scholarly journals Evaluation of Antibacterial and Cytotoxic Properties of a Fluorinated Diamond-Like Carbon Coating for the Development of Antibacterial Medical Implants

Antibiotics ◽  
2020 ◽  
Vol 9 (8) ◽  
pp. 495 ◽  
Author(s):  
Katsutaka Yonezawa ◽  
Masahito Kawaguchi ◽  
Ayumi Kaneuji ◽  
Toru Ichiseki ◽  
Yoshitsugu Iinuma ◽  
...  

Peri-implant infection is a serious complication in surgical procedures involving implants. We conducted an in vitro study to determine whether the use of a fluorinated diamond-like carbon (F-DLC) coating on a titanium alloy surface can prevent peri-implant infection. After applying the F-DLC, we evaluated its antibacterial and cytotoxic properties. The coating groups, containing controlled fluorine concentrations of 5.44%, 17.43%, 24.09%, and 30%, were examined for the presence of Staphylococcus aureus and Escherichia coli according to ISO 22196 for the measurement of antibacterial activity on plastics and other nonporous surfaces. Biological toxicity was evaluated using Chinese hamster V79 cells according to ISO 10993-5 for the biological evaluation of medical devices. In the control group, populations of S. aureus and E. coli substantially increased from 2.4 × 104 to (1.45 ± 1.11) × 106 colony-forming units (CFUs) and from 2.54 × 104 to (4.04 ± 0.44) × 106 CFUs, respectively. However, no bacteria colonies were detected in any F-DLC group with a fluorine concentration of ≥ 17.43%. In the biological toxicity study, an F-DLC coating with a fluorine concentration of 30% showed a colony formation rate of 105.8 ± 24.1%, which did not differ significantly from the colony formation rate of 107.5 ± 31.1% in the nontoxic control group. An F-DLC coating on titanium alloy discs showed excellent in vitro antibacterial activity with no biological toxicity.

2018 ◽  
Vol 30 (10) ◽  
pp. 1342 ◽  
Author(s):  
Zhao-Bo Luo ◽  
Long Jin ◽  
Qing Guo ◽  
Jun-Xia Wang ◽  
Xiao-Xu Xing ◽  
...  

Accumulating evidence suggests that aberrant epigenetic reprogramming and low pluripotency of donor nuclei lead to abnormal development of cloned embryos and underlie the inefficiency of mammalian somatic cell nuclear transfer (SCNT). The present study demonstrates that treatment with the small molecule RepSox alone upregulates the expression of pluripotency-related genes in porcine SCNT embryos. Treatment with the histone deacetylase inhibitor LBH589 significantly increased the blastocyst formation rate, whereas treatment with RepSox did not. Cotreatment with 12.5 μM RepSox and 50 nM LBH589 (RepSox + LBH589) for 24 h significantly increased the blastocyst formation rate compared with that of untreated embryos (26.9% vs 8.5% respectively; P < 0.05). Furthermore, the expression of pluripotency-related genes octamer-binding transcription factor 4 (NANOG) and SRY (sex determining region Y)-box 2 (SOX2) were found to significantly increased in the RepSox + LBH589 compared with control group at both the 4-cell and blastocyst stages. In particular, the expression of NANOG was 135-fold higher at the blastocyst stage in the RepSox + LBH589 group. Moreover, RepSox + LBH589 improved epigenetic reprogramming. In summary, RepSox + LBH589 increases the expression of developmentally important genes, optimises epigenetic reprogramming and improves the in vitro development of porcine SCNT embryos.


