ABSTRACT
BACKGROUND: External surface resorption is pressure-induced resorption and occurs on the external surface of the root, pressure exerted by impacted teeth, is common causes of external surface resorption. Predictive risk factors of impacted supernumerary tooth-associated root resorption (ISTARR) mentioned in this article include supernumerary teeth and patient factors. To investigate the risk factors of impacted supernumerary tooth-associated root resorption and predict the incidence of root resorption. METHODS: This restrospective study enrolled 324 patients with impacted supernumerary tooth. All Cone-Beam Computed Tomography (CBCT) data and patient information were divided into two groups (without tooth root resorption and with root resorption). CBCT images and patient information (age and gender) of 133 patients had adjacent tooth root resorption and 191 did not. seven variables were analysed using binary logistic regression. RESULTS: Individual analysis of potential risk factors showed that age, crown mesiodistal direction, root formation, and odontotheca of the impacted supernumerary tooth were associated significantly with ISTARR. Binary logistic regression showed that impacted supernumerary tooth with odontotheca (Odd Ratio = 2.926), the crown is in the middle (Odd Ratio = 1.446), located at the middle third of the adjacent tooth root (Odd Ratio = 1.614), complete root development (Odd Ratio = 1.334), and patient's age (Odd Ratio = 1.261) were significantly associated with ISTARR risk. CONCLUSIONS: The risk factors of root resorption can be detected and predicted early according to the features of supernumerary tooth and patient's age. Still, more prospective studies with larger sample size are needed to validate the result.
Subject(s)
Cone-Beam Computed Tomography , Root Resorption , Tooth, Impacted , Tooth, Supernumerary , Humans , Cone-Beam Computed Tomography/methods , Tooth, Supernumerary/diagnostic imaging , Tooth, Supernumerary/complications , Root Resorption/diagnostic imaging , Root Resorption/etiology , Tooth, Impacted/diagnostic imaging , Female , Male , Child , Case-Control Studies , Risk Factors , Retrospective Studies , Adolescent , Risk AssessmentABSTRACT
Esophageal cancer is a highly aggressive malignancy with a low response to standard anti-cancer therapies. There is an unmet need to develop new therapeutic strategies to improve the clinical outcomes of current treatments. Cold atmospheric plasma (CAP) is a promising approach for cancer treatment, and has displayed anticancer efficacy in multiple preclinical models. Recent studies have shown that the efficacy of CAP is positively correlated with intracellular reactive oxygen species (ROS) levels. This suggests that aggressively increasing intracellular ROS levels has the potential to further improve CAP-mediated anticancer efficacy. Glutamine plays an important role in cellular ROS scavenging after being converted to glutathione (GSH, a well-described antioxidant) under physiological conditions, so reducing intracellular glutamine levels seems to be a promising strategy. To test this hypothesis, we treated esophageal cancer cells with CAP while controlling the supply of glutamine. The results showed that glutamine did affect the anticancer effect of CAP, and the combination of CAP stimulation and glutamine deprivation significantly inhibited the proliferation of esophageal cancer cells compared to the control group (p < 0.05). Furthermore, flow cytometric analysis documented a significant increase in more than 10% in apoptosis and necrosis of esophageal cancer cells after this synergistic treatment compared to the control group (p < 0.05). Thus, these results provide the first direct evidence that the biological function of CAP can be modulated by glutamine levels and that combined CAP stimulation and glutamine deprivation represent a promising strategy for the future treatment of esophageal cancer.
