Graphene-Based Aerogels for Biomedical Application.

Aerogels are three-dimensional solid networks with incredibly low densities, high porosity, and large specific surface areas. These aerogels have both nanoscale and macroscopic interior structures. Combined with graphene, the aerogels show improved mechanical strength, electrical conductivity, surface area, and adsorption capacity, making them ideal for various biomedical applications. The graphene aerogel has a high drug-loading capacity due to its large surface area, and the porous structure enables controlled drug release over time. The presence of graphene makes it a suitable material for wound dressings, blood coagulation, and bilirubin adsorption. Additionally, graphene's conductivity can help in the electrical stimulation of cells for improved tissue regeneration, and it is also appropriate for biosensors. In this review, we discuss the preparation and advantages of graphene-based aerogels in wound dressings, drug delivery systems, bone regeneration, and biosensors.

Conducting polymer-based scaffolds for neuronal tissue engineering.

Neuronal tissue engineering has immense potential for treating neurological disorders and facilitating nerve regeneration. Conducting polymers (CPs) have emerged as a promising class of materials owing to their unique electrical conductivity and biocompatibility. CPs, such as poly(3,4-ethylenedioxythiophene) (PEDOT), poly(3-hexylthiophene) (P3HT), polypyrrole (PPy), and polyaniline (PANi), have been extensively explored for their ability to provide electrical cues to neural cells. These polymers are widely used in various forms, including porous scaffolds, hydrogels, and nanofibers, and offer an ideal platform for promoting cell adhesion, differentiation, and axonal outgrowth. CP-based scaffolds can also serve as drug delivery systems, enabling localized and controlled release of neurotrophic factors and therapeutic agents to enhance neural regeneration and repair. CP-based scaffolds have demonstrated improved neural regeneration, both in vitro and in vivo, for treating spinal cord and peripheral nerve injuries. In this review, we discuss synthesis and scaffold processing methods for CPs and their applications in neuronal tissue regeneration. We focused on a detailed literature review of the central and peripheral nervous systems.

Production and Bioconversion Efficiency of Enzyme Membrane Bioreactors in the Synthesis of Valuable Products.

The demand for bioactive molecules with nutritional benefits and pharmaceutically important properties is increasing, leading researchers to develop modified production strategies with low-cost purification processes. Recent developments in bioreactor technology can aid in the production of valuable products. Enzyme membrane bioreactors (EMRs) are emerging as sustainable synthesis processes in various agro-food industries, biofuel applications, and waste management processes. EMRs are modified reactors used for chemical reactions and product separation, particularly large-molecule hydrolysis and the conversion of macromolecules. EMRs generally produce low-molecular-weight carbohydrates, such as oligosaccharides, fructooligosaccharides, and gentiooligosaccharides. In this review, we provide a comprehensive overview of the use of EMRs for the production of valuable products, such as oligosaccharides and oligodextrans, and we discuss their application in the bioconversion of inulin, lignin, and sugars. Furthermore, we critically summarize the application and limitations of EMRs. This review provides important insights that can aid in the production of valuable products by food and pharmaceutical industries, and it is intended to assist scientists in developing improved quality and environmentally friendly prebiotics using EMRs.

Tubular nanomaterials for bone tissue engineering.

Nanomaterial composition, morphology, and mechanical performance are critical parameters for tissue engineering. Within this rapidly expanding space, tubular nanomaterials (TNs), including carbon nanotubes (CNTs), titanium oxide nanotubes (TNTs), halloysite nanotubes (HNTs), silica nanotubes (SiNTs), and hydroxyapatite nanotubes (HANTs) have shown significant potential across a broad range of applications due to their high surface area, versatile surface chemistry, well-defined mechanical properties, excellent biocompatibility, and monodispersity. These include drug delivery vectors, imaging contrast agents, and scaffolds for bone tissue engineering. This review is centered on the recent developments in TN-based biomaterials for structural tissue engineering, with a strong focus on bone tissue regeneration. It includes a detailed literature review on TN-based orthopedic coatings for metallic implants and composite scaffolds to enhance in vivo bone regeneration.

