Polyphenolic Carbon Quantum Dots with Intrinsic Reactive Oxygen Species Amplification for Two-Photon Bioimaging and In Vivo Tumor Therapy.

Recent studies indicate that mitochondrial dysfunctions and DNA damage have a critical influence on cell survival, which is considered one of the therapeutic targets for cancer therapy. In this study, we demonstrated a comparative study of the effect of polyphenolic carbon quantum dots (CQDs) on in vitro and in vivo antitumor efficacy. Dual emissive (green and yellow) shape specific polyphenolic CQDs (G-CQDs and Y-CQDs) were synthesized from easily available nontoxic precursors (phloroglucinol), and the antitumor property of the as-synthesized probe was investigated as compared to round-shaped blue emissive CQDs (B-CQDs) derived from well-reported precursor citric acid and urea. The B-CQDs had a nuclei-targeting property, and G-CQDs and Y-CQDs had mitochondria-targeting properties. We have found that the polyphenol containing CQDs (at a dose of 100 µg mL-1) specifically attack mitochondria by excess accumulation, altering the metabolism, inhibiting branching pattern, imbalanced Bax/Bcl-2 homeostasis, and ultimately generating oxidative stress levels, leading to oxidative stress-induced cell death in cancer cells in vitro. We show that G-CQDs are the main cause of oxidative stress in cancer cells because of their ability to produce sufficient •OH- and 1O2 radicals, evidenced by electron paramagnetic resonance spectroscopy and a terephthalic acid test. Moreover, the near-infrared absorption properties of the CQDs were exhibited in two-photon (TP) emission, which was utilized for TP cellular imaging of cancer cells without photobleaching. The in vivo antitumor test further discloses that intratumoral injection of G-CQDs can significantly augment the treatment efficacy of subcutaneous tumors without any adverse effects on BalB/c nude mice. We believe that shape-specific polyphenolic CQD-based nanotheranostic agents have a potential role in tumor therapy, thus proving an insight on treatment of malignant cancers.

Electrically stimulated 3D bioprinting of gelatin-polypyrrole hydrogel with dynamic semi-IPN network induces osteogenesis via collective signaling and immunopolarization.

In recent years, three-dimensional (3D) bioprinting of conductive hydrogels has made significant progress in the fabrication of high-resolution biomimetic structures with gradual complexity. However, the lack of an effective cross-linking strategy, ideal shear-thinning, appropriate yield strength, and higher print fidelity with excellent biofunctionality remains a challenge for developing cell-laden constructs, hindering the progress of extrusion-based 3D printing of conductive polymers. In this study, a highly stable and conductive bioink was developed based on polypyrrole-grafted gelatin methacryloyl (GelMA-PPy) with a triple cross-linking (thermo-photo-ionically) strategy for direct ink writing-based 3D printing applications. The triple-cross-linked hydrogel with dynamic semi-inner penetrating polymer network (semi-IPN) displayed excellent shear-thinning properties, with improved shape fidelity and structural stability during 3D printing. The as-fabricated hydrogel ink also exhibited "plug-like non-Newtonian" flow behavior with minimal disturbance. The bioprinted GelMA-PPy-Fe hydrogel showed higher cytocompatibility (93%) of human bone mesenchymal stem cells (hBMSCs) under microcurrent stimulation (250 mV/20 min/day). Moreover, the self-supporting and tunable mechanical properties of the GelMA-PPy bioink allowed 3D printing of high-resolution biological architectures. As a proof of concept, we printed a full-thickness rat bone model to demonstrate the structural stability. Transcriptomic analysis revealed that the 3D bioprinted hBMSCs highly expressed gene hallmarks for NOTCH/mitogen-activated protein kinase (MAPK)/SMAD signaling while down-regulating the Wnt/ß-Catenin and epigenetic signaling pathways during osteogenic differentiation for up to 7 days. These results suggest that the developed GelMA-PPy bioink is highly stable and non-toxic to hBMSCs and can serve as a promising platform for bone tissue engineering applications.