Subject(s)
Esophageal Neoplasms , Plasma Gases , Humans , Reactive Oxygen Species , Glutamine/pharmacology , Plasma Gases/pharmacology , Cell Line, Tumor , Esophageal Neoplasms/drug therapyABSTRACT
Workshop cluster 1 (WC1) molecules are part of the scavenger receptor cysteine-rich (SRCR) superfamily and act as hybrid co-receptors for the γδ T cell receptor and as pattern recognition receptors for binding pathogens. These members of the CD163 gene family are expressed on γδ T cells in the blood of ruminants. While the presence of WC1+ γδ T cells in the blood of goats has been demonstrated using monoclonal antibodies, there was no information available about the goat WC1 gene family. The caprine WC1 multigenic array was characterized here for number, structure and expression of genes, and similarity to WC1 genes of cattle and among goat breeds. We found sequence for 17 complete WC1 genes and evidence for up to 30 SRCR a1 or d1 domains which represent distinct signature domains for individual genes. This suggests substantially more WC1 genes than in cattle. Moreover, goats had seven different WC1 gene structures of which 4 are unique to goats. Caprine WC1 genes also had multiple transcript splice variants of their intracytoplasmic domains that eliminated tyrosines shown previously to be important for signal transduction. The most distal WC1 SRCR a1 domains were highly conserved among goat breeds, but fewer were conserved between goats and cattle. Since goats have a greater number of WC1 genes and unique WC1 gene structures relative to cattle, goat WC1 molecules may have expanded functions. This finding may impact research on next-generation vaccines designed to stimulate γδ T cells.
Subject(s)
Goats , T-Lymphocytes , Animals , Cattle/genetics , Membrane Glycoproteins/genetics , Receptors, Antigen, T-Cell, gamma-delta/genetics , Receptors, Antigen, T-Cell, gamma-delta/metabolism , Receptors, Scavenger/metabolism , Ruminants , T-Lymphocyte Subsets/metabolism , T-Lymphocytes/metabolismABSTRACT
Acetylation can be regulated by histone deacetylases (HDACs) and histone acetyltransferases (HATs), and the imbalance between HDACs and HATs can lead to cancers. Trichostatin A (TSA), as one of the typical HDAC inhibitors (HDACis), may regulate the acetylation level and thus prevent the proliferation of cancer cells, in which the metabolic change of glycolysis is involved. However, the dynamic process of glycolysis has not yet been explored, and the mechanism is still elusive. In this work, we constructed GFP-actin-HeLa cells to observe the dynamic change of glycolysis in situ and found that the GFP fluorescence enhanced significantly after TSA treatment, which was consistent with the change of pH in the cells (pHi) depending on the change of lactate originated from glycolysis. Furthermore, we confirmed that the glycolysis was inhibited after TSA treatment, and this process was associated with HIF-1α and c-Myc regulation. As such, this work not only demonstrates the usefulness of the GFP fluorescent probe for monitoring the metabolic process of glycolysis in situ, but also sheds new light on the mechanism of glycolysis suppression in the cancer cells treated with HDACi.
Subject(s)
Glycolysis , Histone Deacetylases , Humans , Acetylation , Fluorescence , HeLa Cells , Histone Deacetylases/metabolismABSTRACT
Human immunodeficiency virus (HIV)/acquired immunodeficiency syndrome (AIDS) disrupts the host microbial balance. During disease progression, the oral microbial environment is altered in untreated people living with HIV/AIDS (PLWHA); however, no studies have reported changes in salivary microbial diversity during different stages of HIV infection. Therefore, in this study, we aimed to assess the relationships between immune dysfunction and changes in saliva microbiota. To this end, we collected saliva samples from 11 HIV-negative individuals and 44 PLWHA during different stages based on the Centers for Disease Control and Prevention criteria (stage 0, early stage during the first 6 months after infection; stages 1, 2, and 3 associated with CD4+ T-lymphocyte counts of ≥500, 200-499, and ≤200 or opportunistic infection, respectively). We analyzed salivary microbial community diversity using polymerase chain reaction amplification and Illumina MiSeq sequencing. We found that HIV-positive individuals had significantly greater alpha-diversity in the microbial community composition compared with HIV-negative controls (P < 0.05) except for AIDS (stage 3); however, the predominant salivary microbiota in the five groups remained similar. Porphyromonas in the four positive groups was the only genus that was significantly less abundant in the HIV-positive groups than in the control group (P < 0.05). There were some consistencies between the general abundance of salivary microbiota and AIDS disease progression. Lots of bacterial abundances in the saliva increased dramatically during the acute HIV infection (stage 0), and some of the negligible and abnormally proliferating bacteria in the asymptomatic stage showed a downward trend. Additionally, in the AIDS stage, partial inhibition was observed. Notably, Porphyromonas was closely related to the immune activation of HIV, showing a decline in abundance once infected with HIV. Solobacterium, which induces inflammation, was negatively correlated with CD4 counts. Overall, our findings provided important insights into changes in salivary microbial diversity in PLWHA.