Chitosan Based Biodegradable Composite for Antibacterial Food Packaging Application.

A recent focus on the development of biobased polymer packaging films has come about in response to the environmental hazards caused by petroleum-based, nonbiodegradable packaging materials. Among biopolymers, chitosan is one of the most popular due to its biocompatibility, biodegradability, antibacterial properties, and ease of use. Due to its ability to inhibit gram-negative and gram-positive bacteria, yeast, and foodborne filamentous fungi, chitosan is a suitable biopolymer for developing food packaging. However, more than the chitosan is required for active packaging. In this review, we summarize chitosan composites which show active packaging and improves food storage condition and extends its shelf life. Active compounds such as essential oils and phenolic compounds with chitosan are reviewed. Moreover, composites with polysaccharides and various nanoparticles are also summarized. This review provides valuable information for selecting a composite that enhances shelf life and other functional qualities when embedding chitosan. Furthermore, this report will provide directions for the development of novel biodegradable food packaging materials.

Water-stable, biocompatible, and highly luminescent perovskite nanocrystals-embedded fiber-based paper for anti-counterfeiting applications.

In this study, we present a promising and facile approach toward the fabrication of non-toxic, water-stable, and eco-friendly luminescent fiber paper composed of polycaprolactone (PCL) polymer and CsPbBr3@SiO2 core-shell perovskite nanocrystals. PCL-perovskite fiber paper was fabricated using a conventional electrospinning process. Transmission electron microscopy (TEM) clearly revealed incorporation of CsPbBr3@SiO2 nanocrystals in the fibers, while scanning electron microscopy (SEM) demonstrated that incorporation of CsPbBr3@SiO2 nanocrystals did not affect the surface and diameter of the PCL-perovskite fibers. In addition, thermogravimetric analysis (TGA) and contact angle measurements have demonstrated that the PCL-perovskite fibers exhibit excellent thermal and water stability. The fabricated PCL-perovskite fiber paper exhibited a bright green emission centered at 520 nm upon excitation by ultra-violet (UV) light (374 nm). We have demonstrated that fluorescent PCL-perovskite fiber paper is a promising candidate for anti-counterfeiting applications because various patterns can be printed on the paper, which only become visible after exposure to UV light at 365 nm. Cell proliferation tests revealed that the PCL-perovskite fibers are cytocompatibility. Consequently, they may be suitable for biocompatible anti-counterfeiting. The present study reveals that PCL-perovskite fibers may pave way toward next generation biomedical probe and anti-counterfeiting applications.

Electrospinning Nanofibers as a Dressing to Treat Diabetic Wounds.

Globally, diabetic mellitus (DM) is a common metabolic disease that effectively inhibits insulin production, destroys pancreatic ß cells, and consequently, promotes hyperglycemia. This disease causes complications, including slowed wound healing, risk of infection in wound areas, and development of chronic wounds all of which are significant sources of mortality. With an increasing number of people diagnosed with DM, the current method of wound healing does not meet the needs of patients with diabetes. The lack of antibacterial ability and the inability to sustainably deliver necessary factors to wound areas limit its use. To overcome this, a new method of creating wound dressings for diabetic patients was developed using an electrospinning methodology. The nanofiber membrane mimics the extracellular matrix with its unique structure and functionality, owing to which it can store and deliver active substances that greatly aid in diabetic wound healing. In this review, we discuss several polymers used to create nanofiber membranes and their effectiveness in the treatment of diabetic wounds.

Rediscovery of poly(ethylene glycol)s as a cryoprotectant for mesenchymal stem cells.

