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1.
Chemosphere ; 238: 124753, 2020 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-31545217

RESUMO

Boscalid is a widely used fungicide in agriculture and has been frequently detected in both environments and agricultural products. However, evidence on the neurotoxic effect of boscalid is scarce. In this study, zebrafish served as an animal model to investigate the toxic effects and mechanisms of boscalid on aquatic vertebrates or higher animals. And we unravelled that boscalid induced developmental defects associated with oxidative stress. Developmental defects, including head deformity, hypopigmentation, decreased number of newborn neurons, structural defects around the ventricle, enlarged intercellular space in the brain, and nuclear concentration, were observed in zebrafish embryos after boscalid exposure at 48 hpf. Interestingly, we found that boscalid might directly induce oxidative stress and alter the activity of ATPase, which in turn disrupted the expression of genes involved in neurodevelopment and transmitter-transmitting signalings and melanocyte differentiation and melanin synthesis signalings. Ultimately, the differentiation of nerve cells and melanocytes were both impacted and the synthesis of melanin was inhibited, leading to morphological abnormalities. Additionally, exposure to boscalid led to less and imbalance motion and altered tendency of locomotor in larval fish. Collectively, our results provide new evidences for a comprehensive assessment of its toxicity and a warning for its residues in environment and agricultural products.


Assuntos
Compostos de Bifenilo/toxicidade , Fungicidas Industriais/toxicidade , Larva/efeitos dos fármacos , Niacinamida/análogos & derivados , Estresse Oxidativo/efeitos dos fármacos , Poluentes Químicos da Água/toxicidade , Peixe-Zebra/embriologia , Animais , Diferenciação Celular/efeitos dos fármacos , Modelos Animais de Doenças , Embrião não Mamífero/efeitos dos fármacos , Humanos , Melaninas/biossíntese , Melanócitos/citologia , Neurônios/citologia , Síndromes Neurotóxicas/patologia , Niacinamida/toxicidade , Peixe-Zebra/metabolismo
2.
Int Endod J ; 53(1): 72-83, 2020 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-31419325

RESUMO

AIM: To comparatively evaluate changes in the proliferation and mineralization abilities of dental pulp stem cells (DPSCs) from juvenile and adult rats in a lipopolysaccharide (LPS)-induced inflammatory microenvironment to provide a theoretical basis for the age-related differences observed in DPSCs during repair of inflammatory injuries. METHODOLOGY: DPSCs were isolated from juvenile (JDPSCs) and adult rats (ADPSCs), and senescence-associated ß-galactosidase staining was used to compare senescence between JDPSCs and ADPSCs. Effects of LPS on JDPSCs and ADPSCs proliferation were investigated by cell counting kit-8 assays and flow cytometry. Alizarin red staining, quantitative reverse transcription polymerase chain reaction and Western blot assay were used to examine the effects of LPS on mineralization-related genes and proteins in JDPSCs and ADPSCs. Immunohistochemistry was used to compare interleukin-1ß (IL-1ß) and osteocalcin (OCN) expression in the pulpitis model. Unpaired Student's t-tests and one-way anova were used for statistical analysis. RESULTS: DPSCs were isolated from juvenile and adult rat dental pulp tissues. At low concentrations (0.1-1 µg mL-1 ), LPS significantly promoted the proliferation of JDPSCs (P < 0.01) and ADPSCs (P < 0.01 or P < 0.05), with the effect being stronger in JDPSCs than in ADPSCs. In addition, mineralized nodules and the expression of mineralization-related genes (OCN, DSPP, ALP, BSP) increased significantly after stimulation with LPS (0.5 µg mL-1 ) in JDPSCs and ADPSCs (P < 0.01 or P < 0.05), and JDPSCs displayed a more obvious increase than ADPSCs. Western blots revealed OCN and ALP expression levels in JDPSCs treated with LPS were significantly upregulated (P < 0.05); meanwhile, ALP expression in ADPSCs increased slightly but significantly (P < 0.05), and OCN expression was not affected. Finally, IL-1ß expression was significantly higher (P < 0.05) and OCN expression was significantly lower (P < 0.05) in the inflamed dental pulp of adult rats than in juvenile rats. CONCLUSIONS: A certain degree of inflammatory stimulation promoted the proliferation and mineralization of DPSCs; however, this effect declined with age. The DPSCs of adult donors in an inflammatory microenvironment have a weaker repair ability than that of juvenile donors, who are better candidates for tissues damage repair.


