The functional regeneration of the dentin-pulp complex is pivotal for tooth preservation,yet the molecular mechanisms governing odontoblast differentiation remain poorly understood.In the current study,we revealed a d...The functional regeneration of the dentin-pulp complex is pivotal for tooth preservation,yet the molecular mechanisms governing odontoblast differentiation remain poorly understood.In the current study,we revealed a distinct NKD1^(+) subpopulation exhibiting secretory odontoblast characteristics,which was specifically induced in dental pulp stem cells(DPSCs) by Wnt3a,but not by Wnt5a or Wnt10a through single-cell transcriptomic profiling.We then found that the NKD1^(+) subpopulation was functional conservation,which were consistently identified in the odontoblast layers of developing tooth germs in both murine and miniature pig models,as well as within the apical open area in human molars.This conserved spatial distribution and co-localization with DSPP strongly indicates that NKD1^(+) cells were active dentin-secreting odontoblasts.Analysis of gene regulatory networks using SCENIC identified MSX1 as a key transcription factor regulating the specification of NKD1^(+) lineage.Mechanistically,Wnt3a orchestrates a tripartite cascade:upregulating NKD1/MSX1 expression,triggering NKD1 membrane detachment,and facilitating direct NKD1-MSX1interaction to promote MSX1 nuclear translocation.CUT&Tag analysis demonstrated MSX1 occupancy at promoters of odontogenic regulato rs,esta blishing its necessity for odontogenic gene activation.Murine pulp exposure models validated that Wnt3a-activated NKD1-MSX1 signaling significantly enhances reparative dentin formation.This study delineates an evolutionarily conserved Wnt3aNKD1-MSX1 axis that resolves stem cell heterogeneity into functional odontoblast commitment,providing both mechanistic insights into dentin-pulp regeneration and a foundation for targeted regenerative therapies.展开更多
Periodontitis is caused by overactive osteoclast activity that results in the loss of periodontal supporting tissue and mesenchymal stem cells(MSCs)are essential for periodontal regeneration.However,the hypoxic period...Periodontitis is caused by overactive osteoclast activity that results in the loss of periodontal supporting tissue and mesenchymal stem cells(MSCs)are essential for periodontal regeneration.However,the hypoxic periodontal microenvironment during periodontitis induces the apoptosis of MSCs.Apoptotic bodies(ABs)are the major product of apoptotic cells and have been attracting increased attention as potential mediators for periodontitis treatment,thus we investigated the effects of ABs derived from MSCs on periodontitis.MSCs were derived from bone marrows of mice and were cultured under hypoxic conditions for 72 h,after which ABs were isolated from the culture supernatant using a multi-filtration system.The results demonstrate that ABs derived from MSCs inhibited osteoclast differentiation and alveolar bone resorption.miRNA array analysis showed that miR-223-3p is highly enriched in those ABs and is critical for their therapeutic effects.Targetscan and luciferase activity results confirmed that Itgb1 is targeted by miR-223-3p,which interferes with the function of osteoclasts.Additionally,DC-STAMP is a key regulator that mediates membrane infusion.ABs and pre-osteoclasts expressed high levels of DC-STAMP on their membranes,which mediates the engulfment of ABs by pre-osteoclasts.ABs with knock-down of DC-STAMP failed to be engulfed by pre-osteoclasts.Collectively,MSC-derived ABs are targeted to be engulfed by pre-osteoclasts via DC-STAMP,which rescued alveolar bone loss by transferring miR-223-3p to osteoclasts,which in turn led to the attenuation of their differentiation and bone resorption.These results suggest that MSC-derived ABs are promising therapeutic agents for the treatment of periodontitis.展开更多
