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Distraction osteogenesis (DO) is widely used for bone tissue engineering technology. Immune regulations play important roles in the process of DO like other bone regeneration mechanisms. Compared with others, the immune regulation processes of DO have their distinct features. In this review, we summarized the immune-related events including changes in and effects of immune cells, immune-related cytokines, and signaling pathways at different periods in the process of DO. We aim to elucidated our understanding and unknowns about the immunomodulatory role of DO. The goal of this is to use the known knowledge to further modify existing methods of DO, and to develop novel DO strategies in our unknown areas through more detailed studies of the work we have done.
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Regeneração Óssea/fisiologia , Osso e Ossos , Osteogênese/fisiologia , Osteogênese por Distração/métodos , Engenharia TecidualRESUMO
Nanomaterial-based drug sustainable release systems have been tentatively applied to bone regeneration. They, however, still face disadvantages of high toxicity, low biocompatibility, and low drug-load capacity. In view of the low toxicity and high biocompatibility of polymer nanomaterials and the excellent load capacity of hollow nanomaterials with high specific surface area, we evaluated the hollow polydopamine nanoparticles (HPDA NPs), in order to find an optimal system to effectively deliver the osteogenic drugs to improve treatment of bone defect. Data demonstrated that the HPDA NPs synthesized herein could efficiently load four types of osteogenic drugs and the drugs can effectively release from the HPDA NPs for a relatively longer time in vitro and in vivo with low toxicity and high biocompatibility. Results of qRT-PCR, ALP, and alizarin red S staining showed that drugs released from the HPDA NPs could promote osteogenic differentiation and proliferation of rat bone marrow mesenchymal stem cells (rBMSCs) in vitro. Image data from micro-CT and H&E staining showed that all four osteogenic drugs released from the HPDA NPs effectively promoted bone regeneration in the defect of tooth extraction fossa in vivo, especially tacrolimus. These results suggest that the HPDA NPs, the biodegradable hollow polymer nanoparticles with high drug load rate and sustainable release ability, have good prospect to treat the bone defect in future clinical practice.
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Animais , Ratos , Regeneração Óssea , Indóis , Nanopartículas , Osteogênese , Preparações Farmacêuticas , PolímerosRESUMO
Epithelial-mesenchymal transition (EMT) is involved in both physiological and pathological processes. EMT plays an essential role in the invasion, migration and metastasis of tumours. Autophagy has been shown to regulate EMT in a variety of cancers but not in head and neck squamous cell carcinoma (HNSCC). Herein, we investigated whether autophagy also regulates EMT in HNSCC. Analyses of clinical data from three public databases revealed that higher expression of fibronectin-1 (FN1) correlated with poorer prognosis and higher tumour pathological grade in HNSCC. Data from SCC-25 cells demonstrated that rapamycin and Earle's balanced salt solution (EBSS) promoted autophagy, leading to increased FN1 degradation, while 3-methyladenine (3-MA), bafilomycin A1 (Baf A1) and chloroquine (CQ) inhibited autophagy, leading to decreased FN1 degradation. On the other hand, autophagic flux was blocked in BECN1 mutant HNSCC Cal-27 cells, and rapamycin did not promote autophagy in Cal-27 cells; also in addition, FN1 degradation was inhibited. Further, we identified FN1 degradation through the lysosome-dependent degradation pathway using the proteasome inhibitor MG132. Data from immunoprecipitation assays also showed that p62/SQSTM1 participated as an autophagy adapter in the autophagy-lysosome pathway of FN1 degradation. Finally, data from immunoprecipitation assays demonstrated that the interaction between p62 and FN1 was abolished in p62 mutant MCF-7 and A2780 cell lines. These results indicate that autophagy significantly promotes the degradation of FN1. Collectively, our findings clearly suggest that FN1, as a marker of EMT, has adverse effects on HNSCC and elucidate the autophagy-lysosome degradation mechanism of FN1.
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Feminino , Humanos , Autofagia , Linhagem Celular Tumoral , Fibronectinas , Lisossomos/metabolismo , Neoplasias Ovarianas , Proteína Sequestossoma-1/metabolismo , Carcinoma de Células Escamosas de Cabeça e PescoçoRESUMO
At the present day, curettage and periodontal surgery comprise the main strategy for the treatment of periodontitis, however, these methods are limited in regenerating cementum. It has been found that some biological factors such asenamel matrix derivative (EMD), transforming growth factor-β (TGF-β) and insulin-like growth factor (IGF) could promote cementum regeneration. In the cementum regenerationstudies, there has been a lack of criteria to distinguish cementum from alveolar bone and other types of cementum. Therefore, this article will briefly review the biological factors that affect the cementum regeneration and the molecular markers used to judge the regenerating cementum.
