Fabrication and Characterization of a Multifunctional Coating to Promote the Osteogenic Properties of Orthopedic Implants.

Titanium-based alloys are used in orthopedic applications as fixation elements, hard tissue replacements in artificial bones, and dental implants. Despite their wide range of applications, metallic implant defects and failures arise due to inadequate mechanical bonding, postoperative clotting problems, aseptic loosening, and infections. To improve the surface bioactivity and reduce the corrosion rate of the Ti6Al4V alloy, multi-layered coatings (HAp, BG, Cs, and Hep) were applied via electrophoretic deposition (EPD). XRD images showed the presence of HAp within the coating. In vitro investigation: cell line NIH-3T3 fibroblasts were seeded on the non-coated and coated Ti6Al4V substrates, and their cellular behavior was evaluated. The results indicated that the HApBGCsHep coating could enhance the adhesion and proliferation of NIH 3T3 cells. In addition, the potentiodynamic polarization results are compatible with the in vitro outcome.

Evaluation of the chemopreventive effects of Hypericum perforatum L on DMBA-applied rat oral mucosa.

OBJECTIVE: Hypericum perforatum L also known as St. John's wort is known to have many beneficial properties for the organism including its antioxidant and anticancer activities. It is also known to have shown antiproliferative and cytotoxic effects against various cancer cell lines. The purpose of this study was to investigate the effects of Hypericum perforatum L on 7,12-dimethylbenz(a)anthracene-induced rat oral squamous cell carcinoma model. DESIGN: The in vitro antioxidant properties of Hypericum perforatum L was determined and an extract was prepared. Thirty Wistar male rats were divided randomly into 4 groups (Control group, DMBA group, HP + DMBA group, HP group). The antioxidant defense mechanisms in tissue and blood samples were evaluated biochemically and immunohistochemically, the carcinomatous changes in connective tissue were investigated immunohistochemically and epithelial changes in the tissue samples were evaluated histopathologically. RESULTS: The extract revealed inhibitory effects on some antioxidant enzymes (catalase, glutathione peroxidase). Immunohistochemical evaluations revealed no invasive changes in the connective tissue. Hypericum perforatum L demonstrated chemopreventive effects although it did not prevent carcinomatous changes altogether. CONCLUSIONS: Hypericum perforatum L is a promising chemopreventive agent and further studies are needed in order to evaluate the full potential of this plant.

Evaluation of antioxidant and antimutagenic activities of aluminum chloride.

PURPOSE: Hemostatic agents are used to control hemorrhage and the gingival crevicular fluid for dental applications. In this study; the antimutagenic and antioxidant activities of aluminum chloride (AlCl3), a topical hemostatic agent used especially in the fields of dermatology and dentistry, were determined. To our knowledge, this is the first study that investigates these properties. MATERIALS AND METHODS: The antioxidant activity was determined by DPPH free radical scavenging and ß-carotene-linoleic acid bleaching assays. The antimutagenic activity was evaluated with the Ames Salmonella/ microsome mutagenicity test using Salmonella typhimurium TA98 and TA100 strains. RESULTS: The total antioxidant activity of AlCl3, determined by ß-carotene bleaching assay was found to be 25.59 ± 2.55% and the DPPH scavenging activity of AlCl3 was determined as 17.49 ± 3.07%. AlCl3 showed not any mutagenicity at the tested concentrations by the AMES test used S. typhimurium TA98 and TA100. This drug demonstrated antimutagenic effects at the test concentrations and the strongest antimutagenic activity was observed on 1.25 mg·mL-1/plate concentration of AlCl3. CONCLUSION: AlCl3 showed potent antimutagenic and antioxidant activities and these properties are significant for dentistry and dermatology.

Antimicrobial characteristics and biocompatibility of the surgical sutures coated with biosynthesized silver nanoparticles.

