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Porphyromonas gingivalis exacerbates ulcerative colitis via Porphyromonas gingivalis peptidylarginine deiminase

International Journal of oral Science 2021年第3期13卷 311-322页

作者： Xida Zhao Jingbo Liu Chong Zhang Ning Yu Ze Lu Shuwei Zhang Yuchao Li Qian Li Junchao Liu Dongjuan Liu Yaping Pan Department of Periodontology School and Hospital of StomatologyChina Medical UniversityLiaoning Provincial Key Laboratory of Oral DiseasesShenyangChina Center for Implant Dentistry School and Hospital of StomatologyChina Medical UniversityLiaoning Provincial Key Laboratory of Oral DiseaseShenyangChina The Forsyth Institute CambridgeMAUSA Department of Emergency and oral Medicine School and Hospital of StomatologyChina Medical UniversityLiaoning Provincial Key Laboratory of Oral DiseasesShenyangChina

Abstract:Ulcerative Colitis(UC)has been reported to be related to Porphyromonas gingivalis(P.gingivalis).Porphyromonas gingivalis peptidylarginine deiminase(PPAD),a virulence factor released by P.gingivalis,is known to induce inflammatory responses.To explore the pathological relationships between PPAD and UC,we used homologous recombination technology to construct a P.gingivalis strain in which the PPAD gene was deleted(Δppad)and aΔppad strain in which the PPAD gene was restored(comΔppad).C57 BL/6 mice were orally gavaged with saline,P.gingivalis,Δppad,or comΔppad twice a week for the entire 40 days(days 0-40),and then,UC was induced by dextran sodium sulfate(DSS)solution for 10 days(days 31-40).P.gingivalis and comΔppad exacerbated DDS-induced colitis,which was determined by assessing the parameters of colon length,disease activity index,and histological activity index,butΔppad failed to exacerbate DDS-induced colitis.Flow cytometry and ELISA revealed that compared withΔppad,P.gingivalis,and comΔppad increased T helper 17(Th17)cell numbers and interleukin(IL)-17 production but decreased regulatory T cells(Tregs)numbers and IL-10 production in the spleens of mice with UC.We also cocultured P.gingivalis,Δppad,or comΔppad with T lymphocytes in vitro and found that P.gingivalis and comΔppad significantly increased Th17 cell numbers and decreased Treg cell numbers.Immunofluorescence staining of colon tissue paraffin sections also confirmed these results.The results suggested that P.gingivalis exacerbated the severity of UC in part via PPAD.

关键词： gingivalis colitis colon

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Porphyromonas gingivalis bacteremia increases the permeability of the blood-brain barrier via the Mfsd2a/Caveolin-1 mediated transcytosis pathway

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International Journal of oral Science 2023年第1期15卷 136-147页

作者： Shuang Lei Jian Li Jingjun Yu Fulong Li Yaping Pan Xu Chen Chunliang Ma Weidong Zhao Xiaolin Tang Department of Periodontics School and Hospital of StomatologyLiaoning Provincial Key Laboratory of Oral DiseaseChina Medical UniversityShenyangChina Department of Pediatric Dentistry School and Hospital of StomatologyLiaoning Provincial Key Laboratory of Oral DiseaseChina Medical UniversityShenyangChina Department of Preventive Dentistry School and Hospital of StomatologyLiaoning Provincial Key Laboratory of Oral DiseaseChina Medical UniversityShenyangChina Department of Developmental Cell Biology Key Laboratory of Cell BiologyChina Medical UniversityShenyangChina

Bacteremia induced by periodontal infection is an important factor for periodontitis to threaten general health. P. gingivalis DNA/virulence factors have been found in the brain tissues from patients with Alzheimer’s disease(AD). The blood-brain barrier(BBB) is essential for keeping toxic substances from entering brain tissues. However, the effect of P. gingivalis bacteremia on BBB permeability and its underlying mechanism remains unclear. In the present study, rats were injected by tail vein with P. gingivalis three times a week for eight weeks to induce bacteremia. An in vitro BBB model infected with P. gingivalis was also established. We found that the infiltration of Evans blue dye and Albumin protein deposition in the rat brain tissues were increased in the rat brain tissues with P. gingivalis bacteremia and P. gingivalis could pass through the in vitro BBB model. Caveolae were detected after P. gingivalis infection in BMECs both in vivo and in vitro. Caveolin-1(Cav-1) expression was enhanced after P. gingivalis infection.Downregulation of Cav-1 rescued P. gingivalis-enhanced BMECs permeability. We further found P. gingivalis-gingipain could be colocalized with Cav-1 and the strong hydrogen bonding between Cav-1 and arg-specific-gingipain(RgpA) were detected.Moreover, P. gingivalis significantly inhibited the major facilitator superfamily domain containing 2a(Mfsd2a) expression. Mfsd2a overexpression reversed P. gingivalis-increased BMECs permeability and Cav-1 expression. These results revealed that Mfsd2a/Cav-1 mediated transcytosis is a key pathway governing BBB BMECs permeability induced by P. gingivalis, which may contribute to P. gingivalis/virulence factors entrance and the subsequent neurological impairments.

