Several features of the implant surface, such as roughness, topography, and composition play a relevant role in implant integration with bone. This study was conducted in order to determine the effects of various thin layer hydroxyapatite (HA) coatings on anodized Ti surfaces on the biological responses of a human osteoblast-like cell line (MG63). MG63 cells were cultured on A (100 nm HA coating on anodized surface), B (500-700 nm HA coating on anodized surface), C ($1{\mu}m$ HA coating on anodized surface), and control (non HA coating on anodized surface) Ti. The morphology of these cells was assessed by SEM. The cDNAs prepared from the total RNAs of the MG63 were hybridized into a human cDNA microarray (1,152 elements). The appearances of the surfaces observed by SEM were different on each of the four dental substrate types. MG63 cells cultured on A, C and control exhibited cell-matrix interactions. It was B surface showing cell-cell interaction. In the expression of several genes were up-, and down-regulated on the different surfaces. The attachment and expression of key osteogenic regulatory genes were enhanced by the surface morphology of the dental materials used.
Gingival hyperplasia is frequently associated with the long-term use of phenytoin for control of convulsive disorder. The purpose of this study was to investigate on the effects of lipopolysaccharides (LPS), ursolic acid and oleanolic acid to phenytoin-induced cell activity in human gingival fibroblast. Human gingival fibroblasts were cultured form the healthy gingiva of orthodontic patients. Gingival fibroblasts were trypsinized and transferred to the weels of microtest plates. Fibroblast were cultured in growth medium added $5{\mu}g/ml$ of phenytoin, $5{\mu}g/ml$ of LPS, $10^{-7}M$ of ursolic acid and oleanolic acid. The passage number of cultured fibroblasts were fifth and eight. Cell morphology was examined by inverted microscope and the cell activity was measured by proliferation assay. Ursolic acid significantly modulated cell morphology into globular shape at the concentrantion of $10^{-7}M$ in the presence of phenytoin and LPS, and the cell activity was significantl decreased by ursolic acid or oleanolic acid regardless of the presence of phenytoin and LPS. These results suggested that the increased phenytoin-induced cell activity might be modulated by ursolic acid regardless of the presence of phenytoin and LPS. These results suggested that the increased phenytoin-induced cell activity might be modulated by ursolic acid or oleanolic acid. Further study is needed to clarify their toxicological effects on cellular modulation and mRNA expression change.
Nitric oxide (NO) occurs in various types of cells in the central nervous system. We studied the distribution and morphology of neuronal nitric oxide synthase (NOS)-containing neurons in the visual cortex of mouse and rabbit with antibody immunocytochemistry. We also compared this labeling to that of calbindin D28K, calretinin, and parvalbumin. Staining for NOS was seen both in the specific layers and in selective cell types. The densest concentration of intense anti-NOS immunoreactive (IR) neurons was found in layer VI, while the weak anti-NOS-IR neurons were found in layer II/III in both animals. The NOS-IR neurons varied in morphology. The large majority of NOS-IR neurons were round or oval cells with many dendrites coursing in all directions. Two-color immunofluorescence revealed that only 16.7% of the NOS-IR cells were double-labeled with calbindin D28K in the mouse visual cortex, while more than half (51.7%) of the NOS-IR cells were double-labeled with calretinin and 25.0% of the NOS-IR cells were double-labeled with parvalbumin in mouse. By contrast, 92.4% of the NOS-IR neurons expressed calbindin D28K while only 2.5% of the NOS-IR neurons expressed calretinin in the rabbit visual cortex. In contrast with the mouse, none of the NOS-IR cells in the rabbit visual cortex were double-labeled with parvalbumin. The results indicate that neurons in the visual cortex of both animals express NOS in specific layers and cell types, which do not correlate with the expression of calbindin D28K, calretinin or parvalbumin between the two animals.
Gingiva is remarkly sensitive to certain drugs. Especially, long term use of phentoin, dihydropyrydine (including nifedipine), cyclosporin and other drugs can be lead to pathologic changes in gingival tissue, especially in terms of proliferation of epithelium and connective tissue. Recent study in terms of proliferation of epithelium and connective tissue. Recent study is focused on the inhibition of drug-induced gingival hyperplasia by using medicaments. The purpose of this study was to investigate on the pharmacological effects of nifedipine, retinoic acid and glycyrrhetini acid to the activity in human gingival fibroblast. Human gingival fibroblasts were cultured from the healthy gingiva of orthodontic patients. Gingival fibroblasts were trypsinized and cultured in growth medium added $5{\mu}g/ml$ of nifedipine, $10^{+7}M$ of retinoic acid and glycyrrhetinic acid. The passage number of cultured fibroblasts were between fifth and eighth. The cell morphology was examined by inverted microscope and the cell acitivity was measured by the MTT assay. Nifedipine at the concentration of $5{\mu}g/ml$ was revealed significantly effective to increase the cell activity and lipopolysaccharide was cofactor to increase cell activity in the presence of nifedipine. However, retinoic acid was significantly effective on the globular change of cell morphology and loss of cell process regardless of the presence of nifedipine and LPS. Cell activity was significantly decreased by the glycyrrhetinic acid at the concentration of $10^-M$ regardless of the presence of nifedipine and LPS. These results suggested that the increased cell activity by nifedipine might be modulated by retinoic acid and glycyrrhetinic acid. Further study is needed to clarify on their toxicological effects during cellular modulation and mRNA expression change.