BACKGROUND: A medium containing dimethyl sulfoxide (DMSO) (10% v/v) is most widely used for cell cryopreservation at -196 °C. However, residual DMSO consistently raises concerns because of its toxicity; thus, its complete removal process is required. METHOD: As biocompatible polymers approved by the Food and Drug Administration for various biomedical applications for humans, poly(ethylene glycol)s (PEGs) with various molecular weights (400, 600, 1 K, 1.5 K, 5 K, 10 K, and 20 K Da) were studied as a cryoprotectant of mesenchymal stem cells (MSCs). Considering the cell permeability difference of PEGs depending on their molecular weight, the cells were preincubated for 0 h (no incubation), 2 h, and 4 h at 37 °C in the presence of PEGs at 10 wt.% before cryopreservation at -196 °C for 7 days. Then, cell recovery was assayed. RESULTS: We found that low molecular weight PEGs (400 and 600 Da) exhibit excellent cryoprotecting properties by 2 h preincubation, whereas intermediate molecular weight PEGs (1 K, 1.5 K, and 5 K Da) exhibit their cryoprotecting properties without preincubation. High molecular weight PEGs (10 K and 20 K Da) were ineffective as cryoprotectants for MSCs. Studies on ice recrystallization inhibition (IRI), ice nucleation inhibition (INI), membrane stabilization, and intracellular transport of PEGs suggest that low molecular weight PEGs (400 and 600 Da) exhibit excellent intracellular transport properties, and thus the internalized PEGs during preincubation contribute to the cryoprotection. Intermediate molecular weight PEGs (1 K, 1.5 K, and 5 K Da) worked by extracellular PEGs through IRI, INI, as well as partly internalized PEGs. High molecular weight PEGs (10 K and 20 K Da) killed the cells during preincubation and were ineffective as cryoprotectants. CONCLUSIONS: PEGs can be used as cryoprotectants. However, the detailed procedures, including preincubation, should consider the effect of the molecular weight of PEGs. The recovered cells well proliferated and underwent osteo/chondro/adipogenic differentiation similar to the MSCs recovered from the traditional DMSO 10% system.

Recent Development of Brain Organoids for Biomedical Application.

Over the years, scientists have studied the behavior and anatomy of many animals to understand the own species. However, despite the continuous efforts, it is often difficult to know for certain how the brain works due to the differences between the brains of animals and the human brain. While the use of animal models for research continues, the origin of human cognition and neurological disorders needs further elucidation. To that end, in vitro organoids that exhibit in vivo characteristics of the human brain have been recently developed. These brain-like organoids enable researchers to dive deeper into understanding the human brain, its neurological structures, and the causes of neurological pathologies. This paper reviews the recent developments in the regeneration of brain-like organoids using Matrigel and other alternatives. Further, gel-free methods that may enhance the regeneration process of organoids are discussed. Finally, the vascularized brain organoid growth and development in both in vitro and in vivo conditions are detailed.

Antibacterial activity of essential oils from different parts of plants against Salmonella and Listeria spp.

Essential oils (EOs) have recently gained popularity as natural food preservatives due to their potent antibacterial activity against food pathogens. In this review, the antibacterial activity of EOs from various plant parts and sources against the most important food pathogens Salmonella and Listeria have been discussed. The antibacterial activity of EOs is attributed to their major and minor low-molecular weight terpenes, terpenoids, phenylpropenes and aliphatic components. The major compounds along with minor components of EO extracted from different parts of various plant species were found to be responsible for antibacterial activity. The combination of EO from different sources presented synergistic anti-listerial and anti-salmonella effects. EO combined with biopolymer and in nanoemulsion form presented significant antibacterial activity. The mode of antibacterial action by EO was complex and involves a series of event that has also been discussed in detail.

Poly(l-alanine-co-l-lysine)-g-Trehalose as a Biomimetic Cryoprotectant for Stem Cells.

