Discovery and characterization of heterogeneous and multipotent fibroblast populations isolated from excised cleft lip tissue.

BACKGROUND Regularly discarded lip tissue obtained from corrective surgeries to close the cleft lip represents an easily accessible and rich source for the isolation of primary fibroblasts. Primary fibroblasts have been described to show compelling similarities to mesenchymal stem cells (MSCs). Hence, cleft lip and palate (CLP) lip-derived fibroblasts could be thought as an intriguing cell source for personalized regenerative therapies in CLP-affected patients. METHODS Initially, we thoroughly characterized the fibroblastic nature of the lip-derived mesenchymal outgrowths by molecular and functional assays. Next, we compared their phenotype and genotype to that of bone marrow-mesenchymal stem cells (BM-MSCs) and of human lung-derived fibroblasts WI38, by assessing their morphology, surface marker expression, trilineage differentiation potential, colony-forming (CFU) capacity, and immunomodulation property. Finally, to better decipher the heterogeneity of our CLP cultures, we performed a single cell clonal analysis and tested expanded clones for surface marker expression, as well as osteogenic and CFU potential. RESULTS We identified intriguingly similar phenotypic and genotypic properties between CLP lip fibroblasts and BM-MSCs, which makes them distinct from WI38. Furthermore, our own data in combination with the complex anatomy of the lip tissue indicated heterogeneity in our CLP cultures. Using a clonal analysis, we discovered single cell-derived clones with increased levels of the MSC markers CD106 and CD146 and clones with variabilities in their commitment to differentiate into bone-forming cells and in their potential to form single cell-derived colonies. However, we were not able to gain clones possessing superior MSC-like capacities when compared to the heterogeneous parental CLP population. Additionally, all clones could still generate contractile forces and retained robust levels of the fibroblast specific marker FSP1, which was not detectable in BM-MSCs. CONCLUSIONS Our results suggest that we isolate heterogeneous populations of fibroblasts from discarded CLP lip tissue, which show a prominently multipotent character in their entirety avoiding the need for elaborate subpopulation selections in vitro. These findings suggest that CLP lip fibroblasts might be a novel potential cell source for personalized regenerative medicine of clinical benefit for CLP patients

Lack of IRF6 Disrupts Human Epithelial Homeostasis by Altering Colony Morphology, Migration Pattern, and Differentiation Potential of Keratinocytes.

Variants within the gene encoding for the transcription factor Interferon Regulatory Factor 6 (IRF6) are associated with syndromic and non-syndromic Cleft Lip/Palate (CLP) cases. IRF6 plays a vital role in the regulation of the proliferation/differentiation balance in keratinocytes and is involved in wound healing and migration. Since a fraction of CLP patients undergoing corrective cleft surgery experience wound healing complications, IRF6 represents an interesting candidate gene linking the two processes. However, Irf6 function has been mainly studied in mice and knowledge on IRF6 in human cells remains sparse. Here, we aimed to elucidate the role of IRF6 in human postnatal skin- and oral mucosa-derived keratinocytes. To do so, we applied CRISPR/Cas9 to ablate IRF6 in two TERT-immortalized keratinocyte cultures, which we used as model cell lines. We show that IRF6 controls the appearance of single cells and colonies, with the latter being less cohesive in its absence. Consequently, IRF6 knockout keratinocytes often moved as single cells instead of a collective epithelial sheet migration but maintained their epithelial character. Lack of IRF6 triggered severe keratinocyte differentiation defects, which were already apparent in the stratum spinosum and extended to the stratum corneum in 3D organotypic skin cultures, while it did not alter their growth rate. Finally, proteomics revealed that most of the differentially expressed proteins in the absence of IRF6 could be associated with differentiation, cell-cell adhesion as well as immune response. Our data expand the knowledge on IRF6 in human postnatal keratinocytes, which will help to better understand IRF6-related pathologies

Inequalities in distribution of dental caries in teenagers of Indaiatuba, São Paulo State, 2004

