Effects of low molecular weight heparins and unfractionated heparin on viability of human umbilical vein endothelial cells.

AIM: The aim of this study was to investigate the effects of bemiparin, nadroparin, enoxaparin, and heparin on viability of human umbilical vein endothelial cells (HUVEC). METHODS: Cultivation of HUVEC was performed in an incubator having 5 % CO(2) at 37 °C and with predefined supplemented medium, until cell monolayers attained confluence which occurred after 7 days. The assays were performed in the exponential growth phase of the cells. The cell viability was assessed using the cleavage of tetrazolium salts added to the culture medium. Heparin sodium, enoxaparin sodium, bemiparin sodium, and nadroparin calcium with concentrations of 100, 10, and 1 IU/100 µL were used for the proliferation assay in which cells were incubated for 24, 48, and 72 h with these drugs. The experiments were conducted in four replicates. RESULTS: Among the study drugs with maximal concentration used in the experiments (100 IU/100 µL), heparin was found to be associated with the lowest viability score in 24 and 48 h, while bemiparin showed the lowest at 72 h. Bemiparin 100 IU/100 µL was significantly associated with lower viability score than that of bemiparin 10 IU/100 µL and bemiparin 1 IU/100 µL at every time interval. Among gradual concentrations of enoxaparin, however, concentration of 1 IU/100 µL was associated with the lowest viability scores at every time point. Heparin 1 IU/100 µL, nadroparin 100 IU/100 µL, and enoxaparin 100 IU/100 µL groups had the highest viability score after 72 h of incubation. CONCLUSION(S): Among low molecular weight heparins (LMWHs), 100 IU/100 µL concentration of bemiparin was associated with a more pronounced effect on reducing viability of HUVEC after 72 h of incubation, while nadroparin 100 IU/100 µL and enoxaparin 100 IU/100 µL showed the least effects. LMWHs differ both from each other and heparin with respect to their effects on cellular viability of HUVEC in dose- and time-dependent manner.

In vitro amoebicidal activity of Salvia staminea and Salvia caespitosa on Acanthamoeba castellanii and their cytotoxic potentials on corneal cells.

BACKGROUND/AIMS: Amoebic keratitis is difficult to treat with total efficacy in some patients because of cysts, which are less susceptible than trophozoites to the usual treatments. We investigated the in vitro effectiveness of methanolic extract of Salvia staminea and Salvia caespitosa against Acanthamoeba castellanii, as well as their cytotoxicity on corneal cells in vitro. METHODS: Extracts were evaluated for their amoebicidal activities using an inverted light microscope. The effect of Savia species, with concentrations ranging between 1.0 and 32.0 mg/mL, on the proliferation of A. castellanii trophozoites and cysts were examined in vitro. For determining the cytotoxicity of Salvia species on corneal cells, agar diffusion tests were performed. RESULTS: According to the results obtained from these tests, S. staminea showed remarkable amoebicidal effect on A. castellanii. In the case of the cytotoxic activity, methanolic extract of S. staminea showed no cytotoxicity on corneal cells with a concentration of 16 mg/mL. CONCLUSIONS: Methanolic extract of S. staminea could be considered a new natural agent against Acanthamoeba. However, further evaluation by in vivo testing is needed to confirm the efficiency of its biological effect.

Influence of storage media containing Salvia officinalis on survival of periodontal ligament cells.

AIM: The purpose of this study was to determine the ability of Salvia officinalis (S. officinalis) extracts to serve as a storage medium for the maintenance of periodontal ligament (PDL) cell viability of avulsed teeth. METHODS AND MATERIALS: PDL cells were obtained from healthy third molars and cultured in Dulbecco's Modi?ed Eagle's Medium (DMEM). Cultures were subjected to 4, 2.5, 1.5, and 0.5% S. officinalis solutions, Hank's balanced salt solution (HBSS), phosphate buffered saline (PBS), and tap water. Tissue culture plates were incubated with experimental media at 37 masculineC for 1, 3, 6, 12 or 24 hours. PDL cell viability was assessed by trypan blue exclusion. Statistical analysis of the data was performed by one-way analysis of variance (ANOVA) complemented by the Tukey's test. The level of significance was 5% (p< 0.05). RESULTS: The results showed 2.5% S. officinalis was a more effective storage medium than the other experimental solutions (p<0.05). Only at 1 hour and 3 hours was there found similar effect between 2.5% S. officinalis and HBSS. At 24 hours, 2.5% S. officinalis was found to be significantly better than the other solutions tested. CONCLUSION: S. officinalis can be recommended as a suitable transport medium for avulsed teeth. CLINICAL SIGNIFICANCE: The findings of this study support the use of S. officinalis as another option for clinicians to use to store and transport avulsed teeth until reimplantation procedures can be done.