Assuntos
Polpa Dentária , Células-Tronco , Afeto , Fosfatase Alcalina , Animais , Diferenciação Celular , Proliferação de Células , Células Cultivadas , Humanos , Ratos
3.
Arch Oral Biol ; 109: 104552, 2020 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-31541843

RESUMO

OBJECTIVES: The aim of this in vitro study was to investigate the behavior of osteoblasts on titanium discs under different concentrations of enamel matrix derivatives (EMD) and dentin matrix derivative (DMD). MATERIALS AND METHODS: MC3T3-E1 osteoblast-like cells were cultivated on coated titanium SLA discs with EMD or DMD at 100 µg/ml, 1 mg/ml, 10 mg/ml and 30 mg/ml or left uncoated. Cell viability, proliferation, adhesion and migration were assessed respectively with MTT, BrdU, DAPI and scratch wound healing assays. Messenger ribonucleic acid of different genes related to osteoblastic differentiation was quantified by means of real-time quantitative PCR. Data were analyzed using student t-test for adhesion and migration assay and ANOVA for proliferation assay (p < 0.05). RESULTS: BrdU incorporation was found in proliferative osteoblasts for both test solutions at all concentrations. Osteoblast migrated and covered approximately 70% of the wound area observed at time zero when exposed to EMD and DMD to all concentrations. The increase of gene expression was dependent on the concentration enhancement of EMD and DMD. Higher concentrations showed proliferation augmentation if compared to lower concentrations. CONCLUSIONS: Roughness surface of Ti SLA can limit cell adhesion independent of the presence EMD or DMD. DMD enhances cell migration of osteoblasts on SLA titanium implants in a concentration-dependent manner.


Assuntos
Proteínas do Esmalte Dentário/química , Implantes Dentários , Dentina/química , Osteoblastos/citologia , Titânio , Células 3T3 , Animais , Diferenciação Celular , Proliferação de Células , Camundongos , Propriedades de Superfície
4.
Arch Oral Biol ; 109: 104557, 2020 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-31557575

RESUMO

OBJECTIVE: To investigate the effects of dental x-ray on proliferation and mineralization in human primary osteoblasts as well as on proliferation and apoptotic potential in human periodontal ligament (PDL) cells. DESIGN: Primary osteoblasts and PDL cells were irradiated with various doses of periapical radiography by repeated exposures and further incubated for 1, 3 or 7 days. Cell proliferation was assayed by BrdU incorporation. The effect of dental x-ray on mineralization in osteoblasts either before or after x-ray exposures was determined by Alizarin red staining. Both mRNA and protein expressions of BCL-2, an anti-apoptotic gene, and BAX, a pro-apoptotic gene, in PDL cells were analyzed by RT-qPCR and immunoblotting analysis, respectively. RESULTS: Neither the proliferative nor the mineralization ability of irradiated osteoblasts was different from that of non-irradiated osteoblasts at any doses or time points. By contrast, there was a significant decrease in the proliferation of PDL cells on day 3 after repeated exposures to dental x-ray for 20 times (P < 0.05), whereas the ratio of BCL-2 to BAX mRNA and protein expressions in these irradiated PDL cells was significantly increased (P < 0.05). CONCLUSIONS: Upon multiple exposures to dental x-ray used in intraoral radiography up to 20 times, there is no effect on the proliferation or the mineralization of osteoblasts, whereas the proliferative and apoptotic potentials of PDL cells are transiently decreased.


Assuntos
Fibroblastos/efeitos da radiação , Osteoblastos/efeitos da radiação , Ligamento Periodontal/citologia , Raios X , Adolescente , Adulto , Apoptose , Diferenciação Celular , Proliferação de Células , Células Cultivadas , Criança , Feminino , Humanos , Masculino , Proteínas Proto-Oncogênicas c-bcl-2/genética , Radiografia Dentária , Adulto Jovem , Proteína X Associada a bcl-2/genética
5.
Arch Oral Biol ; 109: 104570, 2020 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-31568992

RESUMO

OBJECTIVE: The aim of this study is to investigate the effects of 17ß-Estradiol (E2) at different concentrations combined with cyclical compressive stress on the proliferation and differentiation of mandibular condylar chondrocytes (MCCs). DESIGN: MCCs, isolated from female Sprague-Dawley rats, were exposed to E2 at different concentration, cyclical compressive stress or the combination, effects of which on MCCs proliferation and differentiation were detected. RESULTS: E2 at physiological concentration (10-9 mol/L) has lower proliferative effects on MCCs, compared with non-physiological concentration (10-12 mol/L or 10-6 mol/L). For MCCs differentiation, effects of E2 at different concentration are totally opposite: E2 at 10-9 mol/L promotes MCCs differentiation, but at 10-12 mol/L or 10-6 mol/L, it inhibits MCCs differentiation. When combined with E2 at 10-9 mol/L, cyclical compressive stress shows synergistic effect on proliferation and differentiation. However, when combined with E2 at 10-12 mol/L or 10-6 mol/L cyclical compressive stress reverses the inhibition in MCCs differentiation provoked by E2 at 10-12 mol/L or 10-6 mol/L. CONCLUSION: Effects of E2 combined with cyclical compressive stress on MCCs proliferation and differentiation are different, which suggests that orthodontist should take fully consideration of the levels of E2 and adopt comprehensive strategies, so as to achieve better orthodontic effect.