Subject(s)
Acquired Immunodeficiency Syndrome , HIV Infections , Microbiota , CD4 Lymphocyte Count , CD4-Positive T-Lymphocytes , HumansABSTRACT
The SWI/SNF complex is a highly conserved chromatin remodeling complex and can hydrolyze ATP by its catalytic subunit BRG1 or BRM to reconstruct the chromatin. To investigate whether this ATP-dependent chromatin remodeling could affect the DNA conformation, we therefore regulated (knocked down or overexpressed) BRG1/BRM in the cells and applied Fourier transform infrared (FTIR) spectroscopy to probe DNA conformational changes. As a result, we found that BRG1/BRM was indeed associated with the DNA conformational changes, in which knockdown of BRG1/BRM reduced Z-DNA conformation, while overexpression of BRG1/BRM enhanced Z-DNA conformation. This Z-DNA conformational transformation was also verified using the Z-DNA-binding proteins. Therefore, this work has provided a direct analytical tool to probe Z-DNA transformation upon ATP-dependent chromatin remodeling.
Subject(s)
Chromatin Assembly and Disassembly , DNA, Z-Form/chemistry , Nucleic Acid Conformation , Spectroscopy, Fourier Transform Infrared , Adenosine Triphosphate/metabolism , Cell Line, Tumor , DNA Helicases/deficiency , DNA Helicases/genetics , DNA, Z-Form/metabolism , Gene Knockdown Techniques , Humans , Nuclear Proteins/deficiency , Nuclear Proteins/genetics , Transcription Factors/deficiency , Transcription Factors/geneticsABSTRACT
CD63, a member of transmembrane-4-superfamily of tetraspanin proteins and a highly N-glycosylated type III lysosomal membrane protein, is known to regulate malignancy of various types of cancers such as melanoma and breast cancer and serves as a potential marker for cancer detection. Recently, its important role as a classic exosome marker was also emphasized. In this work, via using a magnetic bead-based competitive SELEX (systematic evolution of ligands by exponential enrichment) procedure and introducing a 0.5M NaCl as elution buffer, we identified two DNA aptamers (CD63-1 and CD63-2) with high affinity and specificity to CD63 protein (Kd = 38.71nM and 78.43, respectively). Furthermore, CD63-1 was found to be efficient in binding CD63 positive cells, including breast cancer MDA-MB-231 cells and CD63-overexpressed HEK293T cells, with a medium binding affinity (Kd~ 100 nM) as assessed by flow cytometry. When immunostaining assay was performed using clinical breast cancer biopsy, the CD63-1 aptamer demonstrated a comparable diagnostic efficacy for CD63 positive breast cancer with commercial antibodies. After developing a magnetic bead-based exosome immunoaffinity separation system using CD63-1 aptamer, it was found that this bead-based system could effectively isolate exosomes from both MDA-MB-231 and HT29 cell culture medium. Importantly, the introduction of the NaCl elution in this work enabled the isolation of native exosomes via a simple 0.5M NaCl incubation step. Based on these results, we firmly believe that the developed aptamers could be useful towards efficient isolation of native state exosomes from clinical samples and various theranostic applications for CD63-positive cancers.
