technique (RT-PCR) demonstrated overexpression of ß-tubulin and microtubule-associated protein 2 (MAP2) on PCL/CA/GNR compared to PCL/CA composite. Additionally, evaluated of the maturation and neurogenic differentiation of PC12 cells emphasized overexpression of nestin and ß-tubulin by Immunocytochemistry staining onto PCL/CA/GNR in comparison to PCL/CA composite. Notably, these recently developed hybrid scaffolds could be considered for peripheral nerve injury (PNI) regeneration.

Selective cytotoxicity mechanisms and biodistribution of diamond nanoparticles on the skin cancer in C57 mouse.

The cytotoxicity of diamond nanoparticles (DNs) to various cell lines has been on focus by numerous scientists. The cellular toxicity system of DNs has not been fully understood or explained in skin cancer, at this point. This research was carried out to discover and reveal the potential impacts of DNs on the secluded brain, heart, liver, kidney, and skin in addition to evaluation of their cytotoxicity mechanism under test conditions. Their biological activities, for example cell viability, the level of reactive oxygen species (ROS), lipid peroxidation, cytochrome c release and Apoptosis/Necrosis were evaluated. Additionally, the bio-distribution of these nanomaterials in tissues was examined in the C57 mouse. Relying on the findings of the investigation, DNs were found to increase the ROS level, Malondialdehyde (MDA) content, release of cytochrome c, and cell death in skin significantly compared to other groups. In the C57 mouse, DNs were observed to have accumulated in skin tissue more intensively than they did in other organs. The present study presents for the proof that DNs can completely induce cell death signaling in skin cancer without bringing about a high cytotoxicity in other tissues. Results suggest that DNs can be valuable in recognition of skin cancer.

Labelling of human Wharton's jelly-derived mesenchymal stem cells with gold nanorods by biomimicry method.

Mesenchymal stem cell (MSC)-based cell therapy can provide opportunities for the treatment of various diseases. However, when used in vivo, these cells should be labelled and monitored by a non-invasive method during delivery to the desired locations within the body. This study describes a biomimicry method that effectively labels human Wharton's jelly-derived MSCs (hWJ-MSCs) with a photoacoustics (PA) contrast agent, gold nanorods (GNRs), without the need for transfection agents (TAs). In this method for cell labelling, the hWJ-MSCs were co-incubated with non-adherent cells isolated from fresh umbilical cord for 2 days immediately before incubation with GNRs. Next, hWJ-MSCs were labelled with the GNRs at a concentration of approximately 1010 nanorads/mL (NR/mL) followed by transmission electron microscopy (TEM) and inductively coupled plasma mass spectroscopy (ICP-MS) to verify their labelling effectiveness. The GNRs-labelled MSCs prepared by this method had an intracellular gold (Au) concentration of 3.4 ± 0.4 pg/cell, which is an acceptable amount for cell labelling.

PC12 cells proliferation and morphological aspects: Inquiry into raffinose-grafted graphene oxide in silk fibroin-based scaffold.

Peripheral nerves injuries (PNIs) still associated with both clinical and social problems. Accordingly, tissue engineers' and surgeons' attentions have been drawn for finding efficient solutions. Herein, scaffolds based on silk fibroin (SF)/raffinose-grafted-GO (S.RafGO) nanocomposite were fabricated. Subsequently, PC12 cells growth in term of number and morphology were investigated on neat SF polymer, SF/GO (S.GO), and S.RafGO scaffolds. Characterization via scanning electron microscopy (SEM) exhibited more fibrous structures with few lamellar nanosheets for S.GO; although, S.RafGO showed extended lamellar with lower fibrous structure. Due to the incorporation of GO and raffinose-GO nanosheets into SF structure, electrical conductivity increased ~30 and 40%, respectively. Water contact angle data revealed that S.RafGO is more wettable than SF and S.GO. Real-time PCR technique detected higher expressions of the ß-tubulin, MAP2 genes on S.RafGO scaffolds in comparison with S.GO and the control group. Immunocytochemistry staining studies confirmed the overexpression of neural-specific proteins including nestin, ß-tubulin of S.GO, and S.RafGO nanocomposites in comparison with pure SF scaffolds.

Separating mouse malignant cell line (EL4) from neonate spermatogonial stem cells utilizing microfluidic device in vitro.

BACKGROUND: Some children who have survived cancer will be azoospermic in the future. Performing isolation and purification procedures for spermatogonial stem cells (SSC) is very critical. In this regard, performing the process of decontamination of cancerous cells is the initial step. The major objective of the present study is to separate the malignant EL4 cell line in mice and spermatogonial stem cells in vitro. METHODS: The spermatogonial stem cells of sixty neonatal mice were isolated, and the procedure of co-culturing was carried out by EL4 which were classified into 2 major groups: (1) the control group (co-culture in a growth medium) and (2) the group of co-cultured cells which were separated using the microfluidic device. The percentage of cells was assessed using flow cytometry technique and common laboratory technique of immunocytochemistry and finally was confirmed through the laboratory technique of reverse transcription-polymerase chain reaction (RT-PCR). RESULTS: The actual percentage of EL4 and SSC after isolation was collected at two outlets: the outputs for the smaller outlet were 0.12% for SSC and 42.14% for EL4, while in the larger outlet, the outputs were 80.38% for SSC and 0.32% for EL4; in the control group, the percentages of cells were 21.44% for SSC and 23.28% for EL4 (based on t test (p ≤ 0.05)). CONCLUSIONS: The present study demonstrates that the use of the microfluidic device is effective in separating cancer cells from spermatogonial stem cells.