Assuntos
Condrócitos/citologia , Estradiol/farmacologia , Côndilo Mandibular/citologia , Pressão , Animais , Diferenciação Celular , Proliferação de Células , Células Cultivadas , Feminino , Ratos , Ratos Sprague-Dawley
6.
Arch Oral Biol ; 109: 104574, 2020 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-31585238

RESUMO

Fibroblast growth factors (FGFs) are growth factors that play an important role in tooth development, repair, and regeneration. Of the FGF families, basic fibroblast growth factor (bFGF) has been the most frequently investigated in dentistry. Numerous studies have reported advantages of bFGF, while others did not find any additional benefit. This review gives a comprehensive summary of the potential role of bFGF in dental pulp wound healing and regeneration in connection with cell proliferation and differentiation, angiogenesis, and neural differentiation from both in vitro and in vivo studies. Furthermore, the possible underlying mechanisms associated with bFGF in promoting dental pulp wound healing are discussed in this review.


Assuntos
Polpa Dentária/fisiologia , Fator 2 de Crescimento de Fibroblastos/fisiologia , Animais , Diferenciação Celular , Proliferação de Células , Humanos , Neovascularização Fisiológica , Odontogênese , Regeneração , Cicatrização
7.
Arch Oral Biol ; 109: 104572, 2020 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-31600663

RESUMO

OBJECTIVE: To compare three different xeno-free protocols for neural differentiation of human dental pulp stem cells (DPSC). METHODS: DPSC were treated with three different media to induce neural differentiation namely N1 (DMEM for 5 days), N2 (PSC neural induction media for 7 days) and N3 (neural media with B27 supplement, 40 ng/ml bFGF and 20 ng/ml EGF for 21 days). Cell proliferation (MTS assay), morphology, gene (qPCR for NESTIN, VIMENTIN, TUB-3, ENO2, NF-M and NF-H) and protein expression (flow cytometry) of neurogenic markers were assessed at different time points and compared to untreated cells (DMEM supplemented with 10% FBS). Statistical analysis was performed with global significance level of 5%. RESULTS: N1 and N2 formulations increased the genetic expression of two out of six genes TUB-3, NF-M and TUB-3, NF-H, respectively, whereas N3 elevated the expression of all genes by the late stage. N3 also stimulated protein expression for NESTIN, TUB-3 and NF-H. Cells treated with both N2 and N3 presented neuron-like morphology, decreased proliferation and expression of stemness genes at protocol end point. CONCLUSION: N3 was the most effective formulation in promoting a neurogenic shift in gene and protein expression. Cells provided with the N3 formulation exhibited neuron-like morphology, elaborating axonal-like projections concomitant with cell cycle withdrawal and reduced expression of stemness genes indicating greater commitment to a neurogenic lineage.


Assuntos
Técnicas de Cultura de Células , Diferenciação Celular , Polpa Dentária/citologia , Neurogênese , Células-Tronco/citologia , Células Cultivadas , Meios de Cultura , Humanos , Neurônios/citologia
8.
Arch Oral Biol ; 109: 104582, 2020 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-31605918

RESUMO

OBJECTIVE: The aim of this study was to evaluate the proliferation and odontogenic differentiation of human dental pulp cells (hDPCs) and human umbilical cord mesenchymal stem cells (hUCMSCs) in three-dimensional co-culture system which was established with the help of bone morphogenetic protein-2 (BMP-2) and hydrogel. METHODS: hDPCs and hUCMSCs were cultured in different concentrations of hydrogel to explore the more suitable concentrations for subsequent experiments. hUCMSCs and hDPCs induced by BMP-2 were co-cultured in the hydrogel. MTT assay was used to measure the cell viability. The differentiation into odontoblast-like cells were measured by the mRNA expression of dentin salivary phosphoprotein (DSPP), dentin matrix protein-1 (DMP-1), alkaline phosphatase and osteocalcin. Alizarin red staining was performed for the formation of mineralized nodules. RESULTS: hUCMSCs and hDPCs could grow and proliferate in hydrogel scaffold. The growth rate of cells in lower concentrations hydrogels were higher than that of high concentrations hydrogels (P < 0.05). The study showed that 0.25% hydrogel scaffold was more suitable for subsequent experiments than other groups. Compared with hUCMSCs-monoculture and hDPCs-monoculture, the co-culture groups exhibited more proliferative potential, alkaline phosphatase activity and mineralization nodule formation (P < 0.05). The mRNA expression in co-culture groups were higher than that of hUCMSCs-monoculture, closed to or even higher than that of hDPCs-monoculture. CONCLUSION: 0.25% hydrogel was the suitable concentration in co-culture system for subsequent experiments. The co-culture groups had stronger abilities of odontoblastic differentiation and mineralization than cells-monoculture groups, indicated that the co-culture conditions could regulate cell proliferation and differentiation within a certain range.


