Treated dentin matrix(TDM)is an ideal scaffold material containing multiple extracellular matrix factors.The canonical Wnt signaling pathway is necessary for tooth regeneration.Thus,this study investigated whether the...Treated dentin matrix(TDM)is an ideal scaffold material containing multiple extracellular matrix factors.The canonical Wnt signaling pathway is necessary for tooth regeneration.Thus,this study investigated whether the TDM can promote the odontogenic differentiation of human dental pulp stem cells(hDPSCs)and determined the potential role of Wnt/β-catenin signaling in this process.Different concentrations of TDM promoted the dental differentiation of the hDPSCs and meanwhile,the expression of GSK3βwas decreased.Of note,the expression of the Wnt/β-catenin pathway-related genes changed significantly in the context of TDM induction,as per RNA sequencing(RNA seq)data.In addition,the experiment showed that new dentin was visible in rat mandible cultured with TDM,and the thickness was significantly thicker than that of the control group.In addition,immunohistochemical staining showed lower GSK3βexpression in new dentin.Consistently,the GSK3βknockdown hDPSCs performed enhanced odotogenesis compared with the control groups.However,GSK3βoverexpressing could decrease odotogenesis of TDM-induced hDPSCs.These results were confirmed in immunodeficient mice and Wistar rats.These suggest that TDM promotes odontogenic differentiation of hDPSCs by directly targeting GSK3βand activating the canonical Wnt/β-catenin signaling pathway and provide a theoretical basis for tooth regeneration engineering.展开更多
It is still a huge challenge for bone regenerative biomaterial to balance its mechanical,biological and biodegradable properties.In the present study,a new composite material including treated dentin matrix(TDM)andα-...It is still a huge challenge for bone regenerative biomaterial to balance its mechanical,biological and biodegradable properties.In the present study,a new composite material including treated dentin matrix(TDM)andα-calcium sulphate hemihydrate(α-CSH)was prepared.The optimal composition ratio between TDM andα-CSH was explored.The results indicate that both components were physically mixed and structurally stable.Its compressive strength reaches up to 5.027±0.035 MPa for 50%TDM/α-CSH group,similar to human cancellous bone tissues.Biological experiments results show that TDM/α-CSH composite exhibits excellent biocompatibility and the expression of osteogenic related genes and proteins(ALP,RUNX2,OPN)is significantly increased.In vivo experiments suggest that the addition of TDM for each group(10%,30%,50%)effectively promotes cell proliferation and osteomalacia.In addition,50%of the TDM/α-CSH combination displays optimal osteoconductivity.The novel TDM/α-CSH composite is a good candidate for certain applications in bone tissue engineering.展开更多
The invasion of etched dentinal tubules(DTs)by external substances induces dentin hypersensitivity(DH).The deep and compact occlusion of DTs is highly desirable for treating DH but still challenging due to the limited...The invasion of etched dentinal tubules(DTs)by external substances induces dentin hypersensitivity(DH).The deep and compact occlusion of DTs is highly desirable for treating DH but still challenging due to the limited penetrability and mineralization capacities of most current desensitizers.Matrix vesicles(MVs)participate in the regulation of ectopic mineralization.Herein,ectopic MV analogs are prepared by employing natural cell membranes to endow mineral precursors with natural biointerfaces and integrated biofunctions for stimulating dentin remineralization.The analogs quickly access DTs(>20μm)in only 5 min and further penetrate deep into the interior of DTs(an extraordinary~200μm)in 7 days.Both in vitro and in vivo studies confirm that the DTs are efficiently sealed by the newly formed minerals(>50μm)with excellent resistance to wear and acid erosion,which is significantly deeper than most reported values.After repair,the microhardness of the damaged dentin can be recovered to those of healthy dentin.For the first time,cell membrane coating nanotechnology is used as a facile and efficient therapy for in-depth remineralization of DTs in treating DH with thorough and long-term effects,which provides insights into their potential for hard tissue repair.展开更多
