Probiotics

Probiotics in dentistry: Review of the current status

Mallikarjuna K, Shilpi Gupta, Siddharth Singh, Brajesh Dadarya, Priyanka Dausage, Puneet Gupta

International Journal of Contemporary Dentistry. 2013 Sep 18;4(1).

CONTENTS• INTRODUCTION• HISTORY• DEFINITION• PREBIOTIC VS PROBIOTIC• COMPOSITION• MECHANISM OF ACTION• COMERCIALLY USED• HEALTH BENEFITS• PROBIOTICS IN ORAL HEALTH• ROLE IN DENTAL CARIES, PDL DS, HALITOSIS• VEHICLES FOR DELIVERY• RECENT ADVANCES• SAFETY ASPECTS• CONCLUSION

HISTORY

Probiosis = “for life”

“Live microorganisms which, when administered in adequate amounts in food or as dietary supplement confer a health benefit on

the host”. (WHO/FAO 2001)

Living microbes or as food ingredients containing living microbes, that beneficially influence health of the host when used in

adequate numbers.

Microbial preparations or components of microbial cells that have beneficial effect on health and well being.

• “non-digestible food ingredient that beneficially affects the host by selectively stimulating the growth and/or activity of one or a limited number of bacteria in the colon

prebiotic

• live micro-organisms which when administered in adequate numbers confer a health benefit on the host

probiotic• a food or food

supplement will include both the live cells of the beneficial bacteria and the selective substrate

synbiotic

Micro-organisms commonly used

as Probiotics

Lactobacillus rhamnosus GG, ATCC 53103 (LGG)

Control inflammatory

bowel diseases

Alleviates food allergy symptoms

in infants

Strengthened innate

immunity

Lower serum cholesterol

Improved lactose

tolerance

Control of irritable bowel

syndrome

Suppresion of endogenous pathogens

Suppresion of exogenous pathogens

Reduction in risk factors for colon cancer

probiotics

Normalised intestinal microbiota composition

immunomodulation

Metabolic effects

Colonisation resistance

Supply of SCFA and vitamins to colonic epithelium

Lower level of toxicogenic reactions in gut

Bile salt deconjugation & secretion

INDICATIONS• Proven indication

1.Rota virus diarrhoea

2.Reduction of antibiotic associated side effect

• Possible indication

1.Dental caries and periodontal health

2.Food allergies and lactose intolerance

3.Atopic eczema

4.Prevention of vaginitis

5.Urogenital infections

6.Irritable bowel disease

7.Cystic fibrosis

8.Traveller's diarrhoea

9.Enhance oral vaccine administration

10.Helicobacter pylori infection

11.Various cancers

IDEAL PROPERTIES OF PROBIOTIC

• Capable of surviving and metabolising in gut

• Contain large number of viable cells

• Non pathogenic and nontoxic

• Exert beneficial effect on host

Remain viable during storage and

use

Have good sensory

properties

Should not have ability to transfer antibiotic resistance

genes

Be isolated from same

species as its intended host

Functional properties of oral probiotics

Inhibition of pathogen adhesion

Producing antimicrobial compounds

Aggregation & coaggregation

Inhibit pathogen induced

proinflammatory cytokine secretion

Enhance immunomodulator

y effect

Adhesion to gingival or epithelial cells in presence of saliva

Competes for adhesion sites

aggregate Competes for

nutrients & growth factors

Produce antimicrobial compounds including acids

Enhances host immune responses

Inhibit pathogen induced

production of pro-inflammatory

cytokines

Decrease MMP production

Inhibition of adhesion & enhanced clearance

Inhibition of growth of pathogens & other effects on dental plaque ecology

Influences on local & systemic immune responses

Antagonism against pathogens Reduction of inflammation & tissue destruction

Production of antimicrobial

substances

Competition of binding sites

Modulation of immunity

PROBIOTICS

Aggregation to oral biofilm

Adhesion to oral mucosa

Reinforcing oral epithelium

Modulation of inflammatory

response

Resistance to pathogenic colonisation

Prevention of periodontal disease Prevention of caries

Selecting oral probiotics

EXAMPLES OF PROBIOTIC DIETARY SUPPLEMENTS

Brand name (manufacturer) Active Ingredients

Accuflora Pro-Biotic Acidophilus (Northwest Natural Products) Lactobacillus acidophilus, Lactobacillus

rhamnosus, Bifidobacterium bifidum, Lactobacillus sallivarius, Streptococcus thermophilus