Assuntos
Diferenciação Celular , Polpa Dentária/citologia , Hidrogéis , Células-Tronco Mesenquimais/citologia , Fosfatase Alcalina/genética , Proliferação de Células , Células Cultivadas , Técnicas de Cocultura , Proteínas da Matriz Extracelular/genética , Humanos , Odontoblastos/citologia , Osteocalcina/genética , Fosfoproteínas/genética , Sialoglicoproteínas/genética , Cordão Umbilical/citologia
9.
Arch Oral Biol ; 109: 104584, 2020 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-31630006

RESUMO

OBJECTIVES: To investigate whether rutin could protect human periodontal ligament stem cells (hPDLSCs) from TNF-α induced damage to osteogenic differentiation in inflammatory environment and detect the underlying mechanism. MATERIALS AND METHODS: hPDLSCs were identified by flow cytometery. TNF-α was used to stimulate hPDLSCs to establish an inflammation model in vitro. Alkaline phosphatase (ALP) staining, ALP activity test, and Alizarin Red staining were used to detect the changes of osteogenic differentiation ability. The mRNA and protein levels of osteogenic genes were evaluated by RT-PCR and Western Blot. The expression of mTOR was also detected by Western Blot. RESULTS: hPDLSCs were positive to MSCs specific surface markers. The inflammatory environment in vitro could be established by stimulating hPDLSCs with TNF-α (20 ng/mL). TNF-α (20 ng/mL) could decrease the ALP activity and mineralization ability of hPDLSCs and down-regulate the expression of osteogenic genes in inflammatory environment. Moreover, rutin could affect TNF-α (20 ng/mL) induced damage to osteogenic differentiation of hPDLSCs in a dose-dependent manner, 10 µmol/L rutin could significantly reverse the damage caused by TNF-α. In addition, rutin inhibited TNF-α-activated mTOR signal transduction by inhibiting the phosphorylation of mTOR, similar to the effects of rapamycin(a specific mTOR inhibitor). CONCLUSIONS: Rutin could protect hPDLSCs from TNF-α induced damage to osteogenic differentiation in inflammatory environment, and rutin is expected to become a new candidate drug for the treatment of bone defect of periodontitis.


Assuntos
Osteogênese , Ligamento Periodontal/citologia , Rutina/farmacologia , Transdução de Sinais , Células-Tronco/efeitos dos fármacos , Serina-Treonina Quinases TOR/metabolismo , Fator de Necrose Tumoral alfa/efeitos adversos , Fosfatase Alcalina/metabolismo , Diferenciação Celular , Células Cultivadas , Humanos , Inflamação , Células-Tronco/citologia
10.
Arch Oral Biol ; 109: 104579, 2020 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-31634727

RESUMO

OBJECTIVES: To investigate the effect and mechanism of calcium on LS8 cell differentiation, especially on phosphatidylinositol 3 kinase (PI3K) /protein kinase B(AKT) pathway. MATERIALS AND METHODS: Ameloblast-like LS8 cell line was used and additional 0-3.5 mmol/L calcium chloride was treated for 24 h, 48 h. Cell viability and morphological changes, cell cycle and associated regulatory proteins were analyzed. RESULTS: No significant effects on morphological changes were observed. Decreased cell viability and increased S phase cells were accompanied by the significant decrease of cyclin A and cyclin B proteins, and significant increase of cyclin D protein in LS8 cells. Additionally, kallikrein-4 and amelotin expressions were significantly increased. Finally, the levels of PI3K, AKT, p-AKT and forkhead box O3 (FOXO3) significantly downregulated after calcium treatment in LS8 cells. CONCLUSIONS: Calcium inhibit proliferation and promotes differentiation in LS8 cells, this is closely related to the downregulation of PI3K/AKT signal in LS8 cells.