目的研究参与两种粘接剂形成的牙本质粘接界面退变过程中基质金属蛋白酶(MMPs)的种类。方法制备Single Bond 2和Clearfi l S3 Bond微剪切粘接试件,无菌人工唾液中分别浸泡0、6个月测试粘接强度,场发射扫描电镜(FE-SEM)观察界面断裂模式...目的研究参与两种粘接剂形成的牙本质粘接界面退变过程中基质金属蛋白酶(MMPs)的种类。方法制备Single Bond 2和Clearfi l S3 Bond微剪切粘接试件,无菌人工唾液中分别浸泡0、6个月测试粘接强度,场发射扫描电镜(FE-SEM)观察界面断裂模式。制备4 mm×3 mm×1 mm牙本质片,分为对照组、Single Bond 2组和Clearfi l S3Bond组,无菌人工唾液浸泡0、6个月后,液相芯片技术测定各组MMP-1、-2、-3、-8、-9含量。结果老化6个月后,各组粘接强度显著下降。与对照组相比,老化过程中不同粘接剂组MMP-1、-3含量无明显变化,Single Bond 2组MMP-2、-8、-9含量显著降低,Clearfi l S3 Bond组MMP-8、-9含量显著降低。结论老化6个月后两种粘接试件均发生粘接界面退变,牙本质源MMP-2、-8、-9的含量显著降低,提示这些MMPs可能参与了牙本质粘接界面的退变。展开更多
目的:探讨不同粘结条件下健康成人牙本质中基质金属蛋白酶2 (MMP-2)和基质金属蛋白酶9 (MMP-9)表达水平,为临床应用基质金属蛋白酶(MMPs)抑制剂提高粘结效果提供依据。方法:收集20颗成人新鲜离体磨牙,液氮冷却条件下将其研磨为牙本质粉...目的:探讨不同粘结条件下健康成人牙本质中基质金属蛋白酶2 (MMP-2)和基质金属蛋白酶9 (MMP-9)表达水平,为临床应用基质金属蛋白酶(MMPs)抑制剂提高粘结效果提供依据。方法:收集20颗成人新鲜离体磨牙,液氮冷却条件下将其研磨为牙本质粉;模拟口内条件,对牙本质分别进行自酸蚀粘结(Single Bond Universal)和全酸蚀粘结(35%磷酸凝胶+Adpter Single Bond 2)处理,以不进行任何处理的牙本质作为阴性对照组,10%磷酸溶液(自酸蚀粘结)或10%磷酸溶液+35%磷酸凝胶处理的脱矿牙本质粉(全酸蚀粘结)处理的牙本质作为阳性对照组,只进行Single Bond Universal (自酸蚀粘结)处理和35%磷酸凝胶+Adpter Single Bone 2 (全酸蚀粘结)处理的牙本质作为空白对照组;在粘结过程中分别使用MMPs抑制剂氯己定(CHX)和米诺环素(MI)预处理,作为CHX预处理组和MI预处理组;采用10%次氯酸钠进行牙本质老化,作为老化组,并在老化组基础上加入CHX和MI作为CHX预处理老化组和MI预处理老化组。采用明胶酶谱法进行聚丙烯酰胺凝胶电泳,孵育染色脱色后,采用凝胶图像分析系统分析条带,计算各组牙本质中MMP-2和MMP-9表达水平。结果:在自酸蚀粘结条件下,与空白对照组比较,CHX预处理组牙本质中MMP-2和MMP-9表达水平降低(P<0.05),MI预处理组牙本质中MMP-2和MMP-9表达水平降低(P<0.05);与空白老化对照组比较,CHX预处理老化组牙本质中MMP-2和MMP-9表达水平降低(P<0.05),MI预处理老化组牙本质中MMP-2和MMP-9表达水平降低(P<0.05)。在全酸蚀粘结条件下,与空白对照组比较,CHX预处理组和MI预处理组牙本质中MMP-2表达水平降低(P<0.05);与空白老化对照组比较,CHX预处理老化组与MI预处理老化组牙本质中MMP-2表达水平降低(P<0.05)。结论:在牙本质粘结过程中,CHX和MI能够通过降低MMP-2和MMP-9表达水平减缓酶解反应,从而增强粘结强度。展开更多
Fractures are frequently occurring diseases that endanger human health. Crucial to fracture healing is cartilage formation, which provides a bone-regeneration environment. Cartilage consists of both chondrocytes and e...Fractures are frequently occurring diseases that endanger human health. Crucial to fracture healing is cartilage formation, which provides a bone-regeneration environment. Cartilage consists of both chondrocytes and extracellular matrix (ECM). The ECM of cartilage includes collagens and various types of proteoglycans (PGs), which play important roles in maintaining primary stability in fracture healing. The PG form of dentin matrix protein 1 (DMP1-PG) is involved in maintaining the health of articular cartilage and bone. Our previous data have shown that DMP1-PG is richly expressed in the cartilaginous calluses of fracture sites. However, the possible significant role of DMP1-PG in chondrogenesis and fracture healing is unknown. To further detect the potential role of DMP1-PG in fracture repair, we established a mouse fracture model by using a glycosylation site mutant DMP1 mouse (S89G-DMP1 mouse). Upon inspection, fewer cartilaginous calluses and down-regulated expression levels of chondrogenesis genes were observed in the fracture sites of S89G-DMP1 mice. Given the deficiency of DMP1-PG, the impaired IL-6/JAK/STAT signaling pathway was observed to affect the chondrogenesis of fracture healing. Overall, these results suggest that DMP1-PG is an indispensable proteoglycan in chondrogenesis during fracture healing.展开更多