2020 ◽  
Vol 151 ◽  
pp. 01054
Author(s):  
Ekowati Handharyani ◽  
Lina N. Sutardi ◽  
Aulia A. Mustika ◽  
Andriani Andriani ◽  
Sri Yuliani

Previous in vitro studies showed that nanoparticle extract of turmeric, zedoary, and garlic exhibit antibacterial activity against Mycoplasma gallisepticum (M. gallisepticum) which causes chronic respiratory disease (CRD) in chicken. This research aimed to determine the antibacterial activity of nanoparticles of Curcuma longa, Curcuma zedoaria, and Allium sativum extract to CRD infected chicken. In vivo test of antibacterial activity of turmeric, zedoary, and garlic nanoparticle in combination was conducted on chicken infected by M. gallisepticum and Escherichia coli (E.coli). Antibiotic control used was enrofloxacin. As many as 75 chickens were divided into 5 groups containing 15 chickens each. Group one consisted of healthy chickens (positive control); group two consisted of chickens that have been inoculated by bacteria (negative control); group three (treatment) were chickens inoculated by bacterium and given extract nanoparticle combination on day 7 of infection for 7 days; group four (prevention) were chickens inoculated by bacterium and given combination of extract nanoparticles on day 5 before infection for 14 days; group five were chickens inoculated with bacterium and given enrofloxacin antibiotics for 7 days. In vivo research results showed increased body weight and performance indicated by improvements in clinical signs, and gross pathology changes. The combination of three extract nanoparticles showed the best activity in controlling CRD in chicken, both as preventive and curative means.


2019 ◽  
Vol 7 (6) ◽  
pp. 1032-1036 ◽  
Author(s):  
I Gusti Agung Ayu Dharmawati ◽  
Tjokorda Gde Bagus Mahadewa ◽  
I Putu Eka Widyadharma

AIM: The purpose of this study was to determine the antibacterial activity of Lumbricus rubellus earthworms through inhibitory zone diameter to the growth of the bacterium Phorphyromonas gingivalis as the cause of periodontitis. METHODS: This was an experimental study with randomised posttest-only control group design. The study was conducted at the Microbiology Research Center laboratory at the Faculty of Dentistry, Airlangga University, Indonesia. The study was conducted in vitro, the sample size was calculated using the Federer formula as many as four agar plates containing bacteria Phorphyromonas gingivalis, with each plate given five different treatments: control (ethanol), Lumbricus rubellus earthworm extract (ECT) with concentrations of 50%, 25%, 12.5%, and 6.25% respectively. The data in the form of inhibition zone diameter (measured in millimetres) obtained were tested using One-Way ANOVA. RESULTS: The mean diameter of the inhibitory zone extract of Lumbricus rubellus earthworm on the growth of Phorphyromonas gingivalis bacteria in the treatment group had significant differences (p < 0.05). The mean inhibition zones between controls and the ECT treatment group (ECT 50%, ECT 25%, ECT 12.5%) were statistically different (p < 0.05), in contrast with ECT 6.25% (p > 0.05) which did not show significant difference with the control group (p > 0.05). CONCLUSION: Lumbricus rubellus earthworm extract with a concentration of 50% has the largest diameter of the inhibitory zone on the growth of the Phorphyromonas gingivalis bacteria. The 6.25% earthworm extract showed no antibacterial activity against the growth of Phorphyromonas gingivalis bacteria.


2020 ◽  
Vol 102-B (6_Supple_A) ◽  
pp. 158-162
Author(s):  
Quentin Griseti ◽  
Christophe Jacquet ◽  
Pierre Sautet ◽  
Matthew P. Abdel ◽  
Sébastien Parratte ◽  
...  

