**4. Collection of samples**

We recommended that patients brush their teeth three times a day using the Bass modified technique with toothpaste containing fluoride after placement of the appliances.

Clinical, Salivary and Bacterial Markers on the Orthodontic Treatment 173

Unstimulated saliva (seconds) Male 39.85 + 19.17 38.64 + 14.90

 pb 0.3903 0.0026\* Stimulated saliva (mL/min) Male 1.72 + 0.65 1.76 + 0.80

 pb 0.0019\* 0.0835 Buffer capacity Male 8.78 + 2.19 8.71 + 2.16

 pb 0.0381\* 0.1247 Salivary pH Male 7.68 + 0.17 7.74 + 0.09

pb 0.1672 0.7039

pa value between before vs. after placement of appliances, based on paired t-test

Table 1. Distribution of salivary markers by gender in the study (n = 34)

not observed in the bacterial counts distribution (Table 2).

We observed changes in the CFU of SM after the placement of appliances. Before treatment 14/34 subjects had high values (>105); after one month of banding, 16/34 had high values. In the first sample 7/34 subjects had high levels (>105) of *LB*, for the second sample we found 20/34 subjects in these same level, although statistically significant differences were

The acidity of the initial plaque no registered significant modifications after the placement of appliances (*P* = 0.5467); however, we found differences between genders in the initial sample (*P* = 0.0430); the pH between 5.8-7.2 predominating in females, and lowest values (pH 5.4 ± 0.3) in males. The second sample showed more subjects with lowest values (pH 5.4

Differences were observed in the gingival bleeding before and after orthodontic treatment (*P* = 0.0305), with an increased the bleeding in saliva in the second sample. It has to be considered that in the beginning of the study most of the subjects were in the intermediate

Stages

Before After

Female 45.65 + 19.03 57.35 + 17.43 0.4073

Female 1.06 + 0.48 1.36 + 0.50 0.0001\*

Female 6.90 + 2.69 7.70 + 1.59 0.0359\*

Female 7.53 + 0.38 7.73 + 0.09 0.0246\*

Pa

Marker Gender

Salivary markers

Data shown as mean ± SD

**4.3 Bacterial markers** 

**4.4 Plaque pH** 

± 0.3) in both genders (Table 2).

**4.5 Occult blood in saliva** 

\*P < 0.05

pb value between genders, based on paired t-test

Recommendations to avoid retentive, cariogenic, or hard foods during orthodontic treatment were provided verbally.

For take the tests is necessary that the patients avoided eating or drinking and no toothbrushing at least 2 hours before taking the samples in both time points.

In our experience, with thirty-four subjects, where 14 males (mean age, 16.2 ± 3.4 years) and 20 females (mean age, 17.2 ± 6.3 years), we selected patients didn't have any systemic diseases, use of antibiotics at least 15 days before initiating the study, active caries, and on mixed dentition phase were excluded.

Two samples were taken from each patient, one before beginning the orthodontic treatment and the other 1 month after placement of the appliances, because this is the time for the appointment to change the first arch wire. The orthodontics attachments were placed in both arches.

Dental and skeletal diagnoses were obtained from each patient, defined by Angle's molar class and the subspinale-nasion-supramentale cephalometric angle (ANB). Fifteen patients had molar relation class I, fourteen had class II, four had class III, and in one patient it was not possible to determine the relationship due to absent first molars.

As for the skeletal diagnosis, fourteen were class I, nineteen were class II, and one were class III. The dental and skeletal diagnoses were not associated or determined the behavior of any of the studied variables (p>0.05) (data not shown).

### **4.1 Clinical markers**

The mean DMFS index of the subjects was 6.47, although it was greater in females (8.70) that in males (3.28), these differences were not statistically significant (P = 0.1352. The mean O´Leary's plaque index was 44.6 %, males presented a slightly plaque percentage (50.84%) than females (40.15%; P = 0.1809) (data not shown).

