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骨形成蛋白体外对培养的人牙髓组织的诱导作用 被引量:6
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作者 高玉好 诹访文彦 +2 位作者 太田义邦 方一如 杨连甲 《第四军医大学学报》 1994年第5期328-331,共4页
骨形成蛋白可促进牙髓细胞增殖和诱导修复性牙本质形成.作者应用体外组织培养技术,分别通过光镜和透射电镜观察了牛骨形成蛋白对体外培养的人牙髓组织的诱导作用.结果发现,培养3d后可见增殖的星形细胞,其胞浆少,细胞小器不发达... 骨形成蛋白可促进牙髓细胞增殖和诱导修复性牙本质形成.作者应用体外组织培养技术,分别通过光镜和透射电镜观察了牛骨形成蛋白对体外培养的人牙髓组织的诱导作用.结果发现,培养3d后可见增殖的星形细胞,其胞浆少,细胞小器不发达;培养7d后,可见透明基质中埋有一些软骨样细胞,该细胞胞浆丰富,细胞小器发达,并含有丰富的分泌小泡.说明骨形成蛋白可促进牙髓细胞从低分化状态向高分化状态分化. 展开更多
关键词 骨形成蛋白 牙髓 组织培养 骨诱导
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Self-assembly of differentiated dental pulp stem cells facilitates spheroid human dental organoid formation and prevascularization
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作者 Fei Liu Jing Xiao +4 位作者 Lei-Hui Chen Yu-Yue Pan Jun-Zhang Tian Zhi-Ren Zhang Xiao-Chun Bai 《World Journal of Stem Cells》 SCIE 2024年第3期287-304,共18页
BACKGROUND The self-assembly of solid organs from stem cells has the potential to greatly expand the applicability of regenerative medicine.Stem cells can self-organise into microsized organ units,partially modelling ... BACKGROUND The self-assembly of solid organs from stem cells has the potential to greatly expand the applicability of regenerative medicine.Stem cells can self-organise into microsized organ units,partially modelling tissue function and regeneration.Dental pulp organoids have been used to recapitulate the processes of tooth development and related diseases.However,the lack of vasculature limits the utility of dental pulp organoids.AIM To improve survival and aid in recovery after stem cell transplantation,we demonstrated the three-dimensional(3D)self-assembly of adult stem cell-human dental pulp stem cells(hDPSCs)and endothelial cells(ECs)into a novel type of spheroid-shaped dental pulp organoid in vitro under hypoxia and conditioned medium(CM).METHODS During culture,primary hDPSCs were induced to differentiate into ECs by exposing them to a hypoxic environment and CM.The hypoxic pretreated hDPSCs were then mixed with ECs at specific ratios and conditioned in a 3D environment to produce prevascularized dental pulp organoids.The biological characteristics of the organoids were analysed,and the regulatory pathways associated with angiogenesis were studied.RESULTS The combination of these two agents resulted in prevascularized human dental pulp organoids(Vorganoids)that more closely resembled dental pulp tissue in terms of morphology and function.Single-cell RNA sequencing of dental pulp tissue and RNA sequencing of Vorganoids were integrated to analyse key regulatory pathways associated with angiogenesis.The biomarkers forkhead box protein O1 and fibroblast growth factor 2 were identified to be involved in the regulation of Vorganoids.CONCLUSION In this innovative study,we effectively established an in vitro model of Vorganoids and used it to elucidate new mechanisms of angiogenesis during regeneration,facilitating the development of clinical treatment strategies. 展开更多
关键词 human dental pulp stem cells Prevascularized organoids Integrated analyses ANGIOGENESIS Forkhead box protein O1
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Magnetic bioassembly platforms towards the generation of extracellular vesicles from human salivary gland functional organoids for epithelial repair 被引量:5
