Biomedical subjects

Takao Aoyagi

Publications and source records attributed to Takao Aoyagi.

8 recordsLinked to original sources

PubMed2006

Importance of bound water in hydration-dehydration behavior of hydroxylated poly(N-isopropylacrylamide).

In this study, a differential scanning calorimetric analysis was performed to investigate the role of water existing around the polymer chains on their thermoresponsive behaviors using the novel functional temperature-sensitive copolymer, poly(N-isopropylacrylamide-co-2-hydroxyisopropylacrylamide) (poly(NIPAAm-co-HIPAAm)). The HIPAAm comonomers were incorporated into the polymeric chains as hydrophilic parameters, and then the hydration states of poly(NIPAAm-co-HIPAAm) with various HIPAAm compositions were examined. Bound water, which is affected by the polymeric chains to some extent, was produced by adding the copolymers to the water, and the temperature due to the melting of the bound water decreased as the HIPAAm content increased. On the basis of this result, we considered that the interaction between the bound water and the polymeric chains is reinforced by the increasing HIPAAm composition. In addition, the cloud points of the copolymers shifted to a higher temperature, and the endothermic enthalpy for the phase transition decreased with increasing the HIPAAm content, suggesting that the number of water molecules disassociated from the polymeric chains decreased. Based on these results, we postulate that the changes in the interaction between the thermosensitive polymer and bound water exert a strong influence on its phase transition and/or separation, such as the cloud point or dehydration behavior.

Acrylamides↗

PubMed2006

Stimuli-responsive properties of N-isopropylacrylamide-based ultrathin hydrogel films prepared by photo-cross-linking.

To develop stimuli-responsive ultrathin polymer films on a solid substrate, a novel photo-cross-linkable polymer with both temperature- and pH-responsive properties was prepared. The photoreactive 4-aminobenzophenone (BP) was introduced onto the side groups of poly(N-isopropylaclylamide-co-2-carboxyisopropylaclylamide) [poly(NIPAAm-co-CIPAAm)]. This copolymer was designed for highly random sequences of comonomers. After the formation of spin-coated polymer films on a solid substrate, UV-light irradiation started the cross-linking reaction. The spin-coating processes and stability of the polymer films were quantitatively monitored by a quartz crystal microbalance (QCM), and the thickness was estimated using an atomic force microscope (AFM). These measurements confirmed the formation of a very plain polymer film, and the film thickness was precisely controlled by the concentration of the polymer solution used for spin coating. Moreover, the obtained films showed a high stability due to the cross-liking reaction and UV irradiation. Cyclic voltammetry using potassium ferricyanide revealed that the ions could permeate the photo-cross-linked ultrathin polymer films. The permeability of the ultrathin hydrogel films was dramatically changed by varying the pH and temperature of the aqueous media. These observations suggest that the preparation of isopropylacrylamide-based stimuli-responsive ultrathin hydrogel films is possible.

Journal Article↗

PubMed2006

Hydroxylated poly(N-isopropylacrylamide) as functional thermoresponsive materials.

In this study, we developed a poly(N-isopropylacrylamide)-based thermoresponsive polymeric material with a high content of hydroxyl groups. We newly designed the functional monomer, N-(2-hydroxyisopropyl)acrylamide (HIPAAm), considering maintaining the continuous and repeated structure of the isopropylamide group after copolymerization and the monomer reactivity ratios. The thermoresponsive polymer was derived by conventional radical copolymerization of HIPAAm with N-isopropylacrylamide (NIPAAm) in high yield. Estimation of monomer reactivity ratios, r(1) and r(2), supported the almost random sequence of the comonomers. The obtained copolymers showed a very sensitive phase transition and/or separation in response to temperature in aqueous media although they have many hydrophilic parts, and their thermoresponsive behavior was not affected by the pH. Furthermore, the cloud points of these copolymers closely depended on the HIPAAm content and could be easily controlled by adding salts. HIPAAm is expected to regulate the phase transition and/or separation temperature of the NIPAAm-based copolymers while maintaining their desirable sensitive thermoresponse. Differential scanning calorimetric analysis showed that dehydration of the polymer chains occurring in phase transition became incomplete with increasing HIPAAm content. Moreover, it was found that poly(NIPAAm-co-HIPAAm) having a high content of the HIPAAm unit showed liquid-liquid phase separation involving coacervation. The sizes of the coacervate droplets were relatively monodisperse and very minimal. Poly(NIPAAm-co-HIPAAm) is valuable for use in biomedical fields such as bioseparation.