Cellulose nanocrystals vs. cellulose nanospheres: A comparative study of cytotoxicity and macrophage polarization potential.

Nanocellulose application has been increasing owing to its appealing physicochemical properties. Monitoring of the crystallinity, surface topography, and reactivity of this high-aspect-ratio nanomaterial is crucial for efficient tissue engineering. Controlling macrophage polarization phenotype remains a challenge in regenerative medicine and tissue engineering. Herein, we monitored the effects of shape-regulated (rod and spherical) nanocellulose on the macrophage modulatory potential of RAW 246.7 cells in vitro. Spherical nanocellulose (s-NC) exhibited higher thermal stability and biocompatibility than rod nanocellulose. Macrophage polarization was profoundly affected by nanocellulose topography and incubation period. M2 polarization was observed in vitro after 1 day of treatment with s-NC, followed by M1 polarization after treatment for longer periods. Transcriptome analysis similarly revealed that M1 polarization was dominant after 1 day h of incubation with both nanocellulose types. These findings demonstrate that macrophage polarization can be controlled by selecting suitable nanocellulose shape and incubation time for desired applications.

Transcriptomic Changes toward Osteogenic Differentiation of Mesenchymal Stem Cells on 3D-Printed GelMA/CNC Hydrogel under Pulsatile Pressure Environment.

Biomimetic soft hydrogels used in bone tissue engineering frequently produce unsatisfactory outcomes. Here, it is investigated how human bone-marrow-derived mesenchymal stem cells (hBMSCs) differentiated into early osteoblasts on remarkably soft 3D hydrogel (70 ± 0.00049 Pa). Specifically, hBMSCs seeded onto cellulose nanocrystals incorporated methacrylate gelatin hydrogels are subjected to pulsatile pressure stimulation (PPS) of 5-20 kPa for 7 days. The PPS stimulates cellular processes such as mechanotransduction, cytoskeletal distribution, prohibition of oxidative stress, calcium homeostasis, osteogenic marker gene expression, and osteo-specific cytokine secretions in hBMSCs on soft substrates. The involvement of Piezo 1 is the main ion channel involved in mechanotransduction. Additionally, RNA-sequencing results reveal differential gene expression concerning osteogenic differentiation, bone mineralization, ion channel activity, and focal adhesion. These findings suggest a practical and highly scalable method for promoting stem cell commitment to osteogenesis on soft matrices for clinical reconstruction.

3D-printable chitosan/silk fibroin/cellulose nanoparticle scaffolds for bone regeneration via M2 macrophage polarization.

Biopolymers-induced immune microenvironment exhibited prominent effects on bone regeneration. Osteo-immunomodulatory responses of cellulose nanoparticles incorporated chitosan hydrogel scaffolds have not yet been reported. The objective of this study was to monitor the synergistic effects of silk fibroin and cellulose nanoparticles on the immune-modulatory behavior of chitosan biopolymer scaffolds. 3D-printed biodegradable cellulose nanoparticles-reinforced chitosan/silk fibroin (CS/SF/CNPs) scaffolds were fabricated and characterized by different spectroscopic techniques. The improved rheological and recovery strength was observed in CS/SF/CNPs hydrogels than pure polymer hydrogels. A significant shift from M1 â†’ M2 macrophages polarization occurred in the CS/SF/CNPs scaffolds treated groups than the control after 3 days of incubation, showing its immune-modulatory potential. Osteo-immunomodulatory effects of the fabricated scaffolds were analyzed on human bone marrow-derived mesenchymal stem cells (hBMSCs), with macrophages-derived conditioned media (M-CM). Enhanced bone regeneration was observed in the calvaria defect rat model, indicating that the fabricated scaffolds are promising materials for bone-healing applications.

Biological Importance, Therapeutic Benefit and Analytical Aspects of Bioactive Flavonoid Pectolinarin in the Nature.

