618
0.2% and Chlorhexidine 0.2%: Short-Term Effect
Rinsing With Delmopinol
Salivary Microbiology, Plaque, and Gingivitis on
Bruno
Collaert, * Stig Edwardsson,f Rolf Attström, * Jan C. Hase, * Mikael Âstrom, * and Rolf Moverà
The aim of this short-term study was to compare the effect of delmopinol HCl 0.2% and Chlorhexidine digluconate 0.2% rinses on the development of dental plaque, the healing of experimental gingivitis, and the salivary microbiology. As part of a larger study protocol, 45 healthy males enrolled in an oral hygiene program to upgrade their oral health. For this portion of the study, participants had their teeth professionally cleaned on day 0. The participants then abstained from standard mechanical oral hygiene procedures, but applied a placebo solution twice daily for 2 weeks. At the end of this period the subjects received a second professional cleaning and were then assigned to 2 treatment groups: Group 1 rinsed with 10 ml of delmopinol HCl 0.2% and Group 2 rinsed with 10 ml of Chlorhexidine digluconate 0.2% for 1 minute twice daily for the next 2 weeks and continued to refrain from mechanical oral hygiene procedures. At the end of the placebo and active treatment periods 1) saliva samples were taken and cultivated on a series of media; 2) the degree of gingivitis was assessed with gingival crevicular fluid (GCF) and gingivitis index (GI); and 3) the plaque index was assessed and the stainable buccal plaque extension was analyzed planimetrically. No changes in the
salivary microbiological counts were detected for the subjects rinsing with delmopinol. Subjects rinsing with Chlorhexidine showed significant reductions of anaerobes, aerobes, and S. mutans in saliva. The amounts of GCF and GI were reduced largely to the same extent in both treatment groups. Mean plaque extension was reduced by 52% after delmopinol and 88% after Chlorhexidine rinsing. The results show that rinsing with delmopinol for a period of 14 days 1) does not give rise to detectable changes in the cultivable salivary microflora; 2) reduces plaque regrowth less effectively than Chlorhexidine; and 3) promotes the healing of an experimentally induced gingivitis largely to the
same extent as
Chlorhexidine. J Periodontol 1992; 63:618-625.
Key Words: Chlorhexidine/therapeutic
delmopinol/therapeutic prevention; gingivitis/therapy; saliva/microbiology. use;
Delmopinol, ( + / )-3-(4-propylheptyl)-4-morpholinehydrochloride)5 (Fig. 1) has recently been developed as a plaque control agent.1 Delmopinol has been tested in dose-response study during which rinsing with 3 different concentrations (0.05%, 0.1%, 0.2%) were compared to Chlorhexidine digluconate 0.2% when used for 14 days in the absence of mechanical oral hygiene procedures.2 The results showed that the experimentally induced gingivitis healed largely to the same extent in subjects rinsing twice ethanol
-
'Department of Periodontology, University, Malmö, Sweden, department of Microbiology.
Center for Oral Health
*Biosurface AB. §Decapinol, Biosurface AB, Malmö, Sweden.
Sciences, Lund
use; dental
plaque/
daily with 0.1% or 0.2% delmopinol as in subjects rinsing with Chlorhexidine 0.2%. Furthermore, a statistically significant dose-response effect was noted between the amount of plaque and gingivitis, and the dose of delmopinol used. Two weeks of rinsing with delmopinol in the absence of oral hygiene appeared to change the composition of the supragingival plaque to one predominated by coccoid cells.3 The mode of action of delmopinol is not known in detail. Current information shows that delmopinol has a low antimicrobial capacity relative to that of Chlorhexidine.4 The plaque reducing effect therefore seems to be related more to the physico-chemical interaction of delmopinol with tooth and bacterial surfaces. Significant in this respect might be that delmopinol is a surface active agent, which may inter-
Volume 63 Number 7
COLLAERT, EDWARDSSON, ATTSTROM, HASE, ÄSTRÖM, MOVERT day-14
I oral
day 0 hygiene period
\
day 14 delmopinol
placebo period
day HCl
619 28
0.2%|
Chlorhexidine 0 2%
O
professional toothcleaning
*
clinical measurements
salivary microbiology
Figure 2. Experimental design. Figure 1. Structural formula of delmopinol hydrochloride.