Acrylic Resins↗

PubMed2006

Analysis of the formation mechanism for thermoresponsive-type coacervate with functional copolymers consisting of N-isopropylacrylamide and 2-hydroxyisopropylacrylamide.

We now report the formation mechanism of the thermoresponsive-type coacervate with the novel functional temperature-sensitive polymer, poly(N-isopropylacrylamide-co-2-hydroxyisopropylacrylamide) (poly(NIPAAm-co-HIPAAm)), synthesized in our laboratory. The effects of introducing the hydrophilic comonomer (HIPAAm) into the copolymer chains and adding salts on the behaviors of the coacervate droplets induced in the poly(NIPAAm-co-HIPAAm) aqueous solutions were investigated. Not only the particle sizes of the coacervate droplets but also the cloud points of the copolymer solutions could be modulated by the HIPAAm content incorporated in the copolymers. Moreover, the particle sizes of the coacervate droplets were also changed by adding salts. Namely, the particle sizes increased with the decreasing HIPAAm composition and increasing NaCl concentration. In addition, the 1H NMR and differential scanning calorimetric measurements suggested that as the HIPAAm content decreased or NaCl concentration increased, dehydration of the copolymers induced in the phase transition and/or separation became much easier. Therefore, on the basis of the findings obtained from these measurements, we determined that the particle sizes of the coacervate droplets induced in the temperature-sensitive polymers increased as the number of the water molecules, which are dissociated from the polymeric chains during the phase transition and/or separation, increased. Besides, to examine the separation of the model solutes, the aqueous two-phase separation with the coacervate droplets of poly(NIPAAm-co-HIPAAm) was carried out. The partitions of Methyl Orange as a model solute under both acidic (pH 2) and basic (pH 12) conditions were performed. The amount of Methyl Orange partitioned into the coacervate droplets at pH 12 is much greater than that at pH 2, which indicated that the coacervate droplets could recognize a slight difference in the polarity or structure between the model solutes.

Acrylamides↗

PubMed2005

Temperature-responsive cross-linked poly(epsilon-caprolactone) membrane that functions near body temperature.

The objective of this study is to develop a sensitive temperature-responsive material that would function near body temperature. To achieve this purpose, we compounded 2-branched and 4-branched poly(epsilon-caprolactone) macromonomers to modulate the transition temperatures of the resulting cross-linked materials. The temperature-responsive properties were studied using differential scanning calorimetry and X-ray diffraction measurements. As a result, the mixing ratios of each macromonomer or the total macromonomer concentrations were very dominant in modulating the transition temperatures. The materials could successfully control the permeation of the model drug, prednisolone, near body temperature.

Anti-Inflammatory Agents↗

PubMedDate not supplied

Copolymerization of 2-carboxyisopropylacrylamide with N-isopropylacrylamide accelerates cell detachment from grafted surfaces by reducing temperature.

