Nhakhoa HappySmilesLand 25A1 đường Trần Duy Hưng & 138A phố Khương Thượng, Hà Nội - Chuyên làm răng Implant & răng sứ Cercon, Katana, eMax - Hotline: 0982.0982.88
NGHIÊN CỨU VỀ VI KHUẨN TRÊN MIỆNG NGƯỜI KHỎE MẠNH

JCM

 

Journal of Clinical Microbiology, November 2005, p. 5721-5732, Vol. 43, No. 11
0095-1137/05/$08.00+0     doi:10.1128/JCM.43.11.5721-5732.2005
All Rights Reserved.

Defining the Normal Bacterial Flora of the Oral Cavity

Xác định các chủng vi khuẩn bình thường trong khoang miệng

Jørn A. Aas,1,2* Bruce J. Paster,1,3 Lauren N. Stokes,1 Ingar Olsen,2 and Floyd E. Dewhirst1

Department of Molecular Genetics, The Forsyth Institute,1 Faculty of Dentistry, University of Oslo, Oslo, Norway,2 Department of Oral and Developmental Biology, Harvard School of Dental Medicine, Boston, Massachusetts3

Received 10 June 2005/ Returned for modification 2 August 2005/ Accepted 12 August 2005


    ABSTRACT  TÓM TẮT
 Top
 Abstract
 Introduction
 Materials and Methods
 Results and Discussion
 References
 
More than 700 bacterial species or phylotypes, of which over 50% have not been cultivated, have been detected in the oral cavity. Our purposes were (i) to utilize culture-independent molecular techniques to extend our knowledge on the breadth of bacterial diversity in the healthy human oral cavity, including not-yet-cultivated bacteria species, and (ii) to determine the site and subject specificity of bacterial colonization. Nine sites from five clinically healthy subjects were analyzed. Sites included tongue dorsum, lateral sides of tongue, buccal epithelium, hard palate, soft palate, supragingival plaque of tooth surfaces, subgingival plaque, maxillary anterior vestibule, and tonsils. 16S rRNA genes from sample DNA were amplified, cloned, and transformed into Escherichia coli. Sequences of 16S rRNA genes were used to determine species identity or closest relatives. In 2,589 clones, 141 predominant species were detected, of which over 60% have not been cultivated. Thirteen new phylotypes were identified. Species common to all sites belonged to the genera Gemella, Granulicatella, Streptococcus, and Veillonella. While some species were subject specific and detected in most sites, other species were site specific. Most sites possessed 20 to 30 different predominant species, and the number of predominant species from all nine sites per individual ranged from 34 to 72. Species typically associated with periodontitis and caries were not detected. There is a distinctive predominant bacterial flora of the healthy oral cavity that is highly diverse and site and subject specific. It is important to fully define the human microflora of the healthy oral cavity before we can understand the role of bacteria in oral disease.

Xác định được hơn 700 chủng vi khuẩn, trong đó hơn 50% chưa thể nuôi cấy được, đã được tìm thấy trong khoang miệng. Mục đích của chúng tôi là ứng dụng kỹ thuật nuôi cấy phân tử tự do nhằm tăng thêm hiểu biết độ lớn và tính đa dạng của vi khuẩn trong khoang miệng người, bao gồm cả các chủng vi khuẩn chưa thể nuôi cấy được và để xác định các vị trí ưu thế trong miệng đối với từng loại vi khuẩn. Chín vị trí trong khoang miệng được phân tích từ 5 người khoẻ mạnh được đưa vào nghiên cứu. Các vị trí đó bao gồm bề mặt lưỡi, mặt bên của lưỡi, biểu mô má, vòm miệng cứng, vòm miệng mềm, mảng bám thân răng, mảng bám dưới lợi và Amidal. Gen 16S rARN từ mẫu được khuếch đại, nhân bản và cấy chuyển vào E.coli. Lần lượt các gen 16S rARN được dùng để định dạng các chủng hoặc các nhóm có liên quan. Trong 2589 mẫu nhân bản, 141 chủng có ưu thế được phát hiện, trong số đó có 60% chưa thể cấy được. 13 dưới loài được nhận dạng. Các chủng từ các vị trí thông thường thường thuộc nhóm Gemella, Granulicatella, StreptococcusVeillonella                                         n
    INTRODUCTION
 Top
 Abstract
 Introduction
 Materials and Methods
 Results and Discussion
 References
 