The tooth root cementum is a thin, mineralized tissue covering the root dentin that is present primarily as acellular cementum on the cervical root and cellular cementum covering the apical root. While cementum shares...The tooth root cementum is a thin, mineralized tissue covering the root dentin that is present primarily as acellular cementum on the cervical root and cellular cementum covering the apical root. While cementum shares many properties in common with bone and dentin, it is a unique mineralized tissue and acellular cementum is critical for attachment of the tooth to the surrounding periodontal ligament （PDL）. Resources for methodologies for hard tissues often overlook cementum and approaches that may be of value for studying this tissue. To address this issue, this report offers detailed methodology, as well as comparisons of several histological and immunohistochemical stains available for imaging the cementum-PDL complex by light microscopy. Notably, the infrequently used Alcian blue stain with nuclear fast red counterstain provided utility in imaging cementum in mouse, porcine and human teeth. While no truly unique extracellular matrix markers have been identified to differentiate cementum from the other hard tissues, immunohistochemistry for detection of bone sialoprotein （BSP）, osteopontin （OPN）, and dentin matrix protein 1 （DMP1） is a reliable approach for studying both acellular and cellular cementum and providing insight into developmental biology of these tissues. Histoloeical and immunohistochemical aooroaches Drovide insight on developmental biology of cementum.展开更多
基金This study was funded by the National Natural Science Foundation of China(grant numbers 31670994,U1904145,and 81901039)Nature Science Foundation of Henan province(grant number 182300410340)and Union project of Medical and Technology Research Program of Henan Province(grant number LHGJ20190191).
文摘Treated dentin matrix(TDM)is an ideal scaffold material containing multiple extracellular matrix factors.The canonical Wnt signaling pathway is necessary for tooth regeneration.Thus,this study investigated whether the TDM can promote the odontogenic differentiation of human dental pulp stem cells(hDPSCs)and determined the potential role of Wnt/β-catenin signaling in this process.Different concentrations of TDM promoted the dental differentiation of the hDPSCs and meanwhile,the expression of GSK3βwas decreased.Of note,the expression of the Wnt/β-catenin pathway-related genes changed significantly in the context of TDM induction,as per RNA sequencing(RNA seq)data.In addition,the experiment showed that new dentin was visible in rat mandible cultured with TDM,and the thickness was significantly thicker than that of the control group.In addition,immunohistochemical staining showed lower GSK3βexpression in new dentin.Consistently,the GSK3βknockdown hDPSCs performed enhanced odotogenesis compared with the control groups.However,GSK3βoverexpressing could decrease odotogenesis of TDM-induced hDPSCs.These results were confirmed in immunodeficient mice and Wistar rats.These suggest that TDM promotes odontogenic differentiation of hDPSCs by directly targeting GSK3βand activating the canonical Wnt/β-catenin signaling pathway and provide a theoretical basis for tooth regeneration engineering.
基金the National Natural Science Foundation of China(grant numbers 31670994,U1904145 and 81901039)Nature Science Foundation of Henan Province(grant numbers 182300410340,222300420569)+4 种基金Henan Medical Science and Technology Research Project(grant number SBGJ202002073,SBGJ202002075)Science and Technology Department of Science and Technology Research(grant number 182102310456)Henan Province Middle-Aged And Young Health Science and Technology Innovation Leader Training Project(grant number YXKC2021014)Scientific Research and Innovation Team of The First Affiliated Hospital of Zhengzhou University(grant number QNCXTD2023021)Union project of Medical and Technology Research Program of Henan Province(grant number LHGJ20190191).
