Publisher：Materials Chemistry and Physics  

This study aims to investigate the adaptive strategies employed by dental pulp stem cells (DPSCs) in response to oral disease conditions, focusing on their survival and proliferation. DPSCs possess regenerative potential and play a crucial role in the repair and regeneration of dental tissues. The study explores the coping mechanisms of DPSCs under oral disease conditions using a chitosan-based biomaterial reinforced with calcium zirconium nanoparticles (CZNP). The investigation assesses the viability and proliferation of DPSCs in the presence of oral disease conditions and evaluates the performance of the chitosan-CZNP biomaterial. The study presents the characterization and performance evaluation of drug-loaded biomaterials with varying weight percentages of CZNP. Parameters such as porosity, drug release, fracture toughness, tensile strength, degradation rate, and apatite formation are examined. The results demonstrate that increasing the weight percentage of CZNP leads to higher porosity and drug release while maintaining or enhancing fracture toughness, tensile strength, and apatite formation. Notably, the biomaterial containing 7.5 wt% CZNP exhibits the highest drug release, fracture toughness, and apatite formation. These findings suggest that the chitosan-CZNP biomaterial effectively supports the survival and proliferation of DPSCs under oral disease conditions. An artificial neural network (ANN) is employed to predict the properties of samples with different weight percentages and apatite formation. The ANN successfully estimates the degradation rate, tensile strength, fracture toughness, drug release, and porosity of the samples. This research enhances our understanding of the coping mechanisms of DPSCs and highlights the potential of chitosan-CZNP biomaterials in oral tissue engineering and regenerative medicine. Further investigations are warranted to explore the long-term effects and clinical applications of these biomaterials in the treatment of oral diseases.

DOI： 10.1016/j.matchemphys.2024.129610

Web of Science

Scopus

Language：English   Publisher：Genes to Cells  

An autism-associated gene Shank3 encodes multiple splicing isoforms, Shank3a-f. We have recently reported that Shank3a/b-knockout mice were more susceptible to kainic acid-induced seizures than wild-type mice at 4 weeks of age. Little is known, however, about how the N-terminal and ankyrin repeat domains (NT-Ank) of Shank3a/b regulate multiple molecular signals in the developing brain. To explore the functional roles of Shank3a/b, we performed a mass spectrometry-based proteomic search for proteins interacting with GFP-tagged NT-Ank. In this study, NT-Ank was predicted to form a variety of complexes with a total of 348 proteins, in which RNA-binding (n = 102), spliceosome (n = 22), and ribosome-associated molecules (n = 9) were significantly enriched. Among them, an X-linked intellectual disability-associated protein, Nono, was identified as a NT-Ank-binding protein. Coimmunoprecipitation assays validated the interaction of Shank3 with Nono in the mouse brain. In agreement with these data, the thalamus of Shank3a/b-knockout mice aberrantly expressed splicing isoforms of autism-associated genes, Nrxn1 and Eif4G1, before and after seizures with kainic acid treatment. These data indicate that Shank3 interacts with multiple RNA-binding proteins in the postnatal brain, thereby regulating the homeostatic expression of splicing isoforms for autism-associated genes after birth.

DOI： 10.1111/gtc.13142

Web of Science

Scopus

PubMed

Language：English   Publisher：Scientific Reports  

GNAO1 encodes G protein subunit alpha O1 (Gαo). Pathogenic variations in GNAO1 cause developmental delay, intractable seizures, and progressive involuntary movements from early infancy. Because the functional role of GNAO1 in the developing brain remains unclear, therapeutic strategies are still unestablished for patients presenting with GNAO1-associated encephalopathy. We herein report that siRNA-mediated depletion of Gnao1 perturbs the expression of transcripts associated with Rho GTPase signaling in Neuro2a cells. Consistently, siRNA treatment hampered neurite outgrowth and extension. Growth cone formation was markedly disrupted in monolayer neurons differentiated from iPSCs from a patient with a pathogenic variant of Gαo (p.G203R). This variant disabled neuro-spherical assembly, acquisition of the organized structure, and polarized signals of phospho-MLC2 in cortical organoids from the patient’s iPSCs. We confirmed that the Rho kinase inhibitor Y27632 restored these morphological phenotypes. Thus, Gαo determines the self-organizing process of the developing brain by regulating the Rho-associated pathway. These data suggest that Rho GTPase pathway might be an alternative target of therapy for patients with GNAO1-associated encephalopathy.

DOI： 10.1038/s41598-024-68062-x

Web of Science

Scopus

PubMed

Language：English   Publisher：Dmm Disease Models and Mechanisms  

Pathogenic variants in ATP1A3, the gene encoding the α3 subunit of the Na⁺/K⁺-ATPase, cause alternating hemiplegia of childhood (AHC) and related disorders. Impairments in Na⁺/K⁺-ATPase activity are associated with the clinical phenotype. However, it remains unclear whether additional mechanisms are involved in the exaggerated symptoms under stressed conditions in patients with AHC. We herein report that the intracellular loop (ICL) of ATP1A3 interacted with RNA-binding proteins, such as Eif4g (encoded by Eif4g1), Pabpc1 and Fmrp (encoded by Fmr1), in mouse Neuro2a cells. Both the siRNA-mediated depletion of Atp1a3 and ectopic expression of the p.R756C variant of human ATP1A3-ICL in Neuro2a cells resulted in excessive phosphorylation of ribosomal protein S6 (encoded by Rps6) and increased susceptibility to heat stress. In agreement with these findings, induced pluripotent stem cells (iPSCs) from a patient with the p.R756C variant were more vulnerable to heat stress than control iPSCs. Neurons established from the patient-derived iPSCs showed lower calcium influxes in responses to stimulation with ATP than those in control iPSCs. These data indicate that inefficient protein synthesis contributes to the progressive and deteriorating phenotypes in patients with the p.R756C variant among a variety of ATP1A3-related disorders.

DOI： 10.1242/dmm.050574

Web of Science

Scopus

PubMed

Language：English   Publisher：Biochemical and Biophysical Research Communications  

Melatonin entrainment of suprachiasmatic nucleus-regulating circadian rhythms is mediated by MT1 and MT2 receptors. Melatonin also has neuroprotective and mitochondrial activating effects, suggesting it may affect neurodevelopment. We studied melatonin's pharmacological effects on autism spectrum disorder (ASD) neuropathology. Deciduous tooth-derived stem cells from children with ASD were used to model neurodevelopmental defects and differentiated into dopaminergic neurons (ASD-DNs) with or without melatonin. Without melatonin, ASD-DNs had reduced neurite outgrowth, mitochondrial dysfunction, lower mitochondrial Ca²⁺ levels, and Ca²⁺ accumulation in the endoplasmic reticulum (ER) compared to control DNs from typically developing children-derived stem cells. Melatonin enhanced IP3-dependent Ca²⁺ release from ER to mitochondria, improving mitochondrial function and neurite outgrowth in ASD-DNs. Luzindole, an MT1/MT2 antagonist, blocked these effects. Thus, melatonin supplementation may improve dopaminergic system development in ASD by modulating mitochondrial Ca²⁺ homeostasis via MT1/MT2 receptors.

DOI： 10.1016/j.bbrc.2023.09.050

Web of Science

Scopus

PubMed

Language：Japanese  

Type：Competition, symposium, etc. 

Number of participants：500

Type：Competition, symposium, etc. 

Audience：General,　Scientific,　Company,　Civic organization,　Governmental agency

Type：Other

Audience：General,　Scientific,　Company,　Civic organization,　Governmental agency

Type：Lecture