BACKGROUNDS: Plants and their derived products have been used in the traditional system of medicine for the treatment of various forms of human disorders since very ancient times. In the traditional system of medicine and modern allopathic medicine, numerous phytoconstituents have been used for the preparation of various types of formulation. Flavonoidal class phytochemicals are the main active phytoconstituents of plants, fruit, vegetables and beverages. Flavonoidal class phytochemicals are more referred as "nutraceuticals" due to their important pharmacological activities in the mammalian body. METHODS: In order to understand the beneficial health effects of flavonoidal class chemical, the present work summarized the health beneficial aspects of pectolinarin. Present work summarized the medicinal importance, pharmacological activities and analytical aspects of pectolinarin with various experimental models and advance analytical methods. However, all the collected scientific information's have been analyzed in the present work for their health beneficial potential. RESULTS: From the analysis of all the collected scientific information in the present work, it was found that pectolinarin is an important phytochemical present in numerous medicinal plants but especially found in Cirsium japonicum, which is an important medicinal herb of Korea, China and Japan. Pharmacological activities data analysis signified the health beneficial potential of pectolinarin for their anti-rheumatoid arthritis, analgesic, anti-inflammatory, hepatoprotective, anti-diabetic, anti-tumor, anti-dengue, antiviral, neuroprotective and antidepressant activity. However, the effectiveness of pectolinarin in central nervous system, bone, liver and cancerous disorders have been also reported in the literature. Analysis of present scientific information revealed the health beneficial potential of pectolinarin in modern medicine due to their numerous pharmacological activities in different parts of biological systems. Due to their biological importance in food and human health, a better understanding of their biological activities indicates their potentials as therapeutic agents. CONCLUSION: Scientific data of the present work signified the biological potential and therapeutic benefit of pectolinarin.

Graphene Oxide-Based Stimuli-Responsive Platforms for Biomedical Applications.

Graphene is a two-dimensional sp2 hybridized carbon material that has attracted tremendous attention for its stimuli-responsive applications, owing to its high surface area and excellent electrical, optical, thermal, and mechanical properties. The physicochemical properties of graphene can be tuned by surface functionalization. The biomedical field pays special attention to stimuli-responsive materials due to their responsive abilities under different conditions. Stimuli-responsive materials exhibit great potential in changing their behavior upon exposure to external or internal factors, such as pH, light, electric field, magnetic field, and temperature. Graphene-based materials, particularly graphene oxide (GO), have been widely used in stimuli-responsive applications due to their superior biocompatibility compared to other forms of graphene. GO has been commonly utilized in tissue engineering, bioimaging, biosensing, cancer therapy, and drug delivery. GO-based stimuli-responsive platforms for wound healing applications have not yet been fully explored. This review describes the effects of different stimuli-responsive factors, such as pH, light, temperature, and magnetic and electric fields on GO-based materials and their applications. The wound healing applications of GO-based materials is extensively discussed with cancer therapy and drug delivery.

Naturally-derived protein extract from Gryllus bimaculatus improves antioxidant properties and promotes osteogenic differentiation of hBMSCs.

Naturally-derived proteins or peptides are promising biopolymers for tissue engineering applications owing to their health-promoting activity. Herein, we extracted proteins (~90%) from two-spotted cricket (Gryllus bimaculatus) and evaluated their osteoinductive potential in human bone marrow-derived mesenchymal stem cells (hBMSCs) under in vitro conditions. The extracted protein isolate was analyzed for the amino acid composition and the mass distribution of the constituent peptide fraction. Fourier transform infrared (FTIR) spectroscopy was used to determine the presence of biologically significant functional groups. The cricket protein isolate (CPI) exhibited characteristic protein peaks in the FTIR spectrum. Notably, an enhanced cell viability was observed in the presence of the extracted proteins, showing their biocompatibility. The CPI also exhibited antioxidant properties in a concentration-dependent manner. More significant mineralization was observed in the CPI-treated cells than in the control, suggesting their osteoinductive potential. The upregulation of the osteogenic marker genes (Runx2, ALP, OCN, and BSP) in CPI treated media compared with the control supports their osteoinductive nature. Therefore, cricket-derived protein isolates could be used as functional protein isolate for tissue engineering applications, especially for bone regeneration.