Assuntos
Ameloblastos/enzimologia , Cálcio/farmacologia , Fosfatidilinositol 3-Quinase/metabolismo , Proteínas Proto-Oncogênicas c-akt/metabolismo , Transdução de Sinais , Ameloblastos/efeitos dos fármacos , Animais , Diferenciação Celular , Linhagem Celular , Proliferação de Células , Regulação para Baixo , Camundongos
11.
Zhonghua Kou Qiang Yi Xue Za Zhi ; 54(12): 841-846, 2019 Dec 09.
Artigo em Chinês | MEDLINE | ID: mdl-31874485

RESUMO

Objective: To investigate the effect of PR domain zinc finger protein 9 (PRDM9), one of the histone methylated transferases, on osteogenic differentiation ability of periodontal ligament mesenchymal stem cells (PDLSC). Methods: PDLSC with PRDM9 gene knocked down by PRDM9 shRNA using recombinant lentiviral vector were allocated into the PRDM9sh group, and the transfected shRNA was as the control group. The gene expression efficiency was evaluated by reverse transcription polymerase chain reaction (RT-PCR). Alkaline phosphatase activity (ALP), alizarin red staining, mineralization and osteocalcin, which belongs to osteogenic differentiation markers detected by RT-PCR and Western blotting to detect the osteogenic differentiation ability of stem cells from periodontal ligaments in vitro. In vivo, PRDM9sh and control group cells was transplanted into the dorsal dermal to explore the osteogenesis. The area percentage of new osteogenic tissue was calculated by image pro software and statistically analyzed. Results: RT-PCR results showed that the relative expression of PRDM9 gene in PRDM9sh (0.460±0.017) was significantly lower than that in control group (1.000±0.107) (P<0.05). The results of ALP activity determined at 5 days postinduction in a significant decrease in PRDM9sh cells (0.762±0.063) compared with control group (1.225±0.058) (P<0.01). Alizarin red staining induced by osteogenesis at 2 weeks and 3 weeks showed that the staining of PRDM9sh was significantly lighter than that in control group. Quantitative calcium analysis results showed that the calcium ion concentration induced by osteogenesis at 2 weeks and 3 weeks [(0.071±0.004), (0.075±0.001)] in PRDM9sh was significantly lower than that in control group at 2 weeks and 3 weeks [(0.282±0.006), (0.485+0.004)] (P<0.01). RT-PCR results showed that the relative expression of osteocalcin mRNA in PRDM9sh (1.059±0.148) was significantly lower than that in control group at 2 weeks (2.542±0.190) (P<0.01). Western blotting results showed that osteocalcin expression in PRDM9sh was significantly lower than that in control group at 1 and 2 weeks after osteogenesis induction. Animal transplantation experiments results indicated that PRDM9 significantly inhibited the osteogenesis of PDLSC in vivo, and the proportion of osteogenic area calculated showed that the osteogenic capacity of PRDM9sh [(3.8±2.41)%] was significantly lower than that in control group [(24.54±7.06)%](P<0.05). Conclusions: Depletion of PRDM9 repressed the osteogenic differentiation of stem cells from periodontal ligament in vitro and in vivo.


Assuntos
Diferenciação Celular , Histona-Lisina N-Metiltransferase/genética , Osteogênese , Ligamento Periodontal/citologia , Células-Tronco/citologia , Animais , Células Cultivadas , Técnicas de Silenciamento de Genes
12.
Sheng Wu Yi Xue Gong Cheng Xue Za Zhi ; 36(6): 1038-1042, 2019 Dec 25.
Artigo em Chinês | MEDLINE | ID: mdl-31875380

RESUMO

Circular RNA (circRNA) is a type of single-stranded RNA that binds in a closed loop structure by covalent bond. It is highly expressed and has diverse functions in the eukaryotic transcriptome, and it also has the potential to regulate the process of cell differentiation. Stem cells are important seed cells and common research tools in the field of tissue engineering, which have multi-directional differentiation potential and low immunogenicity. Its clinical application for the treatment of diseases has broad prospects, and the research on their differentiation mechanism has gradually penetrated to the molecular level. A number of studies have shown that circRNA participates in stem cell differentiation and plays a key role through a variety of pathways. This article focuses on the expression changes of circRNA during stem cell differentiation and its research advancement in regulating the differentiation mechanism of various stem cells. The review also prospects its possible role in tissue regeneration and repair, in order to further study the molecular mechanism of circRNA involved in stem cell differentiation and provide ideas for clinical practice of stem cells in biomedical engineering.


Assuntos
Diferenciação Celular , Engenharia Tecidual , RNA , Células-Tronco , Transcriptoma
13.
Zhongguo Gu Shang ; 32(11): 1026-1033, 2019 Nov 25.
Artigo em Chinês | MEDLINE | ID: mdl-31870051