Acidophilus Pearls ( Enzymatic therapy) Lactobacillus acidohilus, bifidobacterium longum

Align (Proctor and Gamble) Bifidobacterium infantis

BioBeads Probiotic Acidophilus complex (Natrol ,Inc

Lactobacillus acidophilus, Bifidobacterium bifidum, Bifidobacterium longum ,Lactobacillus rhamnosus

Chewable Probiotic Acidophilus with Bifidus (Nature’s Bounty ,Inc)

Lactobacillus acidophilus, Lactobacillus bifidus

Complete Probiotic Multi Strain (WN Pharmaceuticals Ltd)

Bifidobacterium breve, Bifidobacterium longum ,Lactobacillus acidophilus ,Lactobacillus rhamnosus

Culturelle (Amerifit Brands) Lactobacillus GG

Culturelle for Kids (Amerifit Brands) Lactobacillus GG

Flora –Q (Kenwood Therapeutics) Lactobacillus acidophilus, Bifidobacterium , Lactobacillusparacasei, Streptococcus thermophilus

Florastor (Biocodex ,Inc) Saccharomyces boulardiilyo

Philips’ Colon health Probiotic Caps(Bayer Consumer Health

Lactobacilli gasseri, Bifidobacterium bifidum,bifidobacterium longum

Primadophilus kids (nature ‘s Way Products ,Inc)

Bifidobacteria ,Lactobacilli

Primadophilus Probifia Pearls (Nature ‘ S Way Products, Inc)

Bifidobacterium longum ,bifidobacteriumbifidum, Bifidobacterium breve ,bifidobacterium infantis

RepHresh Pro –B Probiotic Feminine Supplement (Lil Drug Store Products)

Lactobacillus rhamnosus (GR-1),Lactobacillus reuteri(RC-14)

Sustenex daily Probiotic (Ganeden Biotech ,Inc)

Bacillus coagulans

Women’s Probiotic With Cranberry (WN Pharmaceuticals Ltd)

Bifidobacterium longum, Lactobacillus acidophilus ,Lactobacillus casel ,Lactobacillus fermentum ,Lactobacillus rhamnosus, cranberry powdered extract (vaccinium macrocapon)

Different means of probiotic administration for oral health purposes

1. Improvement of intestinal barrier integrity and upregulation of mucin production

2. Involvement in binding of oral microorganisms to proteins (biofilm formation), action on plaque formation and on its complex ecosystem by competing and intervening with bacteria-to-bacteria attachments

3. Killing or inhibition of growth of pathogens through production of bacteriocins or other products, such as acid or peroxide, which are antagonistic towards pathogenic bacteria. Involvement in binding of oral micro-organisms to proteins (biofilm formation)

Role of probiotics in dental caries

Improvement of intestinal barrier integrity and upregulation of mucin production

Lactobacillus reuteri ATCC 55739

Bifidobacterium DN-173 010

L. reuteri MF2-3 (ATCC PTA-

4964)

L. reuteri CF27F (ATCC PTA-4965

L. reuteri FJ3 (ATCC PTA5290)

L. reuteri FJ1 “Prodentis”

(ATCC PTA5289)

Involvement in binding of oral microorganisms to proteins (biofilm formation), action on plaque formation and on its complex ecosystem by competing and intervening with bacteria-to-bacteria attachments

non-acid producing S.

