RESUMO
We revealed that the encapsulation of enzyme-immobilized silica particles in hollow-type spherical bacterial cellulose (HSBC) gels enables the use of the inside of HSBC gels as a reaction field. The encapsulation of horseradish peroxidase (HRP)-immobilized silica particles (Si-HRPs, particle size: 40-50 µm) within HSBC gels was performed by using a BC gelatinous membrane produced at the interface between Komagataeibacter xylinus suspension attached onto an alginate gel containing Si-HRPs and silicone oil. After the biosynthesis of the BC gelatinous membrane, formed from cellulose nanofiber networks, the alginate gel was removed via immersion in a phosphate-buffered solution. Si-HRP encapsulated HSBC gels were reproducibly produced using our method with a yield of over 90%. The pore size of the network structure of the BC gelatinous membrane was less than 1 µm, which is significantly smaller than the encapsulated Si-HRPs. Consequently, the encapsulated Si-HRPs could neither pass through the BC gelatinous membrane nor leak from the interior cavity of the HSBC gel. The activity of the encapsulated HRPs was detected using the 3,3',5,5'-tetramethylbenzidine (TMB)-H2O2 system, demonstrating that this method can encapsulate the enzyme without inactivation. Since HSBC gels are composed of a network structure of biocompatible cellulose nanofibers, immune cells cannot enter the hollow interior, thus, the enzyme-immobilized particles encapsulated inside the HSBC gel are protected from immune-cell attacks. The encapsulation technique demonstrated in this study is expected to facilitate the delivery of enzymes and catalysts that are not originally present in the in vivo environment.
RESUMO
To reduce the risk of the adsorption of granular activated carbon in the gastrointestinal tract, we successfully produced a hollow-type spherical bacterial cellulose (HSBC) gel containing activated carbon with a particle size of 6 µm. In this study, the aim of which was to develop an effective formulation, we evaluated the stability of activated-carbon-encapsulating HSBC gels under various pH conditions. Activated-carbon-encapsulating HSBC gels (ACEGs) retained the activated carbon without leaking when subjected to agitation in acidic or basic environments. The saturated adsorption amount, calculated using the Langmuir adsorption isotherm, was affected by the target adsorbate and pH conditions. These results indicate that ACEGs can adsorb uremic toxins and their precursors similarly to conventional uremic toxin adsorbents while preventing direct contact between the encapsulated activated carbon and the gastrointestinal tract. Compared to powdered activated carbon, the ACEG is less likely to be adsorbed in the gastrointestinal tract. Therefore, the proposed ACEG is a promising new formulation that will contribute to the treatment of renal failure and improve patients' compliance with medication.
RESUMO
In this study, the introduction of a positive charge on the surface of a shape memory material was investigated to enhance cell affinity. To achieve this, the direct chemical modification of a material surface was proposed. Sheet-type, crosslinked poly(caprolactone-co-α-bromo-ɤ-butyrolactone) (poly(CL-co-BrBL)) were prepared, and the direct reaction of amino compounds with bromo groups was conducted on the material surface with a positive charge. Branched poly(CL-co-BrBL) was prepared, followed by the introduction of methacryloyl groups to each chain end. Using the branched macromonomers, stable and sheet-type materials were derived through UV-light irradiation. Then, the materials were soaked in an amino compound solution to react with the bromo groups under various conditions. Differential scanning calorimetry and surface analysis of the modified materials indicated that 10 vol% of N, N-dimethylethylenediamine in n-hexane and 1 h soaking time were optimal to maintain the inherent thermal properties. The achievement of increased luminance and a positive zeta potential proved that the direct modification method effectively introduced the positive charge only on the surface, thereby enhancing cell affinity.
RESUMO
For reducing side effects and improvement of swallowing, we studied the encapsulation of activated carbon formulations with a hollow-type spherical bacterial cellulose (HSBC) gel using two kinds of encapsulating methods: Methods A and B. In Method A, the BC gelatinous membrane was biosynthesized using Komagataeibacter xylinus (K. xylinus) at the interface between the silicone oil and cell suspension containing activated carbon. In Method B, the bacterial cellulose (BC) gelatinous membrane was formed at the interface between the cell suspension attached to the alginate gel containing activated carbon and the silicone oil. After the BC gelatinous membrane was biosynthesized by K. xylnus, alginate gel was removed by soaking in a phosphate buffer. The activated carbon encapsulated these methods could neither pass through the BC gelatinous membrane of the HSBC gel nor leak from the interior cavity of the HSBC gel. The adsorption ability was evaluated using indole, which is a precursor of the uremic causative agent. From curve-fitting, the adsorption process followed the pseudo-first-order and intra-particle diffusion models, and the diffusion of the indole molecules at the surface of the encapsulated activated carbon within the HSBC gel was dominant at the initial stage of adsorption. It was observed that the adsorption of the encapsulated activated carbon by the intraparticle diffusion process became dominant with longer adsorption times.