Subject(s)
Aptamers, Nucleotide/chemistry , Breast Neoplasms/diagnosis , Exosomes/metabolism , SELEX Aptamer Technique/methods , Tetraspanin 30/chemistry , Breast Neoplasms/immunology , Breast Neoplasms/metabolism , Chromatography, Affinity , Exosomes/chemistry , Exosomes/immunology , Female , Humans , Immunoenzyme Techniques , Tetraspanin 30/immunology , Tumor Cells, CulturedABSTRACT
BACKGROUND: The endocannabinoid system is involved in the regulation of periodontal tissue homeostasis. Synthetic cannabinoid methanandamide (Meth-AEA) has improved stability and affinity to cannabinoid receptors compared to its endogenous analog anandamide. In the present study, we investigated the effect of methanandamide on the production of pro-inflammatory mediators in primary human periodontal ligament cells (hPdLCs). METHODS: hPdLCs were treated with Meth-AEA for 24 h, and the resulting production of interleukin (IL)-6, IL-8, and monocyte chemotactic protein (MCP)-1 was measured in the absence or the presence of Porphyromonas gingivalis lipopolysaccharide (LPS). Additionally, the effect of Meth-AEA on the proliferation/viability of hPdLCs was measured by the MTT method. RESULTS: Methanandamide at a concentration of 10 µM significantly inhibited P. gingivalis LPS induced production of IL-6, IL-8, and MCP-1. Basal production of IL-6 and IL-8 was slightly enhanced by 10 µM Meth-AEA. No effect of Meth-AEA on the basal production of MCP-1 was observed. Meth-AEA in concentrations up to 10 µM did not affect the proliferation/viability of hPdLCs, but significantly inhibited it at a concentration of 30 µM. CONCLUSION: Our study suggests that the inflammatory response in periodontal ligament cells could be influenced by the activation of the cannabinoid system, which might be potentially involved in the progression of periodontal disease.
Subject(s)
Arachidonic Acids/pharmacology , Lipopolysaccharides , Periodontal Ligament/drug effects , Porphyromonas gingivalis/drug effects , Cell Proliferation/drug effects , Cells, Cultured , Humans , Interleukin-6ABSTRACT
After infection with T. brucei AnTat 1.1, C57BL/6 mice lost splenic B2 B cells and lymphoid follicles, developed poor parasite-specific antibody responses, lost weight, became anemic and died with fulminating parasitemia within 35 days. In contrast, infected C57BL/6 mice lacking the cytotoxic granule pore-forming protein perforin (Prf1-/-) retained splenic B2 B cells and lymphoid follicles, developed high-titer antibody responses against many trypanosome polypeptides, rapidly suppressed parasitemia and did not develop anemia or lose weight for at least 60 days. Several lines of evidence show that T. brucei infection-induced splenic B cell depletion results from natural killer (NK) cell-mediated cytotoxicity: i) B2 B cells were depleted from the spleens of infected intact, T cell deficient (TCR-/-) and FcγRIIIa deficient (CD16-/-) C57BL/6 mice excluding a requirement for T cells, NKT cell, or antibody-dependent cell-mediated cytotoxicity; ii) administration of NK1.1 specific IgG2a (mAb PK136) but not irrelevant IgG2a (myeloma M9144) prevented infection-induced B cell depletion consistent with a requirement for NK cells; iii) splenic NK cells but not T cells or NKT cells degranulated in infected C57BL/6 mice co-incident with B cell depletion evidenced by increased surface expression of CD107a; iv) purified NK cells from naïve C57BL/6 mice killed purified splenic B cells from T. brucei infected but not uninfected mice in vitro indicating acquisition of an NK cell activating phenotype by the post-infection B cells; v) adoptively transferred C57BL/6 NK cells prevented infection-induced B cell population growth in infected Prf1-/- mice consistent with in vivo B cell killing; vi) degranulated NK cells in infected mice had altered gene and differentiation antigen expression and lost cytotoxic activity consistent with functional exhaustion, but increased in number as infection progressed indicating continued generation. We conclude that NK cells in T. brucei infected mice kill B cells, suppress humoral immunity and expedite early mortality.