文摘It is still a huge challenge for bone regenerative biomaterial to balance its mechanical,biological and biodegradable properties.In the present study,a new composite material including treated dentin matrix(TDM)andα-calcium sulphate hemihydrate(α-CSH)was prepared.The optimal composition ratio between TDM andα-CSH was explored.The results indicate that both components were physically mixed and structurally stable.Its compressive strength reaches up to 5.027±0.035 MPa for 50%TDM/α-CSH group,similar to human cancellous bone tissues.Biological experiments results show that TDM/α-CSH composite exhibits excellent biocompatibility and the expression of osteogenic related genes and proteins(ALP,RUNX2,OPN)is significantly increased.In vivo experiments suggest that the addition of TDM for each group(10%,30%,50%)effectively promotes cell proliferation and osteomalacia.In addition,50%of the TDM/α-CSH combination displays optimal osteoconductivity.The novel TDM/α-CSH composite is a good candidate for certain applications in bone tissue engineering.
基金the National Natural Science Foundation of China(Nos.51925304,51903175,and 51973133).
文摘The invasion of etched dentinal tubules(DTs)by external substances induces dentin hypersensitivity(DH).The deep and compact occlusion of DTs is highly desirable for treating DH but still challenging due to the limited penetrability and mineralization capacities of most current desensitizers.Matrix vesicles(MVs)participate in the regulation of ectopic mineralization.Herein,ectopic MV analogs are prepared by employing natural cell membranes to endow mineral precursors with natural biointerfaces and integrated biofunctions for stimulating dentin remineralization.The analogs quickly access DTs(>20μm)in only 5 min and further penetrate deep into the interior of DTs(an extraordinary~200μm)in 7 days.Both in vitro and in vivo studies confirm that the DTs are efficiently sealed by the newly formed minerals(>50μm)with excellent resistance to wear and acid erosion,which is significantly deeper than most reported values.After repair,the microhardness of the damaged dentin can be recovered to those of healthy dentin.For the first time,cell membrane coating nanotechnology is used as a facile and efficient therapy for in-depth remineralization of DTs in treating DH with thorough and long-term effects,which provides insights into their potential for hard tissue repair.
文摘目的研究参与两种粘接剂形成的牙本质粘接界面退变过程中基质金属蛋白酶(MMPs)的种类。方法制备Single Bond 2和Clearfi l S3 Bond微剪切粘接试件,无菌人工唾液中分别浸泡0、6个月测试粘接强度,场发射扫描电镜(FE-SEM)观察界面断裂模式。制备4 mm×3 mm×1 mm牙本质片,分为对照组、Single Bond 2组和Clearfi l S3Bond组,无菌人工唾液浸泡0、6个月后,液相芯片技术测定各组MMP-1、-2、-3、-8、-9含量。结果老化6个月后,各组粘接强度显著下降。与对照组相比,老化过程中不同粘接剂组MMP-1、-3含量无明显变化,Single Bond 2组MMP-2、-8、-9含量显著降低,Clearfi l S3 Bond组MMP-8、-9含量显著降低。结论老化6个月后两种粘接试件均发生粘接界面退变,牙本质源MMP-2、-8、-9的含量显著降低,提示这些MMPs可能参与了牙本质粘接界面的退变。
文摘目的:探讨不同粘结条件下健康成人牙本质中基质金属蛋白酶2 (MMP-2)和基质金属蛋白酶9 (MMP-9)表达水平,为临床应用基质金属蛋白酶(MMPs)抑制剂提高粘结效果提供依据。方法:收集20颗成人新鲜离体磨牙,液氮冷却条件下将其研磨为牙本质粉;模拟口内条件,对牙本质分别进行自酸蚀粘结(Single Bond Universal)和全酸蚀粘结(35%磷酸凝胶+Adpter Single Bond 2)处理,以不进行任何处理的牙本质作为阴性对照组,10%磷酸溶液(自酸蚀粘结)或10%磷酸溶液+35%磷酸凝胶处理的脱矿牙本质粉(全酸蚀粘结)处理的牙本质作为阳性对照组,只进行Single Bond Universal (自酸蚀粘结)处理和35%磷酸凝胶+Adpter Single Bone 2 (全酸蚀粘结)处理的牙本质作为空白对照组;在粘结过程中分别使用MMPs抑制剂氯己定(CHX)和米诺环素(MI)预处理,作为CHX预处理组和MI预处理组;采用10%次氯酸钠进行牙本质老化,作为老化组,并在老化组基础上加入CHX和MI作为CHX预处理老化组和MI预处理老化组。采用明胶酶谱法进行聚丙烯酰胺凝胶电泳,孵育染色脱色后,采用凝胶图像分析系统分析条带,计算各组牙本质中MMP-2和MMP-9表达水平。结果:在自酸蚀粘结条件下,与空白对照组比较,CHX预处理组牙本质中MMP-2和MMP-9表达水平降低(P<0.05),MI预处理组牙本质中MMP-2和MMP-9表达水平降低(P<0.05);与空白老化对照组比较,CHX预处理老化组牙本质中MMP-2和MMP-9表达水平降低(P<0.05),MI预处理老化组牙本质中MMP-2和MMP-9表达水平降低(P<0.05)。在全酸蚀粘结条件下,与空白对照组比较,CHX预处理组和MI预处理组牙本质中MMP-2表达水平降低(P<0.05);与空白老化对照组比较,CHX预处理老化组与MI预处理老化组牙本质中MMP-2表达水平降低(P<0.05)。结论:在牙本质粘结过程中,CHX和MI能够通过降低MMP-2和MMP-9表达水平减缓酶解反应,从而增强粘结强度。