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作者 Ajjima Chansaenroj Christabella Adine +9 位作者 Sawanya Charoenlappanit Sittiruk Roytrakul Ladawan Sariya Thanaphum Osathanon Sasitorn Rungarunlert Ganokon Urkasemsin Risa Chaisuparat Supansa Yodmuang Glauco R.Souza Joao N.Ferreira 《Bioactive Materials》 SCIE 2022年第12期151-163,共13页
Salivary glands(SG)are exocrine organs with secretory units commonly injured by radiotherapy.Bio-engineered organoids and extracellular vesicles(EV)are currently under investigation as potential strategies for SG repa... Salivary glands(SG)are exocrine organs with secretory units commonly injured by radiotherapy.Bio-engineered organoids and extracellular vesicles(EV)are currently under investigation as potential strategies for SG repair.Herein,three-dimensional(3D)cultures of SG functional organoids(SGo)and human dental pulp stem cells(hDPSC)were generated by magnetic 3D bioassembly(M3DB)platforms.Fibroblast growth factor 10(FGF10)was used to enrich the SGo in secretory epithelial units.After 11 culture days via M3DB,SGo displayed SG-specific acinar epithelial units with functional properties upon neurostimulation.To consistently develop 3D hDPSC in vitro,3 culture days were sufficient to maintain hDPSC undifferentiated genotype and phenotype for EV generation.EV isolation was performed via sequential centrifugation of the conditioned media of hDPSC and SGo cultures.EV were characterized by nanoparticle tracking analysis,electron microscopy and immunoblotting.EV were in the exosome range for hDPSC(diameter:88.03±15.60 nm)and for SGo(123.15±63.06 nm).Upon ex vivo administration,exosomes derived from SGo significantly stimulated epithelial growth(up to 60%),mitosis,epithelial progenitors and neuronal growth in injured SG;however,such biological effects were less distinctive with the ones derived from hDPSC.Next,these exosome biological effects were investigated by proteomic arrays.Mass spectrometry profiling of SGo exosomes predicted that cellular growth,development and signaling was due to known and undocumented molecular targets downstream of FGF10.Semaphorins were identified as one of the novel targets requiring further investigations.Thus,M3DB platforms can generate exosomes with potential to ameliorate SG epithelial damage. 展开更多
关键词 Salivary gland HYPOSALIVATION human dental pulp stem cells Magnetic bioassembly ORGANOIDS EXOSOME
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Genetic modification of miR-34a enhances efficacy of transplanted human dental pulp stem cells after ischemic stroke 被引量:1
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作者 Jianfeng Wang Peibang He +7 位作者 Qi Tian Yu Luo Yan He Chengli Liu Pian Gong Yujia Guo Qingsong Ye Mingchang Li 《Neural Regeneration Research》 SCIE CAS CSCD 2023年第9期2029-2036,共8页
Human dental pulp stem cells(hDPSCs) promote recovery after ischemic stro ke;however,the therapeutic efficacy is limited by the poor survival of transplanted cells.For in vitro expe riments in the present study,we use... Human dental pulp stem cells(hDPSCs) promote recovery after ischemic stro ke;however,the therapeutic efficacy is limited by the poor survival of transplanted cells.For in vitro expe riments in the present study,we used oxygen-glucose deprivation/reoxygenation in hDPSCs to mimic cell damage induced by ischemia/reperfusion.We found that miRNA-34a-5p(miR-34a) was elevated under oxygen-glucose deprivation/reoxygenation conditions in