In vitro evaluation of the amoebicidal activity of garlic (Allium sativum) extract on Acanthamoeba castellanii and its cytotoxic potential on corneal cells.

Free-living protozoa of the genus Acanthamoeba can cause one of the most severe, potentially sight-threatening infections of the eye, the so-called A. keratitis. A. keratitis is difficult to treat because, under adverse conditions, the amoeba encyst and medical therapy is often less effective against cysts than against trophozoites. The aim of this study was to investigate evaluate the in vitro effect of the nonpolar subfraction of the methanol extract of garlic (Allium sativum) on the growth of A. castellanii trophozoites and cysts and also its cytotoxicity on corneal cells in vitro. Extract was evaluated for its amoebicidal activity, using an inverted light microscope. The effect of the nonpolar extract with the concentrations, ranging from 0.78 to 62.5 mg/mL on the proliferation of A. castellanii trophozoites and cysts, were examined in vitro. For the determination of cytotoxicity of the extract on corneal cells, agar diffusion tests were performed. The present study demonstrates the in vitro effectiveness of the garlic against the A. castellanii growth curve. Evaluations revealed that garlic inhibits trophozoite growth in dose- and time-dependent ways. In the case of the cyctotoxic acitivities, it showed no cytotoxicity for the cornea cells in the concentration of 3.90 mg/mL. These findings indicate that nonpolar subfraction of the methanol extracts of garlic has amoebicidal, as well as its cysticidal, properties on Acanthamoeba trophozoites and cysts. Garlic alone, and in combination with other amoebicidal agents, may be used in clinical practices after further investigations.

Evaluation of in vitro effect of Morus rubra (red mulberry) on survival of periodontal ligament cells.

OBJECTIVE: The purpose of this study was to determine the ability of the juice of Morus rubra fruit to serve as a temporary storage medium for the maintenance of periodontal ligament (PDL) cell viability of avulsed teeth. STUDY DESIGN: PDL cells were obtained from healthy third molars and cultured in Dulbecco's Modified Eagle's Medium (DMEM). Cultures were subjected to 4.0%, 2.5%, 1.5%, and 0.5% of the juice of M. rubra fruit, Hank's balanced salt solution (HBSS), phosphate-buffered saline (PBS), and tap water. Tissue culture plates were incubated with experimental media at 37 degrees C for 1, 3, 6, 12, or 24 hours. PDL cell viability was assessed by trypan blue exclusion. Statistical analysis of the data was accomplished using 1-way analysis of variance complemented by the Tukey test. The level of significance was 5% (P < .05). RESULTS: The efficacy of 4.0% and 2.5% M. rubra at 3, 6, and 12 hours was found to be significantly better than HBSS (P < .05). At 24 hours, 4% M. rubra was found to be similar to HBSS, but 2.5% M. rubra was found to be significantly worse than HBSS (P < .005). The results showed that juice of the fruit sample of M. rubra studied at a concentration of 4% was a more effective storage medium than other groups. CONCLUSION: Juice of the fruit of M. rubra can be recommended as a suitable transport medium for avulsed teeth.

The effect of propolis in experimental Acanthamoeba keratitis.

PURPOSE: To examine the effect of propolis in a rat model of Acanthamoeba keratitis and to determine its in vitro cytotoxicity in cultured corneal epithelial cells. METHODS: Eighteen Wistar albino rats were used. Cultured corneal epithelial cells obtained from two healthy rats for in vitro cytotoxicity of propolis. Corneal stromal inoculation was performed in 16 rats with amoebic culture containing 1 x 10(6) amoeba/mL. Rats with Acanthamoeba keratitis 5 days later after the inoculation were divided randomly into four groups, and eight eyes of each group were treated with study drugs. The propolis, chlorhexidine (CHX), propolis plus CHX and control eyes were treated with topical propolis, 0.002% CHX, propolis plus 0.002% CHX and lubricant eye drops, respectively. The study drugs were instilled every one hour for 10 days. All eyes were examined and keratitis graded by slit-lamp biomicroscopy on days 2, 5 and 10 during the administration of the study drugs. After the completion of keratitis grading, all the 16 rats were humanely killed and their corneas were excised and used for Acanthamoeba culture to evaluate presence of Acanthamoeba growth after treatment 14 days later. RESULTS: Concentrations of propolis higher than 7.81 mg/mL cause damage to corneal epithelial cells in the experiment of in vitro cytotoxicity of propolis on corneal epithelial cells. The keratitis grade on day 2 in the CHX eyes was significantly lower than that in the control eyes (P < 0.05). The keratitis grades on days 5 and 10 in the propolis, CHX and propolis plus CHX eyes were significantly lower compared with those on days 5 and 10 in the control eyes (P < 0.05). In the propolis eyes, the keratitis grade on day 5 was significantly lower than that on day 2 (P < 0.05), and it was significantly lower on day 10 compared with that on day 5 (P < 0.05). In the CHX and propolis plus CHX eyes, the keratitis grade on day 10 was significantly lower compared with that on days 2 and 5 (P < 0.05). In the control eyes, there was no significant difference in the keratitis grades on days 2, 5 and 10 (P > 0.05). The culture positivity at Acanthamoeba growth after treatment experiment in the propolis, CHX and propolis plus CHX eyes was significantly lower than that in the control eyes (P < 0.05). CONCLUSIONS: We suggest that propolis had amoebicidal properties in this rat model of Acanthamoeba keratitis. Further investigations to evaluate the antimicrobial activity of the individual fractions of the resin could yield more information about its mechanism of action in treating this disease.