RESUMO

OBJECTIVE: To construct and compare macrophage by gene modification of Wnt5a which co-cultured by human bone marrow-derived mesenchymal stem cells (MSCs) with two dimension (2D) and bone marrow aspirates in vitr transwell, in order to investigate the effect of Wnt5a signaling on cartilage homeostasis through regulation of macrophage pro-inflammatory responses. METHODS: Macrophages, MSCs and bone marrow aspirates specimens were extracted from 6 patients with severe knee deformities undergoing total knee arthroplasty(2 males, 4 females, aged from 58 to 71 years old) from September 2015 to December 2018. The synovial tissues of knee joints were exposed to type II collagenase and obtained single cell suspensions, and the purity of macrophages was determined by Ficoll gradient centrifugation and anti-CD14 antibody flow cytometry. The macrophages were transduced with IFN-γ combined with TNF-α for 48 h, and rAAV-lacZ or Wnt5a transfected for 24 h, then co-cultured model by human bone marrow-derived mesenchymal stem cells (MSCs) with two dimension (2D) and bone marrow aspirates in vitro transwell. HE staining, toluidine blue staining, X-gal staining and anti-Wnt5a, anti-II, X collagen immunohistochemical staining and enzyme-linked immunosorbent assay were applied to detect cell morphology and proliferation (cellularity), viral transfection efficiency respectively. RESULTS: Results of anti-CD68 immunohistochemistry showed macrophage of patients with osteoarthritis increased obviously. Anti-CD 14 flow cytometry confirmed that percentage of isolated synovial macrophages was 90.31%. The level of monocyte chemotactic protein-1 in supernatants was significantly increased after stimulation with IFN-γ and TNF-α, indicating that the macrophages were activated and at proinflammatory condition. After transduction with rAAV-lacZ for 3 days, X-gal staining indicated that lacZ gene transfer could efficiently transduce the activated macrophages with the efficiency over 97.50% for at least 21 days. After transfection macrophages by rAAV-Wnt5a stimulated expression of Wnt5a, and enhanced expression of Wnt5a between two models and inhibited the secretion of MCP-1. The expression of MCP-1 in Wnt5a group was 14.76 and 61.51 pg/ml respectively. In addition, rAAV-Wnt5a gene transfer could promote cell proliferation, chondrogenic differentiation and cartilage matrix synthesis. CONCLUSIONS: Under the condition of macrophage and MSCs 2D monolayer or aspirates transwe ll co-culture, rAAV-mediated over-expression of Wnt5a could promote maintenance of cartilage homeostasis and chondrogenic differentiation of MSCs via macrophage inflammatory response, macrophages may affect cartilage homeostasis and MSCs chondrogenesis through Wnt5a signaling pathway.


Assuntos
Condrogênese , Macrófagos , Idoso , Células da Medula Óssea , Diferenciação Celular , Células Cultivadas , Condrócitos , Técnicas de Cocultura , Feminino , Humanos , Inflamação , Masculino , Pessoa de Meia-Idade , Proteína Wnt-5a
14.
Sheng Wu Gong Cheng Xue Bao ; 35(12): 2238-2256, 2019 Dec 25.
Artigo em Chinês | MEDLINE | ID: mdl-31880133

RESUMO

Dynamic variations of the cell microenvironment can affect cell differentiation, cell signaling pathways, individual growth, and disease. Optogenetics combines gene-encoded protein expression with optical controlling, and offers a novel, reversible, non-invasive and spatiotemporal-specific research tool to dynamically or reversibly regulate cell signaling pathways, subcellular localization and gene expression. This review summarizes the types of optogenetic components and the involved cellular signaling pathways, and explores the application and future prospects of the light-controlled cell signaling pathways.


Assuntos
Optogenética , Diferenciação Celular , Luz , Proteínas , Transdução de Sinais
15.
J Contemp Dent Pract ; 20(10): 1171-1178, 2019 Oct 01.
Artigo em Inglês | MEDLINE | ID: mdl-31883252

RESUMO

AIM: Aim of the study was to investigate the effect of PRP and MTA individually and combined on in vitro human bone marrow mesenchymal stem cells' (MSCs) proliferation and osteo/odontogenic differentiation potential. MATERIALS AND METHODS: MSCs were cultured in vitro with MTA, 5% PRP, 10% PRP, MTA with 5%PRP, and MTA with 10% PRP. Fetal calf serum (FCS) was used as control. Cell viability and proliferative efficiency were tested with cell adhesion and MTT assay. Osteo/odontogenic differentiation was assessed and quantified with alizarin red staining. RESULTS: MTA alone, MTA with 5% PRP, and MTA with 10% PRP showed significantly high proliferation at day 7 and 14 when compared to the control group. Enhanced differentiation and the highest calcium deposition was observed in MTA with the 10% PRP group. CONCLUSION: Within limitations of the in vitro environment, results imply an increased proliferation and induction of MSCs into osteo/odontogenic differentiation by the combination rather than a mere sealing of PRP by MTA. CLINICAL SIGNIFICANCE: PRP and MTA have the potential for true regeneration of the pulp tissue. Moreover, the combination of PRP and MTA can be utilized to expand the MSCs to generate adequate numbers for clinical applications, without xenogenic contamination. How to cite this article: Vanka A, Vishwakarma SK, Bhat MK, et al. Osteo/odontogenic Differentiation of Human Mesenchymal Stem Cells with Platelet-rich Plasma and Mineral Trioxide Aggregate. J Contemp Dent Pract 2019;20(10):1171-1178.