hDPSCs.Inhibition of miR-34a facilitated the prolife ration and antioxidant capacity and reduced the apoptosis of hDPSCs.Moreove r,dual-luciferase reporter gene assay showed WNT1and SIRT1 as the targets of miR-34a.In miR-34a knockdown cell lines,WNT1 suppression reduced cell prolife ration,and SIRT1 suppression decreased the antioxidant capacity.Togethe r,these results indicated that miR-34a regulates cell prolife ration and antioxidant stress via targeting WNT1 and SIRT1,respectively.For in vivo expe riments,we injected genetically modified hDPSCs(anti34a-hDPSCs) into the brains of mice.We found that anti34a-hDPSCs significantly inhibited apoptosis,reduced cerebral edema and cerebral infarct volume,and improved motor function in mice.This study provides new insights into the molecular mechanism of the cell prolife ration and antioxidant capacity of hDPSCs,and suggests a potential gene that can be targeted to improve the survival rate and efficacy of transplanted hDPSCs in brain after ischemic stroke. 展开更多
关键词 antioxidant capacity HO-1 human dental pulp stem cells ischemic stroke MIR-34A Nrf2 proliferation SIRT1 WNT1 β-catenin
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An amelogenin-based peptide hydrogel promoted the odontogenic differentiation of human dental pulp cells 被引量:2
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作者 Xinxin Li Zhaoxia Yu +6 位作者 Shihui Jiang Xiaohua Dai Guanhua Wang Yue Wang Zhimou Yang Jie Gao Huiru Zou 《Regenerative Biomaterials》 SCIE EI 2022年第1期511-521,共11页
Amelogenin can induce odontogenic differentiation of human dental pulp cells(HDPCs),which has great potential and advantages in dentine-pulp complex regeneration.However,the unstability of amelogenin limits its furthe... Amelogenin can induce odontogenic differentiation of human dental pulp cells(HDPCs),which has great potential and advantages in dentine-pulp complex regeneration.However,the unstability of amelogenin limits its further application.This study constructed amelogenin self-assembling peptide hydrogels(L-gel or D-gel)by heating-cooling technique,investigated the effects of these hydrogels on the odontogenic differentiation of HDPCs and explored the underneath mechanism.The critical aggregation concentration,conformation,morphology,mechanical property and biological stability of the hydrogels were characterized,respectively.The effects of the hydrogels on the odontogenic differentiation of HDPCs were evaluated via alkaline phosphatase activity measurement,quantitative reverse transcription polymerase chain reaction,western blot,Alizarin red staining and scanning electron microscope.The mechanism was explored via signaling pathway experiments.Results showed that both the L-gel and D-gel stimulated the odontogenic differentiation of HDPCs on both Day 7 and Day 14,while the D-gel showed the highest enhancement effects.Meanwhile,the D-gel promoted calcium accumulation and mineralized matrix deposition on Day 21.The D-gel activated MAPK-ERK1/2 pathways in HDPCs and induced the odontogenic differentiation via ERK1/2 and transforming growth factor/smad pathways.Overall,our study demonstrated that the amelogenin peptide hydrogel stimulated the odontogenic differentiation and enhanced mineralization,which held big potential in the dentine-pulp complex regeneration. 展开更多
关键词 AMELOGENIN peptide hydrogel odontogenic differentiation human dental pulp cells
