Tough mechanical properties are generally required for tissue substitutes used in regeneration of damaged tissue, as these substitutes must be able to withstand the external physical force caused by stretching. Gelatin, a biopolymer derived from collagen, is a biocompatible and cell adhesive material, and is thus widely utilized as a component of biomaterials. However, the application of gelatin hydrogels as a tissue substitute is limited owing to their insufficient mechanical properties. Chemical cross-linking is a promising method to improve the mechanical properties of hydrogels. We examined the potential of the chemical cross-linking of gelatin hydrogels with carboxy-group-modified polyrotaxanes (PRXs), a supramolecular polymer comprising a poly(ethylene glycol) chain threaded into the cavity of α-cyclodextrins (α-CDs), to improve mechanical properties such as stretchability and toughness. Cross-linking gelatin hydrogels with threading α-CDs in PRXs could allow for freely mobile cross-linking points to potentially improve the mechanical properties. Indeed, the stretchability and toughness of gelatin hydrogels cross-linked with PRXs were slightly higher than those of the hydrogels with the conventional chemical cross-linkers 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide (EDC)/N-hydroxysuccinimide (NHS). In addition, the hysteresis loss of gelatin hydrogels cross-linked with PRXs after repeated stretching and relaxation cycles in a hydrated state was remarkably improved in comparison with that of conventional cross-linked hydrogels. It is considered that the freely mobile cross-linking points of gelatin hydrogels cross-linked with PRXs attenuates the stress concentration. Accordingly, gelatin hydrogels cross-linked with PRXs would provide excellent mechanical properties as biocompatible tissue substitutes exposed to a continuous external physical force.

The molecular mobility of cyclic molecules (e.g. α-cyclodextrins) threaded along a linear polymer chain (e.g. poly(ethylene glycol)) in polyrotaxanes is a unique feature for biomaterials with dynamic functionality. Surfaces with molecular mobility can be obtained by introducing polyrotaxanes. The molecular mobility of polyrotaxane-based surfaces can be modulated by changing the number of threaded cyclic molecules and modifying their functional groups. Biological ligands modified with α-cyclodextrins exhibit increased multivalent interactions with their receptors due to the molecular mobility of the latter. Furthermore, polyrotaxane-based surfaces not only improve the initial response of cells via multivalent interactions, but also affect cytoskeleton formation and the inherent quality of cells, including differentiation. Such polyrotaxane surfaces can emerge as new biointerfaces that can adapt to the dynamic biological nature.

The cytocompatibility of biological and synthetic materials is an important issue for biomaterials. Gelatin hydrogels are used as biomaterials because of their biodegradability. We have previously reported that the mechanical properties of gelatin hydrogels are improved by cross-linking with polyrotaxanes, a supramolecular compound composed of many cyclic molecules threaded with a linear polymer. In this study, the ability of gelatin hydrogels cross-linked by polyrotaxanes (polyrotaxane–gelatin hydrogels) for cell cultivation was investigated. Because the amount of polyrotaxanes used for gelatin fabrication is very small, the chemical composition was barely altered. The structure and wettability of these hydrogels are also the same as those of conventional hydrogels. Fibroblasts adhered on polyrotaxane–gelatin hydrogels and conventional hydrogels without any reduction or apoptosis of adherent cells. From these results, the polyrotaxane–gelatin hydrogels have the potential to improve the mechanical properties of gelatin without affecting cytocompatibility. Interestingly, when cells were cultured on polyrotaxane–gelatin hydrogels after repeated stress deformation, the cells were spontaneously oriented to the stretching direction. This cellular response was not observed on conventional hydrogels. These results suggest that the use of a polyrotaxane cross-linking agent can not only improve the strength of hydrogels but can also contribute to controlling reorientation of the gelatin.