RESUMO
A hollow-type spherical bacterial cellulose (HSBC) gel prepared using conventional methods cannot load particles larger than the pore size of the cellulose nanofiber network of bacterial cellulose (BC) gelatinous membranes. In this study, we prepared a HSBC gel encapsulating target substances larger than the pore size of the BC gelatinous membranes using two encapsulating methods. The first method involved producing the BC gelatinous membrane on the surface of the core that was a spherical alginate gel with a diameter of 2 to 3 mm containing the target substances. With this method, the BC gelatinous membrane was biosynthesized using Gluconacetobacter xylinus at the interface between the cell suspension attached onto the alginate gel and the silicone oil. The second method involved producing the BC gel membrane on the interface between the silicone oil and cell suspension, as well as the spherical alginate gel with a diameter of about 1 mm containing target substances. After the BC gelatinous membrane was biosynthesized, an alginate gel was dissolved in a phosphate buffer to prepare an HSBC gel with the target substances. These encapsulated substances could neither pass through the BC gelatinous membrane of the HSBC gel nor leak from the interior space of the HSBC gel. These results suggest that the HSBC gel had a molecular sieving function. The HSBC gel walls prepared using these methods were observed to be uniform and would be useful for encapsulating bioactive molecules, such as immobilized enzymes in HSBC gel, which is expected to be used as a drug carrier.
Assuntos
Cápsulas/química , Celulose/análogos & derivados , Gluconacetobacter xylinus/química , Microgéis/química , Alginatos/química , Membranas Artificiais , Silicones/químicaRESUMO
Cell sheet tissue engineering is a concept for creating transplantable two-dimensional (2D) and three-dimensional (3D) tissues and organs. This review describes three elements of cell sheet tissue engineering in terms of the chemical and physical effects of material surfaces and the interfacial properties of cell sheets: preparation, harvesting/manipulation and transplantation of cell sheets. An essential technology for the preparation of cell sheets is the use of a temperature-responsive cell culture surface, where the surface of tissue culture polystyrene (TCPS) dish is modified with thin layer of temperature-responsive polymer, poly(N-isopropylacrylamide) (PIPAAm). PIPAAm-immobilized TCPS allows cultured cells to be harvested as a contiguous cell sheet with extracellular matrices (ECMs) by reducing the temperature, while chemical and physical disruption impair ECMs, cell-cell junction, and membrane proteins. Ligand-immobilized and porous hydrophilic PIPAAm-grafted surfaces are able to accelerate cell sheet preparation and harvesting, respectively. In addition, the manipulation of harvested cell sheets with the aid of cell sheet manipulator facilitates the formation of 3D tissues. Cell sheet-based tissues and their transplantation are in seven clinical settings such as heart, cornea, esophagus, periodontal, middle chamber of ear, knee cartilage, and lung. In order to create thick and large 3D tissues and organs, large production of differentiated parenchymal cells from induced pluripotent stem (iPS) cells and vascularization within 3D tissues are key issues. © 2019 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 107A: 955-967, 2019.
Assuntos
Técnicas de Cultura de Células/métodos , Transplante de Células , Engenharia Tecidual/métodos , Animais , Ensaios Clínicos como Assunto , HumanosRESUMO
We developed a novel cultivating system for hollow-type spherical bacterial cellulose (HSBC) gel production without any molds or template. It consisted of floating aqueous medium droplet containing Gluconacetobacter xylinus (G. xylinus) at the boundary of two non-mixed silicone oil layers. The fibrils of bacterial cellulose (BC) were produced at the interface of water and oil phases. Fibril layers effectively thickened layer-by-layer and eventually formed a shell structure. The size of the HSBC gel can be controlled by the volume of dropped cell suspension. For cell suspensions of 50 µL and 10 µL, HSBC gels of approximately 4.0 mm and 2.5 mm were obtained, respectively. The shell of the HSBC gel is the gelatinous membrane formed by well-organized fibril networks; they comprised type-I crystal structure of cellulose. Additionally, we studied release profile of the fluorescein isothiocyanate-dextran (FITC-Dex) and observed that it released rapidly from the HSBC gels compared to from the BC gels obtained by the static culture method. The release behavior from HSBC gel agreed satisfactorily with Higuchi model. Therefore, the shell of HSBC gel is surely a thin gelatinous membrane of BC, and would be useful as a drug release device.
