Effect of Asiasari radix on osteoblastic differentiation of stem cells derived from gingiva.

OBJECTIVE: To examine the dose-dependent impact of Asiasari Radix (A. radix) on the cell viability, differentiation and mineralization of stem cells derived from gingiva. METHODS: Stem cells that were derived from gingiva were grown in the presence of A. radix at final concentrations that ranged from 0.001 to 10 µg/mL. The morphology of the cells was viewed under an inverted microscope and the analysis of cell proliferation was performed by using Cell Counting Kit-8 (CCK-8) on day 1. The alkaline phosphatase activity test was used to assess differentiation and Alizarin red S staining was used to assess mineralization of treated cells. RESULTS: The control group showed spindleshaped, fibroblast-like morphology and the shapes of the cells in 0.001, 0.01, 0.1, 1 and 10 µg/mL of A. radix were similar to that of the control group at day 1. The cultures growing in the presence of 0.001 µg/mL of A. radix at day 1 showed an increase in the CCK-8 value (P < 0.05). Cultures growing in the presence of 0.001 µg/mL of A. radix presented the highest value for alkaline phosphatase activity (P > 0.05). Mineralized extracellular deposits were observed after Alizarin Red S staining and the cultures grown in the presence of 0.001 µg/mL of A. radix showed the highest value for quantitative results for bound dye (P < 0.05). CONCLUSION: Within the limits of this study, A. radix influenced the proliferation of stem cells derived from the gingiva and low concentrations of A. radix might enhance osteogenic differentiation of the stem cells.

Effects of Cimicifugae Rhizoma on the osteogenic and adipogenic differentiation of stem cells.

Cimicifugae Rhizoma, a herb with a long history of use in traditional Oriental medicine is reported to have anti-inflammatory, antioxidant, anti-complement and anticancer effects. The aim of the present study was to evaluate the effects of Cimicifugae Rhizoma extracts on the osteogenic and adipogenic differentiation of human stem cells derived from gingiva. Stem cells derived from gingiva were grown in the presence of Cimicifugae Rhizoma at final concentrations of 0.1, 1 and 10 µg/ml. Cell proliferation analyses were performed at day 15. For osteogenic differentiation experiments, the stem cells were cultured in osteogenic media containing ß-glycerophosphate, ascorbic acid-2-phosphate and dexamethasone, and osteogenic differentiation was evaluated by analysis of osteocalcin expression at 21 days. For adipogenic differentiation experiments, the stem cells were grown in adipogenic induction medium, and the adipogenic differentiation was evaluated by analysis of adipocyte fatty acid-binding protein at day 14. The cultures grown in the presence of 0.1 µg/ml Cimicifugae Rhizoma showed a significant increase in cellular proliferation at day 15 compared with the control group. The relative osteogenic differentiation in the presence of Cimicifugae Rhizoma for the 0.1, 1 and 10 µg/ml groups was 171.5±13.7, 125.6±28.7 and 150.5±9.0, respectively, when that of the untreated control group on day 21 was considered to be 100%. The relative adipogenic differentiation at day 14 of the 0.1, 1 and 10 µg/ml groups in the presence of Cimicifugae Rhizoma was 97.5±15.0, 102.9±12.8 and 87.0±6.8%, respectively when that of the untreated control group on day 14 was considered to be 100%. Within the limits of this study, Cimicifugae Rhizoma increased the proliferation of stem cells derived from the gingiva, and low concentrations of Cimicifugae Rhizoma may increase the osteogenic differentiation of stem cells.

The role of immobilized rennet on carbon cloth in flavor development during ripening of Gouda cheese.

Rennet-free Gouda (RFG) cheese was prepared to investigate the influence of rennet on the non-volatile and volatile profiles of cheese and was characterized by HPLC and GC/MS analyses. Chymosin, a major protease in rennet, was immobilized onto oxidized and chemically modified carbon cloth. The chymosin immobilization efficiency was 60.4%, and the milk-clotting activity used as an index of the stability of the immobilized chymosin decreased by around 20% in 2 weeks. However, the activity was maintained at 70-80% from 2 weeks to 32 weeks and was more stable than that of chymosin solution alone. Non-volatile (organic acids) and volatile profiles of the RFG cheese and rennet-containing normal Gouda cheese were not significantly different during ripening with a few exceptions. Therefore, it can be concluded that cheese flavor is developed by lactic acid fermentation, irrespective of the presence of rennet.

Anti-inflammatory and cytotoxic effects of methanol, ethanol, and water extracts of Angelicae Dahuricae Radix.