jects
fere with the adhesion forces between microbes and the
surroundings.1 Changes in the oral microflora could be beneficial if known pathogens and pathogens associated with gingival infection
reduced or absent. But when such alterations in the indigenous oral flora are induced by the use of chemotherapeutic agents for controlling plaque and gingivitis, attention should be paid to the possible emergence of opportunistic pathogens.5 Microorganisms present in the oral cavity usually establish a balance between those present in saliva and those adhering to shedding or non-shedding surfaces, depending on the affinity of the respective microorganisms to the oral surfaces.6 Saliva can also be considered a medium for the spread of microorganisms from one oral site to the are
other.7
The aim of this short-term study was to detect possible changes in the cultivable salivary microflora after rinsing with delmopinol. Plaque and gingivitis clinical status was monitored during the study to verify earlier obtained results. A comparison was made between rinsing with delmopinol hydrochloride 0.2% and Chlorhexidine digluconate 0.2%. The experimental model used attempted to imitate a clinical condition at which treatment of gingivitis is normally
initiated.
MATERIAL AND METHODS Test Solutions The placebo solution was an aqueous solution containing ethanol 2%, a herb flavor 0.02%, and sodium saccharine 0.01%. The active solutions were an unflavored aqueous 0.2% delmopinol HCl solution, sodium hydroxide pH 5.7, and 0.2% Chlorhexidine digluconate.11 =
Subjects
Forty-five healthy male volunteers (18 to 40 years), who had participated in a previous 6-week study identical to the present study design which monitored the effect of topical application of 3 concentrations of delmopinol HCl on the salivary microflora, plaque, and gingivitis,3 took part in this study. After informed consent was obtained, the sub'Hibutane
Dental, ICI-Pharma, Gothenburg, Sweden.
were admitted on the following basis: 1) complete dentition except for third molars and/or one premolar in each quadrant extracted on orthodontic indication; 2) no periodontal pockets exceeding 4 mm; 3) no crowns or bridges; 4) no buccal and large interproximal restorations; 5) no extensive exposed root surfaces; 6) no caries cavities; and 7) no antibiotic treatment during the preceding 6 months. Analyses were based on results from 39 participants (1 participant withdrew and 5 others were excluded because of non-compliance with the study protocol). All were excluded from analyses before breaking the randomization code.
Study Design
This 6-week study was unsupervised, computer randomized, and performed double-blind with parallel groups (Fig. 2). During the first 2 weeks all participants underwent 3 professional tooth cleanings and instructions in oral hygiene procedures, which resulted in clinically healthy gingiva. On day 0, start of the experimental gingivitis period, the subjects received thorough professional tooth cleaning. During the following 2 weeks they abstained from all mechanical oral hygiene measures, but carefully applied 2 ml of a placebo solution onto their teeth with a soft paint brush twice daily (morning and evening). The placebo was applied because, as mentioned above, the subjects were part of a larger study evaluating the use of topical application of delmopinol.3 At the end of the placebo period all subjects exhibited a mild gingivitis. On day 14 after receiving professional tooth cleaning, the subjects were randomly assigned to 1 of 2 treatment groups. During the next 2 weeks they continued to refrain from all mechanical oral hygiene procedures. Instead they rinsed twice daily (morning and evening) with 10 ml of 0.2% delmopinol (18 subjects) or 0.2% Chlorhexidine (21 subjects). The subjects were urged not to change their dietary habits for the duration of the study, and not to eat or drink for 30 minutes after rinsing. The objective of using this short-term experimental model was to establish a confirmed gingivitis baseline for the measurement of therapeutic effect. Such a baseline ought to be more clinically relevant than clinically healthy gingiva as the starting point for evaluation of plaque control agents. Gingival inflammation assessment. Gingival crevicular fluid (GCF) was obtained at the end of each 2-week period
620
J Periodontol July 1992
DELMOPINOL AS A PLAQUE CONTROL AGENT
from 4 teeth out of the
incisors, canine, and premolare of