Bone regeneration is a major clinical challenge.The main obstacles to bone injury repair are local blood flow disorders and hypoxic microenvironments.Mesenchymal stem cell(MSC)therapy has notable advantages in promoti...Bone regeneration is a major clinical challenge.The main obstacles to bone injury repair are local blood flow disorders and hypoxic microenvironments.Mesenchymal stem cell(MSC)therapy has notable advantages in promoting bone-tissue regeneration.In this study,we established a mouse model of skull bone injury treated with bone marrow mesenchymal stem cells(BMSCs).We found that local BMSC transplantation stimulated vascularized bone regeneration and matrix metalloproteinase(MMP)10 was the major regulatory protein.Local hypoxic microenvironment-induced mitochondrial permeability increased,resulting in cytoplasmic Zn^(2+)accumulation,which is a key factor in activating the JAK1/STAT1/MMP-10 pathway.The cytoplasmic Zn^(2+)enrichment caused ZRT/IRT-like protein 6(ZIP6)inhibition was the key initiating factor in this process.Based on these findings,we designed and engineered CD90@ZIF-8-ICG,with an outer membrane chimeric CD90 antibody containing indocyanine green(ICG),to achieve increased intracellular zinc ion content by targeted delivery of the particles into local MSCs,so that local MMP-10 production and angiogenesis are regulated at the early stage of healing.ICG provided BMSCs with continuous photothermal stimulation in response to the laser intervention,which successfully achieved stable improvement of bone-defect regeneration.This study innovatively describes the regulatory importance of intracellular zinc ion homeostasis and ZIP proteins in the function of transplanted MSCs,as well as the related efficiency strategy development,which elucidates MSC therapy treatment mechanisms and provides strategies for the design and development of stem-cell-based biomaterials.展开更多
The chronic inflammation in periodontitis suppresses the osteogenic potential of human periodontal ligament stem cells(hPDLSCs),posing a significant challenge to endogenous bone regeneration.To address this,we develop...The chronic inflammation in periodontitis suppresses the osteogenic potential of human periodontal ligament stem cells(hPDLSCs),posing a significant challenge to endogenous bone regeneration.To address this,we developed an osteogenic and protein-delivery composite hydrogel system based on metformin carbon dots(MCDs)to enhance the osteogenic potential of hPDLSCs under inflammatory conditions.We successfully synthesized a novel Gel/MCDs@IGF-1 composite hydrogel(Gel)that exhibited excellent biocompatibility and sequentially released MCDs and insulin-like growth factor 1(IGF-1).First,MCDs were synthesized using a onestep hydrothermal method.MCDs promote the osteogenic differentiation of hPDLSCs under lipopolysaccharide(LPS)-induced inflammatory conditions by activating the PI3K/AKT signaling pathway,and alleviate inflammation.Next,MCDs and IGF-1 were assembled into MCDs@IGF-1 complexes through supramolecular interactions,facilitating efficient IGF-1 delivery and reducing its degradation by trypsin.Furthermore,in vitro and in vivo studies demonstrated that the Gel/MCDs@IGF-1 composite hydrogel effectively recruited stem cells,exerted early anti-inflammatory effects,increased the osteogenesis of hPDLSCs under inflammatory conditions,and significantly promoted alveolar bone regeneration in a Sprague-Dawley(SD)rat model of periodontitis.In conclusion,MCDs,with their dual roles in promoting osteogenesis and protein delivery,are a promising candidate nanoplatform for periodontitis therapy.Additionally,the MCDs-based sequential release hydrogel system presents a novel material strategy for the treatment of periodontitis.展开更多
Delayed wound healing in diabetes is a global challenge,and the development of related drugs is a clinical problem to be solved.In this study,purpurolide C(PC),a small-molecule secondary metabolite of the endophytic f...Delayed wound healing in diabetes is a global challenge,and the development of related drugs is a clinical problem to be solved.In this study,purpurolide C(PC),a small-molecule secondary metabolite of the endophytic fungus Penicillium purpurogenum,was found to promote diabetic wound healing.To investigate the key regulation targets of PC,in vitro RNA-seq,molecular docking calcula-tions,TLR4-MD2 dimerization SDS-PAGE detection,and surface plasmon resonance(SPR)were per-formed,indicating that PC inhibited inflammatory macrophage activation by inhibiting both TLR4-MD2 dimerization and MYD88 phosphorylation.Tlr4 knockout in vivo attenuated the promotion effect of PC on wound healing.Furthermore,a delivery system consisting of macrophage liposome and GelMA-based microneedle patches combined with PC(PC@MLIP MN)was developed,which overcame the poor water solubility and weak skin permeability of PC,so that successfully punctured the skin and delivered PC to local tissues,and accurately regulated macrophage polarization in diabetic wound management.Overall,PC is an anti-inflammatory small molecule compound with a well-defined structure and dualtarget regulation,and the PC@MLIP MN is a promising novel biomaterial for the management of diabetic wound.展开更多