Aims The aim of this study was to compare the ability of tantalum, 3D porous titanium, antibiotic-loaded bone cement, and smooth titanium alloy to inhibit staphylococci in an in vitro environment, based on the evaluation of the zone of inhibition (ZOI). The hypothesis was that there would be no significant difference in the inhibition of methicillin-sensitive or methicillin-resistant Staphylococcus aureus (MSSA/MRSA) between the two groups. Methods A total of 30 beads made of three different materials (tantalum/3D porous titanium and smooth titanium alloy) were bathed for one hour in a solution of 1 g vancomycin in 20 ml of sterile water for injection (bath concentration: 50 mg/mL). Ten 1 cm3 cylinders of antibiotic-loaded cement were also created by mixing standard surgical cement with 1 g of vancomycin in standardized sterile moulds. The cylinders were then placed on agar plates inoculated with MSSA and MRSA. The ZOIs were measured each day and the cylinders were transferred onto a new inoculated plate. Results For MSSA and MRSA, no inhibitory effect was found in the control group, and antibiotic-loaded smooth titanium alloy beads showed a short inhibitory effect until day 2. For MSSA, both tantalum and 3D porous titanium beads showed significantly larger mean ZOIs than cement beads (all p < 0.01) each day until day 7 for tantalum and until day 3 for 3D porous titanium. After six days, antibiotic-loaded cement had significantly larger mean ZOIs than the 3D porous titanium (p = 0.027), but no significant difference was found with tantalum (p = 0.082). For MRSA, both tantalum and 3D porous titanium beads had significantly larger mean ZOIs than antibiotic-loaded cement each day until day 6 for tantalum (all p < 0.01) and until day 3 for 3D porous titanium (all p < 0.04). Antibiotic-loaded cement had significantly larger mean ZOIs than tantalum and 3D porous titanium from day 7 to 9 (all p < 0.042). Conclusion These results show that porous metal implants can deliver local antibiotics over slightly varying time frames based on in vitro analysis. Cite this article: Bone Joint J 2020;102-B(6 Supple A):158–162.


2012 ◽  
Vol 24 (5) ◽  
pp. 656 ◽  
Author(s):  
Islam M. Saadeldin ◽  
Ok Jae Koo ◽  
Jung Taek Kang ◽  
Dae Kee Kwon ◽  
Sol Ji Park ◽  
...  

Kisspeptin (Kp) is best known as a multifunctional peptide with roles in reproduction, the cardiovascular system and cancer. In the present study the expression of kisspeptin hierarchy elements (KISS1, GNRH1 and LHB) and their receptors (KISS1R, GNRHR and LHCGR, respectively) in porcine ovary and in cumulus–oocyte complexes (COCs) were investigated, as were its effects on the in vitro maturation (IVM) of oocytes and their subsequent ability to sustain preimplantation embryo competence after parthenogenetic electrical activation. Kp system elements were expressed and affected IVM of oocytes when maturation medium was supplemented with 10–6 M Kp. Oocyte maturation, maternal gene expression (MOS, GDF9 and BMP15), blastocyst formation rate, blastocyst hatching and blastocyst total cell count were all significantly increased when oocytes were matured in medium containing Kp compared with the control group (without Kp). A Kp antagonist (p234) at 4 × 10–6 M interfered with this hierarchy but did not influence the threshold effect of gonadotrophins on oocyte maturation. FSH was critical and permissive to Kp action on COCs by increasing the relative expression of KISS1R. In contrast, Kp significantly increased apoptosis, the expression of pro-apoptotic gene, BAK1, and suppressed trophoblast outgrowths from hatched blastocysts cultured on feeder cells. The present study provides the first functional evidence of the Kp hierarchy in porcine COCs and its role in enhancing oocyte maturation and subsequent developmental competence in an autocrine–paracrine manner. However, Kp supplementation may have a harmful impact on cultured hatched blastocysts reflecting systemic or local regulation during the critical early period of embryonic development.


Zygote ◽  
2011 ◽  
Vol 20 (1) ◽  
pp. 27-32 ◽  
Author(s):  
Byung Chul Jee ◽  
Jun Woo Jo ◽  
Jung Ryeol Lee ◽  
Chang Suk Suh ◽  
Seok Hyun Kim ◽  
...  

SummaryWe performed this study to investigate the effect of histone deacetylase inhibition during extended culture of in vitro matured mouse oocytes. In vitro matured mouse (BDF1) oocytes were cultured in vitro for 6, 12, and 24 h, respectively, and then inseminated. During in vitro culture for 6 and 12 h, two doses of trichostatin A (TSA), a histone deacetylase inhibitor, were added (100 nM and 500 nM) to the culture medium and the oocytes were then inseminated. During the 24-h in vitro culture, two doses of TSA were added (100 nM and 500 nM) to the medium and the oocytes were activated with 10 mM SrCl2. After the 6-h culture, the fertilization rate was similar to that of the control group, but the blastocyst formation rate was significantly decreased. After the 12-h culture, both the fertilization and blastocyst formation rates were significantly decreased. After the 24-h culture, total fertilization failure occurred. In the oocytes cultured for 6 and 12 h, the fertilization and blastocyst formation rates did not differ between the TSA-supplemented and control groups. Although extended culture of the mouse oocytes significantly affected their fertilization and embryo development, TSA supplementation did not overcome their decreased developmental potential.