The objectives of this work was to know the dental caries prevalence and treatment need, besides the distribution of the dental caries in a population of 12 year-old teenagers with high incidence of dental caries. 309 adolescents of public schools of Indaiatuba, São Paulo State, in 2004, were examined. They were selected by process systematic random and the exams followed the World Health Organization criteria. The percentile of school caries free was of 38.8%. Among the adolescents examined, 36.6% (n=113) presented dental caries. DMFT was of 2.50 (IC95%=2.07-2.93). The decayed component was the highest (61.0%). The coefficient of Gini was 0.602 and SiC value was of 5.97 (IC95%=5.32-6.62); in this group especially, 74.7% (n=74) presented dental caries activity in the moment of the exam. Thus, it can be concluded that the adolescents of this city presented low dental caries experience; however, their needs were not assisted, once most of DMFT was formed of the decayed component at 12 years old. In the group SiC, it was verified that this came quite high, being recommended actions addressed to this specific group of adolescents.Os objetivos deste trabalho foram conhecer a prevalência da cárie dentária e a necessidade de tratamento odontológico, além da distribuição da cárie dentária na população que apresenta índices de cárie mais elevados em escolares de doze anos. Foram examinados 309 adolescentes, de escolas públicas de Indaiatuba (SP), em 2004, selecionados mediante processo amostral aleatório sistemático e os exames seguiram os critérios da Organização Mundial de Saúde. O percentual de escolares livres de cárie foi de 38,8%. Dentre os escolares examinados, 36,6% (n=113) apresentaram atividade de cárie. O CPOD foi de 2,50 (IC95%=2,07-2,93). O componente cariado foi o mais prevalente (61,0%). O coeficiente de Gini foi 0,602 e o valor do índice SiC foi de 5,97 (IC95%=5,32-6,62); neste grupo em especial, 74,7% (n=74) apresentaram atividade de cárie no momento do exame. Desta forma, pode-se concluir que os escolares deste município apresentaram baixa experiência de cárie; entretanto, não apresentaram suas necessidades atendidas, uma vez que a maior parte do CPOD era formada do componente cariado, aos doze anos de idade. No grupo SiC, verificou-se que este apresentou-se bastante elevado, sendo recomendadas ações direcionadas a este grupo específico de adolescentes.2173218

[oral Health Among Preschool Children In Brazil, 1999].

To assess dental decay, gum diseases and fluorosis prevalence and evaluate treatment needs of public preschool children. A cross-sectional epidemiological survey on oral health was conducted in 2,805 public preschool children aged 5 and 6 years in Piracicaba, Brazil. Children were examined by a team of ten trained dentists. It was applied a consensus technique and intra and inter agreement among examiners was assessed using the percentage of agreement. Of the total, 44.3% children were free of dental decay at the age of 5 and 38.5% at the age of 6. The deciduous decay teeth index (dmf-t) was 2.64 at the age of 5 and 3.07 at the age of 6, and 31.9% and 37.9%, respectively, presented dmf-t greater than 3. The largest percent of treatment need (45.3%) was surface restoration. Regarding gum health, 68.6% and 72.6% of children aged 5 and 6 years presented gum bleeding. The prevalence of fluorosis was 2.6% at the age of 5 and 6.1% at the age of 6. It is necessary to develop appropriate actions and strategies for higher risk groups.37247-5

Oral health among preschool children in Brazil, 1999

OBJECTIVE: To assess dental decay, gum diseases and fluorosis prevalence and evaluate treatment needs of public preschool children. METHODS: A cross-sectional epidemiological survey on oral health was conducted in 2,805 public preschool children aged 5 and 6 years in Piracicaba, Brazil. Children were examined by a team of ten trained dentists. It was applied a consensus technique and intra and inter agreement among examiners was assessed using the percentage of agreement. RESULTS: Of the total, 44.3% children were free of dental decay at the age of 5 and 38.5% at the age of 6. The deciduous decay teeth index (dmf-t) was 2.64 at the age of 5 and 3.07 at the age of 6, and 31.9% and 37.9%, respectively, presented dmf-t greater than 3. The largest percent of treatment need (45.3%) was surface restoration. Regarding gum health, 68.6% and 72.6% of children aged 5 and 6 years presented gum bleeding. The prevalence of fluorosis was 2.6% at the age of 5 and 6.1% at the age of 6. CONCLUSIONS: It is necessary to develop appropriate actions and strategies for higher risk groups.OBJETIVO: Verificar a prevalência de cárie, doença gengival e fluorose e dimensionar as necessidades de tratamento dos pré-escolares. MÉTODOS: Estudo transversal realizado a partir de um levantamento epidemiológico de saúde bucal em 2.805 crianças de 5 e 6 anos, matriculadas em pré-escolas municipais de Piracicaba, SP. As crianças foram examinadas por uma equipe de dez dentistas, treinados e calibrados. Foi empregada a técnica de consenso, aferindo-se o erro intra e inter examinadores pelo cálculo de percentagem de concordância. RESULTADOS: Obteve-se 44,3% de crianças livres de cárie aos 5 anos e 38,5% aos 6 anos de idade. O índice de cárie para a dentição decídua (ceo-d) foi 2,64 aos cinco anos e 3,07 aos seis anos, sendo que 31,9% e 37,9%, respectivamente, apresentaram ceo-d maior que 3. O maior percentual de necessidade de tratamento (45,3%) foi a indicação de restauração de uma superfície dentária. Quanto à saúde gengival, 68,6% e 72,6% das crianças aos 5 e 6 anos apresentaram sangramento gengival. A prevalência de fluorose observada foi de 2,6% aos 5 e 6,1% aos 6 anos. CONCLUSÕES: Há necessidade da implementação de ações e estratégias adequadas aos grupos de maior risco.24725