mutans strain BCS3-L1

mutacin 1140

significantly less cariogenic than the parent strain JH1140 due to the delete of lactic acid dehydrogenase

open reading frameantibiotic

GTF-C GENE

decrease in extracellular matrix component from 51 to 33 percent of the biofilm volume

Killing or inhibition of growth of pathogens through production of bacteriocins or other products, such as

acid or peroxide, which are antagonistic towards pathogenic bacteria. Involvement in binding of oral micro-organisms to proteins (biofilm formation):

L. acidophilus La5

Weaker inhibition capacity

MS StrainsLactobacilli strains

Calgar et al (2007)

Hasslof et al

Total growth inhibition

L. rhamnosus LB21 L. paracasei F19

L. reuteri PTA 5289

L. reuteri ATCC 55730

Slight growth inhibition

Bifidobacterium animalis subsp. Lactis DN-173010

Cildir et al

S.mutans

Suzuki N et al

E. faecium WB2000 Biofilm formation strepococci

Heng et al

S. salivarius M18broad-spectrum inhibitory activity caries-causing Streptococcus mutans

Twetman et al

3-4 yr childrenDuration : 7 months

Singh et al

Bifidobacterium lactis Bb-12 ATCC27536 Lactobacillus acidophilus La-5

B. adolescentis SPM1005

Lactobacillus rhamnosus ATCC53103 - LGG

Glavina et al

S. mutans Lactobacillus spp. salivary counts

Bosch et al

Lactic acid bacteria

• Oral lymphoid foci – interdental papillae –local immune modulation.

• Glycoprotein – carbohydrate cell surface interaction mediate by inter species interactions

ROLE OF PROBIOTICS IN PERIODONTAL DISEASE

Lactobacilli rhamnosus Fusobacterium nucleatum

• Probiotics stimulate apoptosis of tumor cells through end product formation. inhibit apoptosis of mucosal cells.

• Protect epithelium barrier by maintaining tight junction protein expression and prevent apoptosis of mucous membrane.

Proinflammatory cytokine

reutrin reutricyclin

PDL DS HALITOSIS CANDIDA ALBICANS

New Approaches to achieve Probiotic Effects

Interference with

signaling

mechanisms

quorum sensing mechanism Competence Stimulating

Peptide (CSP)

Targeted

antimicrobial

therapy via a novel

STAMP technology

streptococcus rattus strain

JH145

Streptococcus uberis strain

KJ2

New Evora

kids Oral Probiotic

Chew

Plidenta Pro-t-action toothpaste

Acts in 10 secs

Tasteless, odourless

Lactobacillus paracasei

Safety aspectsShould not be pathogenic

should not have an ability to transfer antibiotic resistance genes

should not have any growth-stimulating effects on bacteria causing

diarrhea

maintain genetic stability in oral microflora

Mishra et al

2016

AIM

To compare the antimicrobial efficacy of probiotics in reducing salivary Candida albicans counts with commonly used antimicrobial agents like 0.2% chlorhexidine and herbal rinse.

METHOD

60 subjects6-4 years9 monthsGroup A: probiotic mint tablet by mixing the tablet with 5 ml of bottled or filtered water . Group B: 0.2% chlorhexidine digluconate oral rinse. Group C: herbal oral rinse.

RESULT

change in mean log10 colony-forming unit (CFU)/ ml of C. albicans in groups A : 0.43 ± 0.72, B : 0.68 ± 1.05 C : 0.22 ± 0.66 CFU/ml respectively

CONCLUSION

probiotic rinse was equally effective as 0.2% chlorhexidine digluconate rinse in reducing C. albicans counts after 1 week of intervention.

HERBAL : least effective.

AIM METHOD RESULT CONCLUSION

Srivastava et al

2016

To estimate the role of probiotic curd on salivary pH and Streptococcus mutans count, over a period of 7 days.

60 volunteersAge:20-25yrsTest Group :30 subjects consumed 100ml of probiotic curd daily for 7 days. Control Group : 30 subjects consumed 100ml of regular curd for 7 days.