Concentration of salivary nitrate is approximately 10-fold to that of serum. Many circumstances such as acute stress could promote salivary nitrate secretion and nitrite formation. However, whether other conditions ca...Concentration of salivary nitrate is approximately 10-fold to that of serum. Many circumstances such as acute stress could promote salivary nitrate secretion and nitrite formation. However, whether other conditions can also be used as regulators of salivary nitrate/nitrite has not yet been explored. The present study was designed to determine the influence of exposure to different music on the salivary flow rate and nitrate secretion and nitrite formation. Twenty-four undergraduate students(12 females and 12 males) were exposed to silence, rock music, classical music or white noise respectively on four consecutive mornings. The unstimulated salivary flow rate and stimulated salivary flow rate were measured. Salivary ionic(Na+, Ca2+Cl-,and PO3-4) content and nitrate/nitrite levels were detected. The unstimulated salivary flow rate was significantly increased after classical music exposure compared to that after silence. Salivary nitrite levels were significantly higher upon classical music and white noise stimulation than those under silence in females. However, males were more sensitive only to white noise with regard to the nitrite increase. In conclusion, this study demonstrated that classical music stimulation promotes salivary nitrite formation and an increase in saliva volume was observed. These observations may play an important role in regulating oral function.展开更多
基金supported by the National Natural Science Foundation of China(82170951,82470961)the Beijing Natural Science Foundation (7222079)+4 种基金the Beijing Hospital Authority"Dengfeng"Talent Training Plan (DFL 20221301)the Beijing Stomatological HospitalCapital Medical University Young Scientist Program (No.YSP202401)the Laboratory for Clinical Medicine and the Central Laboratory of Capital Medical University for their technical support and fundingthe Japan China Sasakawa Medical Fellowship for their generous support and funding。
文摘The functional regeneration of the dentin-pulp complex is pivotal for tooth preservation,yet the molecular mechanisms governing odontoblast differentiation remain poorly understood.In the current study,we revealed a distinct NKD1^(+) subpopulation exhibiting secretory odontoblast characteristics,which was specifically induced in dental pulp stem cells(DPSCs) by Wnt3a,but not by Wnt5a or Wnt10a through single-cell transcriptomic profiling.We then found that the NKD1^(+) subpopulation was functional conservation,which were consistently identified in the odontoblast layers of developing tooth germs in both murine and miniature pig models,as well as within the apical open area in human molars.This conserved spatial distribution and co-localization with DSPP strongly indicates that NKD1^(+) cells were active dentin-secreting odontoblasts.Analysis of gene regulatory networks using SCENIC identified MSX1 as a key transcription factor regulating the specification of NKD1^(+) lineage.Mechanistically,Wnt3a orchestrates a tripartite cascade:upregulating NKD1/MSX1 expression,triggering NKD1 membrane detachment,and facilitating direct NKD1-MSX1interaction to promote MSX1 nuclear translocation.CUT&Tag analysis demonstrated MSX1 occupancy at promoters of odontogenic regulato rs,esta blishing its necessity for odontogenic gene activation.Murine pulp exposure models validated that Wnt3a-activated NKD1-MSX1 signaling significantly enhances reparative dentin formation.This study delineates an evolutionarily conserved Wnt3aNKD1-MSX1 axis that resolves stem cell heterogeneity into functional odontoblast commitment,providing both mechanistic insights into dentin-pulp regeneration and a foundation for targeted regenerative therapies.