Surgical sutures play important role during the wound healing of the surgical sites which are known to be sensitive to microbial infections. Silver nanoparticles (AgNPs) have been recently used as promising agents against multiple-drug resistant microorganisms. This study was designed to coat the sutures with silver nanoparticles obtained via a green synthesis approach. Microbial-mediated biological synthesis of AgNPs were carried out ecofriendly using Streptomyces sp. AU2 cell-free extract and deposited on silk sutures through an in situ process. Sutures coated with biosyntehsized AgNP (bio-AgNP coated sutures) were characterized using Scanning Electron Microscopy (SEM) and elemantal analysis were carried out using Energy Dispersive X-ray Spectroscopy (EDS). The silver amount released by the bio-AgNP coated sutures was calculated by Inductively Coupled Plasma-Mass Spectroscopy (ICP-MS) throughout a degradation process. Antimicrobial potential of the bio-AgNP coated sutures was determined against common pathogenic microorganisms Candida albicans, Escherichia coli and Staphylococcus aureus. To determine the biocompatibility/cytotoxicty of the bio-AgNP coated sutures, the MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium) assay was used through an indirect test method; that the elutions obtained by the extraction of the sutures at 1, 4, 8 and 10. days and were placed in contact with 3T3 fibroblast cell culture. To best of our knowledge, this is the first report about coating of the nonabsorbable silk sutures with silver nanoparticles biosynthesized using a microbial extract.

Characterisation of Streptomyces violascens OC125-8 lipase for oily wastewater treatment.

In this study, the lipase-producing bacterium Streptomyces violascens (GenBank number MF621564) was identified, and the extracellular S. violascens OC125-8 lipase produced by this strain was characterised for use in wastewater treatment. The lipase was partially purified by ammonium sulphate precipitation at a final yield of 3.28-fold purification and a recovery of 56%. The S. violascens OC125-8 lipase exhibited optimum catalytic activity at 40 °C and pH 8.0; it was stable at 30-40 °C with more than 86% residual activity after 1 h; it was also stable over a relatively broad pH range of pH 7.0-11.0, retaining 83.3-100% activity. V max and K m values were calculated as 0.61 µmol/min/mg and 0.259 mM, respectively. Enzyme activity significantly increased in the presence of Fe2+ ion but was inhibited by Ca2+, Mn2+, Cu2+ and Mg2+. The addition of a serine protease inhibitor, phenylmethylsulfonyl fluoride (PMSF), strongly inhibited enzyme activity while ethylenediaminetetraacetic acid (EDTA), a metal chelating agent, had no inhibitory effect. The enzyme was fairly stable in the presence of surfactants as well as sodium perborate. Examination of commercial detergent tolerance revealed that the lipase was strongly stable in Tursil (88%), Pril (97%) and Fairy (98.5%), while the lipase was activated in Omo (113.4%) and Ariel (128.3%). Moreover, the lipase showed highest activity towards olive oil (100%), sunflower oil (90%) and burned sunflower oil (55%), while corn oil (44%) and burned olive oil (15%) were less hydrolysed by the enzyme. These properties demonstrate that S. violascens OC125-8 lipase is an ideal choice for oily wastewater management.

The Effects of Hypericum perforatum L. on the Proliferation, Osteogenic Differentiation, and Inflammatory Response of Mesenchymal Stem Cells from Different Niches.

The aim of this study is to demonstrate and compare the differentiation, proliferation, migration and inflammatory behavior of dental pulp- and bone marrow-derived mesenchymal stem cells (DP-MSCs and BM-MSCs) in response to a Hypericum perforatum ethanol extract. Using xCELLigence, a real-time monitoring system, a dose of 10 µg/mL was found to be the most efficient concentration for vitality. The IC50 values and doubling time were calculated. The results showed that H. perforatum L. was able to accelerate osteogenic differentiation in DP-MSCs, but calcium granulation was impaired in BM-MSCs. H. perforatum L.-induced migration increased when compared to the TNF-α-induced migration in a Transwell migration assay, and the IL-6 cytokine levels between cells also differed. It can be suggested that tissue memory is an important factor in MSCs, and that they differ in their response to external factors. In conclusion, H. perforatum L. can be considered an excellent osteoinductive agent for DP-MSCs but should not be used for BM-MSCs. Tissue-specific osteoinductive agents should be discussed in future studies.