maxillary quadrant. All supragingival plaque was carefully removed without disturbing the gingival margin. The one
buccal surface was dried with air directed from the buccal gingival fold towards the tooth. A 1 mm wide absorbent paper strip was gently inserted at the mesio-buccal aspect of the tooth into the gingival crevice until resistance was felt and left in place for 3 minutes.8 The strips were immediately stained with ninhydrin. The height of the stained area was subsequently measured. The gingivitis index (GI) was recorded as presence or absence of bleeding after gentle probing with a WHO probe to the bottom of the gingival crevice at 6 sites around each tooth: mesio-buccal, buccal, distobuccal, mesio-lingual, lingual, and disto-lingual.9 The GI was measured around all teeth (incisors, canines, premolars, and molars) in the 2 quadrants contralateral to the side used for GCF sampling. The GI was expressed as the percentage of bleeding sites. Plaque assessment. Supragingival plaque was disclosed with erythrosine. The plaque index (PI) according to Turesky10 was scored for the same teeth as those used for the GI. After staining, the subject rinsed with 10 ml of tap water for 10 seconds. Periodic identical color slides for planimetrie analyses11 were taken of the disclosed plaque on canines and premolars used for GI and PI recordings. The slides were enlarged 6.5 times and projected onto white paper. The tooth surface covered with stainable plaque was contoured and measured through computerized digitization. In 50% of the participants, the GCF-flow samples were obtained from the first quadrant, while the GI, PI, and plaque extension were measured in the second and third quadrant. In the other half of the subjects the opposite quadrant in the same jaw was used. The allocation of the subjects to one of the two partial mouth registration groups was randomized so that they were equally represented in the two treatment groups. Extrinsic tooth-staining assessment. At the end of both the placebo and the delmopinol treatment period, a periodic standardized photograph was taken before plaque disclosure of the same quadrants used for the plaque extension measurements. The photographs were compared to each other, with the placebo photo considered baseline. Differences in extrinsic tooth staining were scored as follows: 0 no difference in the degree of staining; 1 more slightly staining only detectable when photographs were compared; 2 visibly more staining, clinically detectable; and 3 profound dark brown staining (esthetically compromising). Because of a technical failure, data from 8 subjects are
Table 1: Culture
Media, Cultivation Procedures, and Identification
Medium and Cultivation Procedures
Identification
Brucella blood agar (BBL) with 5% human erythrocytes, 0.5% laked human erythrocytes and 5 mg/1 menadione (7 days
Total number of anaerobically cultivable
anaerobically) Blood agar12(5-7 days aerobically)
N2) C02 agar* (4 days aerobically, poured plate technique) Staphylococcus medium 110* (2 days aerobically) Mac Conkey agar* (4 days aerobically) Sabouraud Dextrose agar* (4 days aerobically) MSB-agar13 (days
in 5%
Selective lactobacilli
and
microorganisms
Total number of aerobically cultivable
microorganisms streptococci
Mutans
Lactobacilli Salt-tolerant
staphylococci Enteric bacteria Yeasts
*Difco, Detroit, MI.
collected at the end of the placebo and active treatment periods. The two samples were obtained from each participant at the same time of the day. The salivary flow was stimulated by tongue and cheek motion. Saliva (2 to 3 ml) was collected in a sterile vial with glass beads. The samples were processed within 1 hour after collection. The vial was placed in a Vortex mixer for 30 seconds. The culture media listed in Table 1 were inoculated from appropriate dilutions of the homogenized sample. Colony forming units (CFU) on Brucella blood agar resembling black pigmenting PorphyromonasIPrevotella spp.14,15 and Fusobacterium spp.16 were Gram stained and their presence was recorded. CFU with doubtful morphology on MSB-agar were isolated and their capacity to ferment mannitol was studied. The salttolerant Gram-positive cocci growing on 110 agar were tested for the presence of catalase and DNase,# the latter test dividing the isolates into those resembling Staphylococcus aureus and S. epidermidis. CFU of various morphology growing on MacConkey agar identified as Gram-negative rods were considered enteric bacteria. CFU on Sabouraudagar being blastospores in Gram stained smears were considered to be yeasts.