RESUMO
The stability and bio-distribution of genes or drug complexes with poly(ethylene oxide)-poly(propylene oxide)-poly(ethylene oxide) (PEO-PPO-PEO, Pluronic F-68) polymeric micelles (PM) are essential for an effective nanosized PM delivery system. We used Förster resonance energy transfer (FRET) pairs with PM and measured the FRET ratio to assess the stability of PM in vitro and in vivo on the cornea. The FRET ratio reached a plateau at 0.8 with 3% PM. Differential scanning calorimetry measurement confirmed the complex formation of FRET pairs with PM. Confocal imaging with the fluorophores fluorescein isothiocyanate isomer I (FITC) and rhodamine B base (RhB) also showed the occurrence of FRET pairs in vitro. The fluorophores were mixed with 3% PM solution or the FITC-labeled PEO-PPO-PEO polymers (FITC-P) were mixed with RhB-labeled plasmids (RhB-DNA). In addition, the in vitro corneal permeation of FRET pair complexes with PM reached a 0.8 FRET ratio. One hour after eye drop administration, FRET pairs colocalized in the cytoplasm, and surrounded and entered the nuclei of cells in the cornea, and the polymers were located in the corneal epithelial layers, as detected through anti-PEG immunohistochemistry. Furthermore, fluorescence colocalization in the cytoplasm and cell nucleus of the corneal epithelium was confirmed in tissues where RhB or RhB-DNA complexed with FITC-P was found to accumulate. We demonstrate that at a concentration of 3%, PM can encapsulate FRET pairs or RhB-DNA and retain their integrity within the cornea 1 h after administration, suggesting the feasibility and stability of PEO-PPO-PEO polymers as a vehicle for drug delivery.
Assuntos
Córnea/química , Sistemas de Liberação de Medicamentos/métodos , Soluções Oftálmicas/química , Plasmídeos/química , Polietilenoglicóis/química , Propilenoglicóis/química , Animais , Córnea/efeitos dos fármacos , Córnea/metabolismo , Portadores de Fármacos/química , Portadores de Fármacos/metabolismo , Sistemas de Liberação de Medicamentos/instrumentação , Transferência Ressonante de Energia de Fluorescência , Interações Hidrofóbicas e Hidrofílicas , Masculino , Camundongos Endogâmicos BALB C , Camundongos Nus , Soluções Oftálmicas/metabolismo , Soluções Oftálmicas/farmacologia , Plasmídeos/metabolismo , Polietilenoglicóis/metabolismo , Propilenoglicóis/metabolismoRESUMO
A novel shape-memory cell culture platform has been designed that is capable of simultaneously tuning surface topography and dimensionality to manipulate cell alignment. By crosslinking poly(ε-caprolactone) (PCL) macromonomers of precisely designed nanoarchitectures, a shape-memory PCL with switching temperature near body temperature is successfully prepared. The temporary strain-fixed PCLs are prepared by processing through heating, stretching, and cooling about the switching temperature. Temporary nanowrinkles are also formed spontaneously during the strain-fixing process with magnitudes that are dependent on the applied strain. The surface features completely transform from wrinkled to smooth upon shape-memory activation over a narrow temperature range. Shape-memory activation also triggers dimensional deformation in an initial fixed strain-dependent manner. A dynamic cell-orienting study demonstrates that surface topographical changes play a dominant role in cell alignment for samples with lower fixed strain, while dimensional changes play a dominant role in cell alignment for samples with higher fixed strain. The proposed shape-memory cell culture platform will become a powerful tool to investigate the effects of spatiotemporally presented mechanostructural stimuli on cell fate.