Poly(l-alanine-co-l-lysine)-graft-trehalose (PAKT) was synthesized as a natural antifreezing glycopolypeptide (AFGP)-mimicking cryoprotectant for cryopreservation of mesenchymal stem cells (MSCs). FTIR and circular dichroism spectra indicated that the content of the α-helical structure of PAK decreased after conjugation with trehalose. Two protocols were investigated in cryopreservation of MSCs to prove the significance of the intracellularly delivered PAKT. In protocol I, MSCs were cryopreserved at -196 °C for 7 days by a slow-cooling procedure in the presence of both PAKT and free trehalose. In protocol II, MSCs were preincubated at 37 °C in a PAKT solution, followed by cryopreservation at -196 °C in the presence of free trehalose for 7 days by the slow-cooling procedure. Polymer and trehalose concentrations were varied by 0.0-1.0 and 0.0-15.0 wt %, respectively. Cell recovery was significantly improved by protocol II with preincubation of the cells in the PAKT solution. The recovered cells from protocol II exhibited excellent proliferation and maintained multilineage potentials into osteogenic, chondrogenic, and adipogenic differentiation, similar to MSCs recovered from cryopreservation in the traditional 10% dimethyl sulfoxide system. Ice recrystallization inhibition (IRI) activity of the polymers/trehalose contributed to cell recovery; however, intracellularly delivered PEG-PAKT was the major contributor to the enhanced cell recovery in protocol II. Inhibitor studies suggested that macropinocytosis and caveolin-dependent endocytosis are the main mechanisms for the intracellular delivery of PEG-PAKT. 1H NMR and FTIR spectra suggested that the intracellular PEG-PAKTs interact with water and stabilize the cells during cryopreservation.

Folic acid pretreatment and its sustained delivery for chondrogenic differentiation of MSCs.

Dietary uptake of folic acid (FA) improves cartilage regeneration. In this work, we discovered that three days of FA treatment is highly effective for promoting chondrogenic differentiation of tonsil-derived mesenchymal stem cells (TMSCs). In a three-dimensional pellet culture, the levels of typical chondrogenic biomarkers, sulfated glycosaminoglycan, proteoglycan, type II collagen (COL II), SRY box transcription factor 9 (SOX 9), cartilage oligomeric matrix protein (COMP), and aggrecan (ACAN) increased significantly in proportion to FA concentration up to 30 µM. At the mRNA expression level, COL II, SOX 9, COMP, and ACAN increased 3.6-6.0-fold with FA treatment at 30 µM compared with the control system that did not receive FA treatment, and the levels with FA treatment were 1.6-2.5 times greater than those in the kartogenin-treated positive control system. FA treatment did not increase type I collagen α1 (COL I α1), an osteogenic biomarker which is a concern with most chondrogenic promoters. At the high FA concentration of 100 µM, significant decreases in chondrogenic biomarkers were observed, which might be related to DNA methylation. A thermogel system incorporating TMSCs and FA provided sustained release of FA over several days, similar to the FA treatment. The thermogel system confirmed the efficacy of FA in promoting chondrogenic promotion of TMSCs. The increased nuclear translocation of core-binding factor ß subunit (CBFß) and the runt-related transcription factor 1 (RUNX1) expression after FA treatment, together with molecular docking studies, suggest that the chondrogenic enhancement mechanism of FA is mediated by CBFß and RUNX1.

Pralatrexate Sustainably Released from Polypeptide Thermogel Is Effective for Chondrogenic Differentiation of Mesenchymal Stem Cells.