Enhanced osteogenic potential of unzipped carbon nanotubes for tissue engineering.

Carbon nanotubes (CNTs) have attracted significant interest for various applications owing to their superior physicochemical properties. The unzipping of multi-walled carbon nanotubes was accomplished by strong acid treatment. The solution of unzipped carbon nanotubes (u-CNTs) was homogeneous and stable. The u-CNTs were characterized by Fourier transform infrared spectroscopy, X-ray diffraction, and thermogravimetric analysis. The dimensions and morphologies of the synthesized u-CNTs were examined by transmission electron microscopy and scanning electron microscopy. The u-CNTs exhibited increased zeta potential and diameter compared with pure CNTs. A decrease in the thermal stability was observed in the u-CNTs compared with pure CNTs. The u-CNTs exhibited better biocompatibility than pure CNTs in the presence of bone marrow-derived mesenchymal stem cells, showing improved biocompatibility. The u-CNT-treated media generated lower amounts of reactive oxygen species than pure CNTs. Enhanced mineralization was observed in the u-CNT-treated groups compared with the pure CNTs and the control, indicating its better osteogenic potential. The upregulation of osteogenic-associated gene markers in u-CNT groups compared with pure CNTs confirms their superior osteogenic potential. Thus, u-CNTs are potential candidates for tissue engineering applications, especially bone tissue.

Mushroom-Derived Bioactive Molecules as Immunotherapeutic Agents: A Review.

Mushrooms with enhanced medicinal properties focus on finding such compounds that could modulate the human body's immune systems. Mushrooms have antimicrobial, antidiabetic, antiviral, hepatoprotective, antitumor, and immunomodulatory properties due to the presence of various bioactive components. ß-glucans are the major constituent of the mushroom cell wall and play a significant role in their biological activity. This review described the techniques used in the extraction of the active ingredients from the mushroom. We highlighted the structure of the bioactive polysaccharides present in the mushrooms. Therapeutic applications of different mushrooms were also described. It is interesting to note that mushrooms have the potential sources of many bioactive products that can regulate immunity. Thus, the development of functional medicinal food based on the mushroom is vital for human welfare.

Effects of Cirsium setidens (Dunn) Nakai on the osteogenic differentiation of stem cells.

Cirsium setidens (Dunn) Nakai, commonly known as gondre, is a perennial herb that grows predominantly in South Korea. It contains several bioactive phytochemicals with antioxidant, anti­cancer, anti­tumor and anti­inflammatory properties. The present study aimed to investigate the effects of methanolic extracts of gondre on osteogenic differentiation of human periodontal ligament stem cells (hPDLSCs). As characterized by nuclear magnetic resonance spectroscopy and matrix­assisted laser deposition/ionization (time­of­flight) mass spectrometry, the methanol extract of gondre was found to be enriched with pectolinarin. After 48 h, enhanced viability of hPDLSCs was observed in the presence of gondre compared with under control conditions, suggesting the biocompatibility of gondre. Notably, biocompatibility was markedly affected by gondre concentration in cultured media. Relatively high cell viability was observed in medium containing 0.05% gondre. Furthermore, mineralization was significantly higher in hPDLSCs in the presence of gondre compared with that in control cells, indicating their mineralization potential. Increased expression of various transcription markers, such as collagen 1, runt­related transcription factor 2, bone sialoprotein and alkaline phosphatase, was also detected when hPDLSCs were stimulated with gondre compared with in the control groups, further confirming the superior osteogenic potential of gondre extract for tissue engineering applications, particularly in bone tissues.

Evaluation of the Sensing Potential of Stem Cell-Secreted Proteins via a Microchip Device under Electromagnetic Field Stimulation.