Ankaferd Blood Stopper with antibiofilm potential successfully inhibits the extracellular matrix degradation enzymes and promotes wound healing of 3T3 fibroblasts in vitro

Background/aim: The potential inhibitory effects of Ankaferd Blood Stopper (ABS) against biofilm formation of oral microorganisms and its capacity for collagenase, hyaluronidase, and elastase inhibitions that have important roles in wound healing have been investigated. Materials and methods: The wound healing potential was determined by its inhibition ability on collagenase, hyaluronidase, and elastase enzyme activities and was evaluated via scratch wound healing assay on murine 3T3 fibroblasts. The antibiofilm activity was tested against eight oral microorganisms using the crystal violet staining method. Results: At 10% ABS successfully inhibited the biofilm formation of the tested microorganisms. Enzyme inhibition analysis revealed that 3% ABS significantly inhibited all three enzymes related to wound healing. The scratch assay showed that wound closure was faster than that of the control for the 3% ABS/plate. Conclusion: The findings of the present study indicated that ABS has effective wound healing potential with its strong antibiofilm activity against oral cavity microorganisms.

Functional denture soft liner with antimicrobial and antibiofilm properties.

BACKGROUND/PURPOSE: Denture soft liners, especially used for elders who have poor disinfection habits, provide a favourable environment for accumulation and colonization of microorganisms. This in vitro study is aimed to investigate the effectiveness of natural carvacrol incorporation into soft lining material on the inhibition of oral pathogens. MATERIALS AND METHODS: Antimicrobial susceptibility of carvacrol was primarily determined by disc diffusion method. Soft lining material was prepared as recommended by the manufacturer and 10 µL carvacrol was added aseptically to the soft liner discs. Inhibition zones for the control discs without carvacrol (C) and carvacrol-incorporated discs (CL) were determined by disc diffusion method. The biofilm inhibition percentages of carvacrol on soft liner was determined by MTT assay and also observed by Scanning Electron Microscopy (SEM). RESULTS: Carvacrol displayed great antimicrobial activity for yeast, Gram-negative and Gram-positive strains. The highest inhibition zone of carvacrol (41.33 ± 1.53 mm) was measured for Bacillus subtilis strain which is followed by Candida albicans and Streptococcus sanguis (34.00 ± 1.73 mm and 32.33 ± 0.58 mm, respectively). The inhibition zones were also similar for soft liner discs with carvacrol, with the highest inhibition zones against B. subtilis, Streptococcus mutans and C. albicans (43.67 ± 0.58 mm, 40.33 ± 0.58 mm and 38.33 ± 1.15 mm, respectively). Incorporation of carvacrol into the soft liner decreased (98.03 ± 0.2%) of the biofilm formation for C. albicans. CONCLUSION: Carvacrol-incorporation obviously decreased the colonization and plaque formation of oral pathogens, especially C. albicans accumulation. Carvacrol may be useful as a promising agent for antibacterial and antifungal management for denture soft lining materials.

Effects of Syzygium aromaticum, Cinnamomum zeylanicum, and Salvia triloba extracts on proliferation and differentiation of dental pulp stem cells.

INTRODUCTION: Hypersensitivity, local irritative and cytotoxic effects are known for the chemical components of Syzygium aromaticum and Cinnamomum zeylanicum contained in dental materials. However, there is no intimate data in dentistry using the whole extracts of these plants and introducing new ones. Salvia triloba is a well-known anti-inflammatory plant that correspondingly could be used in several dental traumas. OBJECTIVES: We aimed to show and compare the effect of S. aromaticum, C. zeylanicum, and S. triloba extracts on dental pulp stem cells (DPSCs) proliferation, differentiation, and immune responses. MATERIAL AND METHODS: Using xCELLigence, a real time monitoring system, we obtained a growth curve of DPSCs with different concentrations of the Extracts. A dose of 10 µg/mL was the most efficient concentration for vitality. Osteogenic differentiation and anti-inflammatory activities were determined by using an ELISA Kit to detect early and late markers of differentiation. RESULTS: The level of osteonectin (ON, early osteogenic marker) decreased, which indicated that the osteogenic differentiation may be accelerated with addition of extracts. However, the level of osteocalcin (OCN, late osteogenic marker and sign of calcium granulation) differed among the extracts, in which S. aromaticum presented the highest value, followed by S. triloba and C. zeylanicum. Surprisingly, the determined calcium granules were reduced in S. aromaticum and S. triloba. In response to tumor necrosis factor alpha (TNF-α), S. triloba-treated DPSCs showed the most reduced level of IL-6 cytokine level. We suggest C. zeylanicum as a promising osteogenic inducer and S. triloba as a potent anti-inflammatory agent, which could be used safely in biocomposite or scaffold fabrications for dentistry. CONCLUSIONS: Because calcium granule formation and cell viability play a critical role in hard tissue formation, S. aromaticum in dentistry should be strictly controlled, and the mechanism leading to reduced calcium granule formation should be identified.