Folic acid was reported to significantly improve chondrogenic differentiation of mesenchymal stem cells. In a similar mechanism of action, we investigated clinically approved antifolates by the U.S. Food and Drug Administration as chondrogenic-promoting compounds for tonsil-derived mesenchymal stem cells. A poly(ethylene glycol)-poly(l-alanine) thermogelling system was used as a three-dimensional cell culture matrix, where stem cells and antifolates could be incorporated simultaneously during a heat-induced in situ sol-to-gel transition. The antifolates could be supplied over several days by the sustained release of the drug from the thermogel. Initially, seven antifolates were prescreened based on cell viability and expression of a typical chondrogenic biomarker of type II collagen (COL II) at the mRNA level. Then, dapsone, pralatrexate, and trimethoprim were selected as candidate compounds in the second round screening, and detailed studies were carried out on the mRNA and protein expression of various chondrogenic biomarkers including COL II, SRY box transcription factor 9, and aggrecan. Three-dimensional cultures of stem cells in the thermogel in the absence of a chondrogenic promoter compound and in the presence of kartogenin (KGN) were performed as a negative control and positive control, respectively. The chondrogenic biomarkers were significantly increased in the selected antifolate-incorporating systems compared to the negative control system, without an increase in type I collagen (an osteogenic biomarker) expression. Pralatrexate was the best compound for inducing chondrogenic differentiation of the stem cells, even better than the positive control (KGN). Nuclear translocation of the core-binding factor ß subunit (CBFß) and enhanced nuclear runt-related transcription factor 1 (RUNX1) by antifolate treatment suggested that the chondrogenesis-enhancing mechanism is mediated by CBFß and RUNX1. An in silico modeling study confirmed the mechanism by proving the high binding affinity of pralatrexate to a target protein of filamin A compared with other antifolate candidates. To conclude, pralatrexate was rediscovered as a lead compound, and the polypeptide thermogel incorporating pralatrexate and mesenchymal stem cells can be a very effective system in promoting chondrogenic differentiation of stem cells and might be used in injectable tissue engineering for cartilage repair.

PLGA Core-Shell Nano/Microparticle Delivery System for Biomedical Application.

Core-shell particles are very well known for their unique features. Their distinctive inner core and outer shell structure allowed promising biomedical applications at both nanometer and micrometer scales. The primary role of core-shell particles is to deliver the loaded drugs as they are capable of sequence-controlled release and provide protection of drugs. Among other biomedical polymers, poly (lactic-co-glycolic acid) (PLGA), a food and drug administration (FDA)-approved polymer, has been recognized for the vehicle material. This review introduces PLGA core-shell nano/microparticles and summarizes various drug-delivery systems based on these particles for cancer therapy and tissue regeneration. Tissue regeneration mainly includes bone, cartilage, and periodontal regeneration.

Size and Shape Control of Ice Crystals by Amphiphilic Block Copolymers and Their Implication in the Cryoprotection of Mesenchymal Stem Cells.

Precise control over the size and shape of ice crystals is a key factor to consider in designing antifreezing and cryoprotecting molecules for cryopreservation of cells. Here, we report that a poly(ethylene glycol)-poly(l-alanine) (PEG-PA) block copolymer exhibits excellent cryoprotecting properties for stem cells and antifreezing properties for water. As the molecular weight of PA increased from 500, 760, and 1750 Da (P1, P2, and P3) at the same PEG molecular weight of 5000 Da, the ß-sheet content decreased and α-helix content increased. Comparing P2 (PEG-PA; 5000-760) and P4 (PEG-PA: 1000-750), ß-sheets increased as the PEG block length decreased. The critical micelle concentration of the PEG-PA block copolymers was in a range of 0.5-3.0 mg/mL and was proportional to the hydrophobicity of the PEG-PA block copolymers. The P1, P2, and P3 self-assembled into spherical micelles, whereas P4 formed micelles with cylindrical morphology. The difference in the block copolymer structure affected ice recrystallization inhibition (IRI) activity and cryopreservation of cells. IRI activity was assayed via mean largest grain size (MLGS), and interactions between polymers and ice crystal surfaces were studied by dynamic ice-shaping studies. The MLGS decreased to 58 → 53 → 45 → 35 → 23% of that of PBS, as the polymer (PEG-PA 5000-500) concentration increased from 0.0 (PBS; control) → 1.0 → 5.0 → 10 → 30 → 50 mg/mL. The MLGS of PEG 5k solutions (negative control) decreased to 74 → 71 → 64 → 44 → 37% of that of PBS in the same concentration range. P3 and P4 with a longer hydrophobic PA block developed elongated ice crystals at above 30 mg/mL. The dynamic ice-shaping study exhibited that ice crystals became needle-shaped, as the hydrophobicity of the polymer increased as in P2-P4. The cell recovery in the P1 system after cryopreservation at -196 °C for 7 days was 87% of that of the dimethyl sulfoxide (DMSO) 10% system (positive control). The cell recovery was 48% for the P2 system and drastically decreased to less than 30% of that of the DMSO 10% system in the P3, P4, PEG 5k, PEG 1k, PVA 80H, and PVA 100H systems. Current studies suggest that IRI activity, round ice crystal shaping, and membrane stabilization activity of P1 cooperatively provide excellent cell recovery among the candidate systems. Recovered stem cells exhibited excellent proliferation and multilineage differentiation into osteocytes, chondrocytes, and adipocytes. To conclude, the PEG-PA (5000-500) block copolymer is suggested to be a promising antifreezing cryoprotectant for stem cells.

