OBJECTIVES: Dental caries is a widespread multifactorial disease, caused by the demineralization of hard dental tissues. Susceptibility to dental caries is partially genetically conditioned; this study was aimed at finding an association of selected single nucleotide polymorphisms (SNPs) in genes encoding proteins involved in amelogenesis with this disease in children. MATERIALS AND METHODS: In this case-control study, 15 SNPs in ALOX15, AMBN, AMELX, KLK4, TFIP11, and TUFT1 genes were analyzed in 150 children with primary dentition and 611 children with permanent teeth with/without dental caries from the European Longitudinal Study of Pregnancy and Childhood (ELSPAC) cohort. RESULTS: Dental caries in primary dentition was associated with SNPs in AMELX (rs17878486) and KLK4 (rs198968, rs2242670), and dental caries in permanent dentition with SNPs in AMELX (rs17878486) and KLK4 (rs2235091, rs2242670, rs2978642), (p ≤ 0.05). No significant differences between cases and controls were observed in the allele or genotype frequencies of any of the selected SNPs in ALOX15, AMBN, TFIP11, and TUFT1 genes (p > 0.05). Some KLK4 haplotypes were associated with dental caries in permanent dentition (p ≤ 0.05). CONCLUSIONS: Based on this study, we found that although the SNPs in AMELX and KLK4 are localized in intronic regions and their functional significance has not yet been determined, they are associated with susceptibility to dental caries in children. CLINICAL RELEVANCE: AMELX and KLK4 variants could be considered in the risk assessment of dental caries, especially in permanent dentition, in the European Caucasian population.
- MeSH
- amelogeneze * genetika MeSH
- amelogenin genetika MeSH
- dítě MeSH
- lidé MeSH
- longitudinální studie MeSH
- studie případů a kontrol MeSH
- zubní kaz * genetika epidemiologie MeSH
- Check Tag
- dítě MeSH
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
Úvod a cíl: Apikální periodontitida (AP) je zánětlivé onemocnění zubních periradikulárních tkání vyvolané bakteriální infekcí. Ke stanovení bakterií osídlujících kořenové kanálky jsou využívány různé metodické přístupy, analýza bakteriomu celého kořenového systému zubu postiženého AP je však zatím stále výzvou. Cílem naší přehledové práce bylo vytvořit literární rešerši zaměřenou na postupy pro odběr vzorku a metodiku pro analýzu bakteriomu a poté navrhnout vhodný metodický přístup k účelu studia etiopatogeneze tohoto onemocnění. Metodika: Po vyhledávání v databázi PubMed jsme do rešerše vybrali pouze publikační výstupy typu původní práce, ve kterých byla analyzována bakteriální DNA lidských zubů. Výsledky: Metodicky se studie mezi sebou velmi liší, a to jak způsobem odběru vzorku, izolací DNA, tak i samotnou analýzou bakteriální DNA. Častým způsobem odběru vzorku je využití sterilních endodontických papírových čepů. Tento způsob odběru vzorku je sice vhodný v rámci klinické praxe, avšak pro komplexní analýzu prostředí kořenového systému je považován za nedostatečný, a to kvůli samotné morfologii zubu a přítomnosti ramifikací. Jiným způsobem odběru vzorku je resekce kořenového hrotu pomocí sterilních fréz a následné rozemletí apexu nebo provedení stěru sterilními endodontickými papírovými čepy. Ke stanovení bakteriomu je tak využívána pouze apikální část zubu, tudíž bakterie, které kolonizují koronární část zubu a podílejí se na etiopatogenezi onemocnění, nemohou být analyzovány. V recentních studiích je využívána metoda, při které je celý extrahovaný zub postižený AP nadrcen na jemný homogenní prach pomocí kryogenního mletí. Z prachu nadrceného zubu je možné stanovit komplexní bakteriom kořenového systému i dřeňové dutiny, a proto se tento způsob z pohledu přípravy vzorku pro experimentální studii jeví jako optimální. Nejčastěji je při izolaci DNA využívána efektivní kolonková metoda různými purifikačními soupravami a k následné analýze DNA slouží většinou metodiky založené na principu polymerázové řetězové reakce. Sekvenování variabilních oblastí genu pro 16S rRNA je v dnešní době již zlatým standardem pro kategorizaci bakterií a charakterizaci bakteriálních komunit. Závěr: Ke studiu bakteriomu AP se jeví jako nejvhodnější použít vzorky extrahovaných zubů a bezprostředně je zamrazit bez dalších preanalytických kroků. Nadrcený zub je při dodržení sterilních podmínek při kryogenním mletí vhodnou matricí pro izolaci mikrobiální DNA komerčně dostupnými kity. V současné době je pro stanovení bakteriomu a získání informace o relativní abundanci bakteriálních rodů, a to z analytického i ekonomického hlediska, sekvenování nové generace nejlepší volbou.