Angelicae Dahuricae Radix has been used for the treatment of headaches, rhinitis, and colds in traditional medicine. Methanol, ethanol, and water extracts of Angelicae Dahuricae Radix were collected. A statistically significant reduction in the cellular viability of the mouse leukemic monocyte macrophage cell line was noted after treatment with water extracts of Angelicae Dahuricae Radix. Stimulation with lipopolysaccharides (LPS) for 24 h led to a robust increase in nitric oxide production, but Angelicae Dahuricae Radix at 400 µg/mL concentration significantly suppressed nitric oxide produced by the LPS-stimulated RAW 264.7 cells in 70% ethanol, absolute ethanol, 70% methanol, absolute methanol, and boiling water groups (P < 0.05). Pretreatment with absolute ethanol extract of Angelicae Dahuricae Radix suppressed the LPS-stimulated inducible nitric oxide synthase, interleukin-1ß, and cycloxygenase-2 expression. Angelicae Dahuricae Radix showed significant cytotoxic effects on the human adenocarcinoma cell line and keratin-forming cell line. (J Oral Sci 58, 125-131, 2016).

Evaluation of the effects of Cimicifugae Rhizoma on the morphology and viability of mesenchymal stem cells.

Cimicifugae Rhizoma is a traditional herbal medicine used to treat various diseases in Korea, China and Japan. Cimicifugae Rhizoma is primarily derived from Cimicifuga heracleifolia Komarov or Cimicifuga foetida Linnaeus. Cimicifugae Rhizoma has been used as an anti-inflammatory, analgesic and antipyretic remedy. The present study was performed to evaluate the extracts of Cimicifugae Rhizoma on the morphology and viability of human stem cells derived from gingiva. Stem cells derived from gingiva were grown in the presence of Cimicifugae Rhizoma at final concentrations that ranged from 0.001 to 1,000 µg/ml. The morphology of the cells was viewed under an inverted microscope and the analysis of cell proliferation was performed using a Cell Counting kit-8 (CCK-8) assay on days 1, 3, 5 and 7. Under an optical microscope, the control cells exhibited a spindle-shaped, fibroblast-like morphology. The shapes of the cells in the groups treated with 0.001, 0.01, 0.1, 1 and 10 µg/ml Cimicifugae Rhizoma were similar to the shapes in the control group. Significant alterations in morphology were noted in the 100 and 1,000 µg/ml groups when compared with the control group. The cells in the 100 and 1,000 µg/ml groups were rounder, and fewer cells were present. The cultures that were grown in the presence of Cimicifugae Rhizoma at a concentration of 0.001 µg/ml on day 1 had an increased CCK-8 value. The cultures grown in the presence of Cimicifugae Rhizoma at a concentration of 10 µg/ml on day 7 had a reduced CCK-8 value. Within the limits of this study, Cimicifugae Rhizoma influenced the viability of stem cells derived from the gingiva, and its direct application onto oral tissues may have adverse effects at high concentrations. The concentration and application time of Cimicifugae Rhizoma should be meticulously controlled to obtain optimal results.

Evaluation of the effects of Angelicae dahuricae radix on the morphology and viability of mesenchymal stem cells.

Angelicae dahuricae radix is a traditional herbal medicine used to treat various diseases in China and Korea, such as colds, headaches, rhinitis and psoriasis. Angelicae dahuricae radix has been used as an anti-inflammatory, analgesic, antipyretic and antioxidant remedy. This study was performed in order to evaluate the effects of the extracts of Angelicae dahuricae radix on the morphology and viability of mesenchymal stem cells derived from the gingiva. Mesenchymal stem cells derived from the gingiva were grown in the presence of Angelicae dahuricae radix at final concentrations that ranged from 0.001 to 100 µg/ml. The morphology of the cells was viewed under an inverted microscope, and the analysis of cell proliferation was performed with cell counting kit-8 (CCK-8) on days 1, 3 and 7. The cells in the control group had spindle-shaped, fibroblast-like morphology at days 1, 3 and 7 under optical microscopy. The shapes of the cells in 0.001, 0.01, 0.1, 1, 10 and 100 µg/ml Angelicae dahuricae radix were similar to the shapes of the cells in the control group. The relative values of the CCK-8 assays of 0.001, 0.01, 0.1, 1, 10, and 100 µg/ml Angelicae dahuricae radix were 102.5 ± 0.6, 133.3 ± 9.6, 148.4 ± 20.5, 147.7 ± 12.6, 132.3 ± 27.7 and 101.1 ± 4.6%, respectively, when the CCK-8 result of the control group on day 1 was considered to be 100%. There was a marginal increase in cell proliferation at 0.1 and 1 µg/ml groups at day 1; however, this did not achieve statistical significance (P=0.052). The relative values of the CCK-8 assays of 0.001, 0.01, 0.1, 1, 10 and 100 µg/ml Angelicae dahuricae radix were 96.5 ± 1.3, 89.3 ± 0.9, 90.3 ± 3.0, 84.8 ± 12.2, 92.3 ± 4.5 and 86.8 ± 11.7%, respectively, when the CCK-8 result of the control group on day 3 was considered to be 100% (P>0.05). The relative values of the CCK-8 assays of 0.001, 0.01, 0.1, 1, 10 and 100 µg/ml Angelicae dahuricae radix day 7 were 94.9 ± 22.3, 102.8 ± 22.1, 127.4 ± 7.4, 130.4 ± 1.3, 129.2 ± 10.8 and 124.8 ± 9.1%, respectively, when the CCK-8 result of the control group on day 7 was considered to be 100%, but there were no statistically significant differences among the groups (P>0.05). Within the limits of this study, Angelicae dahuricae radix at the tested concentrations did not produce statistically significant differences in the viability of stem cells derived from the gingiva.