Poly(l-Ala-co-l-Lys) Exhibits Excellent Ice Recrystallization Inhibition Activity.

The control of ice recrystallization is very important in cryo-technological fields such as the food industry, biopharmaceuticals, and cell storage. Ice recrystallization inhibition (IRI) compounds are therefore designed to limit the growth of ice crystals, decrease the crystal size, and control the crystal shape. To improve the IRI activity of cryo-systems, various synthetic polymers such as biomimetic polypeptides from polar fish, facially amphiphilic polymers, polyampholytes, poly(vinyl alcohol) derivatives, and block copolymers with hydrophilic-hydrophobic balance have been developed. Except for graphene oxide, poly(vinyl alcohol) has thus far exhibited the best performance among these polymers. Herein, poly(l-alanine-co-l-lysine) (PAK) was shown to exhibit a similar IRI activity to that of poly(vinyl alcohol). Moreover, in contrast to the needle-shaped ice crystals generated by the aqueous PVA solution, the PAK solution was shown to generate cubic-to-spherical shaped ice crystals. Furthermore, neither poly(l-alanine-co-l-aspartic acid) (PAD) nor poly(ethylene glycol) (PEG) with a similar molecular weight provided any significant IRI activity. Examination by FTIR and circular dichroism spectroscopies indicated that the PAK forms α-helices, whereas the PAD forms random coils in water. Further, a dynamic ice shaping study suggested that PAK strongly interacts with ice crystals, whereas PAD and PEG only weakly interact. These results suggest that PAK is an important compound with superior IRI activity and that this activity is dependent upon the functional groups and secondary structure of the polypeptides.

Highly luminescent biocompatible CsPbBr3@SiO2 core-shell nanoprobes for bioimaging and drug delivery.

The encapsulation of lead halide perovskite nanocrystals (PNCs) with an inert protective layer against moisture and the environment is a promising approach to overcome hinderances for their practical use in optoelectronic and biomedical applications. Herein, a facile method for synthesizing highly luminescent and biocompatible CsPbBr3@SiO2 core-shell PNCs with a controlled SiO2 thickness, which are suitable for both cell imaging and drug delivery, is reported. The synthesized CsPbBr3@SiO2 core-shell PNCs exhibit bright green emission at 518 nm upon excitation of 374 nm. Interestingly, a significant increase in the photoluminescence intensity is observed with an increase in the SiO2 shell thickness, which varies with the increasing reaction time. Cytotoxicity results indicate that the CsPbBr3@SiO2 core-shell PNCs are nontoxic, making them suitable for in vitro cell imaging using HeLa cells. Furthermore, doxorubicin physically adsorbed on the surface of CsPbBr3@SiO2 core-shell PNCs is efficiently released in cells when the drug-loaded perovskite nanoprobes are injected in the cells, indicating that these core-shell nanoparticles can be used for drug loading and delivery. The results of this study suggest that the CsPbBr3@SiO2 core-shell PNCs can pave the way for new biomedical applications and processes.

Composite Hydrogel of Methacrylated Hyaluronic Acid and Fragmented Polycaprolactone Nanofiber for Osteogenic Differentiation of Adipose-Derived Stem Cells.