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低浓度洗涤血小板促进人牙髓细胞增殖的作用机制探讨 被引量:3
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作者 段建民 汪维健 菊地宽高 《实用口腔医学杂志》 CAS CSCD 北大核心 2006年第5期675-678,共4页
目的:探讨低浓度洗涤血小板促进人牙髓细胞增殖的作用机制。方法:使用从成年男性志愿者静脉采集、制备的洗涤血小板(washed platelet,WPLT)作用于人牙髓细胞,人牙髓细胞增殖和产生的前列腺素E2(PGE2)采用细胞和PGE2测定试剂盒测定;表达... 目的:探讨低浓度洗涤血小板促进人牙髓细胞增殖的作用机制。方法:使用从成年男性志愿者静脉采集、制备的洗涤血小板(washed platelet,WPLT)作用于人牙髓细胞,人牙髓细胞增殖和产生的前列腺素E2(PGE2)采用细胞和PGE2测定试剂盒测定;表达的细胞内环氧酶-2(COX-2)mRNA采用Realtime RT-PCR方法测定。结果:50ml/LWPLT诱导人牙髓细胞COX-2mRNA的表达在作用后3h达到最高峰,而诱导PGE2的产生快于IL-1β;50ml/LWPLT诱导的人牙髓细胞增殖可以被消炎痛和地塞米松抑制,而添加PGE2可以逆转这种抑制作用;WPLT诱导的PGE2量随着其浓度的成倍增加呈现出明显的增多趋势,而PGE2在适当浓度范围内明显促进了人牙髓细胞的增殖。结论:低浓度WPLT可能通过诱导产生的PGE2促进了人牙髓细胞的增殖。 展开更多
关键词 人类 牙髓 细胞 血小板 前列腺素E2 细胞内环氧酶-2
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血小板衍化生长因子受体在人牙髓组织中表达的分析 被引量:1
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作者 陈强 史俊南 +1 位作者 金岩 赵宇 《牙体牙髓牙周病学杂志》 CAS 2004年第2期72-74,共3页
目的 :观察血小板衍化生长因子 (platelet-derivedgrowthfactor,PDGF)受体在人牙髓组织中的表达和分布情况 ,探讨PDGF在牙髓组织中的作用。方法 :制备人牙髓组织标本 ,采用免疫组织化学方法观察PDGFβ受体在人牙髓组织中的表达与分布情... 目的 :观察血小板衍化生长因子 (platelet-derivedgrowthfactor,PDGF)受体在人牙髓组织中的表达和分布情况 ,探讨PDGF在牙髓组织中的作用。方法 :制备人牙髓组织标本 ,采用免疫组织化学方法观察PDGFβ受体在人牙髓组织中的表达与分布情况。 结果 :在人牙髓组织中的牙髓细胞、微血管内皮细胞表达PDGFβ受体。 结论 :在正常牙髓组织中牙髓细胞、微血管内皮细胞表达PDGFβ受体 ,提示PDGF有可能在牙髓组织的生长、发育等正常生理过程和创伤修复过程中起着重要的作用。 展开更多
关键词 血小板衍化生长因子 受体 牙髓组织
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Inductive effect of bovinc bone morphogenetic protein on human dental pulp tissue in vitro
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作者 高玉好 方一如 杨连甲 《Journal of Medical Colleges of PLA(China)》 CAS 1994年第2期108-111,共4页
Bone morphogenetic protein (BMP) can promote the proliferation of dental pulp cells and induce reparative dentin formation. In this study, the inductive effect of BMP derived from bovine bone matrix on cultured human ... Bone morphogenetic protein (BMP) can promote the proliferation of dental pulp cells and induce reparative dentin formation. In this study, the inductive effect of BMP derived from bovine bone matrix on cultured human dental pulp tissue was observed under light microscope and transmission electron microscope. The results showed that. by the third day of culture, the proliferating star-shaped cells appeared with small cytoplasm and poorly-developed organelles; by the 7th day of the culture, the chondroblast-like cells with rich cytoplasm and well-developed organelles were seen embedded in hyaline matrix. This study suggests that BMP can induce dental pulp cells to differentiate from poorly differentiated state to well-differentiated state. 展开更多
关键词 DENTINOGENESIS cell culture human pulp BONE morphogenetic PROTEIN
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Effect of incorporating Elaeagnus angustifolia extract in PCL-PEG-PCL nanofibers for bone tissue engineering 被引量:1
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作者 Vahideh R.Hokmabad Soodabeh Davaran +3 位作者 Marziyeh Aghazadeh Effat Alizadeh Roya Salehi Ali Ramazani 《Frontiers of Chemical Science and Engineering》 SCIE EI CAS CSCD 2019年第1期108-119,共12页