基金grants from National Key R&D Program of China(Grant No.2022YFC2504200)the National Nature Science Foundation of China(81991504 and 81974149 to Y.L.+7 种基金82201052 to X.Y.L.)the Beijing Municipal Administration of Hospitals Clinical Medicine Development of Special Funding Support(ZYLX202121 to Y.L.)the Innovation Research Team Project of Beijing Stomatological Hospital,Capital Medical University(CXTD202202)the Beijing Municipal Administration of Hospitals’Ascent Plan(DFL20181501 to Y.L.)the Beijing Municipal Administration of Hospitals’Youth Programme(QML20181501 to L.J.G.QML20231505 to X.Y.L.)the Beijing Stomatological Hospital,Capital Medical University Young Scientist Program(No.YSP202103 to X.Y.L.)the Innovation Foundation of Beijing Stomatological Hospital,Capital Medical University(21-09-18 to L.J.G.).
文摘Periodontitis is caused by overactive osteoclast activity that results in the loss of periodontal supporting tissue and mesenchymal stem cells(MSCs)are essential for periodontal regeneration.However,the hypoxic periodontal microenvironment during periodontitis induces the apoptosis of MSCs.Apoptotic bodies(ABs)are the major product of apoptotic cells and have been attracting increased attention as potential mediators for periodontitis treatment,thus we investigated the effects of ABs derived from MSCs on periodontitis.MSCs were derived from bone marrows of mice and were cultured under hypoxic conditions for 72 h,after which ABs were isolated from the culture supernatant using a multi-filtration system.The results demonstrate that ABs derived from MSCs inhibited osteoclast differentiation and alveolar bone resorption.miRNA array analysis showed that miR-223-3p is highly enriched in those ABs and is critical for their therapeutic effects.Targetscan and luciferase activity results confirmed that Itgb1 is targeted by miR-223-3p,which interferes with the function of osteoclasts.Additionally,DC-STAMP is a key regulator that mediates membrane infusion.ABs and pre-osteoclasts expressed high levels of DC-STAMP on their membranes,which mediates the engulfment of ABs by pre-osteoclasts.ABs with knock-down of DC-STAMP failed to be engulfed by pre-osteoclasts.Collectively,MSC-derived ABs are targeted to be engulfed by pre-osteoclasts via DC-STAMP,which rescued alveolar bone loss by transferring miR-223-3p to osteoclasts,which in turn led to the attenuation of their differentiation and bone resorption.These results suggest that MSC-derived ABs are promising therapeutic agents for the treatment of periodontitis.
基金supported by grants from the National Nature Science Foundation of China(82201053,81991504,81974149,82470936)National Key R&D Program of China(Grant NO.2022YFC2504200)+6 种基金the Innovation Research Team Project of Beijing Stomatological Hospital,Capital Medical University(CXTD202202)the Innovation Foundation of Beijing Stomatological Hospital,Capital Medical University(21-09-18)the Beijing Stomatological Hospital,Capital Medical University Young Scientist Program(YSP202105)the Beijing Municipal Administration of Hospitals Clinical Medicine Development of Special Funding Support(ZYLX202121)Beijing Municipal Administration of Hospitals’Ascent Plan(DFL20181501)Beijing Municipal Administration of Hospitals’Youth Programme(QML20231506,QML20181501)Beijing Municipal Administration of Hospitals Incubating Program(PX2023054).