Saliva samples assessed at baseline, after 1.5hour and 7 days of intervention period using pH meter and Mitis Salivarius Bacitracin agar.

reduction in salivary pH after ½ hour and 1 hour in both the groups.

normal curd:statistically significant (p< 0.05) reduction in salivary pH .

probiotic curd :statistically significant (p< 0.05) increase in salivary pH.

S. mutans colony counts probiotic curd >>normal curd.

Short-term consumption of probiotic curds showed marked salivary pH elevation and reduction of salivary S. mutans counts .

can be used for the prevention of enamel demineralization as a long-term remedy keeping in mind its cost effectiveness.

AIM METHOD RESULTS CONCLUSION

Chinnappa et al

2016

To compare mutans streptococci levels in saliva, before and after consumption of probiotic ice cream and ‑curds.

40 caries free children12-14yrsGroup I : 100ml probiotic icecreamGroup II : 100 ml plain icecreamGroup III : 100ml probiotic curdGroup IV: 100 ml plain curd Duration : 7 days• Saliva samples

assessed : • at baseline• 1 hour after

consumption • after 7 days

intervention period

using Mitis salivarius Bacitracin agar.

reduction in salivary mutans streptococci (MS) counts after 1 hour.

After 7 days, probiotic ice cream and ‑curd showed a statistically significant ( P < 0.001) reduction in MS counts as compared to the control ice cream and curd.

• Alternative strategy of displacing pathogenic microorganisms by probiotic bacteria

• Can be exploited for the prevention of enamel demineralization.

AIM METHOD RESULTS CONCLUSIONS

Rodriguez et al

2016

To compare milk supplemented with probiotic lactobacilli with standard milk for the increment of caries in preschool children after 10 mo of intervention.

N = 2612-3 YRS OLD

intervention group:150 mL of milk supplemented with Lactobacillus rhamnosus SP1 (107 CFU/mL).Control group: standard milk.

After 10 mo of probiotic intake, caries prevalence : 54.4% : probiotic group 65.8% : control group.

% of new individuals with cavitated lesions (ICDAS 5-6) in the control group (24.3%) was significantly higher than that in probiotic group (9.7%).

The probiotic group showed an increment of 0.58 ± 1.17 new lesions compared with 1.08 ± 1.70 new lesions observed in the control group.

regular long-term intake of probiotic-supplemented milk may reduce caries development in high-caries preschool children.


Nimish et al

2015

To evaluate the effect of probiotics on the salivary levels of Streptococcus mutans.

50 subjects18-30 yrs 2 ml of unstimulated saliva collected before and after 7 days of administration of probiotic (65 ml once daily) .Yakult probiotic: Lactobacilli casei strain was used.

statistically significant reduction in the salivary levels of S. mutans after the use of probiotics.

non-invasive method for combating infection and dental carie

AIM METHOD RESULTS CONCLUSIONS

JOSE et al

2013

to evaluate and compare the effects of the systemic consumption of probiotic curd and the topical application of probiotic toothpaste on the Streptococcus mutans levels in the plaque of orthodontic patients.

60 ortho pts.Grp A : CONTROLGrp B: probiotic curdGrp C: brush twice daily with probiotic toothpaste.

Plaque specimens: labial surfaces surrounding orthodontic brackets of the maxillary lateral incisors using a 4-pass technique.

S. mutans : evaluated using RT-PCR.

reductions in S mutans concentration in groups 2 and 3.

consumption of probiotic curd and the use of probiotic toothpaste cause a significant decrease in the S mutans levels in the plaque around brackets in orthodontic patients.


Hedayati-Hajikand et al

2015

BMC oral health

To evaluate the effect of probiotic chewing tablets on early childhood caries development in preschool children living in a low socioeconomic multicultural area.

138 healthy 2-3-year-old children .

TEST GROUP: one chewing tablet per day containing three strains of live probiotic bacteria (ProBiora3®)

PLACEBO GROUP : identical tablets without bacteria.

