Anti-microbial coating solutions were prepared by dissolving a fatty acid (palmitic or lauric acid) and chlorhexidine in 99.8% ethanol (C. Roth GmbH, Germany (CR)). In order to prepare the solutions for suture coatings having a mass content of 5% (w/w), 395.0 mg of the corresponding fatty acid and the antiseptic drug were dissolved in 10.0 ml (7.9 g) ethanol. In particular, two groups of coating types were produced. Group I (CL ): Chlorhexidine diacetate (Sigma-Aldrich GmbH, Germany) in lauric acid as drug carrier (CR). Group II (CP ): Chlorhexidine diacetate in the drug carrier palmitic acid (CR). For each type of coating three solutions with 20%, 40% or 60% (w/w) drug concentration were formulated. The percentage of drug was related to the 5% total mass of substances (drug plus drug carrier) dissolved in coating solutions. Components were prepared by weighing, using the precision balance Atilon ATL-224 (Acculab Inc., Massachusetts, USA).
Chlorhexidine diacetate
Manufactured by Merck Group
Sourced in United Kingdom, Germany
Chlorhexidine diacetate is a chemical compound used as a disinfectant and antiseptic agent in various laboratory and medical applications. It is a salt of chlorhexidine and acetic acid, and it exhibits antimicrobial properties against a wide range of bacteria, fungi, and some viruses.
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Lab products found in correlation
Piperazine, by Merck Group (3 mentions)
1 6 hexanediol diacrylate, by Merck Group (2 mentions)
Sodium alginate, by Merck Group (2 mentions)
Triton x 100, by Merck Group (2 mentions)
Cyclohexane, by Merck Group (2 mentions)
Phosphate buffer solution (pbs), by Merck Group (1 mentions)
Ab198228, by Abcam (1 mentions)
Ab21286, by Abcam (1 mentions)
Ab8448, by Abcam (1 mentions)
Ab150681, by Abcam (1 mentions)
In vitro toxicology assay kit ldh based, by Merck Group (1 mentions)
Alp colorimetric assay kit, by Abcam (1 mentions)
Hoechst 33342 trihydrochloride, by Thermo Fisher Scientific (1 mentions)
Calcein am, by Merck Group (1 mentions)
N hexanol, by Merck Group (1 mentions)
Propidium iodide, by Merck Group (1 mentions)
Alizarin red s, by Merck Group (1 mentions)
Glutaraldehyde, by Merck Group (1 mentions)
Ab150077, by Abcam (1 mentions)
Gentamicin sulphate, by Merck Group (1 mentions)
6 protocols using chlorhexidine diacetate
1
Antimicrobial Suture Coatings with Fatty Acids
2
Synthesis and Characterization of Biomimetic Bone Scaffolds
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Triton X-100, tetraethyl orthosilicate (TEOS), 3-aminopropyltriethoxysilane (APTS), 1,6 hexanediol diacrylate, piperazine, sodium alginate, chlorhexidine diacetate, sodium acetate trihydrate, phosphate buffer solution (PBS) tablets, glutaraldehyde, Alizarin Red S, Calcein-AM, propidium iodide and “In Vitro Toxicology Assay Kit- LDH based” were purchased from Sigma-Aldrich, UK.
Cyclohexane, n-hexanol, ammonium hydroxide (35%), dichloromethane (DCM), diethyl ether, dimethyl-sulfoxide (DMSO), ethanol, methanol, glacial acetic acid and HPLC-grade acetonitrile were purchased from Fishers scientific, UK. Trihydrochloride Hoechst 33342 (Thermo Fisher Scientific, Eugene, OR, USA).
Cemex® (Tecres® SpA, Verona, Italy) bone cement was stored and used as from manufacturer’s guidelines.
Colorimetric ALP assay kit, picro-sirius red stain solution (ab150681), rabbit polyclonal antibodies against collagen I (ab21286), rabbit polyclonal antibodies against osteopontin (ab8448), rabbit polyclonal antibodies against osteocalcin (ab198228) and Alexa Fluor 488-labelled goat anti-rabbit IgG (ab150077) were purchased from Abcam, Cambridge, UK.
All reagents were stored according to manufacturer’s guidelines and used as received.
Cyclohexane, n-hexanol, ammonium hydroxide (35%), dichloromethane (DCM), diethyl ether, dimethyl-sulfoxide (DMSO), ethanol, methanol, glacial acetic acid and HPLC-grade acetonitrile were purchased from Fishers scientific, UK. Trihydrochloride Hoechst 33342 (Thermo Fisher Scientific, Eugene, OR, USA).