Acrylic acid (AAc) has been utilized to introduce reactive carboxyl groups to a temperature-responsive polymer, poly(N-isopropylacrylamide) (PIPAAm). However, AAc introduction shifts the copolymer phase transition temperatures higher and dampens the steep homopolymer phase transition with increasing AAc content. We previously synthesized 2-carboxyisopropylacrylamide (CIPAAm) having both a similar side chain structure to IPAAm and a functional carboxylate group in order to overcome these shortcomings. In the present study, these copolymers, grafted onto cell culture plastic, were assessed for cell adhesion control using their phase transition. AAc introduction to PIPAAm-grafted surfaces resulted in excessive surface hydration and hindered cell spreading in culture at 37 degrees C. In contrast, CIPAAm-containing copolymer-grafted surfaces exhibited relatively weak hydrophobicity similar to both homopolymer PIPAAm-grafted surfaces as well as commercial ungrafted tissue culture polystyrene dish surfaces. Cells adhered and spread well on these surfaces at 37 degrees C in culture. As observed previously on PIPAAm-grafted surfaces, cells were spontaneously detached from the copolymer-grafted surfaces by reducing culture temperature. Cell detachment was accelerated on the CIPAAm copolymer-grafted surfaces compared to pure IPAAm surfaces, suggesting that hydrophilic carboxyl group microenvironment in the monomer and polymer is important to accelerate grafted surface hydration below the lower critical solution temperature, detaching cells.

Acrylamides↗

PubMedDate not supplied

Temperature-responsive cell culture surfaces enable "on-off" affinity control between cell integrins and RGDS ligands.

In this study, specific interactions between immobilized RGDS (Arg-Gly-Asp-Ser) cell adhesion peptides and cell integrin receptors located on cell membranes are controlled in vitro using stimuli-responsive polymer surface chemistry. Temperature-responsive poly(N-isopropylacrylamide-co-2-carboxyisopropylacrylamide) (P(IPAAm-co-CIPAAm)) copolymer grafted onto tissue culture grade polystyrene (TCPS) dishes permits RGDS immobilization. These surfaces facilitate the spreading of human umbilical vein endothelial cells (HUVECs) without serum depending on RGDS surface content at 37 degrees C (above the lower critical solution temperature, LCST, of the copolymer). Moreover, cells spread on RGDS-immobilized surfaces at 37 degrees C detach spontaneously by lowering culture temperature below the LCST as hydrated grafted copolymer chains dissociate immobilized RGDS from cell integrins. These cell lifting behaviors upon hydration are similar to results using soluble RGDS in culture as a competitive substitution for immobilized ligands. Binding of cell integrins to immobilized RGDS on cell culture substrates can be reversed spontaneously using mild environmental stimulation, such as temperature, without enzymatic or chemical treatment. These findings are important for control of specific interactions between proteins and cells, and subsequent "on-off" regulation of their function. Furthermore, the method allows serum-free cell culture and trypsin-free cell harvest, essentially removing mammalian-sourced components from the culture process.

Cell Adhesion↗

PubMedDate not supplied

Immobilization of cell-adhesive peptides to temperature-responsive surfaces facilitates both serum-free cell adhesion and noninvasive cell harvest.

We have developed temperature-responsive cell culture surfaces to harvest intact cell sheets for tissue-engineering applications. Both cost and safety issues (e.g., prions, bovine spongiform encephalopathy) are compelling reasons to avoid use of animal-derived materials, including serum, in such culture. In the present study, synthetic cell-adhesive peptides are immobilized onto temperature-responsive polymer-grafted surfaces, and cell adhesion and detachment under serum-free conditions were examined. The temperature-responsive polymer poly(N-isopropylacrylamide) (PI-PAAm) was functionalized by copolymerization with a reactive comonomer having both a carboxyl group and an isopropylacrylamide group. These copolymers were covalently grafted onto tissue culture-grade polystyrene dishes. Synthetic cell-adhesive peptides were then immobilized onto these surfaces via carboxyl groups. Bovine aortic endothelial cells both adhered and spread on these surfaces even under serum-free conditions at 37 degrees C, similar to those in 10% serum-supplemented culture. Spread cells promptly detached from the surfaces on lowering culture temperatures below the lower critical solution temperature of the polymer, 32 degrees C. These surfaces would be useful for serumfree culture for tissue-engineering applications.

Adsorption↗