Week 1 45 minutes

1. Week 1 45 minutes

2. Your mission…..disclosing the facts The Return of the Red Complex Group • 1. Putative pathogens group indicted for chronic attack against of Perry O’Dontal. Al V. O’larbone retreats. • Despite eviction and removal from subgingival spaces 3 months ago, The Red Complex Group has resurfaced and reestablished an organized community stronghold. Reports indicate similar re infiltration of organisms in up to 30% of adult population. How could this happen? Identify periodontal pathogens as eukarocytic and prokaryotic. Sequence and Explain the stages of biofilm formation. • 2. Endotoxin alert!!! Bacteria are talking to each other. How can this be? How do scientists know this? Explain the phenomenon of cellular communication. • 3. Endotoxin alert!!! Host Immune System has deployed rapid response agents: neutrophils and macrophages move straight in to surround and engulf, destroy gram negative pathogens. Who are the primary responders deployed by the body to fight infection? Identify these rapid responders as eukarocytic or prokaryotic. What is the mode of operation? Why will signs of redness and swelling be seen in the surrounding gums? • 4. Prevention Specialist Reports Mechanical Techniques to Block Biofilm Maturation Interrupting the cascade of events triggered by dental biofilms is an important prevention protocol. Which stage of biofilm development can be interrupted to block initiation of infection and inflammation? Report your rationalewhile you explain what is happening in this stage. What strategies and monitoring mechanisms do you recommend?

3. Healthy Gingiva ... Gingivitis Gingivitis Healthy Gums

4. Consider the etiology of … Periodontitis (Gum Disease involving the supporting structures of the teeth) • Write an answer for the following questions: • Did you know Periodontitis is an infection? • What is the primary cause? • What causes the signs and symptoms: redness, swelling, bleeding, halitosis? • Discuss your answers with a partner. Share with the class.

5. Did you know about 50% of the adult population has some form of Gum Disease?

6. Figure Biofilm lodges in the crevices around the teeth both aboveand below the gingival margin. Accumulation of dental plaque biofilmcan result in tooth decay and periodontal disease. (Figure 2006 Keith Kasnot, MA, CMI, FAMI.)

7. Think about it……What is Biofilm? • Can I see biofilm? • What’s in it? • How does it form? • How long does it take to form? • Does it wash away? • What happens if it’s left undisturbed • Is there a host immune response system? • Why should I care about biofilm? Let’s investigate.

8. Can we see Biofilm? Disclosing the plaque/ biofilm oral invaders!

9. Prior knowledge • Cell biology • Are bacteria prokaryotic or eukaryotic? • http://www.biologycorner.com/worksheets/prokaryote_color_key.html • Review the structure of a cell and note the importance of the cell membrane. What is the importance of the pilus? The flagella? • Cell Models: An Interactive Animation comparative size of cell organisms: http://cellsalive.com/howbig.htm

10. Look at the shapes below. How would you describe what you see?

11. “follow” Flossie Newlit 2012Instructor sends tweets • Instruct class to Tweet “follow” Flossie Newlit2012 • hastag My Smile Plan • Instructor will tweet the following messages at designated times (TBA): • Putative pathogens group indicted for chronic attack against of Perry O’Dontal. Al V. O’larbone retreats. • Biofilm found lodging in the crevices around the teeth both above and below the gingival margin Microbiologist seeks to unravel the building of complex communities of bacteria. • Microbiologist investigates biofilm. Starts asking questions from Slide 8. • Day 1-2Planktonic bacteria settle in • Days Rods, vibrios and spirochettes move in • Haltosis present during signs of biofilm invasions

12. 10-15 minute activity • Have students work in groups of 4. Give each group one of the 4 diagrams of biofilm (slides 13-18). Ask students to describe what they see in their diagram. Ask them to record their descriptions of the figures. Write down words that are unfamiliar to them. Ask them to make inferences about what the graph is describing. Have the groups trade graphs with another group and repeat the process. Ask for 2 groups volunteers to share their observations. • Collect student work and return at start of Week 2 session • Assign readings for next class meeting.