DURATION : 1 YEAR

caries increment (Δds) was significantly lower in the test group when compared with the placebo group, 0.2 vs. 0.8 (p<0.05).

Early childhood caries development could be reduced through administration of these probiotic chewing tablets as adjunct to daily use of fluoride toothpaste in preschool children.


Singh et al

2011

To compare the levels of mutans streptococci and lactobacilli in saliva of school children, before and after consumption of probiotic and control ice-cream.

40 SUBJECTS12-14 yr old children Following ani nitial run-in period of 1 week, children in group I and II were given ice-creams ‘A’ and ‘B’, respectively, for 10 days. Being a cross-over study, the ice-creams were interchanged in the two groups after a 2-week wash-out period. Saliva samples at baseline and follow-up were assessed using Dentocult SM and Dentocult LB kits.

probiotic ice-cream : statistically significant reduction (p-value = 0.003) in salivary mutans streptococci levels with no significant effect on lactobacilli levels.

probiotic ice-cream containing Bifidobacterium lactis Bb-12 ATCC27536 and Lactobacillus acidophilus La-5 can reduce the levels of certain caries-associated micro-organisms in saliva.


Caglar et al

2007

To evaluate the effect of xylitol and probiotic chewing gums on salivary mutans streptococci (MS) and lactobacilli (LB).

80 healthy young21-24 yrs

Groups A : probiotic gum Group B: xylitol gum Group C: probiotic + xylitol gum Group D: placebo gum group gums were taken 3 times daily after meals. intervention period was 3 weeks. • probiotic gums

contained 2 strains of Lactobacilli reuteri (ATCC 55730 at a dose of 1 × 108 CFU/gum and ATCC PTA 5289 at a dose of 1 × 108 CFU/gum),

• pellet of the xylitol gum contained ≈1.0 g xylitol as single sweetener

Significant reduction (p < 0.05) of salivary MS was displayed in group A and B.

No alterations of salivary LB was demonstrated in any group.

Daily chewing on gums containing probiotic bacteria or xylitol reduced the levels of salivary MS in a significant way.


Dorantes et al

2015

To assess the effect of the daily ingestion of a mixture of probiotics on the amount of Streptococcus mutans in the oral cavity of preschool-age patients with a high risk of caries.

40 children4-6yrs with high risk for caries.Experimental Group (A): pts brushed their teeth & used fluoridated toothpaste in addition to consuming probiotics daily,

Control Group (B) : pts who brushed their teeth & used fluoridated toothpaste but did not consume probiotics.

CariScreen®:microorganism count was determined at different times: baseline, 7, 14, 21 and 30 days

Both groups showed similar microbial counts at the beginning of the trial (p>0.05)

significant decrease in the count at the end of the study was found in the experimental group (p<0.05) 15 days after suspending ingestion.

significant reduction of RLU values in preschool children who ingested the tested probiotics in relation to the baseline values and 15 days after ceasing consumption.


Caglar et al

2006

To investigate the effect of the probiotic bacterium Lactobacillus reuteri ATCC 55730 on the levels of salivary mutans streptococci and lactobacilli in young adults when ingested by two different delivery systems.

120 healthy young adults (21–24 years)

Group A : 200 ml of water through a prepared straw containing L. reuteri ATCC 55730 once daily for 3 weeks. Group B : 200 ml water through a placebo straw during the same period.

Group C : one tablet containing L. reuteri ATCC 55730 once daily for 3 weeks

Group D: received placebo tablets without bacteria.

significant reduction of the mutans streptococci levels was recorded after ingestion of the probiotic bacteria via the straw (p<0.05) and the tablets (p<0.01).

short-term daily ingestion of lactobacilli-derived probiotics delivered by prepared straws or lozenges reduced the levels of salivary mutans streptococci in young adults.


Mahantesha et al

2015

Journal of International Oral Health 2015; 7(9):47-50

to compare the effectiveness of probiotic ice cream and drink on salivary Streptococcus mutans levels in children of 6-12 years age group.