Introduction, aim: Apical periodontitis (AP) is an inflammatory disease of the dental periradicular tissues caused by a bacterial infection. Various methodological approaches are used to determine the bacteria inhabiting the root canals, however, the analysis of the entire root system of a tooth affected by AP still remains a challenge. The aim of our study was to perform a literature search focused on sample collection procedures and methodologies for bacteriome analysis, and then propose a suitable methodological approach for the purpose of studying the etiopathogenesis of this disease. Methods: After searching the PubMed database, we selected only publications of the original work type in which the bacterial DNA of human teeth was analyzed, for the search. Results: Methodologically, the studies differ greatly, in terms of sample collection, DNA isolation, and bacterial DNA analysis itself. A common method of sample collection is the use of sterile endodontic paper points. Although this method of sampling is suitable in clinical practice, it is considered insufficient for a comprehensive analysis of the environment of the root canal system, due to the morphology of the tooth itself and the presence of ramifications. Another method of sampling is resection of the root tip using sterile burs and subsequent grinding of the apex or smearing with sterile endodontic paper pins. Only the apical part of the tooth is used to determine the bacteriome, therefore bacteria that colonize the coronal part of the tooth and participate in the etiopathogenesis of the disease cannot be analyzed. In recent studies, a method is used in which the entire extracted tooth affected by AP is ground into a fine homogeneous powder using cryogenic grinding. It is possible to determine the complex bacteriome of the root canal system and the pulp chamber from the dust of a crushed tooth, and therefore this method seems optimal for the sample preparation from an experimental study point of view. Most often, an effective column method with various purification kits is used for DNA isolation, and for subsequent DNA analysis, methodologies based on the principle of the polymerase chain reaction are mostly used. Sequencing the variable regions of the gene for 16S rRNA is nowadays already the gold standard for categorizing bacteria and characterizing bacterial communities. Conclusion: To study the AP bacteriome, it seems most appropriate to use extracted tooth samples and immediate freezing of the sample without further pre-analytical steps. The crushed tooth is a suitable matrix for the isolation of microbial DNA with commercially available kits, provided that sterile conditions are maintained during cryogenic grinding. Currently, next-generation sequencing is the best choice for determining the bacteriome and obtaining information about the relative abundance of bacterial genera, both analytically and economically.
- Klíčová slova
- kryomlýnek,
- MeSH
- DNA izolace a purifikace MeSH
- kavita zubní dřeně mikrobiologie patologie MeSH
- lidé MeSH
- periapikální periodontitida * mikrobiologie patologie terapie MeSH
- sekvenční analýza DNA MeSH
- terapie kořenového kanálku MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- práce podpořená grantem MeSH
- přehledy MeSH
BACKGROUND: This pilot study aimed to investigate how fixed orthodontic appliances simultaneously applied on the upper and lower arches affect the oral environment in the medium term. METHODS: The oral status of 30 orthodontic patients was evaluated using the number of decay-missing-filled teeth (DMFT), plaque (PI), and gingival indices (GI) before bonding of fixed orthodontic appliances (T0) and during the therapy (T1). Besides, the gingival crevicular fluid (GCF) and a dental plaque were collected. Samples were analyzed for selected Candida sp. and for 10 selected oral bacteria using mass spectroscopy and multiplex polymerase chain reaction, respectively. RESULTS: In 60% of patients, deterioration of the oral status (demonstrated by the increase in PI) was recorded (p < 0.05). Moreover, the changes in PI correlated with those of GI (p < 0.001). At the T1 time point, the mean representation of Actinomyces sp. in the total prokaryotic DNA in GCF and dental plaque of individual patients increased compared to T0 (p < 0.05). The probability of finding any of the 7 selected periodontal bacteria combined with Candida sp. was 10 times higher in patients in whom PI deteriorated between T0 and T1 (p < 0.01). CONCLUSIONS: Changes in the oral microbial diversity and an increase in PI were observed in the medium term after bonding of orthodontic appliance. Our study highlights the importance of a complex approach in this type of research as the association between clinical characteristics and combined microbial parameters is higher than when evaluated separately.