Most bone tissue engineering models fail to demonstrate the complex cellular functions of living bone; therefore, most translational studies on bone tissue are performed in live models. To reduce the need for live models, we developed a stimulated microchip model for monitoring protein secretion during osteogenesis using human mesenchymal stem cells (hMSCs). We established a bone microchip system for monitoring the in vitro differentiation and sensing the secreted proteins of hMSCs under a sinusoidal electromagnetic field (SEMF), which ameliorates bone healing in a biomimetic natural bone matrix. A 3 V-1 Hz SEMF biophysically stimulated osteogenesis by activating ERK-1/2 and promoting phosphorylation of p38 MAPK kinases. Exposure to a 3 V-1 Hz SEMF upregulated the expression of osteogenesis-related genes and enhanced the expression of key osteoregulatory proteins. We identified 23 proteins that were differentially expressed in stimulated human bone marrow mesenchymal stem cell secretomes or were absent in the control groups. Our on-chip stimulation technology is easy to use, versatile, and nondisruptive and should have diverse applications in regenerative medicine and cell-based therapies.

Multifunctional bioactive chitosan/cellulose nanocrystal scaffolds eradicate bacterial growth and sustain drug delivery.

Chitosan-based hydrogels have received significant interest in tissue engineering and regenerative medicine applications owing to their superior biocompatibility. However, their applications are restricted owing to their weak mechanical strength. Cellulose nanocrystals (CNCs) are often explored as reinforcing agents to improve the native properties of polymers owing to their superior physicochemical properties. We fabricated a multi-functional hydrogel scaffold of chitosan/CNCs by incorporating different amounts of CNCs into a chitosan (CH) hydrogel. Significant enhancement in the mechanical strength was noted in the CH/CNCs as compared to that in pure CH hydrogel scaffolds. The cytocompatibility of the fabricated scaffolds was monitored in the presence of bone-marrow-derived mesenchymal stem cells (BMSCs). Improved cell viability and mineralization were observed with CH/CNC hydrogel scaffolds than those with pure CH hydrogel scaffolds. Enhanced osteogenic-related gene expression was observed in the CH/CNC hydrogel scaffold environment than that in the control, indicating their osteogenic potential, in addition to enhanced antibacterial activity. Developed composite scaffolds exhibited improved sustained drug release compared to that by pure polymer scaffolds, and this was more sustained in the scaffolds with higher CNC content. Therefore, the fabricated scaffolds may have been used in tissue engineering for osteogenesis, as antibacterial agents, and in sustained drug delivery.

Protaetia brevitarsis seulensis Derived Protein Isolate with Enhanced Osteomodulatory and Antioxidative Property.

The osteogenic differentiation of stem cells is profoundly affected by their microenvironmental conditions. The differentiation behavior of stem cells can be tuned by changing the niche environments. The proteins or peptides that are derived by living organisms facilitate the osteogenic differentiation of stem cells. Here, we have evaluated the osteoinductive and antioxidative potential of the Protaetia brevitarsis seulensis insect-derived protein for human bone marrow-derived mesenchymal stem cells (hBMSCs). The amino acid contents in the isolated protein were determined by an amino acid analyzer. Fourier transform infrared (FTIR) spectroscopy and scanning electron microscopy (SEM) were used to analyze the extract's functional groups and surface morphology. The extracted protein exhibited 51.08% ß-sheet conformation. No adverse effects were observed in extract-treated cells, indicating their biocompatibility. The protein isolate showed an excellent antioxidative property. Besides this, an enhancement in the hBMSCs' mineralization has been observed in the presence of treated protein isolates. Notably, osteogenic marker genes and proteins were effectively expressed in the treated cells. These results indicated that the P. brevitarsis-derived protein isolate can be used as a potential antioxidative biomaterial for bone tissue engineering applications.

Bioactive electrospun nanocomposite scaffolds of poly(lactic acid)/cellulose nanocrystals for bone tissue engineering.