Effects of Syzygium aromaticum, Cinnamomum zeylanicum, and Salvia triloba extracts on proliferation and differentiation of dental pulp stem cells

Abstract Hypersensitivity, local irritative and cytotoxic effects are known for the chemical components of Syzygium aromaticum and Cinnamomum zeylanicum contained in dental materials. However, there is no intimate data in dentistry using the whole extracts of these plants and introducing new ones. Salvia triloba is a well-known anti-inflammatory plant that correspondingly could be used in several dental traumas. Objectives: We aimed to show and compare the effect of S. aromaticum, C. zeylanicum, and S. triloba extracts on dental pulp stem cells (DPSCs) proliferation, differentiation, and immune responses. Material and Methods: Using xCELLigence, a real time monitoring system, we obtained a growth curve of DPSCs with different concentrations of the Extracts. A dose of 10 μg/mL was the most efficient concentration for vitality. Osteogenic differentiation and anti-inflammatory activities were determined by using an ELISA Kit to detect early and late markers of differentiation. Results: The level of osteonectin (ON, early osteogenic marker) decreased, which indicated that the osteogenic differentiation may be accelerated with addition of extracts. However, the level of osteocalcin (OCN, late osteogenic marker and sign of calcium granulation) differed among the extracts, in which S. aromaticum presented the highest value, followed by S. triloba and C. zeylanicum. Surprisingly, the determined calcium granules were reduced in S. aromaticum and S. triloba. In response to tumor necrosis factor alpha (TNF-α), S. triloba-treated DPSCs showed the most reduced level of IL-6 cytokine level. We suggest C. zeylanicum as a promising osteogenic inducer and S. triloba as a potent anti-inflammatory agent, which could be used safely in biocomposite or scaffold fabrications for dentistry. Conclusions: Because calcium granule formation and cell viability play a critical role in hard tissue formation, S. aromaticum in dentistry should be strictly controlled, and the mechanism leading to reduced calcium granule formation should be identified.

Thymbra Spicata Var. Intricata Induces Mesenchymal Stem Cell Proliferation and Osteogenic Differentiation

ABSTRACT A natural agent that maintains mesenchymal stem cell (MSCs) viability, promotes osteogenic differentiation while modulating the immunological response could achieve success in regeneration during healing and may also prevent bone resorption and improve regeneration. We aimed to demonstrate that a Thymbra spicata var. intricata extract could induce proliferation, differentiation, and modulate the immune responses of mesenchymal stem cells (MSCs). Using xCELLigence, a real-time monitoring system, we obtained a growth curve of MSCs. A dose of 10 µg/mL was the most efficient concentration for vitality. Osteogenic differentiation and antiinflammatory activities were determined by using an ELISA Kit to detect early and late markers of differentiation. The Osteonectin (ON, early osteogenic marker) level decreased while the Osteocalcin (OCN, late osteogenic marker) level increased in the T. spicata var. intricata treated group, suggesting that T. spicata var. intricata may accelerate osteogenic differentiation. Reduced level of the IL-6 cytokine in repsonse to TNF-α was evident. T. spicata var. intricata could be a promising osteogenic inducer in dentistry and could be used safely in biocomposites or scaffold fabrications.

The antioxidant and antimutagenic activities of Ankaferd blood stopper,a natural hemostatic agent used in dentistry.