#### **4.2 Salivary markers**

We did not find a statistically significant difference in the unstimulated salivary production before and 1-month after the placement of orthodontic appliances, nevertheless according to time point, in both samples, there were differences between the saliva production of males and females (*P* = 0.0026); specifically, the unstimulated salivary production was lower in women (Table 1).

The placement of orthodontic appliances promoted a major stimulated salivary flow in the subjects, with significant differences in the salivary production before and after treatment (*P* = 0.0001). The salivary stimulated flow rate was greater in males at the beginning (*P* = 0.0019; Table 1). The salivary buffer capacity showed differences after placement of the appliances (*P* = 0.0359) and between genders before the treatment (*P* = 0.0381) females showed lower capacity (Table 1).

Significant differences were observed in the salivary pH before and after treatment (*P* = 0.0246) with an increase of the pH value (Table 1).


Data shown as mean ± SD

pa value between before vs. after placement of appliances, based on paired t-test pb value between genders, based on paired t-test

\*P < 0.05

172 Contemporary Approach to Dental Caries

Recommendations to avoid retentive, cariogenic, or hard foods during orthodontic

For take the tests is necessary that the patients avoided eating or drinking and no

In our experience, with thirty-four subjects, where 14 males (mean age, 16.2 ± 3.4 years) and 20 females (mean age, 17.2 ± 6.3 years), we selected patients didn't have any systemic diseases, use of antibiotics at least 15 days before initiating the study, active caries, and on

Two samples were taken from each patient, one before beginning the orthodontic treatment and the other 1 month after placement of the appliances, because this is the time for the appointment to change the first arch wire. The orthodontics attachments were placed in

Dental and skeletal diagnoses were obtained from each patient, defined by Angle's molar class and the subspinale-nasion-supramentale cephalometric angle (ANB). Fifteen patients had molar relation class I, fourteen had class II, four had class III, and in one patient it was

As for the skeletal diagnosis, fourteen were class I, nineteen were class II, and one were class III. The dental and skeletal diagnoses were not associated or determined the behavior of any

The mean DMFS index of the subjects was 6.47, although it was greater in females (8.70) that in males (3.28), these differences were not statistically significant (P = 0.1352. The mean O´Leary's plaque index was 44.6 %, males presented a slightly plaque percentage (50.84%)

We did not find a statistically significant difference in the unstimulated salivary production before and 1-month after the placement of orthodontic appliances, nevertheless according to time point, in both samples, there were differences between the saliva production of males and females (*P* = 0.0026); specifically, the unstimulated salivary production was lower in

The placement of orthodontic appliances promoted a major stimulated salivary flow in the subjects, with significant differences in the salivary production before and after treatment (*P* = 0.0001). The salivary stimulated flow rate was greater in males at the beginning (*P* = 0.0019; Table 1). The salivary buffer capacity showed differences after placement of the appliances (*P* = 0.0359) and between genders before the treatment (*P* = 0.0381) females

Significant differences were observed in the salivary pH before and after treatment (*P* =

toothbrushing at least 2 hours before taking the samples in both time points.

not possible to determine the relationship due to absent first molars.

of the studied variables (p>0.05) (data not shown).

than females (40.15%; P = 0.1809) (data not shown).

treatment were provided verbally.

mixed dentition phase were excluded.

both arches.

**4.1 Clinical markers** 

**4.2 Salivary markers** 

women (Table 1).

showed lower capacity (Table 1).

0.0246) with an increase of the pH value (Table 1).

Table 1. Distribution of salivary markers by gender in the study (n = 34)

## **4.3 Bacterial markers**

We observed changes in the CFU of SM after the placement of appliances. Before treatment 14/34 subjects had high values (>105); after one month of banding, 16/34 had high values.

In the first sample 7/34 subjects had high levels (>105) of *LB*, for the second sample we found 20/34 subjects in these same level, although statistically significant differences were not observed in the bacterial counts distribution (Table 2).