Subject(s)
B-Lymphocyte Subsets/immunology , Killer Cells, Natural/immunology , Lymphocyte Activation/immunology , Spleen/immunology , Trypanosomiasis, African/immunology , Animals , Blotting, Western , Disease Models, Animal , Enzyme-Linked Immunosorbent Assay , Flow Cytometry , Fluorescent Antibody Technique , Mice , Mice, Inbred C57BL , Mice, Knockout , Trypanosoma brucei brucei/immunologyABSTRACT
BACKGROUND: A number of studies have assessed the prognostic value of the neutrophil-to-lymphocyte ratio (NLR) in oral squamous cell carcinoma (OSCC), but their results regarding the predictive value of NLR in OSCC are inconsistent. We therefore performed a meta-analysis to clarify the association between NLR and clinical outcome in OSCC. METHODS: We searched the MEDLINE and Web of Science to identify potential studies investigating the association between NLR and survival in OSCC. RESULTS: A total of 10 studies, enrolling 2135 patients with OSCC, were included. A higher NLR was a negative predictor for both disease-specific survival (hazard ratio [HR] = 1.93, 95% CI: 1.47-2.54) and overall survival (HR = 1.56, 95% CI: 1.28-1.90). CONCLUSION: This suggests a higher NLR is predictive of a poorer prognosis in OSCC. Because determination of NLR is non-invasive and cost-effective, it could be widely used for predicting prognosis in OSCC.
Subject(s)
Carcinoma, Squamous Cell/blood , Lymphocytes , Mouth Neoplasms/blood , Neutrophils , Humans , Leukocyte Count , Prognosis , Survival RateABSTRACT
Silver nanoparticles (Ag NPs) have well-known antibacterial properties and are widely applied in various medical products and general commodities. Although many studies have addressed the toxicity of Ag NPs to mammalian cells, the direct relationship between the number of Ag NPs in living cells and the corresponding cell toxicity has not yet been explicitly demonstrated. In this work, a simple and reusable microfluidic device composed of a quartz cover slip and a glass plate with etched micro-channel and micro-wells was employed for separating and trapping single living cells. The device was silanized to render the surface hydrophobic. For simplicity, HeLa cells as the model cancer cells were used in the study, which were pipette-loaded into an array of micro wells based on dead-end filling. Surface enhanced Raman spectroscopy (SERS) was then employed to examine the living cancer cells and assessed number and distribution of Ag NPs in the cells. Combined with the cell viability assay, we therefore correlated the number of Ag NPs in the cell with the toxicity to the cell directly.
Subject(s)
Metal Nanoparticles/toxicity , Microfluidics , Silver/toxicity , Anti-Bacterial Agents/chemistry , Anti-Bacterial Agents/toxicity , Cell Survival , HEK293 Cells , HeLa Cells , Humans , Metal Nanoparticles/chemistry , Particle Size , Silver/chemistry , Spectrum Analysis, Raman , Toxicity TestsABSTRACT
A nucleosome is made up of DNA and histones, and acetylation of histones perturbs the interaction of DNA and histones and thus affects the chromatin conformation and function. However, whether or how acetylation induces DNA conformation changes is still elusive. In this work, we applied FT-IR spectroscopy to monitor the DNA signals in cells as the histone acetylation was regulated by trichostatin A (TSA), a reversible inhibitor to histone deacetylases (HDACs). Our results unambiguously demonstrate the significant transformation of B-DNA to Z-DNA upon histone acetylation in the TSA treated HeLa cells. This is the first report providing the explicit experimental evidence for such a B-Z transformation of DNA in the epigenetic states of cells.
Subject(s)
DNA, B-Form/metabolism , DNA, Z-Form/metabolism , Histones/metabolism , Acetylation , DNA, B-Form/chemistry , DNA, Z-Form/chemistry , HeLa Cells , Histones/chemistry , Humans , Hydroxamic Acids/pharmacology , Nucleic Acid Conformation , Spectroscopy, Fourier Transform InfraredABSTRACT
Trichostatin A (TSA) is one of histone deacetylase (HDAC) inhibitor drugs which can suppress the enzymatic activity of deacytylases and promote the acetylation of both histone and nonhistone proteins in cells. Investigation of the effect of TSA on cellular acetylation is critical for better understanding of the antitumor drug's mechanism interacting with cancer cells. As Fourier transform infrared spectroscopy (FT-IR) is a powerful analytical tool which can detect nondestructively and quantitatively biological samples without biotagging and biolabeling, here we employed FT-IR spectroscopy to probe the chemical and structural changes of proteins in the TSA treated cells, and with the aid of fluorescent microscopy, we could scrutinize the time-dependent and dose effects on the acetylation level promoted by TSA. Our results showed that TSA caused an elevated level of cellular acetylation and conformational/structural changes of proteins in the cells, and a higher dosage of TSA caused a higher percent of α-helix structure accompanied by an increment of acetylation level in both histones and cytoskeleton proteins. This work therefore not only validates the usefulness of FT-IR spectroscopy in the quantitative assessment of cellular acetylation but also may open an avenue to the in-depth investigation of the effect of HDAC inhibitor drugs such as TSA on cancer cells.
