Comparison of the aerobic microbes in supragingival plaque  of chronic  and aggressive periodontitis patients

 

Gayathri. M¹, Dr. Gheena. S², Dr. Gopinath³

¹II BDS, Saveetha Dental College and Hospitals, Chennai.

²Reader, Department of Oral Pathology, Saveetha Dental College and Hospitals, Chennai.

³Senior lecturer, Department of Microbiology, Saveetha Dental College and Hospitals,Chennai.

 

ABSTRACT:

Aim: To compare the aerobic microbes in supragingival plaque of Chronic Periodontitis (GCP) and Genera Aggressive periodontitis (GAP) patients. Objective: Microbial aspect of GCP and GAP varies in terms of species, number and hence the effect on the tooth. It is important to differentiate between the types to introduce specific medication. Background: Patients with GAP manifest thin deposits  of dental  plaque with little  or no calculus, patients with GCP usually have  very complex and thick deposits  of polymicrobial  communities  on affected  root  surface. Potential  periodontal  pathogens  are usually a part  in the  biofilm, their biochemical  and physiological  properties  are  modified by their interactions  with other members  of the  biofilm  community. Based on data from previous studies, GAP and GCP have a different microbial manifestation. Reason: From  a  research perspective, it  is  essential  that  these  diseases  be  clearly distinguished in order to gain a complete  understanding of their etiology and pathogenesis.

 

 

 

Periodontitis is a progressive inflammatory disease caused by biofilm containing facultative oral micro biota on tooth surface. It is the second most common disease after dental caries affecting about 30% of human population(1). It is characterised by bleeding of gingiva, spacing of teeth, suppuration, loss of alveolar bone and periodontal attachment and leads to the loss of the tooth(1). It is a multifactorial disease caused by polymorphic microbial flora. The susceptibility of host is affected by oral hygiene, genetic predisposition, modifiers of host response (smoking),  systemic disease ,absence of beneficial bacteria and also due to geographical location.

 

The oral biofilm is complex containing about 700 different species of bacteria, most of which are opportunistic in nature. Under favourable conditions, they invade the periodontium causing aggressive immuno- inflammatory response. This leads to a rapid periodontal tissue destruction. The disease is classified into two types chronic and aggressive which may be either local or generalised. The difference between the types is based on the rate of progression (GAP is rapid while GCP is slower), pattern of destruction (GAP is usually generalised and GCP is usually localised), clinical signs of inflammation (lesser in case of localised periodontitis and abundance of plaque or calculus (thicker and more complicated biofilm in chronic periodontitis)(2). Chronic periodontitis is characterised by cycles of progression and stability. In GAP, the lesion has plasma cell dominated inflammatory infiltrate in the connective tissue with neutrophil migration through the pocket lining epithelium and forming a layer between the tissue and the plaque biofilm. GAP and GCP are different in the microbial perspective too.(3) The chronic periodontitis contains about 75% of gram negative and anaerobic species of bacteria like Porphyromonas gingivalis, P intermedia, Tannerella forsythia, Fusobacterium spp, Aggregati bacter actinomy cetemcomitans, Spirochetes while the number of gram negative bacteria in GAP is lesser. The fungal  involvement like Candida albicans enhance the adherence of the microbes to the periodontal ligament.(3)

 

MATERIALS AND METHODS:

All subjects were verbally informed about the investigation, and they were asked to sign informed consent.

 

Clinical parameters observed included: Gingival Index (GI - Loe and Silness 1963) , Plaque Index (PI - Silness and Loe 1964) , Pocket depth (PD) and Clinical Attachment loss (CAL). The  diagnosis  for the  periodontal  status  was  established for all  subjects  based on the  1999 International Classification of periodontal  diseases  and conditions . The  used clinical  criteria  were  as  follows. For aggressive  periodontitis:  1) rapid attachment  loss  and bone  breakdown were  evident  including at  least  one  incisor and one  first  molar, 2) pocket  depth  ≥4 mm, clinical  attachment  loss  ≥3 mm, presence  of bleeding on probing. For chronic  periodontitis:  extensive  deposits  of plaque  and calculus, more  than 10% of teeth with pocket  depth ≥4 mm, and at  least  one  site  having attachment  loss  ≥3 mm.

 

Inclusion criteria: Patient of age group 40-70 who signed the  informed consent, were  entered into the  study. All  patients had at  least  20 teeth, diagnosed as  having AgP  or ChP.

 

Exclusion criteria: The  exclusion criteria  were patients  medically compromised, or having received periodontal  or antibiotic  treatment  within the  preceding 6 months, patients  under orthodontic  treatment, pregnant  women or during lactation , patients having systemic disease and patients  who need antibiotic  prophylaxis  before  screening .

 

Selection  of Site:

Sampling: removal  of  supragingival  plaque  by means  of sterile  cotton  balls and gp points. supragingival  plaque  were  collected  and  deposited  on tubes  containing  sterile  saline  .

 

Culture: the  sample shall  be  deposited in  a  tube  containing  2 ml  of  saline  (0.9%  NaCl).  The samples  were  mixed  using  a  vortex  for  one  minute  to allow  the  dispersal  of  bacteria.  The samples were then collected in uricol. Samples were  shaken  vigorously  for  30 seconds  using the mixer before being prepared  for analysis.

 

The following culture media used for each sample :

a) Nutrient Agar

b) Mitis salivarius agar

c) Lactobacillus agar.