The oral cavity is comprised of many surfaces, each coated with a plethora of bacteria, the proverbial bacterial biofilm. Some of these bacteria have been implicated in oral diseases such as caries and periodontitis, which are among the most common bacterial infections in humans. For example, it has been estimated that at least 35% of dentate U.S. adults aged 30 to 90 years have periodontitis (1). In addition, specific oral bacterial species have been implicated in several systemic diseases, such as bacterial endocarditis (4), aspiration pneumonia (26), osteomyelitis in children (8), preterm low birth weight (6, 20), and cardiovascular disease (2, 34). Surprisingly, little is known about the microflora of the healthy oral cavity.

By using culture-independent molecular methods, we previously detected over 500 species or phylotypes in subgingival plaque of healthy subjects and subjects with periodontal diseases (21), necrotizing ulcerative periodontitis in human immunodeficiency virus-positive subjects (23), dental plaque in children with rampant caries (3), noma (22), and on the tongue dorsum of subjects with and without halitosis (15). Other investigators have used similar techniques to determine the bacterial diversity of saliva (25), subgingival plaque of a subject with gingivitis (16), and dentoalveolar abscesses (10, 29). Over half of the species detected have not yet been cultivated. Data from these studies have implicated specific species or phylotypes in a variety of diseases and oral infections, but still only limited information is available on species associated with health. Recently, Mager et al. (19) demonstrated significant differences in the bacterial profiles of 40 oral cultivable species on soft and hard tissues in healthy subjects. They also found that the profiles of the soft tissues were more similar to each other than those of supragingival and subgingival plaques.

Our purposes were as follows: (i) to utilize culture-independent molecular techniques to extend our knowledge on the breadth of bacterial diversity in the healthy human oral cavity, including not-yet-cultivated phylotypes, and (ii) to determine the site and subject specificity of bacterial colonization.


    MATERIALS AND METHODS
 Top
 Abstract
 Introduction
 Materials and Methods
 Results and Discussion
 References
 
Subjects. Five subjects, representing both genders, ranging in age from 23 to 55 and with no clinical signs of oral mucosal disease were included in the study. Subjects did not suffer from severe halitosis. The periodontia were healthy in that all periodontal pockets were less than 3 mm deep with no redness or inflammation of the gums. Subjects did not have active white spot lesions or caries on the teeth. Our results were consistent with these clinical observations in that species typically found in caries subjects were not detected (3). The subjects had not used antibiotics for the last 6 months.

Sample collection. Samples from the following nine sites were analyzed for each subject: dorsum of the tongue, lateral sides of the tongue, buccal fold, hard palate, soft palate, labial gingiva and tonsils of soft tissue surfaces, and supragingival and subgingival plaques from tooth surfaces. Microbiological samples of supragingival and subgingival plaque samples were taken with a sterile Gracey curette. Other samples were collected with sterile swab brushes.

Sample lysis. Plaque samples were directly suspended in 50 µl of 50 mM Tris buffer (pH 7.6), 1 mM EDTA, pH 8, and 0.5% Tween 20. Proteinase K (200 µg/ml; Roche Applied Science, Indianapolis, IN) was added to the mixture. The samples were then heated at 55°C for 2 h. Proteinase K was inactivated by heating at 95°C for 5 min. Detection of species is dependent upon obtaining DNA that can be amplified. Thus, a difficult-to-lyse bacterium may not be detected. However, by using our lysis technique, we were able to detect many hard-to-lyse species, such as species of Actinomyces and Streptococcus.