BACKGROUND/AIM: This study investigated the antioxidant and antimutagenic properties of Ankaferd blood stopper (ABS), a plant-based topical hemostatic agent used in Turkey to treat external hemorrhages and bleeding during dental surgery. While previous studies have examined the antimicrobial, antiinflammatory, and anticarcinogenic properties of ABS, to our knowledge, this is the first study to report on the antioxidant and antimutagenic activities of this drug. MATERIALS AND METHODS: Antioxidant activity was evaluated using DPPH radical-scavenging and ß-carotene-linoleic acid tests. Antimutagenic activity was assessed using the Ames Salmonella/microsome mutagenicity test with the bacterial mutant strains Salmonella typhimurium TA98 and TA100. RESULTS: Although ABS demonstrated no free-radical-scavenging activity in DPPH assays at the tested concentrations, ß-carotene-linoleic acid testing found ABS to have a total antioxidant activity rate of 47.06 ± 4.41%. Antimutagenic effects were observed on TA100 at plate concentrations of 5%, 0.5%, and 0.05%, and on TA98 only at a plate concentration of 5%. CONCLUSION: ABS was shown to possess antioxidant and antimutagenic properties that could be of potential value in the fields of medicine and dentistry.

Antioxidant, mutagenic, and antimutagenic activities of Tragopogon longirostis var. longirostis, an edible wild plant in Turkey.

OBJECTIVES: The ethanolic extract of Tragopogon longirostis var. longirostis, a wild edible plant in Anatolia was isolated, and its antioxidant, mutagenic, and antimutagenic properties were investigated. MATERIALS AND METHODS: The antioxidant activity (AA) was determined by the inhibition of 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical, total AA, and phenolic compounds. The mutagenic and antimutagenic activities were investigated by Ames Salmonella/microsome mutagenicity test. RESULTS: The IC50 value for DPPH radicals was 7.84 ± 0.603 mg/mL. The total AA increased with an increase in the concentration of the extracts (1, 5, 10, 20, and 30 mg/mL), containing linoleic acid emulsion. The total phenolic content was 284.71 ± 5.6 mg gallic acid equivalent/g extract. The results showed that the ethanolic extract can be considered safe, because it does not have any mutagenic effect at the tested concentrations. As a result, the ethanolic extract of the leaves exhibited antimutagenic effects at 2.5, 0.25, and 0.025 mg/plate concentrations. CONCLUSIONS: To our knowledge, this is the first study of the antioxidant, mutagenic, and antimutagenic activities of T. longirostis var. longirostis. These activities are an important topic in the food industry, as well as in the medical field.

New Lipase for Biodiesel Production: Partial Purification and Characterization of LipSB 25-4.

The lipolytic activities of 300 Streptomyces isolates were determined in Tributyrin and Rhodamine-B Agar. Lipase activities were also measured with p-nitrophenyl palmitate (p-NPP) as a substrate. The strain of Streptomyces bambergiensis OC 25-4 used in this study was selected among 300 strains of Streptomyces from MUCC as the best lipase producer. The incubation conditions were optimized and the inoculum amount, incubation period, effect of carbon and nitrogen sources, and rates of MgSO4 and CaCO3 were investigated. LipSB 25-4 (the lipase produced by S. bambergiensis OC 25-4 strain) was partially purified with ammonium sulphate precipitation, dialysis, and gel filtration chromatography 2.73-fold and with 92.12 U/mg specific activity. The optimal pH and temperature for LipSB 25-4 were determined as 8.0 and 50°C, respectively. The lipase has high stability in all pH and temperature values used in this study. While LipSB 25-4 was slightly activated in the presence of ß-mercaptoethanol, it was slightly reduced by PMSF. The enzyme conserved approximately 75% of its activity at the end of 60 h, in the presence of methanol and ethanol. Since LipSB 25-4 displays high activity in the thermophilic conditions and stability in the presence of organic solvents, this lipase can catalyse the biodiesel production from olive oil by the transesterification reactions.

Chemical composition of endemic Scorzonera sandrasica and studies on the antimicrobial activity against multiresistant bacteria.

The present study describes the antimicrobial activity and chemical composition of Scorzonera sandrasica Hartvig et Strid (Family Asteraceae), endemic to Turkey. The antimicrobial activity of the hexane, chloroform, ethyl acetate, and ethanol extracts of the aerial parts of S. sandrasica was evaluated against microorganisms, including multiresistant bacteria, using a paper disc diffusion method. The chemical composition of the chloroform extract of the plant was determined by gas chromatography and gas chromatography-mass spectrometry. The major compounds of the chloroform extract of the plant were caryophyllene oxide (19.7%), manoyl oxide (16.5%), and manool (11.3%), respectively. The extracts had antibacterial activity; however, no antifungal activity was observed against the two fungi. In particular, the ethanol and chloroform extracts exhibited significant activity against multiresistant strains of Stenotrophomonas maltophilia.