 

The cultures were prepared by a qualified personnel and left to incubate at 37°C for over 24 hours. The cultures were examined and the bacterial species seen on each culture plate were identified.

 

Statistical analysis:

The percentage distribution of the bacterial species identified was done and the results of the two groups were compared. Graphical representation were done using Microsoft Excel.

 

RESULTS:

Generalised chronic periodontitis:

About eight different gram positive bacteria and a gram negative bacteria , pseudomonas were identified from the culture . In addition, candida species were also found in a few samples. A total of 40 strains were found of which 75% belonged to gram positive , 10% to gram negative, 15% to candida. The percentage distribution of each microbial species is illustrated in the following table. (Table 1)

 

Table 1: Percentage distribution of the microbial species in 10 samples of GCP

Bacterial/Fungal Species	No. of strains (out of 10 patients)	Percentage distribution among 10 patients %
I. Gram positive bacteria
Streptococcus mutans	6	60
Streptococcus mitis	3	30
Streptococcus salivarius	5	50
Streptococcus sanguis	5	50
Staphylococcus aureus	7	70
Lactobacillus	3	30
Enterococcus	5	50
Bacillus spp.	1	10
II. Gram negative bacteria
Pseudomonas	4	40
III. Candida spp.	6	60

 

Generalised aggressive periodontitis:

About six different gram positive bacteria and a gram negative bacteria, pseudomonas were identified from the culture. In addition, candida species were also found in a few samples. A total of 24 strains were found of which 75% belonged to gram positive , 12.5% to gram negative, 12.5% to candida. The percentage distribution of each microbial species is illustrated in the following table. (Table 2)

Table 2: Percentage distribution of the microbial species from 5 samples of GAgP.

Bacterial/Fungal Species	No. of strains isolated (from 5 samples)	Percentage distribution among 5 patients %
I. Gram positive bacteria
Streptococcus mutans	4	80
Streptococcus sanguis	1	20
Streptococcus salivarius	2	40
Staphylococcus aureus	5	100
Enterococcus	4	80
Bacillus spp.	2	40
II. Gram negative bacteria
Pseudomonas	3	60
III. Candida spp.	3	60

 

 

 

 

 

 

 

DISCUSSION:

On comparing GCP and GAP ,it was found that streptococcus mitis and lactobacillus were absent in the latter. The percentage distribution of the species were elevated in GAgP except for Streptococcus sanguis and Streptococcus salivarius. The percentage of Staphylococcus aureus was found to be the highest in the composition. It is important to note that Staphylococcus aureus was found in all samples of GAgP patients. A comparative  table based on the percentage distribution is as follows (table 3):

 

Table 3: comparison of percentage distribution of bacterial strains between GCP and GAP

Bacteria/fungal species	GCP (%)	GAP (%)
I. Gram positive bacteria
Streptococcus mutans	60	80
Streptococcus salivarius	50	40
Streptococcus sanguis	50	20
Staphylococcus aureus	70	100
Enterococcus	50	80
Bacillus spp.	10	40
II. Gram negative bacteria
Pseudomonas	40	60
III. Candida spp.	60	60

 

 

 

 

The table above shows that there is no significant difference in the microbial composition in the supragingival plaque samples of GCP and GAgP. This is in accordance with Faveri et al who also found that only minor differences are present in the bacterial composition.(4) Also in a recent study, Rescala et al reported that no significant microbiological  differences were found between GCP and GAgP for any of the species examined. (5)

 

According to Ann L Griffen et al the presence of bacilli is health associated .Hence we may conclude that patients with systemic diseases are more susceptible to GAP. This conclusion is supported by the presence of candida species which occur in cases of diseases like diabetes mellitus, some medications, and immunosuppressive diseases such as AIDS.(6)

 

CONCLUSION:

From this study it can be concluded that there exist no significant difference in the aerobic bacterial composition of supragingival plaque samples of patients with generalised chronic periodontitis and generalised aggressive periodontitis. Better results could have been obtained with a larger sample size and expanding the study to include anaerobic bacteria as well.

 

ABBREVIATIONS:

GCP- Generalised Chronic Periodontitis

GAP- Generalised Aggressive Periodontitis

Spp.-species

 

REFERENCE:

1)    Microbial  role  in  periodontitis:  Have  we  reached  the  top?  Some  unsung  bacteria other  than  red  complex ;Nupur  Arora ;J  Indian  Soc  Periodontol.  ;2014  Jan-Feb;  18(1):  9–13

2)    What  is  there  in  a  name?.  A  literature  review  on  chronic  and  aggressive periodontitis; Sameera  G.  Nath ;J  Indian  Soc  Periodontol.  ;2011  Oct-Dec;  15(4):  318–322.

3)    Predominant Microflora in Chronic and Generalized Aggressive Periodontitis in a Jordanian Population; Habashneh RA ; OMICS international ; 2014 ; Volume 4 • Issue 2 • 1000193

4)    Microbiological diversity of generalized aggressive periodontitis by 16S rRNA clonal analysis.;Faveri M1;. Oral Microbiol Immunol. 2008 Apr;23(2):112-8.

5)    Immunologic and microbiologic profiles of chronic and aggressive periodontitis subjects.; Rescala B ; J Periodontol. ;2010 Sep;81(9):1308-16.

6)    Candida spp. in periodontal disease: a brief review.Sardi JC; J Oral Sci. 2010 Jun;52(2):177-85.

 

 

 

Accepted on 22.06.2016        © RJPT All right reserved

Research J. Pharm. and Tech 2016; 9(9):1363-1366.