Plants have been used for medicinal purposes for thousands of years but they are still finding new uses in modem times. For example, Elaeagnus angustifolia (EA) is a medicinal herb with antinociceptive, anti-inflammat... Plants have been used for medicinal purposes for thousands of years but they are still finding new uses in modem times. For example, Elaeagnus angustifolia (EA) is a medicinal herb with antinociceptive, anti-inflammatory, antibacterial and antioxidant properties and it is widely used in the treatment of rheumatoid arthritis and osteoarthritis. EA extract was loaded onto poly(ε-caprolactone)- poly(ethylene glycol)-poly(ε-caprolactone)(PCLPEG-PCL/EA) nanofibers and their potential applications for bone tissue engineering were studied. The morphology and chemical properties of the fibers were evaluated using Fourier transform infrared spectroscopy, field emission scanning electron microscopy, contact angle measurements and mechanical tests. All the samples had bead-free morphologies with average diameters ranging from 100 to 200 nm. The response of human cells to the PCL-PEGPCL/ EA nanofibers was evaluated using human dental pulp stem cells (hDPSCs). The hDPSCs had better adhesion and proliferation capacity on the EA loaded nanofibers than on the pristine PCL-PEG?PCL nanofibers. An alizarin red S assay and the alkaline phosphatase activity confirmed that the nanofibrous scaffolds induced osteoblastic performance in the hDPSCs. The quantitative real time polymerase chain reaction results confirmed that the EA loaded nanofibrous scaffolds had significantly upregulated gene expression correlating to osteogenic differentiation. These results suggest that PCL-PEG-PCL/ EA nanofibers might have potential applications for bone tissue engineering. 展开更多
关键词 ELAEAGNUS angustifolia SCAFFOLD ELECTROSPINNING human DENTAL pulp stem cell tissue engineering
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Gelatin-biofermentative unsulfated glycosaminoglycans semi-interpenetrating hydrogels via microbial-transglutaminase crosslinking enhance osteogenic potential of dental pulp stem cells 被引量:1
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作者 Annalisa La Gatta Virginia Tirino +8 位作者 Marcella Cammarota Marcella La Noce Antonietta Stellavato Anna Virginia Adriana Pirozzi Marianna Portaccio Nadia Diano Luigi Laino Gianpaolo Papaccio Chiara Schiraldi 《Regenerative Biomaterials》 SCIE 2021年第3期110-123,共14页
Gelatin hydrogels by microbial-transglutaminase crosslinking are being increasingly exploited for tissue engineering,and proved high potential in bone regeneration.This study aimed to evaluate,for the first time,the c... Gelatin hydrogels by microbial-transglutaminase crosslinking are being increasingly exploited for tissue engineering,and proved high potential in bone regeneration.This study aimed to evaluate,for the first time,the combination of enzymatically crosslinked gelatin with hyaluronan and the newly developed biotechnological chondroitin in enhancing osteogenic potential.Gelatin enzymatic crosslinking was carried out in the presence of hyaluronan or of a hyaluronan–chondroitin mixture,obtaining semi-interpenetrating gels.The latter proved lower swelling extent and improved stiffness compared to the gelatin matrix alone,whilst maintaining high stability.The heteropolysaccharides were retained for 30 days in the hydrogels,thus influencing cell response over this period.To evaluate the effect of hydrogel composition on bone regeneration,materials were seeded with human dental pulp stem cells and osteogenic differentiation was assessed.The expression of osteocalcin(OC)and osteopontin(OPN),both at gene and protein level,was evaluated at 7,15 and 30 days