13. Chart 2 for in class exercise

14. Chart 1 for in class exercise

15. Chart 3 for in class exercise

16. Chart 4 for in class exercise

17. Chart5for in class exercise

18. Assignment to complete for Week 2Read the following articles before the next class. Turn in an outline for your readings be sure to answer the questions at the bottom of this slide. Biofilm Basics http://www.dimensionsofdentalhygiene.com/ddhright.aspx?id=59#.UMVvLHdrmSo Are articles from a reliable source? What has qualified these articles as reliable? Where are biofilms found? Do biofilms affect periodontal health? Do cells communicate with each other? How do we know? Which cells are more susceptible to antibiotics Planktonic (free floating) or cells within a biofilm Once removed will a biofilm return? Make a vocabulary list. Record the definitions.

19. Planktonic Bacteria(free floating) What Are Bacterial Biofilms? https://www.youtube.com/watch?v=lpI4WCM_9pM

20. Week 2Unpacking the complex biofilm formation process. • Students turn in a copy of their homework. • Students work in same groups of 4 as week 1 to review the power point slides and video links. Return their interpretations of the flow charts. • At end of session ask students to reflect on their learning.

21. BIOFILM DEVELOPMENT 1 hour after cleaning Stage 1Initial Adherence “The Acquired Pellicle” SAP (Salivary Acquired Pellicle) Mucinsand Glycoproteins in saliva: alpha amylase proline rich protein, statherin And Bacterial cell fragments. Groups share their observations Discussion Question: What is surprising to you.Name3 interesting facts about mucins. What does gyco refer to in gylcoprotein?

22. BIOFILM DEVELOPMENT Stage 2Lag Phase

23. BIOFILM DEVELOPMENT Days 1-2 Stage 2 Lag Phase Planktonic Bacterial cells settle on a surface; arrange themselves in clusters and attach (sessile colony Days 2-4 filamentous bacterial forms start to attach to sessile colonies formation) Groups share their observations Discussion Question: What changed? What is surprising to you

24. Watch the following videos How long does it take bacteria to grow? Bacterial Growth and multiplication: http://cellsalive.com/ecoli.htm Bacteria Cam minute by minute update of streptococcus cell growth http://www.cellsalive.com/cam2.htm

25. BIOFILM DEVELOPMENT Days 4-14 STAGE 3 Rapid Growth bacterial colonization Co-aggregation of bacteria Days 4-7 filamentous bacterial forms; Days 7-14 vibrios and spirochettes Water insoluble SLIME MATRIX forms from bacterial secretion of polysaccharides. This is the “ fuzzy” feeling on your teeth”

26. BIOFILM FORMATION Days 14-24STAGE 4 STEADY STATE & DETACHMENTBACTERIA COMMUNICATING WITH EACH OTHER

27. Days 14-24 ORANGE AND RED COMPLEX BACTERIA MOVE IN What kind of bacteria are these? vibriosand spirochettes

29. As a group, the class will watch the following video about internal biofilms. Start the video at 4:07 minutes. utube video of internal biofilm start video at 4:07 minutes https://www.youtube.com/watch?v=lpI4WCM_9pM

30. Build a biofilm 15-20 minutes.Create your own model for presentation • “Mystery Bag” You may use the contents to construct a biofilm. Each stage of formation must be included. Record the steps. Take pictures and write or make an audio recording of the stages.( contents of mystery bag: double sided tape, straws, twist ties, tube of blistex, saran wrap and colored legoblocks) • Correct sequencing and description of each stage must be included in your final presentation. The final stage must include red complex bacteria. • Presentation Modes: • Video • Photo Peach: upload pictures, text and add music • Power point • Students can complete these for homework.

31. Week 3

32. Week 3 Do bacteria talk to each other? • Ask volunteers to share their homework assignment---the summary of their biofilm models. • Continue to work in same groups

33. Electron Microscope picture of the red complex gram negative bacteria, phorphyromonagingivalis. Notice the fimbriae and surface blebs that contain endotoxin. Endotoxins solicit the host immune response n

34. Create a gram negative pathogen • Given an oval shaped foam section, blistex, pieces of twist ties, create a gram negative pathogen. Using slide 33, place the fimbria and surface blebs to create your pathogen. • Take a picture of it and record an explanation where the endotoxins are contained and their purpose.