关键词： gingivalis expression inhibited

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Exosomes from bone marrow mesenchymal stem cells are a potential treatment for ischemic stroke

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Neural Regeneration Research 2023年第10期18卷 2246-2251页

作者： Chang Liu Tian-Hui Yang Hong-Dan Li Gong-Zhe Li Jia Liang Peng Wang liaoning provincial key laboratory of Neurodegenerative diseases and Department of Neurobiology Jinzhou Medical UniversityJinzhouLiaoning ProvinceChina Institute of Life Science Jinzhou Medical UniversityJinzhouLiaoning ProvinceChina College of Pharmacy Jinzhou Medical UniversityJinzhouLiaoning ProvinceChina

Exosomes derived from human bone marrow mesenchymal stem cells(MSC-Exo)are characterized by easy expansion and storage,low risk of tumor formation,low immunogenicity,and anti-inflammatory effects.The therapeutic effects of MSC-Exo on ischemic stroke have been widely explored.However,the underlying mechanism remains unclear.In this study,we established a mouse model of ischemic brain injury induced by occlusion of the middle cerebral artery using the thread bolt method and injected MSC-Exo into the tail vein.We found that administration of MSC-Exo reduced the volume of cerebral infarction in the ischemic brain injury mouse model,increased the levels of interleukin-33(IL-33)and suppression of tumorigenicity 2 receptor(ST2)in the penumbra of cerebral infarction,and improved neurological function.In vitro results showed that astrocyte-conditioned medium of cells deprived of both oxygen and glucose,to simulate ischemia conditions,combined with MSC-Exo increased the survival rate of primary cortical neurons.However,after transfection by IL-33 siRNA or ST2 siRNA,the survival rate of primary cortical neurons was markedly decreased.These results indicated that MSC-Exo inhibited neuronal death induced by oxygen and glucose deprivation through the IL-33/ST2 signaling pathway in astrocytes.These findings suggest that MSC-Exo may reduce ischemia-induced brain injury through regulating the IL-33/ST2 signaling pathway.Therefore,MSC-Exo may be a potential therapeutic method for ischemic stroke.

关键词： astrocytes bone marrow mesenchymal stem cells brain injury exosome IL-33 inflammation ischemic stroke neurological function neuron ST2

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A tetrahedral framework nucleic acid based multifunctional nanocapsule for tumor prophylactic mRNA vaccination

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Chinese Chemical Letters 2023年第7期34卷 169-175页

作者： Yuhao Liu Songhang Li Shiyu Lin Sirong Shi Taoran Tian Bowen Zhang Tao Zhang Yunfeng Lin State key laboratory of oral diseases National Clinical Research Center for Oral DiseasesWest China Hospital of StomatologySichuan University Chengdu 610041China key laboratory of oral Biomedical Research of Zhejiang Province Zhejiang UniversityHangzhou 31000China Guanghua School of Stomatology Guangdong Provincial Key Laboratory of StomatologySun Yat-sen UniversityGuangzhou 510055China

Synthetic antigen-encoding mRNA plays an increasingly significant role in tumor vaccine technology owing to its antigen-specific immune-activation. However, its immune efficacy is challenged by inferior delivery efficiency and demand for suitable adjuvants. Here, we develop a novel mRNA nanovaccine based on a multifunctional nanocapsule, which is a dual-adjuvant formulation composed of cytosine-phosphateguanine motifs loaded tetrahedral framework nucleic acid(CpG-tFNA) and an immunopeptide murine β-defensin 2(mDF2β). This m RNA nanovaccine successfully achieves intracellular delivery, antigen expression and presentation of dendritic cells, and proliferation of antigen-specific T cells. In a tumor prophylactic vaccination model, it exerts an excellent inhibitory effect on lymphoma occurrence through cellular immunity. This mRNA nanovaccine has promising prophylactic applications in tumors and many other diseases.