Effects of Asiasari radix on the morphology and viability of mesenchymal stem cells derived from the gingiva.

Medicinal herbs used in traditional Oriental medicine, which have been in use clinically for thousands of years, are attractive sources of novel therapeutics or preventatives. Asiasari radix (A. radix) has been suggested for use in the treatment of dental diseases, including toothache and aphthous stomatitis. The aim of this study was to evaluate the effects of A. radix extracts on the morphology and viability of human stem cells derived from the gingiva. An Asiasarum heterotropoides extract was centrifuged and freeze-dried in a lyophilizer. Stem cells derived from the gingiva were grown in the presence of A. radix at concentrations ranging between 0.1 µg/ml and 1 mg/ml (0, 0.1, 1, 10, 100 and 1,000 µg/ml). Cell morphology was evaluated with an optical microscope and the viability of the cells was quantitatively analyzed with a cell counting kit-8 (CCK-8) assay for up to seven days. The untreated control group exhibited normal fibroblast morphology. The shapes of the cells following 0.1, 1, 10 and 100 µg/ml A. radix treatments were similar to those of the control group. However, a significant change was noted in the 1,000 µg/ml group on day 1, when compared with the untreated group. Furthermore, on day 7, the shapes of the cells following 100 and 1,000 µg/ml A. radix treatments were rounder and fewer cells were present, when compared with those of the control group. The cultures that grew in the presence of A. radix did not exhibit any changes in the CCK­8 assay on day 2; however, significant reductions in cell viability were noticed following 100 and 1,000 µg/ml A. radix treatment on days 5 and 7. Within the limits of this study, A. radix influenced the viability of the stem cells derived from the gingiva. Thus, the direct application of A. radix to oral tissues may produce adverse effects at high doses. Therefore, the concentration and application time of A. radix requires meticulous control to obtain optimal results. These effects require consideration, if the use of A. radix is planned for the treatment of dental diseases.

Utilizing intrabolus pressure and esophagogastric junction pressure to predict transit in patients with Dysphagia.

BACKGROUND/AIMS: High-resolution manometry (HRM), with a greatly increased number of recording sites and decreased spacing between sites, allows evaluation of the dynamic simultaneous relationship between intrabolus pressure (IBP) and esophagogastric junction (EGJ) relaxation pressure. We hypothesized that bolus transit may occur when IBP overcomes integrated relaxation pressure (IRP) and analyzed the relationships between peristalsis pattern and the discrepancy between IBP and IRP in patients with dysphagia. METHODS: Twenty-two dysphagia patients with normal EGJ relaxation were examined with a 36-channel HRM assembly. Each of the 10 examinations was performed with 20 and 30 mmHg pressure topography isobaric contours, and findings were categorized based on the Chicago classification. We analyzed the relationships between peristalsis pattern and the discrepancy between IBP and IRP. RESULTS: Twenty-two patients were classified by the Chicago classification: 1 patient with normal EGJ relaxation and normal peristalsis, 8 patients with intermittent hypotensive peristalsis and 13 patients with frequent hypotensive peristalsis. A total of 220 individual swallows were analyzed. There were no statistically significant relationships between peristalsis pattern and the discrepancy between IBP and IRP on the 20 or 30 mmHg isobaric contours. CONCLUSIONS: Peristalsis pattern was not associated with bolus transit in patients with dysphagia. However, further controlled studies are needed to evaluate the relationship between bolus transit and peristalsis pattern using HRM with impedance.