(n = 50) of 6-12 years age with zero decayed missing filled teeth (dmft)/DMF .Saliva samples were collected before consumptions of probiotic ice cream & probiotic drink. Colony count obtained was recorded as baseline data. For both groups probiotic ice cream and drink was given randomly for 7 days and a washout period of 90 days were given & saliva samples were collected & colony counting done.

GRP A: AMUL icecreamFRP B: YAKULT milk

significant reduction in salivary S. mutans level in both groups after 7 days period.

After washout period only probiotic ice cream showed reduction.

There was no significant difference between probiotic ice cream and drink

Probiotic organisms definitely have a role in reducing the salivary S. mutans level and ice cream would be a better choice than drink.


Nase et al

Caries research 2001

To examine whether milk containing LGG has an effect on caries and the risk of caries in children when compared with normal milk.

594 children, 1–6 years old children received the milk with meals from coded containers 5 days a week in the day-care centres for 7 months. Caries risk calculated : based on clinical and microbiological data, comprising mutans streptococcus levels from dental plaque and saliva.

Risk was classified as high : dmft/DMFT or initial caries score >0, and a mutans streptococcus count >105 CFU/ml.

Less dental caries in the LGG group and lower mutans streptococcus counts at the end of the study. LGG was found to reduce the risk of caries significantly (OR = 0.56, p = 0.01; controlled for age and gender, OR = 0.51, p = 0.004).

Milk containing the probiotic LGG bacteria may have beneficial effects on children’s dental health.


Yousuf et al

2015

evaluate the effectiveness of commercially available freeze dried powdered probiotics on mutans streptococci count among 12-15 year-old Indian schoolchildren.

INVITRO, INVIVO32children , 12-5yrs Group A : 11 children using freeze dried Lactobacillus acidophilus, Bifidobacterium longum, Bifidobacterium bifidum and Bifidobacterium lactis.

Group B : 11 children using freeze dried lactic acid bacillus only.

Group C :11 children using placebo powder. Duration : three weeks

examination and sampling of the subjects were done on days 0 (baseline), seven, 14 and 21.

statistically significant reduction (P<0.05) in salivary mutans streptococci counts was recorded up to the second week.

Oral administration of probiotics showed a short-term effect on reduction of mutans streptococci count and showed a preventive role in caries development.


Oliveira et al

2015

Oral administration of probiotics showed a short-term effect on reduction of mutans streptococci count and showed a preventive role in caries development.

In situ experiment, 10 volunteers wore palatine devices containing 4 blocks of bovine dental enamel over 3 phases, during which 20% sucrose solution, Yakult® (Treatment A), and Batavito® (Treatment B) were dropped on the enamel blocks. Salivary microbial counts obtained and biofilm samples were analyzed after each phase. In vivo experiment, the same 10 volunteers drunk Yakult® (Treatment C) and Batavito® (Treatment D) in two phases.

Treatment B resulted in fewer total cultivable anaerobes and facultative microorganisms in biofilms, higher final microhardness, lower percentage change in surface hardness, and smaller integrated subsurface enamel hardness. In vivo study, Treatment D resulted in a reduction in the counts of all microorganisms.

probiotic fermented milk Batavito®, but not Yakult®, reduced the amount of oral microorganisms and mineral loss in bovine enamel.


Yadav et al

2014

To investigate the effect of probiotic Lactobacillus casei Shirota on the values of mutans .streptococci counts in saliva.

31 CHILDRENDuring the intervention periods children were either given probiotic milk or control milk for 10 days. Pre and post intervention salivary samples were subjected to microbiological evaluation.

Numbers of mutans streptococci were taken by identifying colony morphology.

reduction in children with higher levels of mutans streptococci (105) after intervention was 34% in the probiotic group.

Daily consumption of milk containing probiotic bacteria can reduce the levels of mutans streptococci and may contribute to the prevention of dental caries.

Health & Medicine

Probiotics