=
=
=
missing.
=
The GCF samples, GI, PI, and identical color-slides for plaque extension measurements and extrinsic tooth staining measurements were obtained by the same investigator on day 14 and day 28. There was no statistically significant difference for any of the parameters studied between the placebo period of the two treatment groups. Salivary microbiological evaluation. Saliva samples were
Adverse Experiences The subjects were examined at each visit for adverse reactions in the oral cavity and were questioned about the occurrence of any adverse experiences prior to the clinical examination. After the clinical examination at the last visit the subjects were asked if they had experienced an anesthetic sensation in any part of their oral mucosa after rinsing and how long they thought it had lasted.
Statistical Analyses All statistical analyses were performed with non-parametric methods. The subject was the unit of analysis. Wilcoxon's *Difco, Detroit,
MI.
Volume 63 Number 7 S I S
COLLAERT, EDWARDSSON, ATTSTROM, HASE, ÄSTRÖM, MOVERT
621
%
mm
60
0.6
I
A
0.5
c
50
u
L
40
0.4
R E
30
V
I
c
L A R
0.2
20
0.1
10
F L U I D
DELMOPINOL HCl 0.2%
[ZD PLACEBO
DELMOPINOL HCl 0.2%
CHLORHEXIDINE 0.2%
CHLORHEXIDINE 0.2%
ZZ PLACEBO
ül ACTIVE
Figure 3. Mean GCF-values ( + SE) after 14 days ofplacebo, and 14 days of delmopinol HCl 0.2% and Chlorhexidine 0.2% rinsing, respectively. Significant difference between placebo and active treatment is indicated. *P
0.2% and Chlorhexidine 0.2%: Short-Term Effect
Rinsing With Delmopinol
Salivary Microbiology, Plaque, and Gingivitis on
Bruno
Collaert, * Stig Edwardsson,f Rolf Attström, * Jan C. Hase, * Mikael Âstrom, * and Rolf Moverà
The aim of this short-term study was to compare the effect of delmopinol HCl 0.2% and Chlorhexidine digluconate 0.2% rinses on the development of dental plaque, the healing of experimental gingivitis, and the salivary microbiology. As part of a larger study protocol, 45 healthy males enrolled in an oral hygiene program to upgrade their oral health. For this portion of the study, participants had their teeth professionally cleaned on day 0. The participants then abstained from standard mechanical oral hygiene procedures, but applied a placebo solution twice daily for 2 weeks. At the end of this period the subjects received a second professional cleaning and were then assigned to 2 treatment groups: Group 1 rinsed with 10 ml of delmopinol HCl 0.2% and Group 2 rinsed with 10 ml of Chlorhexidine digluconate 0.2% for 1 minute twice daily for the next 2 weeks and continued to refrain from mechanical oral hygiene procedures. At the end of the placebo and active treatment periods 1) saliva samples were taken and cultivated on a series of media; 2) the degree of gingivitis was assessed with gingival crevicular fluid (GCF) and gingivitis index (GI); and 3) the plaque index was assessed and the stainable buccal plaque extension was analyzed planimetrically. No changes in the
salivary microbiological counts were detected for the subjects rinsing with delmopinol. Subjects rinsing with Chlorhexidine showed significant reductions of anaerobes, aerobes, and S. mutans in saliva. The amounts of GCF and GI were reduced largely to the same extent in both treatment groups. Mean plaque extension was reduced by 52% after delmopinol and 88% after Chlorhexidine rinsing. The results show that rinsing with delmopinol for a period of 14 days 1) does not give rise to detectable changes in the cultivable salivary microflora; 2) reduces plaque regrowth less effectively than Chlorhexidine; and 3) promotes the healing of an experimentally induced gingivitis largely to the
same extent as
Chlorhexidine. J Periodontol 1992; 63:618-625.