Amplification of 16S rRNA genes by PCR and purification of PCR products. The 16S rRNA genes were amplified under standardized conditions using a universal primer set (forward primer, 5'-GAG AGT TTG ATY MTG GCT CAG-3'; reverse primer, 5'-GAA GGA GGT GWT CCA RCC GCA-3') (21). Primers were synthesized commercially (Operon Technologies, Alameda, CA). The PCR primers do not necessarily include all bacterial species. Nevertheless, a wide range of phylogenetic types were obtained in this study and our previous studies by using this universal primer set (3, 15, 21-23). PCR was performed in thin-walled tubes with a GeneAmp PCR system 9700 (ABI, Foster City, CA). One microliter of the lysed sample was added to a reaction mixture (final volume, 50 µl) containing 20 pmol of each primer, 40 nmol of deoxynucleoside triphosphates, and 1 U of Platinum Taq polymerase (Invitrogen, San Diego, CA). In a hot-start protocol, the samples were preheated at 95°C for 4 min, followed by amplification under the following conditions: denaturation at 95°C for 45 s, annealing at 60°C for 45 s, and elongation at 72° for 1.5 min, with an additional 15 s for each cycle. A total of 30 cycles were performed; this was followed by a final elongation step at 72°C for 15 min. The results of the PCR amplification were examined by electrophoresis in a 1% agarose gel. DNA was stained with ethidium bromide and visualized under short-wavelength UV light.

Cloning procedures. Cloning of PCR-amplified DNA was performed with the TOPO TA cloning kit (Invitrogen) according to the instructions of the manufacturer. Transformation was done with competent Escherichia coli TOP10 cells provided by the manufacturer. The transformed cells were then plated onto Luria-Bertani agar plates supplemented with kanamycin (50 µg/ml), and the plates were incubated overnight at 37°C. Each colony was placed into 40 µl of 10 mM Tris. Correct sizes of the inserts were determined in a PCR with an M13 (–20) forward primer and an M13 reverse primer (Invitrogen). Prior to sequencing of the fragments, the PCR-amplified 16S rRNA fragments were purified and concentrated with Microcon 100 (Amicon, Bedford, MA), followed by use of the QIAquick PCR purification kit (QIAGEN, Valencia, CA).

16S rRNA gene sequencing. Purified PCR-amplified 16S rRNA inserts were sequenced using an ABI Prism cycle sequencing kit (BigDye terminator cycle sequencing kit with AmpliTaq DNA polymerase FS, GeneAmp PCR system 9700; ABI). The primers used for sequencing have been described previously (21). Quarter dye chemistry was used with 80 µM primers and 1.5 µl of PCR product in a final volume of 20 µl. Cycle sequencing was performed with a GeneAmp PCR system 9700 (ABI) with 25 cycles of denaturation at 96°C for 10 s, annealing at 55° for 5 s, and extension at 60°C for 4 min. The sequencing reactions were run on an ABI 3100 DNA sequencer (ABI).

16S rRNA gene sequencing and data analysis of unrecognized inserts. A total of 2,589 clones with an insert of the correct size of approximately 1,500 bases were analyzed. The number of clones per subject that were sequenced ranged from 42 to 69, with an average of 57.5 and a standard deviation of 7.1. A sequence of approximately 500 bases was obtained first to determine identity or approximate phylogenetic position. Full sequences of about 1,500 bases were obtained by using five to six additional sequencing primers (15) for those species deemed novel. For identification of closest relatives, the sequences of the unrecognized inserts were compared to the 16S rRNA sequences of over 10,000 microorganisms in our database and over 100,000 sequences in the Ribosomal Database Project (7) and the GenBank databases. Our cutoff for species differentiation was 2%, or approximately 30 bases for a full sequence. The similarity matrices were corrected for multiple base changes at single positions by the method of Jukes and Cantor (13). Similarity matrices were constructed from the aligned sequences by using only those sequence positions for which data were available for 90% of the strains tested. Phylogenetic trees were constructed by the neighbor-joining method of Saitou and Nei (24). TREECON, a software package for the Microsoft Windows environment, was used for the construction and drawing of evolutionary trees (28). We are aware of the potential creation of 16S rRNA chimera molecules assembled during the PCR (18). The percentage of chimeric inserts in 16S rRNA gene libraries ranged from 1 to 15%. Chimeric sequences were identified by using the Chimera Check program in the Ribosomal Database Project, by treeing analysis, or by base signature analysis. Species identification of chimeras was obtained, but the sequences were not examined for phylogenetic analysis.