Subject(s)
Histone Deacetylase Inhibitors/pharmacology , Hydroxamic Acids/pharmacology , Acetylation/drug effects , Cell Proliferation/drug effects , Dose-Response Relationship, Drug , HeLa Cells , Humans , Microscopy, Fluorescence , Spectroscopy, Fourier Transform Infrared , Structure-Activity Relationship , Time FactorsABSTRACT
OBJECTIVE: To isolate and culture human periodontal ligament stem cells (PDLSCs) and observe its ultrastructure. METHODS: The proliferation and growth characteristics of human periodontal ligament cells were observed in primary culture and colony culture. PDLSCs were isolated by magnetic activated cell sorting (MACS) and ultrastructural characterization was observed by electron microscopy. RESULTS: When the cells were cultured at low density, PDLSCs grew in a colony-like manner. With the exception of a small amount of rough endoplasmic reticulum, ribosomes, and mitochondria, relatively few organelles were found in the cytoplasm, suggesting that they had remained undifferentiated. CONCLUSION: PDLSCs showed colony-like growth capacity and had ultrastructural characterization with stem cells. This indicated that PDLSCs could act as the appropriate seed cells for cell-based periodontal tissue regeneration.
Subject(s)
Mesenchymal Stem Cells/ultrastructure , Periodontal Ligament/cytology , Cell Culture Techniques , Cell Differentiation , Cell Separation , HumansABSTRACT
Background: Engaging in anal sexual intercourse markedly increases the risk of developing HIV among men who have sex with men (MSM); oral sexual activities tend to uniquely introduce gut-derived microbes to salivary microbiota, which, combined with an individual's positive HIV status, may greatly perturb oral microecology. However, till date, only a few published studies have addressed this aspect. Methods: Based on 16S rRNA sequencing data of bacterial taxa, MicroPITA picks representative samples for metagenomic analysis, effectively revealing how the development and progression of the HIV disease influences oral microbiota in MSM. Therefore, we collected samples from 11 HIV-negative and 44 HIV-positive MSM subjects (stage 0 was defined by HIV RNA positivity, but negative or indeterminate antibody status; stages 1, 2, and 3 were defined by CD4+ T lymphocyte counts ≥ 500, 200-499, and ≤ 200 or opportunistic infection) and selected 25 representative saliva samples (5 cases/stage) using MicroPITA. Metagenomic sequencing analysis were performed to explore whether positive HIV status changes salivary bacterial KEGG function and metabolic pathway in MSM. Results: The core functions of oral microbiota were maintained across each of the five groups, including metabolism, genetic and environmental information processing. All HIV-positive groups displayed KEGG functions of abnormal proliferation, most prominently at stage 0, and others related to metabolism. Clustering relationship analysis tentatively identified functional relationships between groups, with bacterial function being more similar between stage 0-control groups and stage 1-2 groups, whereas the stage 3 group exhibited large functional changes. Although we identified most metabolic pathways as being common to all five groups, several unique pathways formed clusters for certain groups; the stage 0 group had several, while the stage 2 and 3 groups had few, such clusters. The abundance of K03046 was positively correlated with CD4 counts. Conclusion: As HIV progresses, salivary bacterial function and metabolic pathways in MSM progressively changes, which may be related to HIV promoting abnormal energy metabolism and exacerbate pathogen virulence. Further, infection and drug resistance of acute stage and immune cell destruction of AIDS stage were abnormally increased, predicting an increased risk for MSM individuals to develop systemic and oral diseases.