Assuntos
Células-Tronco Mesenquimais , Plasma Rico em Plaquetas , Compostos de Alumínio , Compostos de Cálcio , Diferenciação Celular , Proliferação de Células , Células Cultivadas , Combinação de Medicamentos , Humanos , Óxidos , Silicatos
16.
Zhonghua Shao Shang Za Zhi ; 35(12): 887-890, 2019 Dec 20.
Artigo em Chinês | MEDLINE | ID: mdl-31877614

RESUMO

Dental stem cell is a kind of stem cell isolated from dental hard tissue or periodontal tissue, including dental pulp stem cell, stem cell from human exfoliated deciduous teeth, stem cell from root apical papilla, periodontal ligament stem cell, dental follicle progenitor cell, and so on. As seed cell, dental stem cell provides safe and efficient cell source for nerve tissue engineering research. The review aims to introduce the characteristics of these dental stem cells in promoting the regeneration and preparation of nerve and the clinical application.


Assuntos
Células-Tronco , Engenharia Tecidual , Diferenciação Celular , Saco Dentário , Humanos , Ligamento Periodontal , Regeneração
17.
Yi Chuan ; 41(12): 1110-1118, 2019 Dec 20.
Artigo em Chinês | MEDLINE | ID: mdl-31857282

RESUMO

Myogenesis is a complex physiological process that is mainly involved in the proliferation of myogenic stem cells to form myoblasts, which then differentiated and fused to form multinucleated myotubes. Many proteins have been found to be involved in myoblast fusion, but none of them are muscle-specific fusion proteins. In recent years, two muscle-specific transmembrane proteins, i.e. Myomaker and Myomerger, have been discovered and identified, which can coordinate and promote the fusion of myoblasts and thus participate in the process of myogenesis. In this review, we summarize the research progress of Myomaker and Myomerger in myogenesis, including their expression patterns and functional domains, as well as their participation in myoblast fusion mechanisms, aiming to provide relevant ideas for in-depth study of the myogenesis process and treatment of diseases related to myoblast fusion.


Assuntos
Proteínas de Membrana , Músculo Esquelético , Mioblastos , Animais , Diferenciação Celular , Fusão Celular , Humanos , Proteínas de Membrana/genética , Proteínas de Membrana/metabolismo , Desenvolvimento Muscular , Proteínas Musculares , Músculo Esquelético/citologia , Mioblastos/citologia
18.
Yi Chuan ; 41(12): 1119-1128, 2019 Dec 20.
Artigo em Chinês | MEDLINE | ID: mdl-31857283

RESUMO

Porcine skeletal muscle development is a complex biological process, and differentiation of skeletal muscle satellite cells is an important part of skeletal muscle development. In recent years, it has been found that lncRNA plays an important role in the differentiation of skeletal muscle satellite cells. Here we investigate the effect of lncRNA TCONS_00815878 on the differentiation of porcine skeletal muscle satellite cells. We first used qRT-PCR to detect the expression levels of TCONS_00815878 in six tissues (heart, spleen, lung, kidney, back muscles and leg muscles) of Yorkshire piglets within seven days of birth. At the same time, the expression levels of TCONS_00815878 at five different time points from the embryonic stage to the postnatal stage (35 d, 45 d, 55 d of embryos, and 7 d, 200 d of postpartum leg muscles) were examined. The expression of the differentiation marker genes MyoD, MyoG and MyHC was examined by knocking down TCONS_00815878 in porcine skeletal muscle satellite cells using antisense oligonucleotides (ASO). The target gene of TCONS_00815878 was predicted by bioinformatics analysis, and the function and pathway of its target gene were predicted online using DAVID software. The results showed that TCONS_00815878 had the highest expression level in pig myocardium and leg muscles. Within seven days after birth, TCONS_00815878 increased in the muscle tissue of pigs, and reached the peak of expression level on the 7th day. During the process of proliferation and differentiation of porcine skeletal muscle satellite cells, the expression level of TCONS_00815878 increased during the differentiation stage and peaked at 30 h of differentiation. After knocking down TCONS_00815878, the expression levels of MyoD, MyoG and MyHC were decreased, but the expression level of MyoD was significantly decreased (P<0.05). In addition, functional predictions revealed that the target gene of TCONS_00815878 is enriched in multiple biological processes, such as glycolysis and pyruvate metabolism, related to skeletal muscle satellite cell differentiation. This study speculates that lncRNA TCONS_00815878 may promote the differentiation of porcine skeletal muscle satellite cells.