The $5-HT_6R$ has been considered as an attractive therapeutic target in the brain due to its exclusive expression in the brain. However, the mechanistic linkage between $5-HT_6Rs$ and brain functions remains poorly understood. Here, we examined the effects of $5-HT_6R$-mediated cell morphological changes using immunocytochemistry, Western blot, and live-cell imaging assays. Our results showed that the activation of $5-HT_6Rs$ caused morphological changes and increased cell surface area in HEK293 cells expressing $5-HT_6Rs$. Treatment with 5-HT specifically increased RhoA-GTP activity without affecting other Rho family proteins, such as Rac1 and Cdc42. Furthermore, live-cell imaging in hippocampal neurons revealed that activation of $5-HT_6Rs$ using a selective agonist, ST1936, increased the density and size of dendritic protrusions along with the activation of RhoA-GTP activity and that both effects were blocked by pretreatment with a selective $5-HT_6R$ antagonist, SB258585. Taken together, our results show that $5-HT_6R$ plays an important role in the regulation of cell morphology via a RhoA-dependent pathway in mammalian cell lines and primary neurons.
Background: The efficacy of ginseng, the representative product of Korea, and its chemical effects have been well investigated. The ginsenoside RG3 has been reported to exhibit apoptotic, anticancer, and antidepressant-like effects. Methods: In this report, the putative effect of RG3 on several cellular function including cell survival, differentiation, development and aging process were evaluated by monitoring each specific marker. Also, mitochondrial morphology and function were investigated in ultraviolet (UV)-irradiated normal human dermal fibroblast cells. Results: RG3 treatment increased the expression of extracellular matrix proteins, growth-associated immediate-early genes, and cell proliferation genes in UV-irradiated normal human dermal fibroblast cells. And, RG3 also resulted in enhanced expression of antioxidant proteins such as nuclear factor erythroid 2-related factor-2 and heme oxygenase-1. In addition, RG3 affects the morphology of UV-induced mitochondria and plays a role in protecting mitochondrial dysfunction. Conclusioin: RG3 restores mitochondrial adenosine triphosphate (ATP) and membrane potential via its antioxidant effects in skin cells damaged by UV irradiation, leading to an increase in proteins linked with the extracellular matrix, cell proliferation, and antioxidant activity.
Khanmohammad, Khadije Rezai;Khalili, Mohammad Bagher;Sadeh, Maryam;Talebi, Ali Reza;Astani, Akram;Shams, Ali;Zare, Fateme
Clinical and Experimental Reproductive Medicine
/
v.48
no.2
/
pp.105-110
/
2021
Objective: Uropathogenic Escherichia coli is known to cause urinary tract infections, and the endotoxin (lipopolysaccharide [LPS]) of this bacterium may cause deficiencies of sperm quality and morphology. In the present study, the effects of LPS on mouse sperm were studied, and the levels of interleukin (IL)-17A and possible changes in testis tissue were evaluated. Methods: LPS of uropathogenic E. coli was extracted using the methanol-chloroform method, followed confirmation using sodium dodecyl sulfate-polyacrylamide electrophoresis. Purified LPS (100 ㎍/kg) or phosphate-buffered saline was injected intraperitoneally into BALB/c mice for 7 days consecutively in the test and control groups, mice were sacrificed on days 3, 7, and 42 after the first injection. Blood was tested for levels of IL-17A using the enzyme-linked immunosorbent assay method. Testis tissue and sperm were collected from each mouse and were studied according to standard protocols. Results: The mean sperm count and motility significantly decreased (p=0.03) at 3, 7, and 42 days after the injections. The level of IL-17A in the test groups increased, but not significantly (p=0.8, p=0.11, and p=0.15, respectively). Microscopic studies showed no obvious changes in the morphology of the testis tissue; however, significant changes were observed in the cellular parenchyma on day 42. Conclusion: LPS can stimulate the immune system to produce proinflammatory cytokines, resulting in an immune response in the testis and ultimately leading to deficiency in sperm parameters and testis tissue damage. In addition, the presence of LPS could significantly impair sperm parameters, as shown by the finding of decreased motility.