Composite hydrogels with electrospun nanofibers (NFs) have recently been used to mimic the native extracellular matrix. In this study, composite hydrogels of methacrylated hyaluronic acid containing fragmented polycaprolactone NFs were used for bone tissue engineering. The composite (NF/hydrogel) was crosslinked under ultraviolet (UV) light. The incorporation of fragmented polycaprolactone NFs increased the compression modulus from 1762.5 to 3122.5 Pa. Subsequently, adipose-derived stem cells incorporated into the composite hydrogel exhibited a more stretched and elongated morphology and osteogenic differentiation in the absence of external factors. The mRNA expressions of osteogenic biomarkers, including collagen 1 (Col1), alkaline phosphatase, and runt-related transcription factor 2, were 3-5-fold higher in the composite hydrogel than in the hydrogel alone. In addition, results of the protein expression of Col1 and alizarin red staining confirmed osteogenic differentiation. These findings suggest that our composite hydrogel provides a suitable microenvironment for bone tissue engineering.

Thermogelling Inclusion Complex System for Fine-Tuned Osteochondral Differentiation of Mesenchymal Stem Cells.

How to control osteochondral differentiation of mesenchymal stem cells at a proper stage is a key issue for articular cartilage regeneration. To solve this problem, injectable scaffolds with different chemical functional groups were designed by introducing one equivalent of α-cyclodextrin (α-CD) carboxylate and α-CD phosphate along poly(ethylene glycol)-poly(l-alanine) (PEG-L-PA) block copolymers. Dynamic light scattering, transmission electron microscopy images, and two-dimensional NMR spectra indicated that the PEG-L-PA block copolymers formed inclusion complexes with α-CD derivatives. Aqueous solutions of PEG-L-PA block copolymers (P), α-CD carboxylate/PEG-L-PA block copolymers (PCC), and α-CD phosphate/PEG-L-PA block copolymers (PCP) underwent sol-to-gel transition as the temperature increased. The storage moduli of P, PCC, and PCP gels ranged from 1000 to 1300 Pa at 37 °C. Tonsil-derived mesenchymal stem cells (TMSCs) were incorporated in situ in the gel during thermogelation of P, PCC, and PCP, which became the three-dimensional cell culture systems with different functional groups. After 21 days of incubation of TMSCs in the P, PCC, and PCP systems, the chondrogenic differentiation biomarker of type II collagen significantly increased in the P system, whereas the osteogenic biomarkers of osteocalcin and runt-related transcription factor 2 significantly increased in the PCP system. Both chondrogenic and osteogenic biomarkers were highly expressed in the PCC system. This study proved that thermogelling inclusion complex systems consisting of PEG-L-PA block copolymers and α-CD derivatives could be an excellent injectable matrix for fine-controlling osteochondral differentiation of mesenchymal stem cells.

Culture of neural stem cells on conductive and microgrooved polymeric scaffolds fabricated via electrospun fiber-template lithography.

We developed polymeric scaffolds that can provide both topographical and electrical stimuli on mouse neural stem cells (mNSCs) for potential use in nerve tissue engineering. In contrast to conventional patterning techniques such as imprinting, soft/photolithography, and three-dimensional printing, microgroove patterns were generated by using aligned electrospun fibers as templates, via a process denoted as electrospun fiber-template lithography. The preparation of polyvinylpyrrolidone fibers, followed by the deposition of poly(lactic-co-glycolic acid) (PLGA) and the removal of the fiber template, produced freestanding PLGA scaffolds with microgrooves having widths of 1.72 ± 0.24 µm. The subsequent deposition of polypyrrole (PPy) via chemical oxidative polymerization added conductivity to the microgrooved PLGA scaffolds. The resultant scaffolds were cytocompatible with mNSCs. The microgroove patterns enhanced the alignment and elongation of mNSCs, and the PPy layer promoted the interaction of cells with the surface by forming more and longer filopodia compared with the nonconductive surface. Finally, the neuron differentiation of mNSCs was evaluated by monitoring the Tuj-1 neuronal gene expression. The presence of both microgrooves and the conductive PPy layer enhanced the neuronal differentiation of mNSCs even without electrical stimulation, and the neuronal differentiation was further enhanced by stimulation with a sufficient electrical pulse (1.0 V).