35. Watch the following video(s) about quorum sensing “how cell communicate” Bacteria in biofilm communicate with each other by a process called quorum sensing. This dynamic, sophisticated communication system enables bacteria to monitor each other’s presence and to modulatetheir gene expression in response to the number of bacteria in a given area of the biofilm. Bonnie Brassler in “TED TALKS” explains this process. Talking bacteria: ( click on the “view” tab) http://www.teachersdomain.org/asset/oer08_vid_bassler/ A more detailed version can be seen at http://www.ted.com/talks/lang/en/bonnie_bassler_on_how_bacteria_communicate.html Discussion Questions • How did Bonnie Brassels confirm that cells are communicating with each other? • How do bacteria within the biofilm signal each other?

36. http://www.nlm.nih.gov/medlineplus/ency/article/000821.htm

37. Mechanisms of action Antaomy of a Splinter describes the rapid response of the body to control damage by trauma or bacteria. Diapedesis, chemotaxis Phagocytosis and Oxidative Burst . Each process is illustrated by clicking on the highlighted action verb. http://www.cellsalive.com/ouch1.htm

38. Neutrophils, Macrophages and Lymphocytes Mode of Operation Visit this page. Read the passage. Watch both videos http://www.bbc.co.uk/schools/gcsebitesize/science/aqa_pre_2011/human/defendingagainstinfectionrev4.shtml

39. Cells of the Immune System • Use the remainder of the session to design a 3 minute presentation about today’s lesson. This must be submitted at the next class meeting. Visit the web sites on the following slide to deepen your understanding of the body’s immune system. • Using photo peach or video thread, identify one of the White Blood Cell Rapid responders and explain their “MO” mode of operation. Be sure to identify your Rapid Responder as either eukaryotic and prokaryotic cells • You can create an avatar to represent the Leukocyte. • Generate 1-2 essential questions for your class mates to answer about your Rapid Responder.

40. The white blood cells and host connective tissue cells within • the developing inflammatory lesion are stimulated to synthesize • and release the following: • proinflammatory cytokines these function as cell phones for the immune response • prostanoids • proteolytic enzymes, eg, • interleukin-1 (IL-1), • interleukin-6 (IL-6), • interleukin-8 (IL-8), • tumor necrosisfactor-alpha (TNF-α), • prostaglandinE2 (PGE2), • matrix metalloproteinases. http://science.howstuffworks.com/life/human-biology/blood2.htm How Blood Works by Carl Bianco, MD http://www.wisc-online.com/objects/ViewObject.aspx?ID=ap14704 https://www.youtube.com/watch?v=0TvTyj5FAaQ

41. Week 4After today’s presentations you will complete the final assignment by responding to prompts 1,2, or 4 for the case investigation, “The Return of the Red Complex Group” for Perry O’Dontal and Al V. Olarbone

42. Immune System Rapid Response team moves in to battle Pathogenic, anaerobic invaders attacking subgingival structures!

43. Student Presentations of Rapid Responder Assignment • Have student groups present to each other for peer review of this assignment. • Choose 4 groups to present to the entire class.

44. Periodontal Infection and Inflammation Bacteria and their metabolicbyproducts fuel a cellular immune response within the affected gums. A movement into the area of bacterial infection by a closely crowded together team of white blood cells: neutrophils macrophages lymphoid cells. Blood vessels dilate extra white blood cells and their actions and byproducts signs and symptoms of inflammation

45. Repeated Return of Red Complex Group in Periodontal Pockets Advances the loss of Alveolar Bone It is the host inflammatory-immune response that leads to the clinical signs of gingivitis and chronic periodontitis, resorptionof tooth supporting alveolar bone, and periodontal pocket formation.