文摘Bone regeneration is a major clinical challenge.The main obstacles to bone injury repair are local blood flow disorders and hypoxic microenvironments.Mesenchymal stem cell(MSC)therapy has notable advantages in promoting bone-tissue regeneration.In this study,we established a mouse model of skull bone injury treated with bone marrow mesenchymal stem cells(BMSCs).We found that local BMSC transplantation stimulated vascularized bone regeneration and matrix metalloproteinase(MMP)10 was the major regulatory protein.Local hypoxic microenvironment-induced mitochondrial permeability increased,resulting in cytoplasmic Zn^(2+)accumulation,which is a key factor in activating the JAK1/STAT1/MMP-10 pathway.The cytoplasmic Zn^(2+)enrichment caused ZRT/IRT-like protein 6(ZIP6)inhibition was the key initiating factor in this process.Based on these findings,we designed and engineered CD90@ZIF-8-ICG,with an outer membrane chimeric CD90 antibody containing indocyanine green(ICG),to achieve increased intracellular zinc ion content by targeted delivery of the particles into local MSCs,so that local MMP-10 production and angiogenesis are regulated at the early stage of healing.ICG provided BMSCs with continuous photothermal stimulation in response to the laser intervention,which successfully achieved stable improvement of bone-defect regeneration.This study innovatively describes the regulatory importance of intracellular zinc ion homeostasis and ZIP proteins in the function of transplanted MSCs,as well as the related efficiency strategy development,which elucidates MSC therapy treatment mechanisms and provides strategies for the design and development of stem-cell-based biomaterials.
基金sponsored by Natural Science Foundation of Chongqing,China(Grant No.CSTB2022BSXM-JCX0047)the National Key Research&Development of Program of China(Grant No.2022YFC2504200)the National Natural Science Foundation of China(Grant No.82122015).
文摘The chronic inflammation in periodontitis suppresses the osteogenic potential of human periodontal ligament stem cells(hPDLSCs),posing a significant challenge to endogenous bone regeneration.To address this,we developed an osteogenic and protein-delivery composite hydrogel system based on metformin carbon dots(MCDs)to enhance the osteogenic potential of hPDLSCs under inflammatory conditions.We successfully synthesized a novel Gel/MCDs@IGF-1 composite hydrogel(Gel)that exhibited excellent biocompatibility and sequentially released MCDs and insulin-like growth factor 1(IGF-1).First,MCDs were synthesized using a onestep hydrothermal method.MCDs promote the osteogenic differentiation of hPDLSCs under lipopolysaccharide(LPS)-induced inflammatory conditions by activating the PI3K/AKT signaling pathway,and alleviate inflammation.Next,MCDs and IGF-1 were assembled into MCDs@IGF-1 complexes through supramolecular interactions,facilitating efficient IGF-1 delivery and reducing its degradation by trypsin.Furthermore,in vitro and in vivo studies demonstrated that the Gel/MCDs@IGF-1 composite hydrogel effectively recruited stem cells,exerted early anti-inflammatory effects,increased the osteogenesis of hPDLSCs under inflammatory conditions,and significantly promoted alveolar bone regeneration in a Sprague-Dawley(SD)rat model of periodontitis.In conclusion,MCDs,with their dual roles in promoting osteogenesis and protein delivery,are a promising candidate nanoplatform for periodontitis therapy.Additionally,the MCDs-based sequential release hydrogel system presents a novel material strategy for the treatment of periodontitis.
基金supported by grants from National Key R&D Program of China(Grant NO.2022YFC2504200 to Yi Liu)the National Nature Science Foundation of China(81991504 and 81974149 to Yi Liu,82073978 to Sheng Lin,82201053 to Yitong Liu,81803397 to Guiyang Xia,82122015 to Junji Xu)+6 种基金the Innovation Research Team Project of Beijing Stomatological Hospital,Capital Medical University(CXTD202202 to Yi Liu,China)the Innovation Foundation of Beijing Stomatological Hospital,Capital Medical University(21-09-18 to Lijia Guo,China)the Beijing Stomatological Hospital,Capital Medical University Young Scientist Program(YSP202105 to Yitong Liu,China)the Beijing Municipal Administration of Hospitals Clinical Medicine Development of Special Funding Support(ZYLX202121 to Yi Liu,China)Beijing Municipal Administration of Hospitals'Ascent Plan(DFL20181501 to Yi Liu,China)Beijing Municipal Administration of Hospitals'Youth Programme(QML20181501 to Lijia Guo,QML20231506 to Yitong Liu,China)Beijing Municipal Administration of Hospitals Incubating Program(PX2023054 to Lijia Guo,China).