Key Words: Chlorhexidine/therapeutic
delmopinol/therapeutic prevention; gingivitis/therapy; saliva/microbiology. use;
Delmopinol, ( + / )-3-(4-propylheptyl)-4-morpholinehydrochloride)5 (Fig. 1) has recently been developed as a plaque control agent.1 Delmopinol has been tested in dose-response study during which rinsing with 3 different concentrations (0.05%, 0.1%, 0.2%) were compared to Chlorhexidine digluconate 0.2% when used for 14 days in the absence of mechanical oral hygiene procedures.2 The results showed that the experimentally induced gingivitis healed largely to the same extent in subjects rinsing twice ethanol
-
'Department of Periodontology, University, Malmö, Sweden, department of Microbiology.
Center for Oral Health
*Biosurface AB. §Decapinol, Biosurface AB, Malmö, Sweden.
Sciences, Lund
use; dental
plaque/
daily with 0.1% or 0.2% delmopinol as in subjects rinsing with Chlorhexidine 0.2%. Furthermore, a statistically significant dose-response effect was noted between the amount of plaque and gingivitis, and the dose of delmopinol used. Two weeks of rinsing with delmopinol in the absence of oral hygiene appeared to change the composition of the supragingival plaque to one predominated by coccoid cells.3 The mode of action of delmopinol is not known in detail. Current information shows that delmopinol has a low antimicrobial capacity relative to that of Chlorhexidine.4 The plaque reducing effect therefore seems to be related more to the physico-chemical interaction of delmopinol with tooth and bacterial surfaces. Significant in this respect might be that delmopinol is a surface active agent, which may inter-
Volume 63 Number 7
COLLAERT, EDWARDSSON, ATTSTROM, HASE, ÄSTRÖM, MOVERT day-14
I oral
day 0 hygiene period
\
day 14 delmopinol
placebo period
day HCl
619 28
0.2%|
Chlorhexidine 0 2%
O
professional toothcleaning
*
clinical measurements
salivary microbiology
Figure 2. Experimental design. Figure 1. Structural formula of delmopinol hydrochloride.
jects
fere with the adhesion forces between microbes and the
surroundings.1 Changes in the oral microflora could be beneficial if known pathogens and pathogens associated with gingival infection
reduced or absent. But when such alterations in the indigenous oral flora are induced by the use of chemotherapeutic agents for controlling plaque and gingivitis, attention should be paid to the possible emergence of opportunistic pathogens.5 Microorganisms present in the oral cavity usually establish a balance between those present in saliva and those adhering to shedding or non-shedding surfaces, depending on the affinity of the respective microorganisms to the oral surfaces.6 Saliva can also be considered a medium for the spread of microorganisms from one oral site to the are
other.7
The aim of this short-term study was to detect possible changes in the cultivable salivary microflora after rinsing with delmopinol. Plaque and gingivitis clinical status was monitored during the study to verify earlier obtained results. A comparison was made between rinsing with delmopinol hydrochloride 0.2% and Chlorhexidine digluconate 0.2%. The experimental model used attempted to imitate a clinical condition at which treatment of gingivitis is normally
initiated.
MATERIAL AND METHODS Test Solutions The placebo solution was an aqueous solution containing ethanol 2%, a herb flavor 0.02%, and sodium saccharine 0.01%. The active solutions were an unflavored aqueous 0.2% delmopinol HCl solution, sodium hydroxide pH 5.7, and 0.2% Chlorhexidine digluconate.11 =
Subjects
Forty-five healthy male volunteers (18 to 40 years), who had participated in a previous 6-week study identical to the present study design which monitored the effect of topical application of 3 concentrations of delmopinol HCl on the salivary microflora, plaque, and gingivitis,3 took part in this study. After informed consent was obtained, the sub'Hibutane
Dental, ICI-Pharma, Gothenburg, Sweden.
were admitted on the following basis: 1) complete dentition except for third molars and/or one premolar in each quadrant extracted on orthodontic indication; 2) no periodontal pockets exceeding 4 mm; 3) no crowns or bridges; 4) no buccal and large interproximal restorations; 5) no extensive exposed root surfaces; 6) no caries cavities; and 7) no antibiotic treatment during the preceding 6 months. Analyses were based on results from 39 participants (1 participant withdrew and 5 others were excluded because of non-compliance with the study protocol). All were excluded from analyses before breaking the randomization code.
