The preserved clinical isolated strain of P. gingivalis was grown at 37 °C for 2 days on Columbia agar plates supplemented with %5 Human blood, 5 μg/ml of Hemin (Sigma Aldrich, Shanghai, China), and 1 μg/ml vitamin K1 (Himedia, Mumbai, India) under anaerobic conditions by using anearopack system (Mitsubishi Gas Chemical, Katsushika, Japan). The largest colonies were then selected and anaerobically sub-cultured at 37 °C for 48 h in nutrient broth (Himedia, Mumbai, India) supplemented with Hemin (Sigma Aldrich) and vitamin K1 (Himedia). After applying PBS, spectrophotometry with 624 nm wavelength was used to make a bacterial concentration of up to 1 × 106 CFU; then, the bacteria were mixed with 4% sodium carboxymethyl cellulose (Panreac, Barcelona, Spain). Finally, 0.03 ml of live P. gingivalis with a concentration of 1 × 106 CFU/ml were locally applied to the rats in the gingival sulcus bottom of the left and right mesial and lingual sides of their incisor teeth. The same procedure was repeated every three days for two weeks.
Vitamin k1
Manufactured by HiMedia
Sourced in China, India
Vitamin K1, also known as phylloquinone, is a fat-soluble vitamin that plays a crucial role in the blood coagulation process. It is an essential nutrient required for the proper functioning of certain proteins involved in blood clotting.
Automatically generated - may contain errors
Lab products found in correlation
Bacitracin, by HiMedia (2 mentions)
Colistin methanesulfonate, by HiMedia (2 mentions)
Hemin, by Merck Group (2 mentions)
Nalidixic acid, by HiMedia (2 mentions)
Columbia agar base, by Merck Group (1 mentions)
Bile salt, by HiMedia (1 mentions)
Hemin, by HiMedia (1 mentions)
Salivary α amylase, by Merck Group (1 mentions)
Pancreatin, by Merck Group (1 mentions)
Acetic acid, by RCI Labscan (1 mentions)
Nutrient broth, by HiMedia (1 mentions)
Anearopack system, by Mitsubishi (1 mentions)
4 protocols using vitamin k1
1
Induction of Periodontal Disease in Rats
2
Anaerobic Media Composition for Microbial Cultures
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The composition of media per 500 ml was as follows: Columbia agar base (21.25%) (code LAB001, UK), 5 µg/mL of hemin (1.5 mg) (Sigma-Aldrich, China), 1 µg/mL vitamin K1 (0.5 mg) (Himedia), 5% human blood (25 ml), colistin methanesulfonate (7.68 mg), bacitracin (5 mg) (Himedia), nalidixic acid (7.5 mg) (Himedia), and distilled water [24 (link)].
Media were prepared and stored under strictly anaerobic conditions to prevent oxidation.
Media were prepared and stored under strictly anaerobic conditions to prevent oxidation.
3
Simulated Biofluids Analysis Protocol
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All chemicals used for the analysis of simulated salivary fluids, simulated gastric fluids and simulated intestinal fluids were of analytical grade and were made available from RCI Labscan (Bangkok, Thailand). Salivary α-amylase and pancreatin were purchased from Sigma-Aldrich (St. Louis, MO, USA). Bile salts, hemin, vitamin K1, cysteine HCl and the appropriate media were purchased from HiMedia (Nashik, India). Standard short chain fatty acids (SCFA), including propionic acid, butyric acid, acetic acid and lactic acid, were of HPLC grade and were obtained from RCI Labscan.
4
Isolation of Chronic Periodontitis P. gingivalis
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This study involves clinical strain of P. gingivalis Gram-negative bacteria isolated from periodontal pocket of patients attended Shorsh dental center in Sulaymaniya city/Iraq in August 2017, subgingival plaque samples were collected from systematically healthy persons suffering from chronic periodontitis, from periodontal pockets with of at least six mm depth with sterilized paper point, then the samples were spread on Colombia agar media (LAB001 UK) supplemented with sheep blood %5, 5μg/ml hemin (sigma Aldrich-china), 1μg/ml vitamin K1 (Himedia), 7.68 mg colistin methanesulfonate (Himedia), 5 mg bacitracin (Himedia), and 7.5 mg nalidixic acid (Himedia) according to [18 ]. Plates were incubated for 7-10 days anaerobically using an anaerobic jar and anaerobic gas packs (AnaeroGen® system Oxoid), patients' consent and approvals were obtained prior to collecting the samples.
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