[The natural course of early gastric cancer].

BACKGROUND/AIMS: Early gastric cancer (EGC) has a relatively long natural course compared to advanced gastric cancer (AGC). But, few studies about the natural course of EGC are currently available in the literature. In this study we investigated the natural course of EGC in patients who did not receive any treatment. METHODS: All patients diagnosed with gastric cancer on endoscopy at Kosin University Gospel Hospital between January 2001 and December 2010 were reviewed. Those among them who had EGC and did not receive any treatment were enrolled, and an analysis was performed. RESULTS: Twenty seven patients were enrolled. Nine of the enrolled patients were women and the median age was 73 years (range, 38-95). Initial endoscopic findings revealed 11 cases of protruded type, 16 cases of flat and depressed type. Histopathologic analysis indicated that there were 14 cases of well and moderately differentiated adenocarcinoma, 6 cases of poorly differentiated adenocarcinoma and signet ring cell carcinoma. Twelve patients underwent follow-up endoscopy and three of them developed AGC within a mean of 9.6 months (range, 5-12 months). Overall median survival time was 40 months and the 5-year survival rate was 45%. CONCLUSIONS: Although there were some differences in EGC progression, patients who did not receive any treatment progressed eventually. We were unable to identify specific factors predictive of median survival time in these patients.

Hepatitis C virus (HCV) genotypes and the influence of HCV subtype 1b on the progression of chronic hepatitis C in Korea: a single center experience.

BACKGROUND/AIMS: There is some controversy regarding whether or not hepatitis C virus (HCV) subtype 1b is more influential than non-1b subtypes on the progression of chronic hepatitis (CH) C to liver cirrhosis (LC) and hepatocellular carcinoma (HCC). METHODS: We retrospectively analyzed 823 patients with chronic HCV infection, including 443 CH patients, 264 LC patients, and 116 HCC patients, who were HCV RNA positive and HBsAg negative. These patients had not received any prior treatment with either interferon alone or a combination of interferon and ribavirin. RESULTS: HCV subtypes 1b (51.6%) and 2a/2c (39.5%) were the two most common genotypes. The proportions of genotypes 2 (2a/2c, 2b, and 2) and 3 were 45.8% and 1.1%, respectively. One case of genotype 4 was found. HCV subtype 1b (47.3%) was less common than the non-1b subtypes (52.7%) in non-LC patients, but its proportion (56.9%) was higher than that of non-1b subtypes (43.1%) in LC patients (P=0.006). The proportions of patients with HCV subtype 1b did not differ significantly between the LC (55.3%) and HCC (60.3%) groups. Older age, male gender, and the relative progression of liver damage (non-LC vs. compensated LC vs. decompensated LC) were significant risk factors for HCC, with odds ratios of 1.081 (95% confidence interval [CI], 1.056-1.106), 5.749 (95% CI, 3.329-9.930), and 2.895 (95% CI, 2.183-3.840), respectively. HCV subtype 1b was not a significant risk factor for HCC (odds ratio, 1.423; 95% CI, 0.895-2.262). CONCLUSIONS: HCV subtypes 1b and 2a/2c were the two most common HCV genotypes. HCV subtype 1b seemed to be more influential than non-1b subtypes on the progression of CH to LC, but not on the development of HCC from LC.

Immobilization of a mediator onto carbon cloth electrode and employment of the modified electrode to an electroenzymatic bioreactor.

5,5'-Dithiobis(2-nitrobenzoic acid) (DTNB) was selected as an electron transfer mediator and was covalently immobilized onto high porosity carbon cloth to employ as a working electrode in an electrochemical NAD(+)-regeneration process, which was coupled to an enzymatic reaction. The voltammetric behavior of DTNB attached to carbon cloth resembled that of DTNB in buffered aqueous solution, and the electrocatalytic anodic current grew continuously upon addition of NADH at different concentrations, indicating that DTNB is immobilized to carbon cloth effectively and the immobilized DTNB is active as a soluble one. The bioelectrocatalytic NAD+ regeneration was coupled to the conversion of L-glutamate into alpha-ketoglutarate by L-glutamate dehydrogenase within the same microreactor. The conversion at 3 mM monosodium glutamate was very rapid, up to 12 h, to result in 90%, and then slow up to 24 h, showing 94%, followed by slight decrease. Low conversion was shown when substrate concentration exceeding 4 mM was tested, suggesting that L-glutamate dehydrogenase is inhibited by alpha-ketoglutarate. However, our electrochemical NAD+ regeneration procedure looks advantageous over the enzymatic procedure using NADH oxidase, from the viewpoint of reaction time to completion.