Nucleotide sequence accession numbers. The complete 16S rRNA gene sequences of clones representing novel phylotypes defined in this study, sequences of known species not previously reported, and published sequences are available for electronic retrieval from the EMBL, GenBank, and DDBJ nucleotide sequence databases under the accession numbers shown in Fig. 1 to 9.


View larger version (59K):
[in this window]
[in a new window]
 
FIG. 1. Bacterial profiles of the buccal epithelium of healthy subjects. Distribution and levels of bacterial species/phylotypes among five subjects are shown by the columns of boxes to the right of the tree as either not detected (clear box), <15% of the total number of clones assayed (shaded box), or ≥15% of the total number of clones assayed (darkened box). The 15% was chosen arbitrarily. GenBank accession numbers are provided. Marker bar represents a 5% difference in nucleotide sequences.

 

 


View larger version (67K):
[in this window]
[in a new window]
 
FIG. 9. Bacterial profiles of the subgingival plaque of healthy subjects. Distribution and levels of bacterial species/phylotypes among five subjects are as described for Fig. 1. Novel phylotypes identified in this study are indicated in bold. GenBank accession numbers are provided. Marker bar represents a 5% difference in nucleotide sequences.

 

 

    RESULTS AND DISCUSSION
 Top
 Abstract
 Introduction
 Materials and Methods
 Results and Discussion
 References

 
It has long been known that oral bacteria preferentially colonize different surfaces in the oral cavity as a result of specific adhesins on the bacterial surface binding to complementary specific receptors on a given oral surface (11, 12). Indeed, the profiles of 40 cultivable bacterial species differed markedly on different oral soft tissue surfaces, saliva, and supragingival and subgingival plaques from healthy subjects (19). The purpose of this study was to define the predominant bacterial flora of the healthy oral cavity by identifying and comparing the cultivable and the not-yet-cultivated bacterial species on different soft tissues and supra- and subgingival plaques.

Based on the analysis of 2,589 16S rRNA clones, the bacterial diversity of the microflora from nine different sites of five clinically healthy subjects was striking—a total of 141 different bacterial taxa representing six different bacterial phyla were detected. The six phyla included the Firmicutes (previously referred to as the low-G+C gram positives, such as species of Streptococcus, Gemella, Eubacterium, Selenomonas, Veillonella, and related ones), the Actinobacteria (previously referred to as the high-G+C gram positives, such as species of Actinomyces, Atopobium, Rothia, and related ones), the Proteobacteria (e.g., species of Neisseria, Eikenella, Campylobacter, and related ones), the Bacteroidetes (e.g., species of Porphyromonas, Prevotella, Capnocytophaga, and related ones), the Fusobacteria (e.g., species of Fusobacterium and Leptotrichia), and the TM7 phylum, for which there are no cultivable representatives. As determined in our previous studies using culture-independent molecular techniques (15, 21, 22), over 60% of the bacterial flora was represented by not-yet-cultivated phylotypes. Thirteen new phylotypes (see Fig. 2 through 9) were identified for the first time in this study. In a comparable ongoing study of caries in permanent teeth, using the same techniques, we detected 224 species in the analysis of 1,275 16S rRNA clones with about 60% as not-yet-cultivable phylotypes (J. A. Aas, S. R. Dardis, A. L. Griffen, L. N. Stokes, A. M. Lee, I. Olsen, F. E. Dewhirst, E. J. Leys, and B. J. Paster. J. Dent. Res. 84:abstract 2805, 2005). Consequently, it is important to identify both the cultivable and not-yet-cultivated bacterial flora in a given environment before we can ascribe association to health or disease status. There is no evidence to suggest that the not-yet-cultivated segment is any less important than the cultivable segment.