46. Summary: • Periodontal Infections are preventable. • Mature biofilms harbor bacteria that cause infections. Biofilms begin to form on teeth with in 1 hour after cleaning and bacteria colonize, organize, cooaggregate and form a protective coating”slime” layer which makes management difficult. • The bacteria talk to each other through a process known as quorum sensing. • The host immune system recognizes foreign body and dispatches its WBC responders. • The action of the host immune system in its fight against infection leads to inflammatory and retreat of supporting structures. • An ounce of prevention is worth a pound of gold!

47. What will you do to maintain the health of your teeth and gums? Healthy Gingiva ... Gingivitis Gingivitis Healthy Gums

48. Your mission…..disclosing the facts Choose 1 of the 3 prompts below. Prepare a 3-5 minute presentation for the class. The Return of the Red Complex Group 1. Putative pathogens group indicted for chronic attack against of Perry O’Dontal. Al V. O’larbone retreats. Despite eviction and removal from subgingival spaces 3 months ago, The Red Complex Group has resurfaced and reestablished an organized community stronghold. Reports indicate infiltration of similar groups in up to 50% of adults. How could this happen? Explain the stages of biofilm formation. 2. Endotoxin alert!!! Bacteria are talking to each other. Explain this phenomenon. 3. Endotoxin alert!!! Host Immune System has deployed rapid response agents: neutrophils and macrophages move straight in to surround and engulf gram negative pathogens. What will happen? Why will signs of redness and swelling be seen in the surrounding gums? 4. Prevention Specialist Reports Mechanical Techniques to Block Biofilm Maturation Interrupting the cascade of events triggered by dental biofilms is an important prevention protocol. Which stage of biofilm development can be interrupted to block initiation of infection and inflammation? Report your rationale. What strategies and monitoring mechanisms do you recommend?

49. Consider the etiology of … Periodontitis (Gum Disease involving the supporting structures of the teeth) • Write an answer for the following questions: • Did you know Periodontitis is an infection? • What is the primary cause? • Can you Prevent the Risk of Periodontitis? How? • Prepare an informational pamphlet that gives evidence based answers and rationales for the questions: • Are mouthrinses effective in removing the cause of periodontal infections ? • Are antibiotics effective in in removing the cause of periodontal infections ?

50. Disclose the facts • Small group recording and reporting of group’s observation, discussion, interpretation of at least 2 diagrammatic summary charts. Summarize your theories and be prepared to share with the class. Before the end of the first session write a reflection of class discussion and turn in your group summary and your personal reflection. • ****Assignment: Review the questions on slide 8. Complete the assigned readings and outline the key points in the articles. Answer the questions on Slide 18. Turn in copies of your outlines and answers to questions by next week’s class session. • Flossie Newlit12 asks class to follow tweets. • Next week’s class meeting: • In Class work in groups to unpack the complexity of biofilms. Work through each slide and visit the corresponding websites. • Mystery Bag (contains double sided tape, yellow dots, legos, straws, pipe cleansers, twisties, saran wrap}, Build a biofilm: Use your cell phone of a camera to Take of picture of each stage of the biofilm construction. This gives us an understanding of the sequence of biofilm formation. Record an explanation at each stage that clearly and accurately examinseand convesycomplex ideas, concepts, and information abut the biofilm. Use idpadioor a tape recorder. • Write a reflection about the class activities and your ability to understand the assigned reading for this week. Turn it in. ***Assignment: Revise your outline. Clarify important points that were unclear to you. • Week 3 Do cells talk to each other ? Begin with slide 25. • Week 4 Immune Response: Who are the members of the rapid response team. What is the MO (mode of operation)? Choose a member of the white blood cell rapid response team Use Photo peach and to personify or create a description for one of the following. Neutrophils: Macrophage: • Summative Assessment: Choose 1 of 4 prompts. Prepare 3-5 minute class presentation.. You can use media technogies such as , I movie, voice thread, • Voice Thread or Photo Peach: upload pictures, text and music to explain the biofilm process and the return of The Red Complex Group End with recommendations to prevent organization of these bacteria. • Construct a sophisticated model and Create a video report • Create a pamphlet • Create a facebook page for Biofilm: Friend protective factors and have them write on your wall. Deny friend request from periodontal pathogens. • Write a scientific article