Poly(lactic acid) (PLA)/cellulose nanocrystal (CNC) composite scaffolds were fabricated using an electrospinning technique to evaluate the influence of CNCs on the biocompatibility and osteogenic potential of PLA. The scaffolds were characterized using Fourier transform infrared spectroscopy (FTIR), X-ray diffraction pattern (XRD), transmission electron microscopy (TEM), and atomic force microscopy (AFM). A significant enhancement of the mechanical properties occurred in the composite scaffolds compared to pure polymer. This is due to the stronger interactions between the polymer chains and CNCs. The composite scaffolds exhibited higher thermal stability compared to pure polymer. Notably, excellent adhesion and proliferation was observed in the presence of the fabricated composite scaffolds, indicating their superior biocompatibility. Higher mineralization was noted on the surface of composite scaffolds. The fabricated scaffolds were significantly covered by the cultured cells and exhibited greater fluorescence intensity vis-à-vis control. Additionally, the fact that higher expression of osteogenic gene markers was observed in composite scaffolds confirms their enhanced osteogenic potential. The bone regeneration potential of the fabricated scaffold was monitored in a rat calvarial defect model after 3 weeks of treatment. The fabricated scaffold demonstrated excellent biocompatibility and superior osteoinductivity. Therefore, the fabricated scaffolds possess potential to be used as a biomaterial for tissue engineering applications.

Influence of Maitake (Grifola frondosa) Particle Sizes on Human Mesenchymal Stem Cells and In Vivo Evaluation of Their Therapeutic Potential.

Maitake (Grifola frondosa) mushroom has received an enormous amount of attention as a dietary supplement due to its high nutritional values. The particle sizes of G. frondosa mushrooms were monitored by a classifying mill. ß-Glucans are the bioactive component of the mushroom, and it was revealed through Fourier transform infrared spectroscopy (FTIR), proton and carbon nuclear magnetic resonance (1H and 13C-NMR), matrix-assisted laser desorption/ionization, and time-of-flight (MALDI-TOF) spectrometry. The biocompatibility of G. frondosa particles, as well as induced osteogenesis of hMSCs, was evaluated through WST-1 assay and alizarin staining (ARS) technique, respectively. Notably, enhanced cell viability was noted in the presence of G. frondosa. Significantly improved calcium deposition has observed from hMSCs with G. frondosa, suggesting to their mineralization potential. The expression of osteogenic related gene markers was examined in the presence of G. frondosa through real-time polymerase chain reaction (qPCR) technique. The upregulation of osteogenic gene markers in the presence of G. frondosa particles was indicating their superior osteogenic potential. Besides, G. frondosa also activated the secretion of various kinds of proteins from the hMSCs indicating their potential for tissue engineering applications. Enhanced secretion of different immunoglobulins was observed in rat serum in the presence of G. frondosa, further demonstrating their therapeutic nature. Therefore, G. frondosa is effective for enhanced osteogenesis and can be utilized as a natural, edible, and osteogenic agent.

Evaluation of the Osteogenic Potential of Stem Cells in the Presence of Growth Hormone under Magnetic Field Stimulation.

The human alveolar bone-derived mesenchymal stem cells (hABMSCs) are considered an attractive source for the development of bone tissues. However, their mechanism of action is still unclear. This work aimed to investigate the potential of the natural human growth hormone (NHGH) derived from stem cells under magnetic field (MF) stimulation for tissue engineering by exploring the paracrine or autocrine effects of hABMSCs in vitro. The secretion of anti-inflammatory cytokines and growth factors from hABMSCs was profoundly affected by the intensity of the applied MFs. The effects of stimulated MFs on vascular endothelial growth factor (VEGF) and bone morphogenetic protein-2 (BMP-2) production were quantified by an ELISA kit. Notably, higher cell metabolic activity was observed in MF stimulation compared to the control, and this was more prominent in 130 mT strength of MF. An enhancement in the production of VEGF and BMP-2 was noted in MFs compared to the control. Moreover, higher accumulation of osteogenesis-related genes has occurred in MFs than the control. Furthermore, a significant enhancement in cell metabolic activity and mineralized nodule formations was spotted in the presence of NHGH via MF stimulation; vis-à-vis, MF stimulation only through autocrine and paracrine effects demonstrated the better osteogenic potential of NHGH in the presence of MFs for tissue engineering applications.