Electrical stimulation induces differentiation of human cardiosphere-derived cells (hCDCs) to committed cardiomyocyte.

Logistic complexities of heart transplantation embossed the necessity of utilizing novel methods, which enable heart regeneration. Human cardiosphere-derived cells (hCDCs) are taken into consideration as a promising cell resource in cell therapy in recent years. In this study, we designed an electrochemical stimulation system, which sends square pulses to the hCDCs and records their electrical response. Morphology, viability and differentiation of hCDCs are monitored at certain time courses of the treatment. Differentiating hCDCs aligned perpendicularly with respect to the direction of applied electric current, and obtained a spindle-like morphology, while they remained viable. At the same time, specific cardiac marker genes including GATA4, cTnT and α-MHC showed a considerable up-regulation. Our findings confirm that hCDCs differentiate to committed cardiomyocytes when hCDCs receive an electrical energy of 0.06 - 0.12 Wh. This amount of electrical energy could be applied to the stem cells using versatile electrical stimulation patterns via commercially available devices.

Polymeric and inorganic nanoscopical antimicrobial fillers in dentistry.

Failure of dental treatments is mainly due to the biofilm accumulated on the dental materials. Many investigations have been conducted on the advancements of antimicrobial dental materials. Polymeric and inorganic nanoscopical agents are capable of inhibiting microorganism proliferation. Applying them as fillers in dental materials can achieve enhanced microbicidal ability. The present review provides a broad overview on the state-of-the-art research in the field of antimicrobial fillers which have been adopted for incorporation into dental materials over the last 5 years. The antibacterial agents and applications are described, with the aim of providing information for future investigations. STATEMENT OF SIGNIFICANCE: Microbial infection is the primary cause of dental treatment failure. The present review provides an overview on the state-of-art in the field of antimicrobial nanoscopical or polymeric fillers that have been applied in dental materials. Trends in the biotechnological development of these antimicrobial fillers over the last 5 years are reviewed to provide a backdrop for further advancement in this field of research.

Metal-Based Nanostructures/PLGA Nanocomposites: Antimicrobial Activity, Cytotoxicity, and Their Biomedical Applications.

Among the different synthetic polymers developed for biomedical applications, poly(lactic-co-glycolic acid) (PLGA) has attracted considerable attention because of its excellent biocompatibility and biodegradability. Nanocomposites based on PLGA and metal-based nanostructures (MNSs) have been employed extensively as an efficient strategy to improve the structural and functional properties of PLGA polymer. The MNSs have been used to impart new properties to PLGA, such as antimicrobial properties and labeling. In the present review, the different strategies available for the fabrication of MNS/PLGA nanocomposites and their applications in the biomedical field will be discussed, beginning with a description of the preparation routes, antimicrobial activity, and cytotoxicity concerns of MNS/PLGA nanocomposites. The biomedical applications of these nanocomposites, such as carriers and scaffolds in tissue regeneration and other therapies are subsequently reviewed. In addition, the potential advantages of using MNS/PLGA nanocomposites in treatment illnesses are analyzed based on in vitro and in vivo studies, to support the potential of these nanocomposites in future research in the biomedical field.

Progress in Conductive Polyaniline-Based Nanocomposites for Biomedical Applications: A Review.

Inherently conducting polymers (ICPs) are a specific category of synthetic polymers with distinctive electro-optic properties, which involve conjugated chains with alternating single and double bonds. Polyaniline (PANI), as one of the most well-known ICPs, has outstanding potential applications in biomedicine because of its high electrical conductivity and biocompatibility caused by its hydrophilic nature, low-toxicity, good environmental stability, and nanostructured morphology. Some of the limitations in the use of PANI, such as its low processability and degradability, can be overcome by the preparation of its blends and nanocomposites with various (bio)polymers and nanomaterials, respectively. This review describes the state-of-the-art of biological activities and applications of conductive PANI-based nanocomposites in the biomedical fields, such as antimicrobial therapy, drug delivery, biosensors, nerve regeneration, and tissue engineering. The latest progresses in the biomedical applications of PANI-based nanocomposites are reviewed to provide a background for future research.

Antimicrobial gum bio-based nanocomposites and their industrial and biomedical applications.

Gum polysaccharides are derived from renewable sources. They are readily available, inexpensive, non-hazardous and eco-friendly. Depending upon the source, gums may be categorized as microbial gums, plant exudate gums or seed gums. Naturally occurring gum carbohydrates find multiple applications in the biomedical arena, compared with synthetic compounds, because of their unique structures and functionalities. Gums and their biocomposites are preferred for sustained drug delivery because they are safe and edible as well as more susceptible to biodegradation. The present review provides a state-of-the-art conspectus on the industrial and biomedical applications of antimicrobial gum-based biocomposites. Different kinds of gums polysaccharides will first be addressed based on their sources. Metal-, carbon- and organic-based nanostructures that are used in gum nanocomposites will then be reviewed with respect to their industrial and biomedical applications, to provide a backdrop for future research.