Supramolecular Surface Coatings with Acetylated Polyrotaxane-Based Triblock Copolymers for Thermal Regulation of Cell Adhesion and Fabrication of Cell Sheets.

Polyrotaxanes (PRXs) containing acetylated α-cyclodextrins exhibit a temperature-dependent phase transition in aqueous solutions across their lower critical solution temperature (LCST) of approximately 26.6 °C. To gain insights into the interactions of acetylated PRXs (Ac-PRXs) with biological components, thermoresponsive supramolecular surfaces were prepared by coating tissue culture polystyrene (TCPS) surfaces with Ac-PRX triblock copolymers, and their surface properties across the LCST were evaluated. The wettability and protein adsorption of Ac-PRX-coated surfaces changed significantly between 10 and 37 °C, whereas the uncoated TCPS and unmodified PRX-coated surfaces did not alter the wettability and protein adsorption at 10 and 37 °C. The adhesion, proliferation, morphology, and adhesion strength of NIH/3T3 cells on Ac-PRX-coated surfaces were found to be similar to those of the uncoated and unmodified PRX-coated surfaces. However, the adhesion strength of NIH/3T3 cells on Ac-PRX-coated surfaces decreased drastically at 10 °C. Consequently, the cells spontaneously detached from the Ac-PRX-coated surfaces without enzymatic treatment. Additionally, when incubating confluent cells at 10 °C, the cells detached from Ac-PRX-coated surfaces as cell sheets while retaining extracellular matrix proteins. The findings of this study provide new directions for the design of thermoresponsive supramolecular biointerfaces for applications in bioseparation and cell manipulation.

Long-term physical therapy for neuropathic pain after cervical spinal cord injury and resting state electroencephalography: a case report.

INTRODUCTION: Neuropathic pain after spinal cord injury is difficult to treat, and it is associated with abnormalities in the function of the thalamus-to-cortex neural circuitry. Aerobic exercise provides immediate improvement in neuropathic pain and is associated with abnormal resting electroencephalography (EEG) findings in patients with spinal cord injury. This study aimed to investigate whether physical therapy, including walking, can improve neuropathic pain and EEG peak alpha frequency (PAF) in the long term in a patient with cervical spinal cord injury. CASE PRESENTATION: A 50-year-old man was admitted with a cervical spinal cord insufficiency injury sustained one week prior. The residual height was C5. Neuropathic pain was observed in the fingers bilaterally. A numerical rating scale (NRS) was evaluated to measure the weekly mean and maximum intensities of pain. Resting EEG was measured, and the PAF was calculated. Each time point was evaluated in 2-week intervals from the time of admission, and the rate of change (Δ) of PAF was calculated based on the initial evaluation. Interventions included 18 weeks of standard physical therapy focusing on gait, with additional intensive gait training (4-10 weeks). The NRS scores for the mean and maximum intensities of pain decreased significantly after 6 weeks, and ΔPAF increased significantly after 4 weeks. Improvement in PAF coincided with the start of intensive gait training. DISCUSSION: PAF shifts to a high frequency during intensive gait training, suggesting the effectiveness of aerobic exercise. Furthermore, there is a close relationship between PAF, pain, and the quantification of pain changes.

Independent Roles of Molecular Mobility and Zeta Potential on Supramolecular Surfaces in the Sequence of RAW264.7 Macrophage Responses.

Surface properties of biomaterials affect the morphologies and inflammatory responses of macrophages. Recently, biomaterial design utilizing these properties has been explored to build a scaffold for balancing the immune system in vivo. In the present study, polyrotaxane surfaces with different functional groups including methyl, amino, and sulfo groups are utilized to clarify the effect of molecular mobility and zeta potential of these surfaces on RAW264.7 macrophage responses. At 24 h post-seeding, the majority of the cells adhere onto each surface, and the initial spreading is suppressed by more negatively-charged polyrotaxane surfaces. From 24 to 48 h of incubation, the spreading areas on the unmodified and methylated surfaces significantly increase, whereas those on the aminated and sulfonated surfaces remain unchanged. These results suggest that the initially cellular spreading process depends on the zeta potential, while the subsequent spreading process is governed by the molecular mobility. After lipopolysaccharide stimulation, the less mobile surfaces induce higher expression of inflammation-related genes than highly mobile surfaces, suggesting that molecular mobility is the main factor modulating the inflammatory activity in macrophages. These findings indicate that the zeta potential and molecular mobility of polyrotaxane surfaces may play independent roles in the sequence of macrophage responses.

Improved epithelial cell-cell adhesion using molecular mobility of supramolecular surfaces.

Cells can sense the surrounding microenvironmental properties including contact with biomaterials. Although in vitro cell fates in response to the physical properties of cell-adhesive materials have been widely reported, their influence on cell-cell adhesion is unclear. Here, we investigated the role of molecular mobility on polyrotaxane surfaces in epithelial cell-cell adhesion. Polyrotaxane surfaces with high mobility induced cytoplasmic yes-associated protein (YAP) localization in epithelial cells, whereas those with low mobility induced nuclear YAP localization, suggesting that YAP localization is switched by the mobility of the polyrotaxane surface. The cytoplasmic YAP localization increased the expression of tight junction-associated genes. A scratch assay revealed that although the epithelial cells on the low mobile surface rapidly initiated their migration, the cells on the highly mobile surface delayed their migration. Thus, this finding suggests that polyrotaxane surfaces with higher mobility induce cytoplasmic YAP localization, leading to stronger cell-cell adhesion. The polyrotaxane biointerface is promising as a powerful tool to improve the physical immune system and repair biological tissues.

Fretibacterium sp. human oral taxon 360 is a novel biomarker for periodontitis screening in the Japanese population.

BACKGROUND: Periodontitis is a common inflammatory disease, leading to bone destruction and tooth loss. Screening for periodontitis is important in preventing the progress of this disease. Various types of bacteria have been examined as potential screening targets, but only culturable pathogenic bacteria have been considered candidates. Recently, the various uncultivable bacteria have been identified in microbiome studies, but the value of these bacteria in periodontitis screening remains unknown. OBJECTIVES: The aim of this study was to evaluate the diagnostic use of uncultivable bacteria Fretibacterium sp. HOT 360 and TM7 sp. HOT 356 for periodontitis screening in the Japanese population. MATERIAL AND METHODS: Stimulated saliva samples were collected from 217 participants (periodontitis group, n = 157; healthy group, n = 60). The two uncultivable bacterial species selected were: Fretibacterium sp. human oral taxon 360 (Fretibacterium sp. HOT 360) and TM7 sp. human oral taxon 356 (TM7 sp. HOT 356). The levels of these two bacterial species were compared with those of Porphyromonas gingivalis (P. gingivalis), a keystone pathogen in periodontitis. These three species of bacteria were then quantified using qualitative real-time polymerase chain reaction (qPCR) with specific primers and Taqman probes. Statistical analysis was performed by SPSS 20.0 software. P value was statistically significant at .05. RESULTS: The populations of uncultivable bacterial species TM7 sp. HOT 356 and Fretibacterium sp. HOT 360 were significantly higher in periodontitis group than in healthy group. Only Fretibacterium sp. HOT 360 showed a significantly positive correlation with such periodontal parameters as probing pocket depth (PPD) and bleeding on probing (BOP). CONCLUSION: These findings indicate that uncultivable bacteria Fretibacterium sp. HOT 360 can be used as a saliva-based diagnostic bacterial biomarker for periodontitis screening.