Subject(s)
HIV Infections , Homosexuality, Male , RNA, Ribosomal, 16S , Saliva , Humans , Male , Saliva/microbiology , Saliva/virology , HIV Infections/microbiology , RNA, Ribosomal, 16S/genetics , Adult , Bacteria/classification , Bacteria/genetics , Bacteria/isolation & purification , Microbiota , Metagenomics , CD4 Lymphocyte Count , Middle Aged , Young Adult , Sexual and Gender MinoritiesABSTRACT
OBJECTIVE: To observe the effect of icariin on human periodontal ligament cells (hPDLCs) differentiation to osteoblast gene expression. METHODS: The fifth generation of the cultured hPDLCs was added with the concentration of 0.01 mg/L icariin, and the added osteogenic medium used as blank control group, alizarin red staining of icariin on human periodontal ligament cells was observed for 21 days; the 2, 4, and 6 days of Q-PCR quantitative analysis of icariin on human periodontal ligament cells were made for osteogenesis gene alkaline phosphatase (ALP), type I collagen and osteocalcin (OC) gene expression. RESULTS: For the 21 days, alizarin red staining icariin group formed more mineralized nodules; on the 2nd, 4th, and 6th days, the group of icariin promoted the expression of ALP and OC mRNA, reached the peak value on day 6, compared with the control group with significant difference (20.15±6.67 vs. 7.90±0.71, 4.13±0.56 vs. 3.55±0.08, P<0.01). The second day, the highest expression of type I collagen appeared, then decreased gradually after, statistically compared with the control group (P<0.05). CONCLUSION: Icariin can promote the human periodontal ligament cells differentiation to osteoblast, and promote the osteogenesis gene expression.
Subject(s)
Cell Differentiation/drug effects , Drugs, Chinese Herbal/pharmacology , Flavonoids/pharmacology , Osteoblasts , Periodontal Ligament , Adolescent , Alkaline Phosphatase/genetics , Alkaline Phosphatase/metabolism , Cells, Cultured , Child , Collagen Type I/genetics , Collagen Type I/metabolism , Drugs, Chinese Herbal/isolation & purification , Epimedium/chemistry , Flavonoids/isolation & purification , Gene Expression Regulation , Humans , Osteoblasts/cytology , Osteoblasts/metabolism , Osteocalcin/genetics , Osteocalcin/metabolism , Osteogenesis/drug effects , Periodontal Ligament/cytology , Periodontal Ligament/metabolism , Plants, Medicinal/chemistry , RNA, Messenger/metabolismABSTRACT
Infrared spectrum stems from the matter's absorption of light in the infrared (IR) light region. Generally, this infrared light absorption is due to the transition of vibrational and rotational energy levels of the involved molecule. Since different molecules have different structures and vibration modes, infrared spectroscopy can therefore be widely applied to analyze the chemical compositions and structures of molecules. Here we describe the method of application of infrared spectroscopy in the investigation of Z-DNA in cells, as infrared spectroscopy can distinguish DNA secondary structures sensitively and the band at 930 cm-1 is specifically attributed to the Z-form DNA. Based on the curve fitting, the relative content of Z-DNA in the cells may be evaluated.
Subject(s)
DNA, Z-Form , Spectrophotometry, Infrared/methods , DNA , VibrationABSTRACT
Z-DNA refers to the left-handed double-helix DNA that has attracted much attention because of its association with some specific biological functions. However, because of its low content and unstable conformation, Z-DNA is normally difficult to observe or identify. Up to now, there has been a lack of unified or standard analytical methods among diverse techniques for probing Z-DNA and its transformation conveniently. In this work, NaCl, MgCl2, and ethanol were utilized to induce d(GC)8 from B-DNA to Z-DNA in vitro, and Fourier transform infrared (FTIR) spectroscopy was employed to monitor the transformation of Z-DNA under different induction conditions. The structural changes during the transformation process were carefully examined, and the DNA chirality alterations were validated by the circular dichroism (CD) measurements. The Z-DNA characteristic signals in the 1450 cm-1-900 cm-1 region of the d(GC)8 infrared (IR) spectrum were observed, which include the peaks at 1320 cm-1, 1125 cm-1 and 925 cm-1, respectively. The intensity ratios of A1320/A970, A1125/A970, and A925/A970 increased with Z-DNA content in the transition process. Furthermore, compared with the CD spectra, the IR spectra showed higher sensitivity to Z-DNA, providing more information about the molecular structure change of DNA. Therefore, this study has established a more reliable FTIR analytical approach to assess BZ DNA conformational changes in solutions, which may help the understanding of the Z-DNA transition mechanism and promote the study of Z-DNA functions in biological systems.