Assuntos
Nanoestruturas/química , Poliésteres/química , Animais , Técnicas de Cultura de Células , Sobrevivência Celular/fisiologia , Camundongos , Microscopia de Força Atômica , Microscopia de Fluorescência , Células NIH 3T3RESUMO
Apoptotic cell death serves important roles in homeostasis by eliminating dangerous, damaged, or unnecessary cells without causing an inflammatory response by externalizing phosphatidylserine to the outer leaflet in the phospholipid bilayer. In this study, we newly designed apoptotic cell membrane-inspired monomer and polymer which have the phosphoryl serine group as the anti-inflammatory functional moiety and demonstrate here for the first time that administration of an apoptotic cell membrane-inspired phosphorylserine polymer can protect murine macrophages (RAW 264.7) from lipopolysaccharide-induced inflammation. Interestingly, statistical copolymers with phosphorylcholine monomer that mimicked more precisely the apoptotic cell membrane result in more effective suppression of macrophage activation. This study provides new insights into the rational design of effective polymeric materials for anti-inflammatory therapies.
RESUMO
The development of stimuli responsive polymers has progressed significantly with novel preparation techniques, which has allowed access to new materials with unique properties. Dual thermoresponsive (double temperature responsive) block copolymers are particularly of interest as their properties can change depending on the lower critical solution temperature (LCST) or upper critical solution temperature (UCST) of each segment. For instance, these block copolymers can change from being hydrophilic, to amphiphilic or to hydrophobic simply by changing the solution temperature without any additional chemicals and the block copolymers can change from being fully solubilized to self-assembled structures to macroscopic aggregation/precipitation. Based on the unique solution properties, these dual thermo-responsive block copolymers are expected to be suitable for biomedical applications. This review is divided into three parts; LCST-LCST types of block copolymers, UCST-LCST types of block copolymers, and their potential as biomedical applications.
RESUMO
Although mechanostructural signals from the surrounding matrix have been known to regulate cell functions, the effects of substrate fluidity are poorly understood. Here, we demonstrate that the adhesion and morphology of cells are regulated by the fluidity on widely used biodegradable polymer substrates, rather than the substrate elasticity. We have designed cell culture films with different elasticity and fluidity using poly(ε-caprolactone-co-D,l-lactide) (CL-DLLA). The elasticity was successfully controlled by adjusting the amorphous-crystal phase transition temperature (Tm) of CL-DLLA without changing the surface wettability; i.e., the CL-DLLA displays more viscous (liquidlike) behavior at 37 °C with increasing DLLA contents. The fluidity was varied by chemically cross-linking the polymer networks. This CL-DLLA system was used to test the effect of variations in a substrate's fluidity on cell behavior. Differences were observed in adhesion, spreading and morphology of NIH 3T3 fibroblasts. Increasing the fluidity decreased cell spread area but enhanced the formation of spheroids. Although direct comparison of the elastic modulus between cross-linked and non-cross-linked samples are difficult, it was found that the substrate stiffness produced little changes in cell spread area, indicating that cells sense more dynamic nature of their surrounding environment. These findings will serve as the basis for new development of tissue engineering scaffolds and engineered stem cell niche as well as investigation of dynamic effects of mechanostructural stimuli on cell fate.
RESUMO
This work describes an intriguing strategy for the creation of arbitrarily shaped hydrogels utilizing a self-healing template (SHT). A SHT was loaded with a photo-crosslinkable monomer, PEG diacrylate (PEGDA), and then ultraviolet light (UV) crosslinked after first shaping. The SHT template was removed by simple washing with water, leaving behind the hydrogel in the desired physical shape. A hierarchical 3D structure such as "Matreshka" boxes were successfully prepared by simply repeating the "self-healing" and "photo-irradiation" processes. We have also explored the potential of the SHT system for the manipulation of cells.
RESUMO
Our study reports a versatile immobilization method of Hemagglutinating Virus of Japan Envelope (HVJ-E) for the generation of viral-mimetic surfaces for hormone resistant prostate cancer cell isolation. HVJ-E has recently attracted much attention as a new type of therapeutic material because hormone resistant prostate cancer cells such as PC-3 cells possess the HVJ-E receptors, GD1a. The HVJ-E was successfully immobilized on precursor films composed of poly-l-lysine and alginic acid via layer-by-layer assembly without changing the biological activity. The monolayer adsorption of HVJ-E particles was confirmed by quartz crystal microbalance, fluorescent and atomic force microscopy analyses. By developing the HVJ-E coating with an affinity based cell trap within a glass capillary tube, we are able to gently isolate PC-3 from LN-Cap cells that represent adenocarcinoma without compromising cell viability. We achieved approximately 100% cell separation efficiency only by 60 seconds of flowing. We believe that the proposed technique offers significant promise for the creation of a hormone resistant cancer cell trap on a broad range of materials.