Cemex® (Tecres® SpA, Verona, Italy) bone cement was stored and used as from manufacturer’s guidelines.
Colorimetric ALP assay kit, picro-sirius red stain solution (ab150681), rabbit polyclonal antibodies against collagen I (ab21286), rabbit polyclonal antibodies against osteopontin (ab8448), rabbit polyclonal antibodies against osteocalcin (ab198228) and Alexa Fluor 488-labelled goat anti-rabbit IgG (ab150077) were purchased from Abcam, Cambridge, UK.
All reagents were stored according to manufacturer’s guidelines and used as received.
3
Powders for Gingival Fibroblast Stimulation
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Two different commercially available powders approved for subgingival use in air-polishing devices were used to stimulate HGF. A glycine-based powder (Perio-Mate Powder, NSK Europe, Eschborn, Germany) and an erythritol-based powder with the addition of 0.3% CHX (Air-Flow Plus, EMS Electro Medical Systems, Nyon, Switzerland) were investigated. Both powders were each dissolved in culture medium at a concentration of 1 g/20 ml. This concentration is based on previous research (Sygkounas et al., 2018) . In order to be able to investigate the individual components of the powder based on erythritol/CHX, a solution of 30 mg/20 ml was prepared with chlorhexidine diacetate (Sigma Aldrich, Munich, Germany) according to the proportion. Correspondingly, a solution with pure erythritol (Sigma Aldrich)
was also prepared at a ratio of 970 mg/20 ml. Unstimulated cells served as control.
was also prepared at a ratio of 970 mg/20 ml. Unstimulated cells served as control.
4
Biofunctionalized Silica Nanoparticle Synthesis
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Triton X-100, Tetraethyl orthosilicate (TEOS), (3-Aminopropyl) triethoxysilane (APTS), sodium alginate, chlorhexidine diacetate, sodium acetate trihydrate, piperazine, gentamicin sulphate, phosphate buffer solution (PBS) tablets, o-phthaldialdehyde reagent were purchased from Sigma-Aldrich, UK.
Cyclohexane, 1-hexanol, ammonium hydroxide 35 %, acetonitrile, ethanol, methanol, glacial acetic acid and 1-propanol were purchased from Fishers, UK. All reagents were stored according to manufacturer's guidelines and used as received.
Cyclohexane, 1-hexanol, ammonium hydroxide 35 %, acetonitrile, ethanol, methanol, glacial acetic acid and 1-propanol were purchased from Fishers, UK. All reagents were stored according to manufacturer's guidelines and used as received.
5
Synthesis and Characterization of Titanium Dioxide Nanoparticles
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Titanium (IV) oxide (anatase, < 25nm, 99.7%), (3-Aminopropyl) triethoxysilane (APTS, 99%), chlorhexidine diacetate, 1,6 hexanediol diacrylate, piperazine, sodium alginate, phosphate buffer solution (PBS) tablets, sodium acetate trihydrate (≥99%), brain heart infusion (BHI) agar and broth, Mueller-Hinton (MB) agar and broth, MTT (3-4, 5-dimethylthiazol 2,5-diphenyltetrazolium bromide and In Vitro Toxicology Assay Kit LDH were purchased from Sigma-Aldrich, UK. Dichloromethane (DCM), diethyl ether, dimethyl-sulfoxide (DMSO), ethanol, xylene, HPLC grade acetonitrile, glacial acetic acid and toluene were purchased from Fisher, UK. RPMI-1640 medium, minimum Essential Medium Eagle (MEM), heat-inactivated foetal bovine serum (FBS) and 1% penicillin-streptomycin (PS) were purchased from Gibco by Life Technologies, US.
All chemicals were reagent grade, stored according to manufacturer's guidelines and used as received.
All chemicals were reagent grade, stored according to manufacturer's guidelines and used as received.
6
Fabrication and Characterization of Chlorhexidine Spheres
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Chlorhexidine spheres were fabricated and characterized as described previously [25] . Briefly, 15mg/ml chlorhexidine diacetate (Sigma-Aldrich, Lot: 19H0417) solution and 0.33 M CaCl2 (Sigma-Aldrich, Lot: SLBF7416 V) solution were mixed at 1:1 ratio by volume at room temperature. The mixture was shaken for 1 min, and then centrifuged at 2000 rpm for 1 min (Eppendorf centrifuge 5417C, Germany). The sedimentation was then washed three times with 0.33 M CaCl2 solution and then centrifuged again. Thereafter, the product was freeze dried (ScanVac Cool Safe Freeze Drying, Denmark) at -107 ℃, 0.009 mBar for 1 day, and was used for all the following experiments. Several reproducible batches (3) were produced for this work.
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