2019 ◽  
Vol 01 (03) ◽  
pp. 131-135
Author(s):  
Shubiao Han ◽  
Wei Han ◽  
Xiaodong Zhang ◽  
Junxia Liu ◽  
Guoning Huang

Background: This study was conducted to evaluate the impact of vitrification on mitochondrial of human IVM oocytes. Methods: A total of 401 immature oocytes were obtained from ovarian stimulated cycles, which were randomly divided into fresh and vitrification groups after IVM. According to the cultured time after thawing, the vitrification groups were divided into 0 hours (0 h), 2 hours (2 h), or 4 hours (4 h) subgroups. Mitochondrial morphology and oxygen consumption were compared among the four groups. After fertilization by ICSI, normal fertilization, cleaved embryos, and blastocyst formation rate were also calculated. Results: The mean gray value of mitochondria structure was significantly decreased in 0 h and 2 h groups when compared to control group (0.48 ± 0.09, 0.50 ± 0.36 vs. 0.61 ± 0.12, respectively; P [Formula: see text] 0.05), and recovered (0.61 ± 0.24 vs. 0.61 ± 0.12, P [Formula: see text] 0.05) in 4 h group. In addition, oxygen consumption was also significantly decreased in 0 h and 2 h groups compared to fresh (2.91 ± 0.77 fmol/s, 3.26 ± 1.34 fmol/s vs. 3.96 ± 1.44 fmol/s, respectively; P [Formula: see text] 0.05), and recovered after 4 h culture (3.96 ± 1.44 fmol/s vs. 4.41 ± 1.38 fmol/s, respectively; P [Formula: see text] 0.05). The percentage of normal fertilization and cleaved embryos were no differences among the four groups, however, blastocyst development rate was significantly lower in 0 h group. Conclusion: These results indicate that during the vitrification process, the oxygen consumption and mitochondrial structure of oocytes may undergo temporary dynamic changes, but appear to recover by 4 hours.


Zygote ◽  
2021 ◽  
pp. 1-7
Author(s):  
Nguyen Thi Men ◽  
Thanh Quang Dang-Nguyen ◽  
Tamas Somfai ◽  
Hiep Thi Nguyen ◽  
Junko Noguchi ◽  
...  

Summary This study was conducted to examine whether the nuclear to cytoplasmic (N/C) ratio had any influence on the timing of embryo compaction and blastocoel formation, as well as formation rate and quality of blastocyst. First, we produced embryos with increased N/C ratio by removal of approximately one-third of the cytoplasm and with decreased N/C ratio by doubling the oocyte cytoplasm with an enucleated oocyte. The initiation of compaction and cavitation in reduced cytoplasm group was significantly earlier (P < 0.05) compared with the control and doubled cytoplasm groups. The rate of blastocysts in the reduced cytoplasm and doubled cytoplasm groups was significantly lower (P < 0.05) compared with the control group. Blastocyst quality in terms of total cell number in the reduced cytoplasm group was significantly lower (P < 0.05) compared with the doubled cytoplasm group, but not different from the control group. Next, we produced embryos with various N/C ratios by oocyte fusion combined with cytochalasin D treatment. The onset of compaction and cavitation in the 2N/2C group (decreased N/C ratio) was significantly delayed (P < 0.05) or had the tendency to be delayed (P = 0.064), respectively, compared with the control group (2N/1C). A significantly higher rate of blastocyst was observed in the 4N/2C group compared with the 1N/1C group (P < 0.05) but not different from the remaining groups. These results demonstrated that an increase in N/C ratio caused an earlier occurrence of morula compaction and blastocyst formation in both in vitro fertilization (IVF) and parthenogenetically activated pig embryos.