关键词： Tetrahedral framework nucleic acid(tFNA) RNA nanovaccine Cytosine-phosphate-guanine(CpG)motifs Murineβ-defensin 2(mDF2β) Tumor immunotherapy

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Targeting DNA-PKcs promotes anti-tumoral immunity via triggering cytosolic DNA sensing and inducing an inflamed tumor immuno-microenvironment in metastatic triple-negative breast cancer

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Genes & diseases 2023年第5期10卷 1809-1811页

作者： Chao Ma Yuanhua Qin Ying Wang Chuanliang Zhu Xingjie Gao Pingping Zhang Yupeng Gu Shuailong Zhang Jintao Lin Jiahui Wang Weifeng Mao College of Basic Medical Sciences Dalian Medical UniversityDalianLiaoning 116044China liaoning provincial key laboratory of Medical Molecular Biology DalianLiaoning 116044China

Triple-negative breast cancer(TNBC)is highly malignant and refractory to immunotherapy through impeding the immune cell infiltration and inflammation in the tumor microenvironment(TME).1 DNA-dependent protein kinase catalytic subunit(DNA-PKcs)is a member of the phosphatidylinositol 3-kinase-related kinase family,which is required for the non-homologous end joining repair.2 The effect of DNA-PKcs on the generation of cytosolic DNA and inflammation response in tumor immuno-environment is not defined.We found a specific DNA-PKcs inhibitor,NU7441,induced cytosolic DNA,stimulator of interferon genes(STING),and retinoic acid-inducible gene I(RIG-I)signals in vitro.In Balb/c immune-competent mice bearing 4T1 TNBC cells,NU7441 impaired the tumor growth and metastasis,and increased the CD45+leukocytes,CD4+T cells,CD8+T cells,and CD1a+antigen-presenting cells,as well as MHC-I and interferon alpha receptor(IFNAR)in TME.However,in Balb/c athymic nude mice without IFNAR and CD8+T cells in TME,NU7741 did not influence tumor growth.These results show that inhibition of DNA-PKcs triggers cytosolic DNA sensing and induces an inflamed TME to promote anti-tumoral immunity,which provides a strategy to alter the inflammation and lymphocyte infiltration in TME to increase the efficacy of immunotherapy in TNBC and other cancers with an immune-suppressive TME.

关键词： immunity inflammation interferon

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MicroRNA-146a-loaded magnesium silicate nanospheres promote bone regeneration in an inflammatory microenvironment

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Bone Research 2024年第1期12卷 153-164页

作者： Jiakang Yang Jing Shuai Lixuen Siow Jingyi Lu Miao Sun Wenyue An Mengfei Yu Baixiang Wang Qianming Chen Stomatology Hospital School of StomatologyZhejiang University School of MedicineZhejiang Provincial Clinical Research Center for Oral DiseasesKey Laboratory of Oral Biomedical Research of Zhejiang ProvinceCancer Center of Zhejiang UniversityHangzhou 310000China

Reconstruction of irregular oral-maxillofacial bone defects with an inflammatory microenvironment remains a challenge,as chronic local inflammation can largely impair bone healing.Here,we used magnesium silicate nanospheres(MSNs)to load microRNA-146a-5p(miR-146a)to fabricate a nanobiomaterial,MSN+miR-146a,which showed synergistic promoting effects on the osteogenic differentiation of human dental pulp stem cells(hDPSCs).In addition,miR-146a exhibited an anti-inflammatory effect on mouse bone marrow-derived macrophages(BMMs)under lipopolysaccharide(LPS)stimulation by inhibiting the NF-κB pathway via targeting tumor necrosis factor receptor-associated factor 6(TRAF6),and MSNs could simultaneously promote M2 polarization of BMMs.MiR-146a was also found to inhibit osteoclast formation.Finally,the dual osteogenic-promoting and immunoregulatory effects of MSN+miR-146a were further validated in a stimulated infected mouse mandibular bone defect model via delivery by a photocuring hydrogel.Collectively,the MSN+miR-146a complex revealed good potential in treating inflammatory irregular oralmaxillofacial bone defects.