of culture.Scanning electron microscopy(SEM)and two-photon microscope observations were performed to assess bone-like extracellular matrix(ECM)deposition and to observe the cell penetration depth.In the presence of the heteropolysaccharides,OC and OPN expression was upregulated and a higher degree of calcified matrix formation was observed.Combination with hyaluronan and chondroitin improved both the biophysical properties and the biological response of enzymatically crosslinked gelatin,fastening bone deposition. 展开更多
关键词 HYDROGELS GELATIN HYALURONAN biotechnological chondroitin bone regeneration human dental pulp stem cells
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Identification and Isolation of Human Dental Pulp Stem Cells
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作者 Xue-Chao YANG Ming-Wen FAN(Ministry Education Key Lab. For Oral Biomedical Engineering, Shool of Stomatology, Wuhan University,Wuhan 430079,China) 《生物医学工程学杂志》 EI CAS CSCD 北大核心 2005年第S1期101-102,共2页
关键词 CELL DPSCs Identification and Isolation of human Dental pulp Stem Cells DSPP
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Human pulp tissue dissolution ability of different extracts of Sapindus mukorossi: An in vitro study
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作者 Öznur Güçlüer Esin Akarsu +2 位作者 Emre Yavuz Kürsat Er Alper Kustarcl 《Chinese Herbal Medicines》 CAS 2020年第2期178-182,共5页
Objective:Due to the many negative properties of sodium hypochlorite used in current root canal treatment,interest in biocompatible natural agents is increasing day by day.The aim of this study was to evaluate whether... Objective:Due to the many negative properties of sodium hypochlorite used in current root canal treatment,interest in biocompatible natural agents is increasing day by day.The aim of this study was to evaluate whether various extract solutions of Sapindus mukorossi have dissolution effects on human pulp tissues.Methods:Primarily powder extracts were obtained by extracting fruit shells of S.mukorossi in different solvents(ethanol,methanol,buthanol and distilled water).The test solutions were prepared and randomly separated into six groups with 10 samples in each group:ethanol extract,methanol extract,butanol extract,distilled water extract of S.mukorossi,sodium hypochlorite(Na OCl)and the control group.Among these,S.mukorossi extracts were separated into two subgroups,depending on their concentration level(50μg/m L and 100μg/m L).The pulp tissues of freshly extracted human molars were used for dissolution test.The weights of the pulpal tissues were measured and recorded for two times after the samples were placed in the solutions.Statistical analysis for all descriptive statistics was performed using SPSS 22(P<0.05).Results:Our results showed that maximum percent yield of preparation was obtained in methanol extract of S.mukorossi.Among all of the groups,the best dissolution capacity was seen in the Na OCl group(positive control group).Among S.mukorossi groups,the best tissue solvent solution was found in SMM group at 50μg/m L and SMB group at 100μg/m L.Conclusion:The different extracts of S.mukorossi had a capacity to dissolve pulp tissue but this capacity was less than Na OCl.Therefore,further studies will enable the creation of a commercial solution for clinical use by increasing the effectiveness of S.mukorossi while combining it with other endodontic irrigation solutions. 展开更多