Study Design
This 6-week study was unsupervised, computer randomized, and performed double-blind with parallel groups (Fig. 2). During the first 2 weeks all participants underwent 3 professional tooth cleanings and instructions in oral hygiene procedures, which resulted in clinically healthy gingiva. On day 0, start of the experimental gingivitis period, the subjects received thorough professional tooth cleaning. During the following 2 weeks they abstained from all mechanical oral hygiene measures, but carefully applied 2 ml of a placebo solution onto their teeth with a soft paint brush twice daily (morning and evening). The placebo was applied because, as mentioned above, the subjects were part of a larger study evaluating the use of topical application of delmopinol.3 At the end of the placebo period all subjects exhibited a mild gingivitis. On day 14 after receiving professional tooth cleaning, the subjects were randomly assigned to 1 of 2 treatment groups. During the next 2 weeks they continued to refrain from all mechanical oral hygiene procedures. Instead they rinsed twice daily (morning and evening) with 10 ml of 0.2% delmopinol (18 subjects) or 0.2% Chlorhexidine (21 subjects). The subjects were urged not to change their dietary habits for the duration of the study, and not to eat or drink for 30 minutes after rinsing. The objective of using this short-term experimental model was to establish a confirmed gingivitis baseline for the measurement of therapeutic effect. Such a baseline ought to be more clinically relevant than clinically healthy gingiva as the starting point for evaluation of plaque control agents. Gingival inflammation assessment. Gingival crevicular fluid (GCF) was obtained at the end of each 2-week period
620
J Periodontol July 1992
DELMOPINOL AS A PLAQUE CONTROL AGENT
from 4 teeth out of the
incisors, canine, and premolare of
maxillary quadrant. All supragingival plaque was carefully removed without disturbing the gingival margin. The one
buccal surface was dried with air directed from the buccal gingival fold towards the tooth. A 1 mm wide absorbent paper strip was gently inserted at the mesio-buccal aspect of the tooth into the gingival crevice until resistance was felt and left in place for 3 minutes.8 The strips were immediately stained with ninhydrin. The height of the stained area was subsequently measured. The gingivitis index (GI) was recorded as presence or absence of bleeding after gentle probing with a WHO probe to the bottom of the gingival crevice at 6 sites around each tooth: mesio-buccal, buccal, distobuccal, mesio-lingual, lingual, and disto-lingual.9 The GI was measured around all teeth (incisors, canines, premolars, and molars) in the 2 quadrants contralateral to the side used for GCF sampling. The GI was expressed as the percentage of bleeding sites. Plaque assessment. Supragingival plaque was disclosed with erythrosine. The plaque index (PI) according to Turesky10 was scored for the same teeth as those used for the GI. After staining, the subject rinsed with 10 ml of tap water for 10 seconds. Periodic identical color slides for planimetrie analyses11 were taken of the disclosed plaque on canines and premolars used for GI and PI recordings. The slides were enlarged 6.5 times and projected onto white paper. The tooth surface covered with stainable plaque was contoured and measured through computerized digitization. In 50% of the participants, the GCF-flow samples were obtained from the first quadrant, while the GI, PI, and plaque extension were measured in the second and third quadrant. In the other half of the subjects the opposite quadrant in the same jaw was used. The allocation of the subjects to one of the two partial mouth registration groups was randomized so that they were equally represented in the two treatment groups. Extrinsic tooth-staining assessment. At the end of both the placebo and the delmopinol treatment period, a periodic standardized photograph was taken before plaque disclosure of the same quadrants used for the plaque extension measurements. The photographs were compared to each other, with the placebo photo considered baseline. Differences in extrinsic tooth staining were scored as follows: 0 no difference in the degree of staining; 1 more slightly staining only detectable when photographs were compared; 2 visibly more staining, clinically detectable; and 3 profound dark brown staining (esthetically compromising). Because of a technical failure, data from 8 subjects are