Severe Early Childhood Caries (sECC) is a multifactorial disease associated with the occurrence of specific oral microorganisms and other environmental, behavioral, and genetic factors. This study aimed to construct a multivariable model including the occurrence of Candida spp. and selected behavioral factors (length of breastfeeding, serving sweet beverages and beginning of brushing child's teeth) to determine their relationships to the occurrence of sECC. In this case-control study 164 children with sECC and 147 children without dental caries were included. MALDI-TOF MS and multiplex qPCR were used to identify Candida spp. and selected bacteria in dental plaque samples, respectively. A questionnaire on oral hygiene, diet, and children's health was filled in by the parents. The constructed multivariable logistic regression model showed an independent influence of the microbial and behavioral factors in sECC etiopathogenesis. The occurrence of C. albicans and C. dubliniensis was associated with higher odds of sECC development (odds ratio, OR: 9.62 and 16.93, respectively), together with breastfeeding of 6 months or less (OR: 2.71), exposure to sweet beverages (OR: 3.77), and starting to brush child's teeth after the 12th month of age (OR: 4.10), all statistically significant (p < 0.01). Considering the high occurrence of C. albicans and C. dubliniensis in dental plaque in children with sECC, we propose them as "keystone pathogens" and risk factors for sECC. The models showed that presence of specific species of Candida in dental plaque may be a better descriptor of sECC than the mentioned behavioral factors.
- MeSH
- Candida albicans MeSH
- Candida MeSH
- dítě MeSH
- lidé MeSH
- náchylnost k zubnímu kazu MeSH
- předškolní dítě MeSH
- Streptococcus mutans MeSH
- studie případů a kontrol MeSH
- zubní kaz * MeSH
- zubní plak * MeSH
- Check Tag
- dítě MeSH
- lidé MeSH
- předškolní dítě MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Toxic epidermal necrolysis (TEN) represents a rare drug-induced autoimmune reaction with delayed-type hypersensitivity that initiates the process of developing massive keratinocyte apoptosis, dominantly in the dermoepidermal junction. Although the etiopathophysiology has not yet been fully elucidated, the binding of Fas ligand (FasL, CD95L) to the Fas receptor (CD95) was shown to play a key role in the induction of apoptosis in this syndrome. The knowledge of the role of immunoglobulin G (IgG) in inhibition of Fas-mediated apoptosis contributed to the introduction of i.v. Ig (IVIg) in the therapy of TEN patients. Despite great enthusiasm for this therapy at the end of the 1990s, subsequent studies in various populations and meta-analyses could not unequivocally confirm the efficacy of the IVIg-based treatment concept. Today, therefore, we are faced with the dilemmas of how to adjust therapy of TEN patients most effectively, which patients could benefit from IVIg therapy and what dose of the preparation should be administrated. The ground-breaking question is: do the host genetic profiles influence the responsiveness and side-effects of IVIg therapy in TEN patients? Based on recent pharmacological, immunological and genetic findings, we suggest that the variability of IVIg therapy outcomes in TEN patients may be related to functional variants in Fas, FasL and Fc-γ receptor genes. This novel concept could lead to improved quality of care for patients with TEN, facilitating personalized therapy to reduce mortality.