Assuntos
Biomimética/métodos , Lisina/química , Terapia Viral Oncolítica/métodos , Neoplasias da Próstata/química , Vírus Sendai/química , Linhagem Celular Tumoral , Separação Celular , Sobrevivência Celular , Humanos , Lisina/metabolismo , Masculino , Neoplasias da Próstata/patologia , Vírus Sendai/metabolismoRESUMO
A dual pH and glucose responsive boronic acid containing nanofiber was constructed for the reversible capture and release of lectins. The effects of surface groups and pH values on selective lectin capture were investigated by fluorescence microscopy. Compared to the pristine nanofibrous membrane, glucose and galactose functionalized nanofiber surfaces showed significantly higher capture of ConA and Jacalin, under alkaline conditions. On the other hand, treatment of the modified nanofibers with an acidic solution resulted in the detachment of both the lectins and glycopolymers from the nanofiber surface. As expected, once the glycopolymers are displaced, no lectins were adhered to the nanofiber surface under alkaline conditions. These functional nanofibers can therefore be easily modified and hence can be used for quick removal of selective proteins or toxins from the solution.
Assuntos
Ácidos Borônicos/química , Concanavalina A/química , Galactose/química , Glucose/química , Lectinas/química , Nanofibras/química , Lectinas de Plantas/química , Concentração de Íons de Hidrogênio , SoluçõesRESUMO
In this study, we present anti-cancer drug containing nanofiber-mediated gene delivery to treat liver cancer. Electro-spun nanofibers have big potential for local delivery and sustained release of therapeutic gene and drugs. We reported a temperature-responsive nanofibers mainly compounded by branched poly(ε-caprolactone) (PCL) macro-monomers and anti-cancer drug paclitaxel. The nanofiber could be administrated into liver tumors to dramatically hinder their growth and prevent their metastasis. As a result, paclitaxel encapsulated PCL (PTX/PCL) nanofibers with diameters of around several tens nanometers to 10 nm were successfully obtained by electro-spinning and observed in scanning electron microscopy (SEM). Nanoparticles composed of disulfide cross-linked branched PEI (ssPEI) and anti-cancer therapeutic gene miRNA-145 were complexed based on the electrostatic interaction and coated over the paclitaxel-loaded nanofiber. MicroRNA 145/ssPEI nanoparticles (MSNs) immobilized on the PTX/PCL nanofiber showed time-dependent sustained release of the microRNA for enhanced uptake in neighboring liver cancer cells without any noticeable cytotoxicity. From this study we are expecting a synergistic effect on the cancer cell suppression since we have combined the drug and gene delivery. This approach uses the nanofibers and nanoparticles together for the treatment of cancer and the detailed investigation in vitro and in vivo must be conducted for the practicality of this study. The polymer is biodegradable and the toxicity issues must be cleared by our approach.
Assuntos
Sistemas de Liberação de Medicamentos/métodos , Técnicas de Transferência de Genes , Terapia Genética/métodos , Neoplasias Hepáticas , Nanofibras/química , Poliésteres , Humanos , Neoplasias Hepáticas/genética , Neoplasias Hepáticas/terapia , MicroRNAs/genética , Paclitaxel/química , Paclitaxel/farmacologia , Poliésteres/química , Poliésteres/farmacologiaRESUMO
We demonstrate here a local- and remote-control of gel disintegration by using photoinduced proton transfer chemistry of photoacid generator (PAG). The gels were prepared by simply mixing two polymers, poly(N-isopropylacrylamide-co-5-methacrylamido-1,2-benzoxaborole) (P(NIPAAm-co-MAAmBO)) and poly(3-gluconamidopropyl methacrylamide) (PGAPMA) via the synergistic interaction of benzoxaborole and diol groups. The o-nitrobenzaldehyde (o-NBA) was then loaded into the gel as a PAG. The benzoxaborole-diol interaction was successfully disintegrated upon UV irradiation due to the local pH decrease inside the gel. When the gel was irradiated to a specific gel region, the synergistic interactions were disintegrated only at the exposed region. Of special interest is that the whole material eventually transitioned from gel to sol state, as the generated protons diffused gradually toward the nonilluminated region. The ability of the proposed gel-sol transition system via photoinduced proton diffusion may be beneficial for not only prompt pH changes within the gel but also the design of predictive and programmable devices for drug delivery.