Reproduction ◽  
2006 ◽  
Vol 132 (4) ◽  
pp. 559-570 ◽  
Author(s):  
Tamás Somfai ◽  
Manabu Ozawa ◽  
Junko Noguchi ◽  
Hiroyuki Kaneko ◽  
Katsuhiko Ohnuma ◽  
...  

We investigated nuclear progression and in vitro embryonic development after parthenogenetic activation of porcine oocytes exposed to cytochalasin B (CB) during in vitro maturation (IVM). Nuclear progression was similar in control oocytes and oocytes matured in the presence of 1 μg/ml CB (IVM-CB group) by 37 h IVM; at this time the proportion of oocytes that had reached or passed through the anaphase-I stage did not differ significantly between the IVM-CB and the control groups (61.3 and 69.9% respectively; P < 0.05). After IVM for 37 h, no polar body extrusion was observed in the IVM-CB group. In these oocytes, the two lumps of homologous chromosomes remained in the ooplasm after their segregation and turned into two irregular sets of condensed chromosomes. By 41 h IVM, the double sets of chromosomes had reunited in 89.5% IVM-CB oocytes and formed a single large metaphase plate, whereas 68.8% of the control oocytes had reached the metaphase-II stage by this time. When IVM-CB oocytes cultured for 46 h were stimulated with an electrical pulse and subsequently cultured for 8 h without CB, 39.0% of them extruded a polar body and 82.9% of them had a female pronucleus. Chromosome analysis revealed that the majority of oocytes that extruded a polar body were diploid in both the control and the IVM-CB groups. However, the incidence of polyploidy in the IVM-CB group was higher than that in the control group (P < 0.05). In vitro development of diploid parthenotes in the control and the IVM-CB groups was similar in terms of blastocyst formation rates (45.8 and 42.8% respectively), number of blastomeres (39.9 and 44.4 respectively), the percentage of dead cells (4.3 and 2.9% respectively), and the frequency of apoptotic cells (7.3 and 6.3% respectively). Tetraploid embryos had a lower blastocyst formation rate (25.5%) and number of cells (26.2); however, the proportion of apoptotic nuclei (7.0%) was similar to that in diploid parthenotes. These results suggest that the proportion of homozygous and heterozygous genes does not affect in vitro embryo development to the blastocyst stage.


2012 ◽  
Vol 521 ◽  
pp. 191-205 ◽  
Author(s):  
K. Navaneetha Pandiyaraj ◽  
Jan Heeg ◽  
Christoph Mewes ◽  
Marion Wienecke ◽  
Torsten Barfels ◽  
...  

Silver containing diamond like carbon films were coated on the surface of polyethylene film (PET) using novel hybrid sputtering method. Polymeric substrates can create soft, flexible, highly absorbent and cost-effective materials by selecting or controlling their molecular structures. The material silver is known to be a potential antibacterial material. The silver containing coating has been potentially recommended for synthesis biomedical materials. In the present work, we discussed the antibacterial activity of the silver containing DLC film coated PET film surfaces which was coated as a function of deposition power level. The surface morphology of the Ag-DLC was analysed by atomic force microscopy (AFM) and scanning electron microscopy (SEM). The atomic concentration and structure of the Ag-DLC was measured by energy dispersive X-ray (EDX) and Raman spectroscopy. The hydrophilicity of the films was measured by contact angle analysis. The antibacterial activity of Ag–DLC films were evaluated by bacterial eradication tests withEscherichia coliat incubation time of one day. In addition, blood compatibility of the Ag-DLC films were studied byIn Vitroblood compatibility tests. It was found that the surface of the obtained Ag-DLC decreases with increasing the deposition power level. The antibacterial and hemocompatibility of the silver containing DLC film increase gradually with increase of deposition power level. Our results revealed that the Ag-incorporated DLC films are potentially useful as biomedical devices having good antibacterial and hemocompatibility.


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