Objectives : Araliae Continentalis Radix(AC) is a medicinal herb belonging to the drug efficacy group treating musculoskeletal disorders(MSD) with anti-inflammatory, analgesic and antipyretic action. However, due to morphologic and onomastic similarity, adulterants(Angelicae Pubescentis Radix: AP, Gypsophilae Oldhamianae Radix: GO, Levistici Officinalis Radix: LO) have been included or replaced the standard. Methods : Multilateral methods were carried out on the identification of AC and its adulterants. Macroscopic and microscopic characteristics were observed by using stereoscope and microscope. For the comparison of chromatogram pattern, standard compounds were analyzed simultaneously using high performance liquid chromatography. Results : 1. The macroscopic identification of original plants was determined by the phyllotaxis type, the inflorescence type, the leaf margin and the color of flowers. The macroscopic identification of herbal materials was examined by oil spots, the cambium, heteromorphic vascular bundles, and the pholem. 2. For the microscopic identification, the fact whether its xylem ray is proliferated or not was first determined. Then medicinal herbs were secondly divided by cellular inclusions, fiber bundles, the distribution of secretary canals and the shape of cambium. 3. AC and its adulterants showed different chromatographic fingerprints. AC was containing continentalic acid and kaurenoic acid. AP was containing osthole and columbianadin. LO was containing osthole and falcarindiol. None of the compounds were found in GO. Conclusions : This recent identification keys of might be helpful to discriminate the pharmacopoeia standard and its adulterants for the right usage in clinics.
Septin4 belong to a family of polymerizing GTP-binding proteins that are required for many cellular functions, such as membrane compartmentalization, vesicular trafficking, mitosis, and cytoskeletal remodeling. Since, Septin4 is expressed specifically in the testis, we aimed to determine the association between Septin4 gene expression with sperm quality, DNA damage, and stress oxidative level in infertile patients. The present study included 60 semen samples that grouped into three groups: normozoospermia (n=20), asthenozoospermia (n=20), astheno-teratozoospermia (n=20). Initially, semen parameters were analyzed by using the World Health Organization protocol. The mRNA expression of Septin4 in sperm was examined using reverse transcription-polymerase chain reaction. Oxidative stress markers, i.e., total antioxidant capacity, superoxide dismutase, catalase, glutathione peroxidase, and malondialdehyde, were determined by ELISA kit. The current study showed a statistically significant highly positive correlation in Septin4 gene expression with sperm motility, normal morphology, viability, capacity, and sperm mitochondrial membrane potential (MMP). However, it showed significant negative correlation with sperm DNA fragmentation. Septin4 had a significant correlation with stress oxidative factor and antioxidant enzyme levels. In conclusion, Septin4 gene expression provides clinical useful information for the diagnosis of male infertility. It might be a marker for discrimination between fertile and infertile patients. The current study showed a statistically significant highly positive correlation in Septin4 gene expression with sperm motility, normal morphology, viability, capacity, and sperm MMP. However, it shows significant negative correlation with sperm DNA fragmentation. Septin4 had a significant correlation with stress oxidative factor and antioxidant enzyme levels.
Journal of the Korean Association of Oral and Maxillofacial Surgeons
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v.34
no.6
/
pp.616-621
/
2008
Osseointegration is a result of bone formation and bone regeneration processes, which takes place at the interface between bone and implant, and it indicates a rigid fixation that can be stably maintained while functional loading is applied inside the oral cavity as well as after implant placement. Although many researches were carried out about osseointegration mechanism, but cellular and molecular events have not been clarified. With recent development of molecular biology, some researches have examined biological determinants, such as cytokine, growth factors, bone matrix proteins, during osseointegration between bone and implant surface, other researches attempted to study the ways to increase bone formation by adhering protein to implant surface or by inserting growth factors during implant placement. Cellular research on the reaction of osteoblast especially to surface morphology (e.g. increased roughness) has been carried out and found that the surface roughness of titanium implant affects the growth of osteoblast, cytokine formation and mineralization. While molecular biological research in dental implant is burgeoning. Yet, its results are insignificant. We have been studying the roles of growth factors during osseointegration, comparing different manifestations of growth factors by studying the effect of osseointegration that varied by implant surface. Of many growth factors, $TGF-{\beta}$, IGF-I, BMP2, and BMP4, which plays a significant role in bone formation, were selected, and examined if these growth factors are manifested during osseointegration. The purpose of this article is to present result of our researches and encourage molecular researches in dental implant.
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