Assuntos
Portadores de Fármacos/química , Processos Fotoquímicos , Polímeros/química , Prótons , Resinas Acrílicas/química , Benzeno/química , Liberação Controlada de Fármacos , Géis , Concentração de Íons de Hidrogênio , Análise Espaço-Temporal , Temperatura , Raios UltravioletaRESUMO
Inactivated Hemagglutinating Virus of Japan Envelope (HVJ-E) was immobilized on electrospun nanofibers of poly(ε-caprolactone) by layer-by-layer (LbL) assembly technique. The precursor LbL film was first constructed with poly-L-lysine and alginic acid via electrostatic interaction. Then the HVJ-E particles were immobilized on the cationic PLL outermost surface. The HVJ-E adsorption was confirmed by surface wettability test, scanning laser microscopy, scanning electron microscopy, and confocal laser microscopy. The immobilized HVJ-E particles were released from the nanofibers under physiological condition. In vitro cytotoxic assay demonstrated that the released HVJ-E from nanofibers induced cancer cell deaths. This surface immobilization technique is possible to perform on anti-cancer drug incorporated nanofibers that enables the fibers to show chemotherapy and immunotherapy simultaneously for an effective eradication of tumor cells in vivo.
RESUMO
The vascularization of tissue engineered products represents a key issue in regenerative medicine which needs to be addressed before the translation of these protocols to the bedside can be foreseen. Here we propose a multistep procedure to prepare pre-vascularized three-dimensional (3D) cardiac bio-substitutes using dynamic cell cultures and highly porous biocompatible gelatin scaffolds. The strategy adopted exploits the peculiar differentiation potential of two distinct subsets of adult stem cells to obtain human vascularized 3D cardiac tissues. In the first step of the procedure, human mesenchymal stem cells (hMSCs) are seeded onto gelatin scaffolds to provide interconnected vessel-like structures, while human cardiomyocyte progenitor cells (hCMPCs) are stimulated in vitro to obtain their commitment toward the cardiac phenotype. The use of a modular bioreactor allows the perfusion of the whole scaffold, providing superior performance in terms of cardiac tissue maturation and cell survival. Both the cell culture on natural-derived polymers and the continuous medium perfusion of the scaffold led to the formation of a densely packaged proto-tissue composed of vascular-like and cardiac-like cells, which might complete maturation process and interconnect with native tissue upon in vivo implantation. In conclusion, the data obtained through the approach here proposed highlight the importance to provide stem cells with complementary signals in vitro able to resemble the complexity of cardiac microenvironment.
RESUMO
We report here that the direction of aligned cells on nanopatterns can be tuned to a perpendicular direction without use of any biochemical reagents. This was enabled by shape-memory activation of nanopatterns that transition from a memorized temporal pattern to the original permanent pattern by heating. The thermally induced shape-memory nanopatterns were prepared by chemically crosslinking semi-crystalline poly(ε-caprolactone) (PCL) in a mold to show shape-memory effects over its melting temperature (Tm = 33°C). Permanent surface patterns were first generated by crosslinking the PCL macromonomers in a mold, and temporary surface patterns were then embossed onto the permanent patterns. The temporary surface patterns could be easily triggered to transition quickly to the permanent surface patterns by a 37°C heat treatment, while surface wettability was independent of temperature. To investigate the role of dynamic and reversible surface nanopatterns on cell alignment on the PCL films before and after a topographic transition, NIH 3T3 fibroblasts were seeded on fibronectin-coated PCL films with a temporary grooved topography (grooves with a height of 300 nm and width of 2 µm were spaced 9 µm apart). Interestingly, cells did not change their direction immediately after the surface transition. However, cell alignment was gradually lost with time, and finally cells realigned parallel to the permanent grooves that emerged. The addition of a cytoskeletal inhibitor prevented realignment. These results clearly indicate that cells can sense dynamic changes in the surrounding environments and spontaneously adapt to a new environment by remodeling their cytoskeleton. These findings will serve as the basis for new development of spatiotemporal tunable materials to direct cell fate.