View larger version (28K):
[in this window]
[in a new window]
 
FIG. 2. Bacterial profiles of the maxillary anterior vestibule of healthy subjects. Distribution and levels of bacterial species/phylotypes among five subjects are as described for Fig. 1. Novel phylotypes identified in this study are indicated in bold. GenBank accession numbers are provided. Marker bar represents a 5% difference in nucleotide sequences.

 

 
Bacterial profiles for each site tested for each subject are shown in nine phylogenetic trees (see Fig. 1 through 9). In these dendrograms, the distribution of bacterial species or phylotypes in each subject can be observed at a glance. For example, it is clear that Streptococcus mitis, S. mitis bv. 2, and Gemella hemolysans are the predominant species of the buccal epithelium (Fig. 1). Each of these species was detected in most or all of the subjects and represented ≥15% of the total number of assayed clones (Fig. 1). Similarly, the predominant bacterial flora for the other eight oral sites was examined. In the maxillary anterior vestibule, S. mitis, Granulicatella spp., and Gemella spp. predominated (Fig. 2). On the tongue dorsum, several species of Streptococcus, such as S. mitis, Streptococcus australis, Streptococcus parasanguinis, Streptococcus salivarius, Streptococcus sp. clone FP015, and Streptococcus sp. clone FN051, Granulicatella adiacens, and Veillonella spp. were the predominant species (Fig. 3). On the lateral tongue surface (Fig. 4), S. mitis, S. mitis bv. 2, Streptococcus sp. clone DP009, Streptococcus sp. clone FN051, S. australis, G. adiacens, G. hemolysans, and Veillonella spp. predominated. It is interesting that there were considerable differences in the bacterial profiles of the tongue dorsum and the lateral tongue surface. For example, S. mitis bv. 2 was well represented at the lateral side of the tongue but not detected on the tongue dorsum. Conversely, S.parasanguinis strain 85-81 was present on the tongue dorsum, but was absent on the lateral side. This was not surprising, because these surfaces are known to be different in ultrastructure and function. For instance, the lateral side of the tongue has a smooth nonkeratinized surface, in contrast to the dorsum of the tongue, which is a keratinized, highly papillated surface with a large surface area and underlying serous glands. These anatomic differences likely influence the ecology of these habitats and create microbial environmental differences.


View larger version (63K):
[in this window]
[in a new window]
 
FIG. 3. Bacterial profiles of the tongue dorsum of healthy subjects. Distribution and levels of bacterial species/phylotypes among five subjects are as described for Fig. 1. Novel phylotypes identified in this study are indicated in bold. GenBank accession numbers are provided. Marker bar represents a 5% difference in nucleotide sequences.

 

 


View larger version (59K):
[in this window]
[in a new window]
 
FIG. 4. Bacterial profiles of the lateral tongue surface of healthy subjects. Distribution and levels of bacterial species/phylotypes among five subjects are as described for Fig. 1. Novel phylotypes identified in this study are indicated in bold. GenBank accession numbers are provided. Marker bar represents a 5% difference in nucleotide sequences.

 

 
On the hard palate, the predominant bacterial species included S. mitis, S. mitis biovar 2, Streptococcus sp. clone FN051, Streptococcus infantis, Granulicatella elegans, G. hemolysans, and Neisseria subflava (Fig. 5). On the soft palate, S. mitis, other cultivable and not-yet-cultivable species of Streptococcus, G. adiacens and G. hemolysans were predominant (Fig. 6). The tonsil bacterial flora (Fig. 7) was rather diverse—some subjects had Prevotella and Porphyromonas spp. (subjects 1 and 5), and others had Firmicutes species (subjects 2, 3, and 4). On the tooth surface, several species of Streptococcus, including Streptococcus sp. clone EK048, S. sanguinis, and S. gordonii, and Rothia dentocariosa, G. hemolysans, G. adiacens, Actinomyces sp. clone BL008, and Abiotrophia defectiva were often detected (Fig. 8). Finally, in subgingival plaque, several species of Streptococcus and Gemella were often found (Fig. 9).