Saúde bucal dos pré-escolares, Piracicaba, Brasil, 1999

OBJECTIVE: To assess dental decay, gum diseases and fluorosis prevalence and evaluate treatment needs of public preschool children. METHODS: A cross-sectional epidemiological survey on oral health was conducted in 2,805 public preschool children aged 5 and 6 years in Piracicaba, Brazil. Children were examined by a team of ten trained dentists. It was applied a consensus technique and intra and inter agreement among examiners was assessed using the percentage of agreement. RESULTS: Of the total, 44.3% children were free of dental decay at the age of 5 and 38.5% at the age of 6. The deciduous decay teeth index (dmf-t) was 2.64 at the age of 5 and 3.07 at the age of 6, and 31.9% and 37.9%, respectively, presented dmf-t greater than 3. The largest percent of treatment need (45.3%) was surface restoration. Regarding gum health, 68.6% and 72.6% of children aged 5 and 6 years presented gum bleeding. The prevalence of fluorosis was 2.6% at the age of 5 and 6.1% at the age of 6. CONCLUSIONS: It is necessary to develop appropriate actions and strategies for higher risk groups.OBJETIVO: Verificar a prevalência de cárie, doença gengival e fluorose e dimensionar as necessidades de tratamento dos pré-escolares. MÉTODOS: Estudo transversal realizado a partir de um levantamento epidemiológico de saúde bucal em 2.805 crianças de 5 e 6 anos, matriculadas em pré-escolas municipais de Piracicaba, SP. As crianças foram examinadas por uma equipe de dez dentistas, treinados e calibrados. Foi empregada a técnica de consenso, aferindo-se o erro intra e inter examinadores pelo cálculo de percentagem de concordância. RESULTADOS: Obteve-se 44,3% de crianças livres de cárie aos 5 anos e 38,5% aos 6 anos de idade. O índice de cárie para a dentição decídua (ceo-d) foi 2,64 aos cinco anos e 3,07 aos seis anos, sendo que 31,9% e 37,9%, respectivamente, apresentaram ceo-d maior que 3. O maior percentual de necessidade de tratamento (45,3%) foi a indicação de restauração de uma superfície dentária. Quanto à saúde gengival, 68,6% e 72,6% das crianças aos 5 e 6 anos apresentaram sangramento gengival. A prevalência de fluorose observada foi de 2,6% aos 5 e 6,1% aos 6 anos. CONCLUSÕES: Há necessidade da implementação de ações e estratégias adequadas aos grupos de maior risco

Dental caries activity in adolescents of city with very low dental caries prevalence: Paulínia, São Paulo, 2004

Os objetivos deste trabalho foram verificar a atividade de cárie, além de conhecer a prevalência de cárie e fluorose e as necessidades de tratamento em escolares de 12 anos de idade do município de Paulínia. Foram examinadas 413 crianças, de escolas públicas e particulares de Paulínia, São Paulo, em 2004, selecionadas mediante processo amostral aleatório sistemático. Para medir a experiência de cárie, necessidades de tratamento e fluorose dentária adotou-se os critérios da OMS (1997). Para a atividade de cárie, optou-se por uma simplificação dos critérios de diagnóstico de Nyvad et al. (1999). Os índices utilizados para cárie dentária foram: CPOD e Significant Caries Index (SiC). Os livres de cárie representaram 60,8% dos escolares. O CPOD foi de 0,9 (IC95%=0,8-1,1). O maior componente do CPOD foi o obturado (52,3%) seguido pelo componente cariado (47,2%). Nas crianças que apresentaram atividade de cárie o CPOD foi de 1,37 (IC95%=1,1-1,7) e nas sem atividade de cárie o CPOD foi de 0,57 (IC95%=0,4-0,7) (pThe objectives of this work were to verify the caries activity and to know the caries and fluorosis prevalence among 12-year-old schoolboys and girls in the municipality of Paulinia. Four hundred and thirteen children from public and private schools selected by means of systematic random sampling process were investigated. In order to assess the caries experience, treatment necessities and dental flourosis, the WHO criteria were adopted (1997). For caries activity, the Nyvad et al. (1999) simplified diagnosis criteria were adopted. The indexes used for dental caries were: CPOD and Significant caries Index (SiC). Those free from caries represented 60.8% of students. The CPOD index was of 0.9 (IC95% = 0.8-1.1). The obturated component (52.3%) was the first for CPOD, followed by the decayed component (47.2%). In children presenting caries activity, the COPD was of 1.37 (IC95%=1.1-1.7) and in those without caries activity, the COPD was of 0.57 (IC95%=0.4-0.7) (