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Objective@#To evaluate the effect of vascular endothelial growth factor (VEGF)-loaded microspheres on dental pulp tissue regeneration and vascularization in vivo.@*Methods@#In vitro release experiment and human umbilical vein endothelial cell migration experiment were conducted with VEGF loaded microspheres. The dental pulp stem cells (DPSC) were co-cultured with VEGF microspheres to observe the compatibility between the cells and the microspheres. DPSC and VEGF loaded microspheres were injected into the root lumen through the apical foramen, which were then transplanted subcutaneously into nude mice. Histological and immunohistochemical features were observed after nine weeks.@*Results@#DPSCs attached and spread on the surface of the microspheres. HE staining showed that the regenerated pulp-like tissue fulfilled the whole apex and middle third of the root. Differentiated odontoblast-like cells aligned with the existing tubular root dentin.@*Conclusions@#VEGF-loaded microspheres promoted the regeneration of pulp-like tissues and formation of blood vessels.
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Objective:To investigate the transport pathway and intracellular distribution of the of fluorescent carbon dots(CDs)synthesized by folic acid and polyethyleneimine(PEI)through the membrane of MC3T3-E1 cells and its effect on the cells,and to clarify the mechanism.Methods:The fluorescent CDs with the function of cell imaging were synthesized by hydrothermal method using folic acid and PEI as the raw materials;MTT assay was applied to screen the best concentration of CDs.The MC3T3-E1 cells were divided into blank control group,folic acid group and CDs group.The biocompatibility of CDs was evaluated by the detection of cell cycle,apoptosis and cellular reactive oxygen species(ROS)level.Nystatin as a kind of caveolae inhibitor and nocodazole as a kind of macropinocytosis inhibitor were used to find out the pathway through which the cells took in the CDs.Using the charcteristic of CDs with blue fluorescence stimulated by ultraviolet ray,the organelle probes were used to observe the distribution of CDs.Results:Compared with blank control group,the cells in different concentrations(100-450 mg·L-1)of CDs groups showed no cytotoxicity at 24 h(P>0.05);at 48 h,the cell proliferation rate was reduced to 68.4% of blank control group when the concentration of CDs reached 350 mg · L -1(P<0.05). Compared with blank control group,the percentages of cells in G0phase and G1phase in CDs group were decreased (P<0.05),and the percentage of cells in S phase was increased(P<0.05);the percentages of cells in G2phase and M phase were increased,but there no was significant differences(P>0.05).Compared with blank control group,the apoptotic rates of the cells in folic acid group and CDs group had no significant differences(P>0.05). Compared with blank control group,the intracellular ROS levels in folic acid group and CDs group were significantly decreased(P<0.05).Compared with blank control group,the uptake amount of CDs in the cells was decreased in nystatin group(P<0.05).The blue fluorescence of CDs overlapped with the red fluorescence of mitochondria under an inverted fluorescence microscope,the blue fluorescence of CDs overlapped with the red fluorescence of lysosomes;they didn't overlap completely with the red fluorescence of the endoplasmic reticulum;the blue fluorescence of CDs overlapped poorly with the red fluorescence of Golgi apparatus.Conclusion:CDs perform well in biocompatibility and they can be distributed to different organelles after taken in by the cells.They can be used as a kind of gene carrier in transgenic therapy.
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Objective:To study the expressions of cell polarity related proteins CDC42 and PAR3 during tooth germ development in the mice,and to discuss their possible roles during tooth development in the mice.Methods:The whole heads were obtained from the mouse embryo on the days 13.5,14.5,16.5 and 18.5 (E13.5,E14.5,E16.5 and E18.5) and the mice on the postnatal days 1 (PN1) and 5 (PN5).The tissues were fixed in paraformaldehyde,decalcified,dehydrated,embedded in paraffin,and sectioned.The histology of tooth germ was observed by HE staining.The expressions of CDC42 and PAR3 during tooth germ development were detected by immunohistochemistry staining.Results:The HE staining results showed that E13.5,E14.5,E16.5 and E18.5were the bud stage,the cap stage,the early and the late bell stage of tooth germ development,respectively;the tooth germ of PN1 mice showed the matured odontoblasts and ameloblasts;the tooth germ of PN5 mice showed the completed tooth crown development.The immunohistochemistry staining results showed that CDC42 expressed in the tooth germ of the mice at E13.5,E14.5 and E16.5;the CDC42 expression at E 18.5 was reduced compared with E13.5,E14.5 and E16.5;CDC42 mainly expressed in the odontoblasts and ameloblasts of the tooth germ of the mice at PN1 and PN5;PAR3 weakly expressed in the tooth germ of the mice at E13.5 and E14.5,and it was increased at E16.5 and E18.5.At PN1 and PN5,the expressions of PAR3 were decreased compared with E18.5.Conclusion:CDC42 and PAR3 partieipat in the mouse tooth development;during the early stage of tooth germ development,they may be involved in the proliferation and migration of mouse dental germ;during the late stage,CD42 and PAR3 may be involved in the differentiation of the odontoblasts and the ameloblasts,especially in the establishment and maintenance of cell polarity.
