Bacterial contamination was performed as previously described by Flörke et al. [24 (link)]. Briefly, immediately after the CAP treatment, cultivation with 10 mL of sterile nutrient solution (BHI, Brain–Heart-Infusion Broth, Carl Roth GmbH + Co. KG, Karlsruhe, Germany) and 100 μL of bacterial culture with E. faecalis (ATCC 29,212) was conducted at 37 °C for 24 h (Heraeus B6060, Heraeus Holding GmbH, Hanau, Germany). Afterward, the boxes (Eppendorf pipette tip reusable boxes Eppendorf AG, Hamburg, Germany) containing the models were sterilized at 121 °C (Autoclave Melag Vacuklav 24, MELAG Medizintechnik oHG, Berlin, Germany). On the first day, the membranes were infected with 200 mL of sterile BHI and 100 μL of the overnight culture and then incubated at 37 °C (Scientific C24 Incubator Shaker, New Brunswick Scientific, Edison, NJ, USA). After 4 h, the optical density was controlled via a spectrophotometer (BioPhotometer 6131, Eppendorf AG, Hamburg, Germany) at 600 nm (OD600), which was set to 0.8. The nutrient solution was exchanged every 24 h with 200 mL of sterile BHI for 6 days.
Scientific c24 incubator shaker
Manufactured by Eppendorf
Sourced in United States
The Scientific C24 Incubator Shaker is a laboratory equipment designed to provide controlled temperature and agitation for various applications. It features a shaking platform that can accommodate a range of sample containers, and the temperature can be adjusted and maintained within a specified range.
Automatically generated - may contain errors
4 protocols using scientific c24 incubator shaker
1
Bacterial Contamination of Dental Implant Membranes
2
Bacterial Biofilm Formation on Implants
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A cultivation with 10 ml sterile nutrient solution (BHI, Brain–Heart-Infusion Broth, Carl Roth GmbH + Co. KG, Karlsruhe, Germany) and 100 μl bacterial culture with E. faecalis (ATCC 29,212) was conducted at 37 °C for 24 h. (Heraeus B6060, Heraeus Holding GmbH, Hanau, Germany). Afterwards, the boxes (Eppendorf pipette tip reusable boxes Eppendorf AG, Hamburg, Germany) containing the models were sterilized at 121 °C (Autoclave Melag Vacuklav 24, MELAG Medizintechnik oHG, Berlin, Germany). The models with implants were placed in the boxes with the implants facing towards the bottom of the box, to enable continuous wetting with bacterial suspension. On the first day, the implants were infected with 200 ml of sterile BHI and 100 μl of the overnight culture and then incubated at 37 °C (Scientific C24 Incubator Shaker, New Brunswick Scientific, Edison, New Jersey, USA). After 4 h, the optical density was controlled via spectrophotometer (BioPhotometer 6131, Eppendorf AG, Hamburg, Germany) at 600 nm (OD600), which was set to 0.8. Nutrient solution was exchanged in every 24 h with 200 ml of sterile BHI for 6 days.
3
Titanium Dental Implant Contamination
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A total of 36 titanium dental implants (3.5 mm in diameter, 9.5 mm in length) with a Ti Grade V surface (Friadent®/Ankylos®, Dentsply Implants, Mannheim, Germany) was inserted into frontal bone (n: 12) and the basis of the corpus mandible (n: 24). Eighteen implants were contaminated via inoculation of Enterococcus faecalis for 7 days. On the first day, the implants were infected with 200 mL of sterile Brain Heart Infusion (BHI) and 100 μL of the overnight culture and then incubated at 37 °C (Scientific C24 Incubator Shaker, New Brunswick Scientific, Edison, NJ, USA). After 4 h, the optical density was controlled via spectrophotometer (BioPhotometer 6131, Eppendorf AG, Hamburg, Germany) at 600 nm (OD600), which was set to 0.8. Nutrient solution was exchanged every 24 h with 200 mL of sterile BHI for 6 days. At the end of the seventh day of incubation, the implant surface was controlled by using chromogenic agar (chromID® VRE, bioMérieux, Nurtingen, Germany) to detect the colored colonies.
4
Quantifying Bacterial Colonization of Biomaterials
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To quantify bacterial colonization of algae-PLA composites and porcine and bovine collagen membranes, colony-forming units (CFU) were used to estimate the number of viable bacteria. A cultivation with 10 ml sterile nutrient solution (BHI, Brain–Heart-Infusion Broth, Carl Roth GmbH + Co. KG, Karlsruhe, Germany) and 100 μl bacterial culture with E. faecalis (ATCC 29,212) was conducted at 37 °C for 24 h. (Heraeus B6060, Heraeus Holding GmbH, Hanau, Germany). Afterwards, the boxes (Eppendorf pipette tip reusable boxes Eppendorf AG, Hamburg, Germany) containing the models were sterilized at 121 °C (Autoclave Melag Vacuklav 24, MELAG Medizintechnik oHG, Berlin, Germany). On the first day, the membranes were infected with 200 ml of sterile BHI and 100 μl of the overnight culture and then incubated at 37 °C (Scientific C24 Incubator Shaker, New Brunswick Scientific, Edison, New Jersey, USA). After 4 h, the optical density was controlled via spectrophotometer (BioPhotometer 6131, Eppendorf AG, Hamburg, Germany) at 600 nm (OD600), which was set to 0.8. Nutrient solution was exchanged in every 24 h with 200 ml of sterile BHI for 6 days. All experiments were conducted in triplicate.
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