关键词： inflammatory healing stimulation

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MBG/PGA-PCL composite scaffolds provide highly tunable degradation and osteogenic features

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Bioactive Materials 2022年第9期7卷 53-67页

作者： Jiangfeng Li Chunyi Wang Guoxing Gao Xing Yin Ximing Pu Bing Shi Yang Liu Zhongbing Huang Juan Wang Jingtao Li Guangfu Yin College of Biomedical Engineering Sichuan UniversityChengdu610065PR China State key laboratory of oral diseases National Clinical Research Center for Oral DiseasesDepartment of Oral and Maxillofacial SurgeryWest China Hospital of StomatologySichuan UniversityChengdu610041PR China State key laboratory of oral diseases National Clinical Research Center for Oral DiseasesDepartment of OrthodonticsWest China Hospital of StomatologySichuan UniversityChengdu610041PR China Shandong provincial key laboratory of Biomedical Polymers Shandong Academy of Pharmaceutical SciencesJinan250101PR China

It remains a challenge to achieve satisfactory balance between biodegradability and osteogenic capacity in biosynthetic bone grafts.In this study,we aimed to address this challenge by incorporating mesoporous bioactive glass(MBG)into poly(caprolactone-co-glycolide)(PGA-PCL)at gradient ratios.MBG/PGA-PCL(PGC/M)scaffolds with MBG incorporation ratio at 0,10%,25%and 40%(PGC/M0-40)were synthesized using a modified solvent casting-particulate leaching method,and their physiochemical and biological properties were comprehensively evaluated.PGC/M scaffolds exhibited highly perforated porous structure with a large-pore size of 300-450μm,with ordered MBGs of around 6.0 nm mesopores size uniformly dispersed.The increase in MBG incorporation ratio significantly improved the scaffold surface hydrophilicity,apatite-formation ability and pH stability,increased the weight loss rate while insignificantly influenced the molecular chains degradation of PGA-PCL component,and facilitated the attachment,spreading,viability and proliferation of rat bone marrow stromal cells(rBMSCs)on scaffolds.Moreover,rBMSCs cultured on PGC/M10-40 scaffolds demonstrated enhanced ALP activity and osteogenesis-related gene expression in a MBG dose-dependent manner as compared with those cultured on PGC/M0 scaffolds.When implanted to the rat cranial bone defect,PGC/M25 and PGC/M40 scaffolds induced significantly better bone repair as compared to PGC/M0 and PGC/M10 scaffolds.Besides,the biodegradability of PGC/M scaffolds correlated with the MBG incorporation ratio.These data suggested this novel PGC/M scaffolds as promising bone repair biomaterial with highly tunable hydrophilicity,bioactivity,cytocompatibility,osteogenic activity as well as biodegradability.

关键词： Mesoporous bioactive glasses Poly(caprolactone-co-glycolide) Osteogenic differentiation Bone repair

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oral microbiota accelerates corrosion of 316L stainless steel for orthodontic applications

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Journal of Materials Science & Technology 2022年第33期128卷 118-132页

作者： Qingfu Wang Mingxing Zhang Chuntian Yang Yi Yang Enze Zhou Pan Liu Daiqiang Jin Dake Xu Lin Wu Fuhui Wang School and Hospital of Stomatology China Medical UniversityLiaoning Provincial Key Laboratory of Oral DiseasesShenyang 110001China Shenyang National laboratory for Materials Science Northeastern UniversityShenyang 110819China Electrobiomaterials Institute Key Laboratory for Anisotropy and Texture of Materials(Ministry of Education)Northeastern UniversityShenyang 110819China Department of oral Implantology Tianjin Stomatological HospitalSchool of MedicineNankai University&Tianjin Key Laboratory of Oral and Maxillofacial Function ReconstructionTianjin 300041China

In this work,microbiologically influenced corrosion(MIC)of 316 L stainless steel(SS)caused by oral microbiota was investigated with HOMINGS 16 S rRNA gene sequencing technology,and electrochemical and surface analysis techniques.The results showed that oral microbiota from different subjects developed multi-species biofilms with significant differences in structure and composition of bacteria strains on the316 L SS coupons.In the presence of oral microbiota,more severe pitting corrosion and faster dissolution of metallic ions including Ni and Cr were observed.The biofilm considerably decreased the pitting potential of 316 L SS from 1268.0±29.1 mV vs.SCE(abiotic control)to less than 500 mV vs.SCE.The corrosion current density in the presence of oral microbiota from subject 1(115.3±83.3 nA cm^(-2))and subject 2(184.4±162.0 nA cm^(-2))was at least 4 times more than that in the abiotic medium(28.0±2.3 nA cm^(-2)).The electroactive microorganisms with the potential to facilitate corrosion via extracellular electron transfer found in oral microbiota may be mainly responsible for the accelerated corrosion.