In vitro effectiveness of Thymus sipyleus subsp. sipyleus var. sipyleus on Acanthamoeba castellanii and its cytotoxic potential on corneal cells.

Acanthamoeba species are an important cause of microbial keratitis that may cause severe ocular inflammation and visual loss. Acanthamoeba keratitis is difficult to treat, without total efficacy in some patients because of cysts which is less susceptible than trophozoites to the usual treatments. We investigated here the in vitro amoebicidal activity of methanolic extract of Thymus sipyleus subsp. sipyleus var. sipyleus from Turkish flora against Acanthamoeba castellanii and also its cytotoxicity on corneal cells in vitro. Extract was evaluated for its amoebicidal activity using an inverted light microscope. The effect of the polar extract with the concentrations ranging from 1.0 to 32.0 mg/ml on the proliferation of A. castellanii trophozoites and cysts were examined in vitro. For the determination of cytotoxicity of the extract on corneal cells, agar diffusion tests were performed. According to results obtained from the tests, the extract evaluated here showed remarkable amoebicidal effect on A. castellanii. In the case of the cytotoxic activities, it showed no cytotoxicity for corneal cells in the concentration of 32 mg/ml. As a result, polar subfraction of the methanolic extract of Thymus sipyleus subsp. sipyleus var. sipyleus could be concluded as a new natural agent for the treatment of Acanthamoeba infections. On the other hand, it still needs to be further evaluated by in vivo test systems to confirm the efficiency of its biological effect.

Effect of propolis on survival of periodontal ligament cells: new storage media for avulsed teeth.

Propolis is a multifunctional material used by bees in the construction and maintenance of their hives. Propolis possesses several biologic activities such as anti-inflammatory, antibacterial, antioxidant, antifungal, antiviral, and tissue regenerative, among others. The purpose of this study was to determine the ability of propolis to serve as a temporary storage medium for the maintenance of periodontal ligament (PDL) cell viability of avulsed teeth. PDL cells were obtained from healthy third molars and cultured in Dulbecco's Modified Eagles Medium (DMEM). Cultures were subjected to 10% propolis solution, 20% propolis solution, long-shelf life light milk with lower fat content (milk), Hank's Balanced Salt Solution, tap water as the negative control, and DMEM as the positive control. Tissue culture plates were incubated with experimental media at 37 degrees C for 1, 3, 6, 12, or 24 hours. PDL cell viability was assessed by trypan blue exclusion. Statistical analysis of the data was accomplished by using one-way analysis of variance complemented by the Tukey test. The level of significance was 5% (p<0.05). The results showed that 10% propolis was a more effective storage medium than other groups. In conclusion, propolis can be recommended as a suitable transport medium for avulsed teeth.

In vitro amoebicidal activity of four Allium species on Acanthamoeba castellanii and their cytotoxic potentials on corneal cells.

Amoebic keratitis is difficult to treat without total efficacy in some patients because of cysts, which is less susceptible than trophozoites to the usual treatments. We investigated the in vitro effectiveness of the methanolic extract of four Allium species from Turkey against Acanthamoeba castellanii and its cytotoxicity on corneal cells in vitro. Extracts were evaluated for their amoebicidal activities using an inverted light microscope. The effect of the Allium species with concentrations ranging from 1.0 to 32.0 mg/ml on the proliferation of A. castellanii trophozoites and cysts were examined in vitro. For the determination of the cytotoxicity of the Allium species on corneal cells, agar diffusion tests were performed. According to the results obtained from the tests, A. scrodoprosum subsp. rotundum showed remarkable amoebicidal effect on A. castellanii, while the others remained inactive. In the case of cyctotoxic activities, the methanolic extracts of A. scrodoprosum showed no cytotoxicity for the cells in the concentration of 32 mg/ml, while the others exerted significant cytotoxic activities between the concentrations of 1 and 16 mg/ml. As a result, the methanolic extract of A. scrodoprosum could be concluded as a new natural agent against Acanthamoeba. On the other hand, it still needs to be further evaluated by in vivo test systems to confirm the efficiency of its biological effect.