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Objective:To study the antimicrobial properties of CaO/ZnO core-shell nanoparticles.Methods:The CaO/ZnO core-shell nanoparticles were prepared via precipitation method.The pH and calcium ion release from the samples which composed of eugenol and nanoparticles were examined respectively.The form of the particles was observed under electron microscope,the ions were analysed by inductively coupled plasma(ICP).The antibacterial activities against Streptococcus mutans,Enterococcus faecalis,Escherichia coli and Staphylococcus aureus were evaluated by agar diffusion test (ADT).Results:CaO/ZnO core-shell nanoparticles were spherical with core-shell structure and with the diameter of 80-90 nm.The calcium ion release and pH were gradually increasing from the nanoparticles in PBS.The antibacterial activity of CaO/ZnO core-shell nanoparticles-eugenol was significantly greater than that of iRoot SP and zinc oxide-eugenol sealer(P<0.01).Conclusion:CaO/ZnO core-shell nanoparticles possess antibacterial activity.
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BACKGROUND: Bone marrow mesenchymal stem cells (BMSCs) are widely used in the field of tissue engineering because of their multi-directional differentiation potential. Micro RNAs play an important role in promoting osteogenic differentiation of BMSCs.OBJECTIVE: To investigate the effect of miR-218 and miR-26a on the osteogenic differentiation of rat BMSCs, and to provide reference for the study on osteogenic differentiation of BMSCs and the clinical application. METHODS: The bone marrow of the femur of Wistar rats was extracted and the BMSCs were isolated and cultured to the 3rd generation. MiR-218, miR-26a and polyethyleneimine (PEI) were mixed in a specific ratio to form a miRNA/PEI complex. Meanwhile, a negative control group was established.RESULTS AND CONCLUSION: (1) Rat BMSCs grew well. (2) The optimal concentration of miRNA mimics was 50 nmol/L by MTT method. (3) The expression of alkaline phosphatase and collagen type I mRNA was significantly increased (P < 0.05). (4) Alkaline phosphatase staining showed that compared with the blank control group and the negative control group, the cytoplasm showed obvious coloring. (5) There were a lot of mineralized nodules shown by alizarin red staining. Therefore, miR-218/PEI complex, miR-26a/PEI complex and miR-218+miR-26a/PEI co-transfection complex could promote the osteogenic differentiation of BMSCs. Under the same conditions, the osteogenesis of miR-26a was slightly better than that of miR-218.
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Objective:To study the lethal effect of vitamin C carbon dots on oral squamous cell carcinoma KB cells, and to clarity its related mechanism.Methods: The KB cells were treated with different concentrations (5, 10, 20, 40 and 80 mg·L-1) of vitamin C carbon dots in vitro as experimental groups, and 0 mg·L-1 vitamin C carbon dots group was used as blank control group.MTT assay was used to detect the proliferation rates of KB cells in various groups;colony formation assay was used to detect the colony formation ability of KB cells;Western blotting was performed to detect the protein expression levels of autophagy related protein LC3 in KB cells in various groups;flow cytometry was used to detect the apoptotic rates of KB cells in various groups.Results: Compared with blank control group, the proliferation rates and colony formation abilities of KB cells in 20, 40 and 80 mg·L-1 carbon dots groups were markedly decreased (P<0.01).Compared with blank control group, the protein expression level of LC3 Ⅱ in 40 mg·L-1 carbon dots group was increased(P<0.05);the apoptotic rate of KB cells was markedly increased(P<0.01).Conclusion: Vitamin C carbon dots can kill the oral squamous cell carcinoma KB cells effectively, suppress the proliferation and impair the colony formation ability of KB cells, which is related to autophagy and apoptosis of KB cells.