关键词： oral microbiota Microbiologically influenced corrosion Stainless steel Electroactive microorganism

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Effects of surface roughening on antibacterial and osteogenic properties of Ti-Cu alloys with different Cu contents

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Journal of Materials Science & Technology 2021年第29期88卷 158-167页

作者： Wei Zhang Shuyuan Zhang Hui Liu Ling Ren Qiang Wang Yang Zhang School and Hospital of Stomatology China Medical UniversityShenyang 110001China Liaoning Provincial Key Laboratory of Oral Diseases Shenyang 110001China Institute of Metal Research Chinese Academy of SciencesShenyang 110016China

In order to reduce implant-related infections and improve early osseointegration,we performed HF+anodic oxidation on the surface of the new antibacterial Ti-Cu alloy to make the titanium-based implants have a micro/submicron structure.On this basis,a series of surface-modified Ti-x Cu alloys with different Cu contents(3,5 and 7 wt%)were fabricated and a comprehensive study was conducted on MC3 T3-E1 cell adhesion,proliferation,apoptosis and its antibacterial activity against Staphylococcus aureus.Results showed that the Ti-x Cu alloys by HF etching+anodized possessed multifunctional characteristics of antibiofilm and antibacterial abilities,excellent biocompatibility and osteogenesis promoting abilities.Increment of Cu content significantly contributed to the antibacterial and osteogenic properties of HF etching+anodized Ti-Cu alloys.Cell proliferation rates of HF etching+anodized Ti-7 Cu alloys were lower than those of Ti-3 Cu and Ti-5 Cu alloys,while the early cell apoptosis rates were higher than Ti-3 Cu and Ti-5 Cu groups.All the above results finally presented that the HF etching+anodized Ti-5 Cu alloy exhibited extremely strong antibacterial properties,good biological compatibility and osteogenic ability,as well as the most excellent ductility and corrosion resistance,providing a great potential application for the future dental implantation.

关键词： Ti-Cu Cu contents HF Antibacterial Osteogenesis

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Remodeling of the osteoimmune microenvironment after biomaterials implantation in murine tibia: Single-cell transcriptome analysis

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Bioactive Materials 2023年第4期22卷 404-422页

作者： Jia Li Congrui Zhao Yangbo Xu Lu Song Yanqi Chen Yuzi Xu Yang Ma Siyuan Wang Antian Xu Fuming He Department of Prosthodontics Stomatology HospitalSchool of StomatologyZhejiang University School of MedicineZhejiang Provincial Clinical Research Center for Oral DiseasesKey Laboratory of Oral Biomedical Research of Zhejiang ProvinceCancer Center of Zhejiang UniversityHangzhouChina

Osseointegration seems to be a foreign body reaction equilibrium due to the complicated interactions between the immune and skeletal systems.The heterogeneity of the osteoimmune microenvironment in the osseointegration of implant materials remains elusive.Here,a single-cell study involving 40043 cells is conducted,and a total of 10 distinct cell clusters are identified from five different groups.A preliminary description of the osteoimmune microenvironment revealed the diverse cellular heterogeneity and dynamic changes modulated by implant properties.The increased immature neutrophils,Ly6C+CCR2hi monocytes,and S100a8hi macrophages induce an aggressive inflammatory response and eventually lead to the formation of fibrous capsule around the stainless steel implant.The enrichment of mature neutrophils,FcgR1hi and differentiated immunomodulatory macrophages around the titanium implant indicates favorable osseointegration under moderate immune response.Neutrophil-depletion mice are conducted to explore the role of neutrophils in osseointegration.Neutrophils may improve bone formation by enhancing the recruitment of BMSCs via the CXCL12/CXCR3 signal axis.These findings contribute to a better knowledge of osteoimmunology and are valuable for the design and modification of‘osteoimmune-smart’biomaterials in the bone regeneration field.

关键词： Osteoimmunology Neutrophil Implant Osseointegration Single-cell transcriptomics

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