Osteogenic potential of human mesenchymal stem cells on eggshells-derived hydroxyapatite nanoparticles for tissue engineering.

Nanocrystalline hydroxyapatite (HAp) was synthesized from biowaste eggshells through sonication followed by the heat treatment. Calcium oxide as a precursor moiety for the synthesis of HAp was obtained through the heat treatment of eggshells at 900°C for 3 hr. The prepared HAp was characterized by Fourier-transform infrared spectroscopy (FTIR), X-ray powder diffraction (XRD), and scanning electron microscopy (SEM). The appearance of the FTIR absorption peaks in between at 516-1031 and 3,636 cm-1 shows phosphate and hydroxyl groups in prepared HAp, respectively. The XRD-patterns indicate the formation of HAp started within 5 min of sonication. The SEM morphologies suggested that the synthesized HAp was highly crystalline and compact. We tested the elemental analysis of the synthesized HAp through X-ray fluorescence spectroscopy and inductively coupled plasma mass spectroscopy. The higher Ca/P ratio has observed in heat-treated HAp. These results show that heat treatment facilitates the formation of highly crystalline and compact HAp. Cytotoxicity and osteogenic potential of human mesenchymal stem cells (hMSCs) were also evaluated in the presence of HAp. No significant cytotoxicity was noted in the presence of HAp, suggested their biocompatibility. Enhanced osteogenesis of hMSCs occurred with HAp powder, confirming the feasibility in the treatment of osteogenesis. Thus, synthesized HAp has the potential to use a biomaterial in tissue engineering applications for bone tissues.

Enhanced Osteogenesis of Human Mesenchymal Stem Cells in Presence of Single-Walled Carbon Nanotubes.

Human mesenchymal stem cells (hMSCs) have attracted significant attention for tissue engineering because of their ability to differentiate into bone cells, chondrocytes, adipocytes, and muscle cells. Single-walled carbon nanotubes (SWCNTs) have been considered as a potential material for tissue engineering applications due to their unique properties, such as high aspect ratio, excellent electrocatalytic activity, and biocompatibility. Here we prepared exfoliated SWCNTs layers through an ultra-sonication process in the acidic medium and evaluated their cytotoxicity using hMSCs. Improved viability and osteogenesis of hMSCs were observed in the presence of exfoliated SWCNTs. Besides, the higher expression of osteogenic differentiation-related genes in the presence of exfoliated SWCNTs further confirmed their enhanced osteogenic nature. These results indicated the potential of SWCNTs as a biomaterial for tissue engineering applications.

Structural Elucidation and Immune-Enhancing Effects of Novel Polysaccharide from Grifola frondosa.

Beta-glucan (ß-glucan) is a macromolecule structure where glucose unit has bonded through ß-glycosidic bond at 1 and 3 positions. It is well known as a natural immunomodulator without exhibiting any side effects via enhancing immunity. Mushroom contains a large amount of ß-glucan and it has anticancerous and antioxidant efficacy. Structure and physical properties of ß-glucan are highly influenced by the types of mushroom. In particular, Grifola frondosa has ß-1, 3 and ß-1, 6 bonds in their structure. It has been noted that ß-glucan content also depends upon the size of mushroom particles. The exact content of ß-glucan and their immunological activity by a particle size of G. frondosa have yet to be fully elucidated. Herein, ß-glucan contents were analyzed according to the particle size of leaf mushroom followed by cell activation and immunoactivity analysis. The highest ß-glucan content was observed at a particle size of 20-30 µm (27.65 ± 0.30 w/w). All samples showed ~ 103% cell activation compared to the control and greater cell activity was observed at higher concentration. The significant increase in cytokines secretion was observed in the presence of 20-30 µm particle size of G. frondosa compared to the control. This study suggested that 20-30 µm size is the suitable size of G. frondosa that can be used as a health supplement and food additive to act as an immune booster, hypotensive agent, and hypoglycemic agent.