Subject(s)
DNA, Z-Form , Nucleic Acid Conformation , Spectrophotometry, Infrared , DNA/chemistry , Spectroscopy, Fourier Transform Infrared , Circular DichroismABSTRACT
OBJECTIVE: To investigate the accuracy of Tooth-Implant digital guide-assisted implantation, explore the influence of periodontitis on the accuracy of the digital guide, and evaluate the effect of the residual abutment looseness after periodontitis treatment on the implant accuracy of the digital guide. METHODS: In this retrospective clinical study, 45 patients who received dental implantation at the Department of Periodontology, Beijing Stomatological Hospital affiliated with Capital Medical University, were selected and grouped. Group A consisted of non-periodontitis patients (n=15) who underwent Tooth-Implant digital guide-assisted implantation surgery. Group B was composed of periodontitis patients (n=15) who received Tooth-Implant digital guide-assisted implantation surgery. Group C included periodontitis patients (n=15) with freehand implantation. Three dental landmarks were identified to compare the planned implant position generated by the Tooth-Implant digital guide before implantation and the actual implant position in the same patient. Differences in implant depth, angle, shoulder and apex were analyzed before and after the implantation. RESULTS: There were statistical differences in implant depth, angle, shoulder, and apex between group B and group C. While statistical significance was found only in the implant angle and depth between group A and group B, not in the implant shoulder or apex. In periodontitis patients treated by Tooth-Implant digital guide-assisted implantation, significant differences were identified in implant depth and shoulder between non-abutment looseness and abutment looseness subgroups, but not in implant angle and apex. Under the digital guide-assisted implantation, no significant differences were found in implant depth, angle, shoulder and apex at different jaw positions, but at different tooth positions, significant differences were identified in implant angle and apex, not in implant depth and shoulder. The accuracy of Tooth-Implant digital guide-assisted implantation was consistent with previous data. CONCLUSIONS: The Tooth-Implant digital guide-assisted implantation has reliable implant accuracy that outperforms freehand implantation. Periodontitis is a factor affecting the accuracy of digital guide in dental implant placement, and this could be due to the looseness of residual abutments after systematic periodontal treatment. Different jaw positions have no impact on the accuracy of digital guide-assisted implantation, but different tooth positions have an impact on the accuracy of implant placement using a digital guide.
ABSTRACT
OBJECTIVE: To investigate the effects of emdogain, enamel matrix derivative (EMD), on the proliferation, cell cycle, mineralization and ultrastructure of human periodontal ligament (PDL) cells in vitro. METHODS: The influence of cell growth on PDL cells was evaluated by Cell Counting Kit-8 (CCK-8) in the presence and absence of emdogain, after 1, 3, and 5 d of culture. DNA synthesis and ultrastructure of PDL cells were observed by flow cytometry(FCM) and transmission electron microscopy (TEM) in the presence and absence of emdogain after 3 d of culture. The increasing of osteogenic capacity was verified by the expression changes of osteogenic differentiation markers of bone sialoprotein (BSP) and osteopontin (OPN) in emdogain-treated PDL cells by immunohistochemicl staining. RESULTS: Incubation of PDL cells with emdogain after 3 d significantly stimulated cell growth and DNA synthesis. Emdogain enhanced the osteogenic potential of PDL cells by high expression of osteogenic differentiation markers of BSP and OPN. CONCLUSION: The data indicate that Emdogain enhances cell proliferation and promotes differentiation of PDL cells, which contributes to periodontal tissue regeneration .