It is known that anaerobic microbes like Porphyromonas gingivalis (P. gingivalis) are involved in the progression and initiation of multiple forms of periodontal disease such as chronic periodontitis. However, in order to culture this gram-negative anaerobic microbes in the laboratory we need a long time (about 72 hours) with a complete absence of oxygen (≤0.001%) to reach optimal cell density. Therefore it is usually treated in an anaerobic chamber. Because this traditional method to culture P. gingivalis is costly and time consuming, we tried to elaborate in this study a new, rapid and less expensive method for culturing the anaerobes, such as P. gingivalis and P. endodontalis. With the use of a rotary shaker under anaerobic conditions, it would seal the bacterial growth media from oxygen exposure using a mineral oil overlay. Additional analysis by western blot helped us to study more about the cell’s biological functions. As a result, we found that the RAW264.7 cells treated by P. gingivalis or P. endodontalis showed three groups of kinase phosphorylation levels. As these kinases are key factors for the bacteria induced inflammation, we are interested to find how to reduce such bacteria (i.e. P. gingivalis and P. endodontalis) induced inflammation via regulation of these related kinase phosphorylation in future studies.
Based on our experiment, we believe that this method can be suitable for growing most species of anaerobic bacteria. Thus, we envision that this new method could be widely used in areas of both research and industry in the near future.
| Identifer | oai:union.ndltd.org:bu.edu/oai:open.bu.edu:2144/37069 |
| Date | 16 July 2019 |
| Creators | Alamry, Nujud |
| Contributors | Tang, Xiaoren, Dibart, Serge |
| Source Sets | Boston University |
| Language | en_US |
| Detected Language | English |
| Type | Thesis/Dissertation |
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