基金Key Project of Chinese National Programs for Research and Development (No.2016YFC1102075, Yao Sun)National Natural Science Foundation of China (Nos.81470715, 81771043, 81822012, Yao Sun+4 种基金81770873, 81722031, Xiaogang Wang81670962, Zuolin Wang)Shanghai Health System (No.2017BR009 , Yao Sun ), Tongji University (Nos.TJ15042119036 and TJ2000219143, Zuolin Wang)Chinese Universities Scientific Fund (No.kx0200020173386, Rui Yue)We would like to appreciate Dr.Chunlin Qin (College of Dentistry, Texas A&M University) for providing the DMP1-N antibody and assistance.We thank Qigang Wang Group, School of Chemical Science and Engineering, Tongji University for providing biom echanical testing machine.We would also like to thank Xiaojuan Yang, Gongchen Li, and Mengmeng Liu for their help in revising the paper.
文摘Fractures are frequently occurring diseases that endanger human health. Crucial to fracture healing is cartilage formation, which provides a bone-regeneration environment. Cartilage consists of both chondrocytes and extracellular matrix (ECM). The ECM of cartilage includes collagens and various types of proteoglycans (PGs), which play important roles in maintaining primary stability in fracture healing. The PG form of dentin matrix protein 1 (DMP1-PG) is involved in maintaining the health of articular cartilage and bone. Our previous data have shown that DMP1-PG is richly expressed in the cartilaginous calluses of fracture sites. However, the possible significant role of DMP1-PG in chondrogenesis and fracture healing is unknown. To further detect the potential role of DMP1-PG in fracture repair, we established a mouse fracture model by using a glycosylation site mutant DMP1 mouse (S89G-DMP1 mouse). Upon inspection, fewer cartilaginous calluses and down-regulated expression levels of chondrogenesis genes were observed in the fracture sites of S89G-DMP1 mice. Given the deficiency of DMP1-PG, the impaired IL-6/JAK/STAT signaling pathway was observed to affect the chondrogenesis of fracture healing. Overall, these results suggest that DMP1-PG is an indispensable proteoglycan in chondrogenesis during fracture healing.
基金supported (in part) by the Intramural Research Program of the National Institute of Arthritis and Musculoskeletal and Skin Diseases of the National Institutes of Health
文摘The tooth root cementum is a thin, mineralized tissue covering the root dentin that is present primarily as acellular cementum on the cervical root and cellular cementum covering the apical root. While cementum shares many properties in common with bone and dentin, it is a unique mineralized tissue and acellular cementum is critical for attachment of the tooth to the surrounding periodontal ligament （PDL）. Resources for methodologies for hard tissues often overlook cementum and approaches that may be of value for studying this tissue. To address this issue, this report offers detailed methodology, as well as comparisons of several histological and immunohistochemical stains available for imaging the cementum-PDL complex by light microscopy. Notably, the infrequently used Alcian blue stain with nuclear fast red counterstain provided utility in imaging cementum in mouse, porcine and human teeth. While no truly unique extracellular matrix markers have been identified to differentiate cementum from the other hard tissues, immunohistochemistry for detection of bone sialoprotein （BSP）, osteopontin （OPN）, and dentin matrix protein 1 （DMP1） is a reliable approach for studying both acellular and cellular cementum and providing insight into developmental biology of these tissues. Histoloeical and immunohistochemical aooroaches Drovide insight on developmental biology of cementum.