Antimicrobial activity and chemical composition of endemic Centaurea cariensis subsp. niveo-tomentosa.

The antimicrobial activity of the n-hexane, chloroform, ethyl acetate and ethyl alcohol extracts of the aerial parts of Centaurea cariensis subsp. niveo-tomentosa was evaluated against microorganisms, including multi-antibiotic resistant bacteria, using the paper disc diffusion method. The chemical composition of the chloroform extract of this plant was determined by gas chromatography and gas chromatography-mass spectrometry. The chloroform extract exhibited significant antibacterial activity for all bacteria tested, which are important pathogens. The major compounds of the chloroform extract were caryophyllene oxide (20.79%), spathulenol (14.73%), beta-eudesmol (9.27%), beta-caryophyllene (6.84%), n-cetylalcohol (6.27%), cubenol (5.23%) and cis-alpha-santalol (4.67%), respectively.

Chemical composition of endemic Centaurea austro-anatolica and studies of its antimicrobial activity against multi-resistant bacteria.

Hexane, chloroform, ethyl acetate and ethanolic extracts of the aerial parts of Centaurea austro-anatolica Hub.-Mor. (Asteraceae) were evaluated against microorganisms, including multiresistant bacteria, using a paper disc diffusion method. The chloroform extract exhibited significant antibacterial activity toward all bacteria tested. The chemical composition of the chloroform extract was determined by gas chromatography (GC) and gas chromatography-mass spectrometry (GC-MS). The major compounds of the extract were caryophyllene oxide (21.32 %), spathulenol (10.86 %), n-tricosanol (9.58 %) and geranyl isovalerate (8.71 %).

In vitro study of antibacterial activity on multi-resistant bacteria and chemical composition of the chloroform extract of endemic Centaurea drabifolia subsp. cappadocica.

The antimicrobial activity of the n-hexane, chloroform, ethyl acetate and ethanol extracts of the aerial parts of C. drabifolia S.M. subsp. cappadocica (DC.) Wagenitz (Asteraceae) was evaluated against microorganisms including multi-antibiotic resistant bacteria using the paper disc diffusion method. The chemical composition of the chloroform extract of this plant was determined by gas chromatography and gas chromatography-mass spectrometry. The chloroform extract exhibited significant antibacterial activity against all the bacteria tested, except Stenotrophomonas maltophilia MU63. The major compounds of the chloroform extract were spathulenol (14.1%), caryophyllene oxide (12.5%), octadecanol (10.2%), ethyl palmitate (7.7%), [Z,Z]-10,12-hexadecadienal (6.0%), 3-hydroxy p-anisaldehyde (5.9%) and pentacosane (5.8%).

The in vitro antimicrobial activities of the essential oils of some Lamiaceae species from Turkey.

In this study, antimicrobial activities of the essential oils obtained by using the hydrodistillation method from some lamiaceous plants--Mentha longifolia (L.) Hudson ssp. longifolia, M. longifolia (L.) Hudson ssp. typhoides (Briq.) Harley var. typhoides, Mentha pulegium L., Salvia fruticosa Miller, Salvia tomentosa Miller, Calamintha nepeta (L.) Savi ssp. glandulosa (Req.) P.W. Ball, Nepeta cadmea Boiss., Lavandula stoechas L. ssp. stoechas, and Ziziphora tenuior L.--were determined by using the disc diffusion method. The plants used in this study were collected from different localities of Mugla Province, Turkey. All the essential oils isolated from the plants were very effective against Gram-positive and Gram-negative bacteria, which included multiple-antibiotic resistant strains, except Pseudomonas aeruginosa ATCC 27853 and Pseudomonas fluorescens MU 87. The essential oils of the plant species, except S. tomentosa and S. fruticosa, were very effective against Candida albicans. The antimicrobial activities varied depending on the species, subspecies, or variety. In fact, essential oils of some plants belonging to the same taxa but collected from different localities showed different levels of antimicrobial activities.