Table 1: Culture
Media, Cultivation Procedures, and Identification
Medium and Cultivation Procedures
Identification
Brucella blood agar (BBL) with 5% human erythrocytes, 0.5% laked human erythrocytes and 5 mg/1 menadione (7 days
Total number of anaerobically cultivable
anaerobically) Blood agar12(5-7 days aerobically)
N2) C02 agar* (4 days aerobically, poured plate technique) Staphylococcus medium 110* (2 days aerobically) Mac Conkey agar* (4 days aerobically) Sabouraud Dextrose agar* (4 days aerobically) MSB-agar13 (days
in 5%
Selective lactobacilli
and
microorganisms
Total number of aerobically cultivable
microorganisms streptococci
Mutans
Lactobacilli Salt-tolerant
staphylococci Enteric bacteria Yeasts
*Difco, Detroit, MI.
collected at the end of the placebo and active treatment periods. The two samples were obtained from each participant at the same time of the day. The salivary flow was stimulated by tongue and cheek motion. Saliva (2 to 3 ml) was collected in a sterile vial with glass beads. The samples were processed within 1 hour after collection. The vial was placed in a Vortex mixer for 30 seconds. The culture media listed in Table 1 were inoculated from appropriate dilutions of the homogenized sample. Colony forming units (CFU) on Brucella blood agar resembling black pigmenting PorphyromonasIPrevotella spp.14,15 and Fusobacterium spp.16 were Gram stained and their presence was recorded. CFU with doubtful morphology on MSB-agar were isolated and their capacity to ferment mannitol was studied. The salttolerant Gram-positive cocci growing on 110 agar were tested for the presence of catalase and DNase,# the latter test dividing the isolates into those resembling Staphylococcus aureus and S. epidermidis. CFU of various morphology growing on MacConkey agar identified as Gram-negative rods were considered enteric bacteria. CFU on Sabouraudagar being blastospores in Gram stained smears were considered to be yeasts.
=
=
=
missing.
=
The GCF samples, GI, PI, and identical color-slides for plaque extension measurements and extrinsic tooth staining measurements were obtained by the same investigator on day 14 and day 28. There was no statistically significant difference for any of the parameters studied between the placebo period of the two treatment groups. Salivary microbiological evaluation. Saliva samples were
Adverse Experiences The subjects were examined at each visit for adverse reactions in the oral cavity and were questioned about the occurrence of any adverse experiences prior to the clinical examination. After the clinical examination at the last visit the subjects were asked if they had experienced an anesthetic sensation in any part of their oral mucosa after rinsing and how long they thought it had lasted.
Statistical Analyses All statistical analyses were performed with non-parametric methods. The subject was the unit of analysis. Wilcoxon's *Difco, Detroit,
MI.
Volume 63 Number 7 S I S
COLLAERT, EDWARDSSON, ATTSTROM, HASE, ÄSTRÖM, MOVERT
621
%
mm
60
0.6
I
A
0.5
c
50
u
L
40
0.4
R E
30
V
I
c
L A R
0.2
20
0.1
10
F L U I D
DELMOPINOL HCl 0.2%
[ZD PLACEBO
DELMOPINOL HCl 0.2%
CHLORHEXIDINE 0.2%
CHLORHEXIDINE 0.2%
ZZ PLACEBO
ül ACTIVE
Figure 3. Mean GCF-values ( + SE) after 14 days ofplacebo, and 14 days of delmopinol HCl 0.2% and Chlorhexidine 0.2% rinsing, respectively. Significant difference between placebo and active treatment is indicated. *P