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Pathogen-associated molecular patterns (PAMPs) are conservative molecules associated with groups of pathogens or their products. These molecules are recognized by relevant receptors. PAMPs induce the expression of inflammatory cytokines through the signal cascade. The role of PAMPs in the initiation and development of periodontitis is recently attracting attention. PAMPs induce the expression of inflammatory mediators after they are recognized in the periodontium. This process damages the periodontal soft tissue and osseous tissue, thus resulting in periodontitis. The results of this study will provide an excellent resolution for the treatment of periodontitis by blocking the pathogenic pathway of PAMPs.
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Humanos , Citocinas , Moléculas com Motivos Associados a Patógenos , Periodontite , PeriodontoRESUMO
Objective:To transfect the non-viral vector polyethylenimine (PEI)mediated miR-2861 mimic into the MC3T3-E1 cell line,and to explore the transfection efficiency of PEI/miR-2861 complex and its effects on the proliferation and osteogenesis differentiation in pre-osteoblasts. Methods:The proper amount of PEI was blended with miR-2861 mimic and negative control (NC)separately in a ratio of N∶P=10∶1,and they were divided into experiment group and NC group. The NC/PEI complex acted as the NC group was used to eliminate the interference of osteogenesis from the addition of double-stranded RNA mimic.MTT assay was used to determine the optimal concentration of PEI/miR-2861 mimic complex.The fluorescence imaging technique and bulge-loop RT-PCR were used to detect the transfection efficiency and mRNA expression of miRNA-2861 in the cells with different concentrations (10,30, 50,and 100 nmol · L-1 ), separately.The osteogenesis ability of MC3T3 cells was identified with RT-PCR and Alizarin red staining with the selected concentration of PEI/miR-2861 by transient transfection.Results:Compared with blank control group,the proliferation rates of MC3T3 cells in 100 nmol·L-1 PEI/miR-2861 group was decreased significantly at 72 h (P < 0. 05 ). With the increasing of transfected concentration the transfection efficiency of miRNA/PEI complex was increased gradually.The results of Alizarin red staining and quantitative analysis showed that calcium deposits were more and bigger in experiment group after induced for 21 d,while both in blank control group and NC group they were less.Conclusion:The miRNA-2861 mimic can be effectively transfected into the MC3T3-E1 cell line and expresses with a high level,which is mediated by PEI as the gene vector.miR-2861 mimic has a certain ability of promoting osteogenesis differentiation of MC3T3-E1 cells.
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OBJECTIVE: To investigate the involvement of ephrinB2 in periodontal tissue remodeling in compression areas during orthodontic tooth movement and the effects of compressive force on EphB4 and ephrinB2 expression in osteoblasts and osteoclasts. METHODS: A rat model of experimental tooth movement was established to examine the histological changes and the localization of ephrinB2 in compressed periodontal tissues during experimental tooth movement. RAW264.7 cells and ST2 cells, used as precursor cells of osteoclasts and osteoblasts, respectively, were subjected to compressive force in vitro. The gene expression of EphB4 and ephrinB2, as well as bone-associated factors including Runx2, Sp7, NFATc1, and calcitonin receptor, were examined by quantitative real-time polymerase chain reaction (PCR). RESULTS: Histological examination of the compression areas of alveolar bone from experimental rats showed that osteoclastogenic activities were promoted while osteogenic activities were inhibited. Immunohistochemistry revealed that ephrinB2 was strongly expressed in osteoclasts in these areas. Quantitative real-time PCR showed that mRNA levels of NFATc1, calcitonin receptor, and ephrinB2 were increased significantly in compressed RAW264.7 cells, and the expression of ephrinB2, EphB4, Sp7, and Runx2 was decreased significantly in compressed ST2 cells. CONCLUSIONS: Our results indicate that compressive force can regulate EphB4 and ephrinB2 expression in osteoblasts and osteoclasts, which might contribute to alveolar bone resorption in compression areas during orthodontic tooth movement.
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Animais , Ratos , Reabsorção Óssea , Expressão Gênica , Imuno-Histoquímica , Modelos Animais , Osteoblastos , Osteoclastos , Reação em Cadeia da Polimerase em Tempo Real , Receptores da Calcitonina , RNA Mensageiro , Técnicas de Movimentação DentáriaRESUMO
A rare case of submandibular venous malformation with multiple phleboliths is reported.The clinical pathology,diagnosis ,treat-ment,causes and differential diagnosis of submandibular gland sialolithiasis were discussed based on related literatures.