Dental caries activity in primary dentition, Indaiatuba, São Paulo, Brazil, 2004

This study aimed to verify caries activity and analyze caries experience, treatment needs, and enamel defects in 5-year-old preschool children in Indaiatuba, São Paulo State, Brazil. 624 children were selected by systematic random sampling in 2004. WHO criteria were used to measure caries experience and treatment needs. Nyvad et al. diagnostic criteria were used to analyze caries activity. Mann-Whitney and qui-square tests were used to analyze the results, with 5% significance. 40.5% of schoolchildren displayed caries activity. The dfmt was 1.62 (95%CI: 1.35-1.89). Among children with dfmt > 0, the index was 3.81 (95%CI: 3.36-4.26), and 91.4% of these children presented caries activity. Care Index was 12.3%. These results suggest that treatment measures and health promotion should continue to be prioritized, since the control measures and caries interruption during the initial stages are well known. Dental care should especially target schoolchildren with more significant caries experience.Os objetivos deste trabalho foram verificar a atividade de cárie, além de conhecer a experiência de cárie (ceod), necessidades de tratamento e defeitos de esmalte em pré-escolares de cinco anos de idade. Examinaram-se 624 crianças de Indaiatuba, São Paulo, Brasil, em 2004, selecionadas por meio de processo amostral aleatório sistemático. Para medir a experiência de cárie e necessidade de tratamento adotou-se os critérios da OMS. Para a atividade de cárie, optou-se por uma simplificação dos critérios de diagnóstico de Nyvad et al. Na análise estatística utilizaram-se os testes Mann Whitney e qui-quadrado com nível de significância de 5%. 40,5% dos pré-escolares apresentaram atividade de cárie. O ceod foi de 1,62 (IC95%: 1,35-1,89). Entre os doentes (ceod > 0) o ceod foi 3,81 (IC95%: 3,36-4,26), ainda neste grupo, 91,4% das crianças apresentaram atividade de cárie. O índice de cuidados foi de 12,3%. Sugere-se que medidas de prevenção e tratamento direcionadas a essa população continuem sendo priorizadas, uma vez que as medidas de controle e paralisação da cárie em seus estágios iniciais são bem conhecidas. Além disso, recomenda-se cuidado direcionado aos pré-escolares com maior experiência de cáries.59360

A Living Cell Repository of the Cranio-/Orofacial Region to Advance Research and Promote Personalized Medicine

The prevalence of congenital anomalies in newborns is estimated to be as high as 6%, many of which involving the cranio-/orofacial region. Such malformations, including several syndromes, are usually identified prenatally, at birth, or rarely later in life. The lack of clinically relevant human cell models of these often very rare conditions, the societal pressure to avoid the use of animal models and the fact that the biological mechanisms between rodents and human are not necessarily identical, makes studying cranio-/orofacial anomalies challenging. To overcome these limitations, we are developing a living cell repository of healthy and diseased cells derived from the cranio-/orofacial region. Ultimately, we aim to make patient-derived cells, which retain the molecular and genetic characteristics of the original anomaly or disease in vitro, available for the scientific community. We report our efforts in establishing a human living cell bank derived from the cranio-/orofacial region of otherwise discarded tissue samples, detail our strategy, processes and quality checks. Such specific cell models have a great potential for discovery and translational research and might lead to a better understanding and management of craniofacial anomalies for the benefit of all affected individuals

Relation between oral health and nutritional condition in the elderly

Oral health is a prerequisite for a good chewing function, which may have an impact on food choices and nutritional well-being. OBJECTIVE: This study was designed to evaluate the relationship between oral health status and nutritional status in the elderly. MATERIAL AND METHODS: In this cross-sectional study, 33 elderly people from the Group for the Elderly Interdisciplinary Geriatrics and Gerontology Program, at Fluminense Federal University, Niteroi, RJ, Brazil, completed a questionnaire to collect information on socioeconomic status, eating habits, physical activity and health habits, undertook a clinical oral examination, blood test, and anthropometric measurements, and were allocated into groups according to age. The oral health status was assessed using the index for decayed, missing and filled teeth (DMFT). The nutritional status was assessed using hemoglobin, hematocrit and albumin concentrations in blood, anthropometric values and the body mass index. RESULTS: Tooth loss was the biggest nuisance to the elderly subjects (57.6%), followed by the use of dentures (30.3%) and ill-fitting dentures (33.3%). 66.6% of patients had difficulty in chewing, and 54.5% reported this to be due to prostheses and 13.6% to the absence of teeth. A significant correlation was found between DMFT and the value of suprailiac skinfold thickness (rho=0.380, p=0.029). CONCLUSION: The results support the temporal association between tooth loss and detrimental changes in anthropometry, which could contribute to increased risk of developing chronic diseases