文摘Delayed wound healing in diabetes is a global challenge,and the development of related drugs is a clinical problem to be solved.In this study,purpurolide C(PC),a small-molecule secondary metabolite of the endophytic fungus Penicillium purpurogenum,was found to promote diabetic wound healing.To investigate the key regulation targets of PC,in vitro RNA-seq,molecular docking calcula-tions,TLR4-MD2 dimerization SDS-PAGE detection,and surface plasmon resonance(SPR)were per-formed,indicating that PC inhibited inflammatory macrophage activation by inhibiting both TLR4-MD2 dimerization and MYD88 phosphorylation.Tlr4 knockout in vivo attenuated the promotion effect of PC on wound healing.Furthermore,a delivery system consisting of macrophage liposome and GelMA-based microneedle patches combined with PC(PC@MLIP MN)was developed,which overcame the poor water solubility and weak skin permeability of PC,so that successfully punctured the skin and delivered PC to local tissues,and accurately regulated macrophage polarization in diabetic wound management.Overall,PC is an anti-inflammatory small molecule compound with a well-defined structure and dualtarget regulation,and the PC@MLIP MN is a promising novel biomaterial for the management of diabetic wound.
基金supported by the Beijing Municipal Government grant(Beijing Laboratory of Oral Health,PXM2021-014226000041)the Beijing Municipal Science and Technology Commission(Z181100001718208)+7 种基金the Beijing Municipal Education Commission(119207020201)the Innovation Research Team Project of Beijing Stomatological Hospital,Capital Medical University(CXTD202201)the Chinese Research Unit of Tooth Development and Regeneration,Academy of Medical Sciences(2019-12M-5031)the National Natural Science Foundation of China(92049201,82030031,81991504,and 92149301)the Beijing Advanced Innovation Center for Big Data-based Precision Medicine(PXM2021_014226_000026)the Beijing Municipal Government(Beijing Scholar Program,PXM2020_014226_000005 and PXM2021_014226_000020)the Beijing Municipal Colleges and Universities High Level Talents Introduction and Cultivate Project-Beijing Great Wall Scholar Program(CIT&TCD 20180332)the National Key Research and development Program(2022YFA1104401)。
基金supported by the National Natural Science Foundation of China (81400527, 91649124, 81300896)the Beijing Municipality Government (Beijing Scholar Program PXM2016_014226_000034, PXM2016_014226_000006, PXM2015_014226_000116, PXM2015_ 014226_000055, PXM2015_014226_000052, PXM2014_014226_000048, PXM2014_014226_000013, PXM2014_014226_000053, Z121100005212004, PXM2013_014226_000055, PXM2013_014226_000021, PXM2013_014226_07_000080 and TJSHG201310025005), the 2016 QNRC001 Young Elite Scientist Sponsorship Program by CAST (2016 QNRC001)+2 种基金the Beijing Municipal Administration of Hospitals’ Youth Programme (QML20151402)Beijing Municipal Natural Science Foundation (7142069)Beijing NOVA program (2015B062 to Junji Xu)
文摘Concentration of salivary nitrate is approximately 10-fold to that of serum. Many circumstances such as acute stress could promote salivary nitrate secretion and nitrite formation. However, whether other conditions can also be used as regulators of salivary nitrate/nitrite has not yet been explored. The present study was designed to determine the influence of exposure to different music on the salivary flow rate and nitrate secretion and nitrite formation. Twenty-four undergraduate students(12 females and 12 males) were exposed to silence, rock music, classical music or white noise respectively on four consecutive mornings. The unstimulated salivary flow rate and stimulated salivary flow rate were measured. Salivary ionic(Na+, Ca2+Cl-,and PO3-4) content and nitrate/nitrite levels were detected. The unstimulated salivary flow rate was significantly increased after classical music exposure compared to that after silence. Salivary nitrite levels were significantly higher upon classical music and white noise stimulation than those under silence in females. However, males were more sensitive only to white noise with regard to the nitrite increase. In conclusion, this study demonstrated that classical music stimulation promotes salivary nitrite formation and an increase in saliva volume was observed. These observations may play an important role in regulating oral function.