The aims of this study were to investigate the feasibility of using a DLP 3D printer to fabricate a crown using scan data before tooth preparation, and to investigate the effect of additional heat curing on the mechanical properties of the urethane dimethacrylate (UDMA)-based 3D printed crown. A silicone fitting test was used to evaluate the internal adaptation of the crown. For ultimate tensile strength (UTS), the specimens were tested after 24 h storage in water at 37 °C or after 10,000 thermal cycles (TC) between 5–55 °C. For shear bond strength (SBS), a PMMA self-curing resin was filled into a Teflon ring mounted onto the polished UDMA specimens. The internal adaptation of the crowns fabricated with cement space was better than those with no cement space. There was no significant difference in UTS between light-curing and additional heat-curing groups after TC. As for the SBS, there was a significant difference after TC between the two groups. Crowns can be fabricated by a DLP 3D printer using pre-preparation scans with a cement space defined in the software. Additional heat curing of the UDMA-based crown reduced residual monomer and improved its mechanical properties.

Thermoresponsive cell-culture polystyrene (PS) surfaces that are grafted with poly(N-isopropylacrylamide) (PIPAAm) facilitate the cultivation of cells at 37 °C and the detachment of cultured cells as a sheet with an underlying extracellular matrix (ECM) by reducing the temperature. However, the ECM and cell detachment mechanisms are still unclear because the detachment of cells from thermoresponsive surfaces is governed by complex interactions among the cells/ECM/surface. To explore the dynamic behavior of serum protein adsorption/desorption, thermoresponsive surfaces that correspond to thermoresponsive tissue-culture PS dishes were formed on sensor chips for quartz crystal microbalance with dissipation (QCM-D) measurements. X-ray photoelectron spectroscopy (XPS) measurements and temperature-dependent frequency and dissipation shifts, Δf and ΔD, using QCM-D revealed that the thermoresponsive polymers were successfully grafted onto oxidized, thin PS films on the surfaces of the sensor chips. Increased amounts of adsorbed bovine serum albumin (BSA) and fibronectin (FN) were observed on the thermoresponsive polymer-grafted surfaces at 37 °C when compared with those at 20 °C because of enhanced hydrophobic interactions with the hydrophobic, thermoresponsive surface. While the calculated masses of adsorbed BSA and FN using QCM-D were 3–5 times more than those that were obtained from radiolabeling, the values were utilized for relative comparisons among the same substrate. More importantly, the thermoresponsive, dynamic behavior of serum protein adsorption/desorption was monitored using the QCM-D technique. Observations of this dynamic behavior revealed that the BSA and FN that were adsorbed at 37 °C remained on both surfaces after decreasing the temperature to 20 °C.

Polyrotaxanes are supramolecular assemblies consisting of cyclic molecules (e.g., α-cyclodextrins) and linear polymer chains (e.g., poly[ethylene glycol]), in which cyclic molecules can move along the polymer chain. Here, we examined the effect of functional groups introduced into the α-cyclodextrins of polyrotaxane on cell responses such as adhesion, proliferation, and differentiation. Polyrotaxane-based triblock copolymers modified with methyl (CH₃, hydrophobic, and nonionic), hydroxy (OH, hydrophilic and nonionic), amino (NH₂, cationic), and sulfo (SO₃H, anionic) groups were coated on the surface of the culture plate to fabricate polyrotaxane surfaces with different surface chemistries. The chemical compositions of each surface were determined via time-of-flight secondary ion mass spectrometry and X-ray photoelectron spectroscopy. The contact angle hysteresis reflecting the molecular mobility and zeta potential of each polyrotaxane surface changed depending on the functional groups. When osteoblast and adipocyte differentiation was induced in human mesenchymal stem cells cultured on each polyrotaxane surface, the cells adhered to the SO₃H-modified polyrotaxane surfaces exhibited osteoblast differentiation, whereas the cells adhered to the OH-, NH₂-, and SO₃H-modified polyrotaxane surfaces preferentially underwent adipocyte differentiation compared with those on the unmodified and CH₃-modified polyrotaxane surfaces. Interestingly, the SO₃H-modified polyrotaxane surfaces promoted both osteoblast and adipocyte differentiation. High molecular mobility and negative charge on the SO₃H-modified polyrotaxane surfaces are expected to contribute to the facilitation of both osteoblast and adipocyte differentiation.