#### **4.4 Plaque pH**

The acidity of the initial plaque no registered significant modifications after the placement of appliances (*P* = 0.5467); however, we found differences between genders in the initial sample (*P* = 0.0430); the pH between 5.8-7.2 predominating in females, and lowest values (pH 5.4 ± 0.3) in males. The second sample showed more subjects with lowest values (pH 5.4 ± 0.3) in both genders (Table 2).

#### **4.5 Occult blood in saliva**

Differences were observed in the gingival bleeding before and after orthodontic treatment (*P* = 0.0305), with an increased the bleeding in saliva in the second sample. It has to be considered that in the beginning of the study most of the subjects were in the intermediate

Clinical, Salivary and Bacterial Markers on the Orthodontic Treatment 175

It is established that orthodontic treatment induces changes in the oral environment, with increasing concentrations of mutans streptococci and lactobacilli, as well as increased blood

This is exacerbated in stages which increase the use of attachments and the consequent

The clinical markers showed that males had lower DMFS index than females, nevertheless,

The stimulated salivary flow increased after the placement of orthodontic appliances. In the present study it was greater in males, which is similar to international reports (Chang et al.,

The mean salivary flow rate for both genders was found in normal parameters: 1-3 mL/min in stimulated saliva and from 0.25-0.35 mL/min in unstimulated saliva. The variability of

The results support a direct and prolonged stimulatory effect after one month of treatment

The saliva buffer capacity presented a significant increase in females after the orthodontic therapy. Males showed higher buffer capacity than females, this difference between genders

The salivary buffer capacity prevents the settling of pathogenic microorganisms in mouth,

Salivary pH demonstrated a significant increase in the 1st month of treatment, as opposed to other studies in which it has been demonstrated that the pH suffers alterations after 3

Males showed a more acid plaque pH at the beginning of the treatment, nevertheless an increase in acidity was demonstrated 1 month with orthodontic appliances in females. The plaque pH had significant differences between genders when the patients already were in

The orthodontic appliances protected the plaque from the tooth brushing action, the mastication, and the salivary fluid. Accumulating more on the cervical region of the brackets or below the arches wire, which is the zone where a major demineralization can be found

The present study showed an increase of CFU after placement of the orthodontic appliances, which has been demonstrated also by Chang (1999). Nevertheless, these changes in the 1st month of study were not statistically significant, as opposed to report by this same author. Before initiating treatment, the majority of patients showed incipient periodontal disease detected trough the Salivaster® test, possibly caused by crowding, which in the majority of patients is the principal motivation for the orthodontic treatment. Likewise, a significant increase of periodontal disease was observed 1 month after initiating orthodontic therapy

the plaque index by O´Leary showed more plaque in males of this study.

salivary flow rate has been established by other researchers (Torres et al, 2006).

being an important risk indicator, because it reveals the response of the host.

concentration in saliva and decreased plaque pH.

1999; Bretz et al., 2001; De Vigna et al., 2008).

with fixed orthodontic appliances on salivary flow.

months of orthodontic treatment (Chang et al., 1999).

have been demonstrated previously.

orthodontic treatment.

(Migale et al., 2009).

difficulty of proper hygiene.

**5. Conclusions** 


level with 1 mg/dL of occult blood in saliva (incipient periodontal disease) and one month later the periodontal disease present increase (Table 2).

NC = negative at culture

Pa value between before vs. after placement of appliances, based on X2 test

Pb value between genders, based on X2 test

\* P < 0.05

Table 2. Distribution of bacterial markers, plaque pH and occult blood in saliva by gender in the study (n = 34)

It is established that orthodontic treatment induces changes in the oral environment, with increasing concentrations of mutans streptococci and lactobacilli, as well as increased blood concentration in saliva and decreased plaque pH.

This is exacerbated in stages which increase the use of attachments and the consequent difficulty of proper hygiene.