Assuntos
Diferenciação Celular , Regulação da Expressão Gênica no Desenvolvimento , Músculo Esquelético , RNA Longo não Codificante , Células Satélites de Músculo Esquelético , Animais , Diferenciação Celular/genética , Proliferação de Células , Células Cultivadas , Perfilação da Expressão Gênica , Técnicas de Silenciamento de Genes , Desenvolvimento Muscular , Músculo Esquelético/citologia , RNA Longo não Codificante/genética , Células Satélites de Músculo Esquelético/citologia , Suínos
19.
Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi ; 35(9): 783-788, 2019 Sep.
Artigo em Chinês | MEDLINE | ID: mdl-31750818

RESUMO

Objective To investigate the function and differentiation of 1/17 type helper T (Th1/17) cells. Methods Bioinformatics analysis was performed using a gene chip dataset (GSE104021) in GEO which contains gene expression data from Th17 cells and Th1/17 cells of healthy human subjects. Taking Th17 cells as the control, R language software was used to analyze the differentially expressed genes (DEGs) between Th17 cells and Th1/17 cells, so as to explore the main functional molecules of Th1/17 cells. After that, gene ontology (GO) analysis and Kyoto encyclopedia of genes and genomes (KEGG) analysis of DEGs were conducted by R language software. Finally, the genes enriched into the target biological process in the GO analysis were selected for protein-protein interaction network (PPI) analysis to explore the differentiation process of Th1/17 cells. Results Analysis of DEGs showed that, compared with Th17 cells, the underexpressed genes in Th1/17 cells were interleukin 17A (IL-17A) and C-C motif chemokine receptor 4 (CCR4). The over-expressed genes were coiled-coil domain-containing 3 (CCDC3), C-C motif chemokine ligand 4 (CCL4), colony stimulating factor 2 receptor beta common subunit (CSF2RB), C-C motif chemokine ligand 5 (CCL5), interferon gamma (IFNG) and epithelial stromal interaction 1 (EPSTI1). In GO analysis, cell component analysis showed that the expression products of these DEGs were mainly located at external side of plasma membrane and the granulocyte-macrophage colony stimulating factor (GM-CSF) receptor complex; biological process analysis showed that the expression products of DEGs were involved in the upregulation of interleukin 23 (IL-23), the chemokine-mediated signaling pathway and the upregulated chemotaxis of natural killer (NK) cells; molecular function analysis showed that the expression products of these DEGs had C-C motif chemokine 5 receptor (CCR5) binding activity, cytokine activity and interferon gamma (IFN-γ) receptor binding activity. The results of KEGG analysis showed that the DEGs were enriched in the cytokine-cytokine receptor interaction, rheumatoid arthritis, inflammatory bowel disease and chemokine signaling pathways. The GO analysis showed that DEGs IL-17A and IFNG were enriched to the biological process of upregulating IL-23 production. PPI showed that IL-17A and IFNG had biological functions of regulating cytokine production and myeloid white blood cell differentiation. Conclusion Bioinformatics analysis showed that the protein products encoded by overexpressed genes CCL4, CSF2RB, CCL5, IFNG and EPSTI1 in Th1/17 cells were potential functional effectors of Th1/17 cells. Th1/17 cells could produce IFN-γ and IL-17A, which act on macrophages and dendritic cells (DCs) derived from myeloid white blood cells, thus promoting the differentiation of macrophages and DCs and the production of IL-23. IL-23 promotes trans-differentiation of Th17 cells into Th1/17 cells.


Assuntos
Diferenciação Celular , Mapas de Interação de Proteínas , Células Th1/citologia , Células Th17/citologia , Biologia Computacional , Ontologia Genética , Humanos
20.
Adv Exp Med Biol ; 1182: 39-77, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31777014

RESUMO

The antitumor effect of Ganoderma (Lingzhi) is closely related to immunoregulation. Based on our research and other references, this article discussed the antitumor effect of Ganoderma mediated by immunological mechanism, including promoting the function of mononuclear-macrophages and natural killers; promoting M1-type macrophage polarization vs M2-type; promoting maturation and differentiation of dendritic cells, increasing its antigen presentation, activating lymphocytes and increasing cytotoxicity of cytotoxin T lymphocyte; promoting production of cytokines; and inhibiting tumor escape from immune surveillance. Also, clinical studies with immunological indexes were reviewed.


Assuntos
Antineoplásicos/farmacologia , Produtos Biológicos/farmacologia , Imunomodulação , Neoplasias/tratamento farmacológico , Reishi/química , Apresentação do Antígeno , Diferenciação Celular , Citocinas/imunologia , Células Dendríticas/imunologia , Humanos , Macrófagos/imunologia , Linfócitos T/imunologia
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