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lab meeting presentation
167
Sandeep Kumar
oral biology
university of Leeds
Supervisors:
Dr E L Mostafa
Dr Jinous Tahmassebi
Dr Farideh Javid
Identification of the abilities of Cannabinoids to address deterrent inflammation during periodontitis and their role during periodontal tissue regeneration.
Title
Sandeep Kumar
Supervisors:
Dr EL Mostafa Raif
Dr Jinous Tahmassebi
Dr Farideh Javid
PhD Second Year
introduction
Periodontium and Periodontitis
Gradual inflammation of periodontal tissues i.e. periodontitis leads to loss of bone, eventually loss of function
commonly found disease, incidence depends on age, sex and socioeconomic conditions.Indirect costs due to oral conditions
worldwide amounted to US$144 billion yearly.
Journal of Dental Research 2017, Vol. 96(4) 380–387
Implications of periodontal diseases
J Can Dent Assoc 2000; 66:594-7
ETIOLOGY:PERIODONTITIS
environmental perturbations in the periodontal tissues
1. trauma
2. idiopathic growth of a keystone pathogen
3. host is genetically susceptible
pathogens elicit inflammation
changes the nutrient foundation of the ecological niche (i.e. periodontal pocket).
The altered nutrient foundation promotes the proportional expansion of pathobionts relative to symbionts, promoting inflammation that ultimately leads to connective tissue and bone destruction.
- oral hygiene maintenance
- local drug delivery -- antibiotics
- systemic antibiotics
- surgical approaches
- anti-inflammatory modulation
periodontitis
NEWER APPROACHES TO ADDRESS PERIODONTITIS
Review
Fabisiak A et al. 2017 Cannabinoids as gastrointestinal anti-inflammatory drugs.
canabinoids:
1. acts as anti-inflammatory agents
2. acts in preventing bone resorption
3. got receptors in human body
4. acts on endocannabinoid system
Ribeiro A et al. 2012 Cannabidiol(CBD), a non-psychotropic plant-derived cannabinoid, decreases inflammation in a murine model of acute lung injury
Li et al. 2013 Anti-Inflammatory Role of Cannabidiol (CBD) and O-1602 in Cerulein-Induced Acute Pancreatitis in Mice
Periodontitis :
1. Is a inflammatory disease
2. causes bone resorption
3. canabinoid receptors are found in periodontal ligament
Li Dehao et al. 2017 Cannabidiol(CBD) administration reduces sublesional cancellous bone loss in rats with severe spinal cord injury
Recently it is also found that the endocannabinoid system does not just respond to the endocannabinoids produced in the body, but also respond to external cannabinoids like the phytocannabinoid cannabidiol, or CBD.
objectives
AIM
To evaluate the ability of cannabinoids to address the chronic periodontal diseases.
1. To investigate the anti-inflammatory effects of cannabinoids.
2. To assess the effect of cannabinoids on osteogenesis.
3. To assess the effect of canabinoids on osteoclastogenesis.
CANNABINOIDS IN PERIODONTITIS
Marcelo H. Napimoga et al. 2014- used CBD and showed anti-inflammatory effects -------- experimental periodontitis in rats
Abidi A A et al. 2018 targeted the endocannabinoid system, showed it could offer a new therapeutic approach for treating chronic inflammation in periodontal disease.
Niyogi et al 2019 showed canabinoids have antibacterial property against periodontal pathogens
Interleukin-8 (IL-8) plays vital role in pathogenesis of periodontal diseases.
IN-VITRO model
LPS/Flagellin ------ TIG-K cells/ periodontal cells --- inflammatory response, at m-RNA level and protein level
inflammatory pathway involved
confirmation of pathway using knockdown and gene silencing
Key Results
il-8 transcription at 3 hours
Results
cytotoxicity (LPS, Flagellin and Canabinoids )
il-8 translation at 24 hours
il-8 translation at 3 hours
TIG- K (GINGIVAL KERATINOCYTES): OBSERSEVED EXPRESSION
Results replicated using Periodontal cells from various donors and also with CBG( canabegerol)
Analysis
AEA (Anandamide)/2AG (2-Arachidonoylglycerol)
CBD (Canabidiol)/ CBG ( Canabegerol)
cell viability
Subtopic 1
CBD
TIG-K viability with test drug
CBG,
AM-251- slective antagonist CB1
O-1602 - Non- Selective Antagonist
AM-630- Selective antagonist CB2
Transcription analysis
Subtopic 2
1 uM
Translation
AT 3 HOURS
AT 24 HOURS
Subtopic 3
1 uM
Conclusion
Pre-treatment with CBD markedly attenuates the expression of IL-8 stimulated TIG-K/periodontal cells with obnoxious stimulus flagellin.
This non-psychotropic cannabis product (CBD) may therefore play a vital role in addressing chronic periodontal problems.
What’s next
To identify other inflammatory cytokines like IL-6, IL-1b and TNF-α at mRNA and protein levels
Genes regulating the mineralization i.e. RANK/RANKL and OPG will be studied. Identify their role in bone mineralization and demineralisation during Periodontitis.