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Proteomic profile of rat periodontal tissues in health and periodontal disease

ABSTRACT: The periodontal ligament tissue plays a crucial role in oral functions by providing structural support to the tooth. Due to the presence of undifferentiated mesenchymal cells, research into its regenerative capacity is ongoing. Pathological conditions can affect these functions and protein composition. The aim of this study was to evaluate through proteomic analysis the protein profile of the periodontal ligament (PDL) tissues of rats submitted to experimental periodontitis induction. The study was approved by the Ethical Committee on Animal Experimentation of the Sao Paulo State University, School of Dentistry at Araraquara, Brazil (protocol number 16/2015) and followed the Animal Research: Reporting of In Vivo Experiments guidelines. A total of eight male adult Holtzman rats, average weight of 300 g, were used and housed in plastic cages properly identified, with a 12-hours lights/dark cycle, under controlled temperature (22-25 °C), receiving water ad libitum and standard laboratory diet (Labina/Purina, Ribeirão Preto, Brazil). The animals were randomly assigned into two groups: control without any intervention (C, n=3) and experimental periodontitis (P, n=5). The animals of the group P were anesthetized with intramuscular injections of 10% ketamine chlorhydrate (0.08 ml/100 g body weight) and 2% xylazine chlorhydrate (0.04 ml/100 g body weight). To induce periodontitis, a cotton thread was placed bilaterally around the upper first molars and maintained with a knot for 20 days. After this period, all animals were euthanized by anesthetic overdose and had their maxillary jaws collected for subsequent analyses.

INSTRUMENT(S):

ORGANISM(S): Rattus Norvegicus (rat)

TISSUE(S): Periodontal Ligament

DISEASE(S): Periodontal Disease

SUBMITTER: Maria Eduarda Lopes

LAB HEAD: Joni Augusto Cirelli

PROVIDER: PXD055817 | Pride | 2025-05-01

REPOSITORIES: Pride

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Project description:The prevalence of obesity has significantly increased worldwide. Therefore, this study aimed to evaluate the influence of obesity in the proteomic profile of periodontal ligament (PDL) tissues of rat first maxillary molars submitted to orthodontic tooth movement. The study was approved by the Ethical Committee on Animal Experimentation of the Sao Paulo State University, School of Dentistry at Araraquara, Brazil (protocol number 16/2015) and followed the Animal Research: Reporting of In Vivo Experiments guidelines. Therefore, for this study, we used ten male adult Holtzman rats (weight average – 300 g, approximately 8 weeks old). Animals were randomly assigned to one of the following experimental groups: 1 – M group: animals received a standard diet and 75 days after that, the first maxillary molar was randomly selected to be submitted to OTM for 15 days (n=5), and 2 – OM group: animals received a high-fat diet to induce obesity, and 75 days after that the first maxillary molar was randomly selected to be subjected to OTM for 15 days (n=5). All animals were kept under controlled temperature (22-25 °C), receiving water ad libitum with a 12-hour light/dark cycle. To induce obesity, animals of the OM group received a high-fat diet. In contrast to the standard diet (Labina/Purina, Ribeirão Preto, Brazil) administered to the M group. The high-fat diet contained around 3.82 kcal/g and was composed of 27.5 % of standard rat chow plus 25 % of peanuts, 25 % of milk chocolate, and 12.5 % sweet biscuits and contained 20 g of protein, 48 g of carbohydrate, 20 g of total fat, and 4 g of fiber per 100 g of diet. This feed was daily replaced and had started 75 days before baseline (M initiation). The procedure to induce OTM was by a closed coil nickel-titanium spring (Sentalloy, GAC Central Slip, New York, EUA) connected the maxillary first molar and central incisors with a steel wire (CrNi, 0.20 mm, 55.01.208; Morelli, Sorocaba, Brazi). Fifteen days after OTM, animals were euthanized by anesthetic overdose.

Project description:Periodontal regeneration study. Periodontitis is a common chronic inflammatory disease which may lead to tooth loss. The ultimate goal of periodontal therapy is complete regeneration of the tissues lost as a result of periodontitis. However, most regenerative clinical procedures are unpredictable, largely because there is a lack of understanding of how the various tissues comprising the periodontium (gingival epithelium, gingival connective tissue, periodontal ligament, aveolar bone and cementum) interact during the regenerative process. The aim of this study was to investigate the molecular mechanisms that are involved in periodontal regeneration. For this purpose, paired sets of gingival, ligament and regenerating cells were isolated from patients that have been treated for periodontitis via a regenerative surgical procedure. A microarray analysis using the Hu133A Affymetrix arrays was used to identify the genes and pathways that were characteristic of the three different tissues. We have identified over 500 transcripts in global comparison and intrapatient comparisons that were differentially expressed between ligament and gingival samples, and approximately 30 transcripts that characterized the regenerating cell population. By using a functional classification based on Gene Ontology we were able to group the transcripts into 12 groups. Proteases/Protease inhibitors were differentially expressed between the ligament and gingiva highlighting high ECM remodelling activity by gingival cells. On the other hand, ligament cells were shown to have increased protein synthesis by increasing their nucleolar and ribosomal gene complement. We were able to identify 57 various DNA binding proteins, among them were some well characterised transcription factors, transcriptional regulators, including DNA polymerases and transcriptional co-activators. Furthermore, we have also identified changes in expression of their targets in the two tissues. This is the first study to characterise the gene expression profile of periodontal regenerating cells and allowed us to gain valuable insights into the cellular and molecular mechanisms that occur during the regenerative process. We have identified signalling pathways which are consistent with clinical observations of the regenerative properties of gingival and periodontal ligament cells. These pathways provide candidate targets that can be manipulated in order to achieve a superior regenerative clinical outcome. Keywords: other

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	Multiconsensus_from_Reports_control_vs_perio.xlsx	Xlsx
	Multiconsensus_from_Reports_legend.docx	Other

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Proteomic profile of rat periodontal tissues in health and periodontal disease

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