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Objective:To study the effect of IL-24 on the differentiation and function of osteoclasts.Methods:Mature osteoclasts were isolated from long bones of neonate rats.Optimal multiplicity of infection(MOI)of AdCMV-EGFP was determined.Then osteo-clasts and RAW264.7 cells were transfected with AdCMV-EGFP and AdCMV-IL-24 respectively.The function of osteoblasts was studied by the observation of bone resorption lacunae,the differentiation of RAW264.7 cells was evaluated by the expression of osteo-clast related genes examined with real-time PCR.Results:Osteoclasts were TRAP positive with more than 2 neclei.MOI=400 was suitable for AdCMV-EGFP transfection.The resorption lacunae area in AdCMV-IL24 transfected cells was larger than that in AdCMV-EGFP transfected cells(P<0.05).Real-time PCR showed that under induced conditions,osteoclastic related genes NFATc1,CTSK and TRAP were changed in RAW264.7 cells transfected with AdCMV-IL-24.Conclusion:IL-24 may promote the differentiation and function of osteoclasts.
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0.05);) Single Bond provided significantly higher shear bond strengths than Adper Prompt in all groups((P
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Objective To establish the method to express leukemia inhibitory factor(LIF) in fetal fibroblasts in order to provide theoretical foundation for establishment of transgenic animal model.Methods Using the fetal of 13.5 d ICR mouse,the primary fetal fibroblasts were cultivated by trypsin enzyme digestion.The lineared eukaryotic expression vector pcDNA3.1-LIF was transferred to the fetal fibroblasts of mouse with liposome induction.The positive cells were selected by G418,genome DNA of the positive cells was extracted and sequenced. Results The primary fetal fibroblasts of mouse were successfully obtained by isolating and culturing,and fetal fibroblasts expressing LIF were established by transferring.The sequencing result demonstrated that the homology of clone plasmid of positive cells was about 99%.Conclusion Eukaryotic expression vector pcDNA3.1-LIF is successfully transferred to the fetal fibroblasts of mouse.
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Objective To evaluate the effects of different loading occasions of orthodontic force on the stability of miniscrew as an anchorage.Methods The titanium miniscrews were respectively implanted into canine and posterior region of both upper and lower jaws of 3 crossbred dogs.The implants were immediately loaded with 200 g orthodontic force or loaded after 2 weeks of implantation.The dogs were sacrificed after 8 weeks.The samples including the microscrew implant and its around bone tissue were extracted,fixed and X-ray photographed.Then the specimens were decalcified,embedded,sectioned and stained with HE and GOMORI.The sections were examined under light microscope.Results The stability of miniscrew in posterior region of jaws was higher than that in canine region in X-ray photograph.There was more bone trabecular formation between miniscrew and jaw bone in non-immediately loaded group than those in immediately loaded group with HE and GOMORI.Conclusion The non-immediately loaded implant as an anchorage can afford more stability than the immediately loaded implant.
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Objective To observe the effects of porous polylactic acid/polyglycolic acid copolymer(PLGA) filling into extraction socket on the regeneration of alveolar bone in rats.Methods Sixty male Wistar rats were divided into experimental group and control group(n=30).PLGA scaffold was immediately implanted in the mandibular incisor root sockets after removal of incisor teeth.Soft X-ray photography,structural observation,light microscopy were used to evaluate the effects of composite on bone healing in a rat tooth extraction socket.Results The relative lengths of residual alveolar ridge in the experimental group 4,8 weeks after operation were shorter than that in the control group,and there were significant differences between the experimental group and the control group(P
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<p><b>OBJECTIVE</b>The purpose of this study was to investigate the risk factors involving lymph node metastasis of squamous cell carcinoma and evaluate if the risk factors would be effective in predicting neck lymph node metastasis.</p><p><b>METHODS</b>Based on the original data of 106 cases with carcinoma of tongue, four related factors, including maximal diameter, degree of differentiation, mode of invasion and histological grade of malignancy were analyzed statistically.</p><p><b>RESULTS</b>The maximal diameter and the grade of tumor cell differentiation showed no significant correlation with cervical lymph node metastasis; but the mode of invasion and the histological grade of malignancy demonstrated a significant correlation.</p><p><b>CONCLUSION</b>The results obtained suggested that the mode of invasion and the histological grade of malignancy were closely related to cervical lymph node metastasis.</p>