关键词 DISSOLUTION herbal extract human pulp tissue NAOCL Sapindus mukorossi Gaertn
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Human dental pulp stem cells: Applications in future regenerative medicine 被引量:21
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作者 Pravin D Potdar Yogita D Jethmalani 《World Journal of Stem Cells》 SCIE CAS 2015年第5期839-851,共13页
Stem cells are pluripotent cells, having a property of differentiating into various types of cells of human body. Several studies have developed mesenchymal stem cells(MSCs) from various human tissues,peripheral blood... Stem cells are pluripotent cells, having a property of differentiating into various types of cells of human body. Several studies have developed mesenchymal stem cells(MSCs) from various human tissues,peripheral blood and body fluids. These cells are then characterized by cellular and molecular markers to understand their specific phenotypes. Dental pulp stem cells(DPSCs) are having a MSCs phenotype and they are differentiated into neuron, cardiomyocytes, chondrocytes, osteoblasts, liver cells and β cells of islet of pancreas. Thus, DPSCs have shown great potentiality to use in regenerative medicine for treatment of various human diseases including dental related problems. These cells can also be developed into induced pluripotent stem cells by incorporation of pluripotency markers and use for regenerative therapies of various diseases. The DPSCs are derived from various dental tissues such as human exfoliated deciduous teeth, apical papilla, periodontal ligament and dental follicle tissue. This review will overview the information about isolation, cellular and molecular characterization and differentiation of DPSCs into various types of human cells and thus these cells have important applications in regenerative therapies for various diseases. This review will be most useful for postgraduate dental students as well as scientists working in the field of oral pathology and oral medicine. 展开更多
关键词 human dental pulp STEM CELLS Mesenchymalstem CELLS DENTIN PLURIPOTENCY STEM cell therapy Molecular MARKERS
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体外持续培养对人牙髓细胞分化能力的影响 被引量:16
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作者 杨雪超 樊明文 《牙体牙髓牙周病学杂志》 CAS 2003年第2期62-65,共4页
目的 :探讨人牙髓细胞在体外持续培养时 ,其分化能力的变化趋势。方法 :体外持续培养人牙髓细胞 ,检测不同代次细胞的碱性磷酸酶活性、钙化能力及Ⅰ型胶原表达 ,并对比分析。结果 :随着体外培养细胞代次的增多 ,在同等刺激分化条件下 ,... 目的 :探讨人牙髓细胞在体外持续培养时 ,其分化能力的变化趋势。方法 :体外持续培养人牙髓细胞 ,检测不同代次细胞的碱性磷酸酶活性、钙化能力及Ⅰ型胶原表达 ,并对比分析。结果 :随着体外培养细胞代次的增多 ,在同等刺激分化条件下 ,人牙髓细胞的碱性磷酸酶活性整体呈下降趋势 ,钙化结节的数目和面积逐渐减少 ,Ⅰ型胶原表达合成逐渐减少。结论 :人牙髓细胞在体外持续培养时 ,分化能力逐渐下降 ,可能与其含有的未分化细胞 (人牙髓干细胞 ) 展开更多
关键词 体外持续培养 人牙髓细胞 分化能力 ALP活性
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应用不同方法进行人牙髓细胞原代培养的比较研究 被引量:12
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作者 刘俊 李玉晶 《北京口腔医学》 CAS 2005年第3期148-152,共5页
目的比较并建立人牙髓细胞原代培养的理想方法。方法分别采用组织块法、酶消化法和组织块消化法进行人牙髓细胞的体外原代培养,倒置显微镜及台盼蓝染色观察并比较三者的培养成活率、细胞活力、细胞形态和数量、培养耗时等,免疫组化法鉴... 目的比较并建立人牙髓细胞原代培养的理想方法。方法分别采用组织块法、酶消化法和组织块消化法进行人牙髓细胞的体外原代培养,倒置显微镜及台盼蓝染色观察并比较三者的培养成活率、细胞活力、细胞形态和数量、培养耗时等,免疫组化法鉴定细胞来源。结果三种方法均可从人牙髓组织分离培养获得外胚间充质来源的牙髓细胞。组织块法耗时较长,4周能进行首次传代,传代成功率50%,细胞形态较单一;酶消化法能在较短时间内获得形态多样的人牙髓细胞,但培养成活率较低,且细胞数量较少,8天时传代成功率为20%;组织块消化法不仅培养成活率(75%)和8天传代成功率(62.5%)较高,而且在较短时间内可多量获得多种形态类型的人牙髓细胞。结论组织块消化法是一种更加科学可行的人牙髓细胞原代培养的优化方法,可为进行人牙髓干细胞的分离培养及特性研究提供实验方法学基础。 展开更多
关键词 人牙髓细胞 细胞培养 方法学