View larger version (68K):
[in this window]
[in a new window]
 
FIG. 5. Bacterial profiles of the hard palate of healthy subjects. Distribution and levels of bacterial species/phylotypes among five subjects are as described for Fig. 1. Novel phylotypes identified in this study are indicated in bold. GenBank accession numbers are provided. Marker bar represents a 5% difference in nucleotide sequences.

 

 


View larger version (67K):
[in this window]
[in a new window]
 
FIG. 6. Bacterial profiles of the soft palate of healthy subjects. Distribution and levels of bacterial species/phylotypes among five subjects are as described for Fig. 1. Novel phylotypes identified in this study are indicated in bold. GenBank accession numbers are provided. Marker bar represents a 5% difference in nucleotide sequences.

 

 


View larger version (68K):
[in this window]
[in a new window]
 
FIG. 7. Bacterial profiles of the tonsils of healthy subjects. Distribution and levels of bacterial species/phylotypes among five subjects are as described for Fig. 1. Novel phylotypes identified in this study are indicated in bold. GenBank accession numbers are provided. Marker bar represents a 5% difference in nucleotide sequences.

 

 


View larger version (62K):
[in this window]
[in a new window]
 
FIG. 8. Bacterial profiles of the tooth surfaces of healthy subjects. Distribution and levels of bacterial species/phylotypes among five subjects are as described for Fig. 1. Novel phylotypes identified in this study are indicated in bold. GenBank accession numbers are provided. Marker bar represents a 5% difference in nucleotide sequences.

 

 
The number of cultivable and not-yet-cultivable species detected in each subject for each site is also shown in Fig. 1 through 9. For example, in Fig. 1, only four species (S. mitis, S.mitis bv. 2, A. defectiva, and G. hemolysans) were detected on the buccal epithelium in subject 5. For convenience, the number of bacterial species detected in each oral site for each subject is summarized in Table 1. Surprisingly, the highest number of different species was found on the tonsils in subject 1, with 28 predominant species; collectively, 57 species were detected on the tonsils (Table 1; Fig. 7). In contrast, the maxillary anterior vestibule had the lowest diversity of the bacterial flora compared with the other sites, with as few as three to nine species detected among the subjects (Table 1; Fig. 2). A common question asked is how many bacterial species are present in the oral cavity of a single individual. The last column of Table 1 indicates the number of predominant species in each healthy individual from all nine different sites; thus, the numbers ranged from a low of 34 i

Nguồn http://jcm.asm.org/cgi/

Các bài viết khác
HSL TEAM
Đăng ký khám / Booking
Họ tên: (*)
Điện thoại: (*)
Email: (*)
Địa điểm khám:
Thời gian khám:
Pick a Date


Lời nhắn - Thư gửi HSL / Message
Dịch vụ / Services
Văn bằng chứng chỉ / Certificates
Bài viết mới nhất
Hỏi đáp nha khoa
Hỏi: Chào bác sĩ Tôi năm nay 22 tuổi, hiện tôi bị răng thưa hàm ...
Đáp: Chào bác sĩ Tôi năm nay 22 tuổi, hiện tôi bị răng thưa hàm trên, vị trí là giữa răng cửa bê...
Hỏi: Chào bác sĩ.em năm nay 20 tuổi.em bị hở hai răng cửa.em sử d...
Đáp: Chào bác sĩ.em năm nay 20 tuổi.em bị hở hai răng cửa.em sử dụng phương pháp chụp sứ 2 răng được không và giá ...
Hỏi: Chao bac si chau nam any 16 tuoi bac si co the cho chau biet...
Đáp: Bạn tham khảo thêm bài này nhé: http://nhakhoahappysmilesland.com.vn/0605000/nan-chinh-rang.html

và b...
Hỏi: Chào Bác sĩ, Cháu năm nay 24t. Cháu hơi mất tự tin vì 2 răng...
Đáp: Chào Bác sĩ, Cháu năm nay 24t. Cháu hơi mất tự tin vì 2 răng cửa hơi hô. Vậy bây giờ cháu muốn đ...
Hỏi: chào bác sỹ! cháu năm nay 20 tuổi, cháu có một cái răng bị s...
Đáp: chào bác sỹ! cháu năm nay 20 tuổi, cháu có một cái răng bị sâu. dạo trước cháu k để ý đế...
Hỏi: thưa bác sĩ đợt vừa rồi cháu mới làm răng thẩm mĩ .cì 2 cái ...
Đáp: thưa bác sĩ đợt vừa rồi cháu mới làm răng thẩm mĩ .cì 2 cái răng của của cháu bị to.nhưng sau khi làm...
Hỏi: Em nam nay 20t,rang cua em moc khong deu nhau va moc chen va...
Đáp: Em nam nay 20t,rang cua em moc khong deu nhau va moc chen vao nhau.Em muon di nieng rang.Em co bj nho rang khog,chj phj khoang bao nhjeu cho ca 2 ham,...
Hỏi: Thưa bác sĩ,cháu bị hô 2 răng cửa hàm trên. Cháu dự ...
Đáp: Thưa bác sĩ,cháu bị hô 2 răng cửa hàm trên. Cháu dự định nắn chỉnh nha bằng răng sứ. Xin bác sĩ cho cháu biết chí ...
Hỏi: cháu năm nay 19t chau muon nắn chỉnh răng thi chi phí khoảng...
Đáp: cháu năm nay 19t chau muon nắn chỉnh răng thi chi phí khoảng bnhiêu ạ

Chào bạn! Cảm ơn ...

Hỏi: Chào bác sĩ, nha khoa chúng ta có dịch vụ lắp ghép răng khển...
Đáp: Chào bác sĩ, nha khoa chúng ta có dịch vụ lắp ghép răng khểnh không ạ! Nếu có thì cho tôi...
Hỏi: Chào Bác Sĩ, cháu có một cái răng hồi bé lên bé hơn các răng...
Đáp: Chào Bác Sĩ, cháu có một cái răng hồi bé lên bé hơn các răng bình thường. Giờ ch&...
Hỏi: chào bác sĩ ! e có 2 chiếc răng khểnh, nhưng bây giờ e muốn ...
Đáp: chào bác sĩ ! e có 2 chiếc răng khểnh, nhưng bây giờ e muốn nhổ đi 1 chiếc bên trái. tại nó hơi mất thẩ...
Hỏi: thua bac si.em nam nay 24 tuoi.em co 2 rang cua hoi ho nen m...
Đáp: thua bac si.em nam nay 24 tuoi.em co 2 rang cua hoi ho nen muon nan chinh lai.vay bac si cho em hoi chi phi nan chinh la bao nhieu a.cam on bs.
...
Hỏi: Thua bac si chau nam nay 22 tuoi, chau bi vau rang cua.xin b...
Đáp: Thua bac si chau nam nay 22 tuoi, chau bi vau rang cua.xin bac si tu van giup chau cac chinh rang nao la tot nhat.Va chi phi khoang bao nhieu khong a....
Hỏi: Chào bác sĩ, hiện nay con tôi 5 tuổi đã mọc đủ 20 răng (10 t...
Đáp: Chào bác sĩ, hiện nay con tôi 5 tuổi đã mọc đủ 20 răng (10 trên, 10 dưới). Hiện trạng răng con tôi bị sâu...
NHA KHOA HAPPYSMILESLAND
  25A1 Trần Duy Hưng - Cầu Giấy & 138A Khương Thượng - Đống Đa - Hà Nội - Hotline: 0982.0982.88
Email: NhakhoaHSL@gmail.com - Youtube Chanel "BacsiThang" - Facebook "BacsiThang NguyenCao"
Copyright © 2011 NhakhoaHappySmilesLand.vn - Thiết kế và phát triển bởi Ovem!Software