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体外人牙髓细胞的矿化特性 被引量:13
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作者 彭彬 樊明文 《中华口腔医学杂志》 CAS CSCD 北大核心 1997年第5期285-287,共3页
为探讨体外人牙髓细胞的生物学特性,采用体外细胞连续培养、钙质染色、X射线能谱分析与透射电镜观察等方法,对体外人恒牙牙髓细胞的矿化特性进行了研究并与人牙龈成纤维细胞(humangingivafibroblasts,HG... 为探讨体外人牙髓细胞的生物学特性,采用体外细胞连续培养、钙质染色、X射线能谱分析与透射电镜观察等方法,对体外人恒牙牙髓细胞的矿化特性进行了研究并与人牙龈成纤维细胞(humangingivafibroblasts,HGFs)进行了比较。结果表明,人牙髓细胞在体外可以复层生长并形成细胞结节,HGFs则无此能力,牙髓细胞的碱性磷酸酶活性亦较HGFs者高;细胞结节钙质染色呈阳性,结节内钙磷含量明显增高;人牙髓细胞有许多与成牙本质细胞相似的超微结构特征,胞间基质中可见致密晶状小体。结果提示,体外连续培养的人牙髓细胞有向成牙本质细胞分化的可能,这可为研究人牙髓细胞的分化和矿化提供有价值的思路。 展开更多
关键词 人牙髓细胞 细胞培养 矿化
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人牙髓干细胞在聚乳酸-聚乙醇酸共聚物支架上粘附与增殖的研究 被引量:17
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作者 张巍巍 李艳萍 +3 位作者 梁鹏 董海波 甘友华 牛玉梅 《口腔医学研究》 CAS CSCD 北大核心 2011年第12期1083-1085,1088,共4页
目的:研究成人牙髓干细胞(HDPSCs)在聚乳酸-聚乙醇酸共聚物(PLGA)支架上粘附与增殖的情况。方法:采用酶消化法分离、培养人牙髓干细胞,免疫组化法及体外诱导分化对细胞进行鉴定。将人牙髓干细胞与PLGA支架材料进行复合培养,扫描电镜(SEM... 目的:研究成人牙髓干细胞(HDPSCs)在聚乳酸-聚乙醇酸共聚物(PLGA)支架上粘附与增殖的情况。方法:采用酶消化法分离、培养人牙髓干细胞,免疫组化法及体外诱导分化对细胞进行鉴定。将人牙髓干细胞与PLGA支架材料进行复合培养,扫描电镜(SEM)观察支架材料形态,细胞粘附、增殖及基质分泌情况;细胞计数检测其增殖力。结果:细胞接种2、5、10d,扫描电镜及细胞计数均显示HDPSCs与PLGA支架材料粘附紧密,生长状态良好,细胞明显增殖(P<0.05),有丰富的细胞外基质形成。结论:PLGA是一种适宜人牙髓干细胞粘附与增殖的支架材料。 展开更多
关键词 成人牙髓干细胞 聚乳酸-聚乙醇酸共聚物 组织工程
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人脱落乳牙牙髓干细胞与人恒牙牙髓干细胞成骨分化及破骨能力的差异 被引量:14
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作者 路博闻 刘娜 +3 位作者 徐璐璐 石海刚 张洋 张维 《南方医科大学学报》 CAS CSCD 北大核心 2016年第2期180-185,共6页
目的比较处于生理性根吸收时期的自然脱落乳牙牙髓干细胞(SHED)和恒牙牙髓干细胞(DPSCs)的成骨分化及破骨能力,探讨两者在成骨和破骨相关分子的表达情况。方法体外分离、培养和纯化处于生理性根吸收时期的自然脱落乳牙牙髓干细胞与恒牙... 目的比较处于生理性根吸收时期的自然脱落乳牙牙髓干细胞(SHED)和恒牙牙髓干细胞(DPSCs)的成骨分化及破骨能力,探讨两者在成骨和破骨相关分子的表达情况。方法体外分离、培养和纯化处于生理性根吸收时期的自然脱落乳牙牙髓干细胞与恒牙牙髓干细胞;两种干细胞在成骨诱导14 d和21 d,分别进行ALP染色、茜素红染色,并通过Real-time PCR检测SHED和DPSCs矿化诱导后的成骨与破骨相关基因表达。结果倒置显微镜下观察,SHED和DPSCs形态均为长梭形;两种干细胞经矿化诱导后,茜素红染色可见SHED的矿化结节计数多于DPSCs(P<0.05),SHED的ALP活性亦强于DPSCs(P<0.05)。RT-PCR检测结果显示,经矿化诱导后两种干细胞均表达成骨与破骨相关基因Runx2、OCN、ALP、OPG和RANKL,但DPSCs的Runx2、OCN和ALP的表达水平均低于SHED(P<0.05),且SHED反应细胞破骨/成骨能力的RANKL/OPG的比值显著高于DPSCs(P<0.05)。结论与DPSCs相比,SHED不仅具有较强的成骨分化能力,还兼有较强的破骨能力,为SHED参与生理性根吸收骨改建的调控机制提供实验依据。 展开更多
关键词 生理性根吸收 人脱落乳牙牙髓干细胞 恒牙牙髓干细胞 成骨分化 破骨细胞
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人牙髓细胞体外培养的方法学研究 被引量:11
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作者 彭彬 樊明文 +1 位作者 边专 陈智 《口腔医学纵横》 CSCD 1997年第4期198-200,共3页
作者观察了组织块法和酶消化法以及DMEM和RPMI1640二种常用培养液在人牙髓细胞体外培养中的效果。结果表明,组织块法培养出的牙髓细胞为单一的成纤维细胞样细胞,用DMEM时,其传代成功率明显高于用RPMI1604;胰蛋白酶消化法、胰蛋白酶... 作者观察了组织块法和酶消化法以及DMEM和RPMI1640二种常用培养液在人牙髓细胞体外培养中的效果。结果表明,组织块法培养出的牙髓细胞为单一的成纤维细胞样细胞,用DMEM时,其传代成功率明显高于用RPMI1604;胰蛋白酶消化法、胰蛋白酶和胶原酶联用可培养出成纤维细胞样细胞,少量的校形细胞和内皮样细胞,但细胞数量较少,生长缓慢,达不到传代要求。DMEM在培养板和组织培养瓶中均较适合牙髓细胞的生长,而RPMI1640效果不理想。提示组织块法和DMEM适合于人牙髓细胞的体外培养。 展开更多
关键词 体外培养 人牙髓细胞 方法学
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载柚皮苷羟基磷灰石胶原支架对人牙髓干细胞体外增殖及成骨向分化的影响 被引量:14
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作者 侯婷婷 潘爽 +3 位作者 李艳萍 何丽娜 肖婉鲁 牛玉梅 《口腔医学研究》 CAS 北大核心 2019年第8期744-747,共4页
目的:探讨载柚皮苷羟基磷灰石胶原复合支架对人牙髓干细胞体外增殖以及成骨向分化的影响。方法:体外扩增人牙髓干细胞与复合支架联合培养。采用CCK-8法检测人牙髓干细胞在支架上的增殖能力,通过检测碱性磷酸酶活性、茜素红染色和蛋白质... 目的:探讨载柚皮苷羟基磷灰石胶原复合支架对人牙髓干细胞体外增殖以及成骨向分化的影响。方法:体外扩增人牙髓干细胞与复合支架联合培养。采用CCK-8法检测人牙髓干细胞在支架上的增殖能力,通过检测碱性磷酸酶活性、茜素红染色和蛋白质印迹法观察支架对细胞体外成骨向分化能力的影响。结果:载柚皮苷羟基磷灰石胶原复合支架能促进牙髓干细胞增殖(P<0.05),并能增加其ALP活性(P<0.05)增强矿化能力,并对成骨相关标记蛋白RUNX-2和BMP-2的表达具有上调作用。结论:载柚皮苷羟基磷灰石胶原支架可提高人牙髓干细胞的增殖能力以及成骨分化的潜能。 展开更多
关键词 柚皮苷 羟基磷灰石胶原支架 人牙髓干细胞 成骨向分化增殖
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