Gene Expression and Accumulation of Fibrillin-1, Fibrillin-2, and Tropoelastin in Cultured Periodontal Fibroblasts

2002 ◽  
Vol 81 (11) ◽  
pp. 771-775 ◽  
Author(s):  
E. Tsuruga ◽  
K. Irie ◽  
T. Yajima
Keyword(s):  
Gene Expression ◽  
Periodontal Ligament ◽  
Northern Blot Analysis ◽  
Elastic System ◽  
Elastic Fibers ◽  
Gingival Fibroblasts ◽  
Periodontal Ligament Fibroblasts ◽  
Human Gingiva ◽  
Fibrillin 1 ◽  
Cell Layers

The elastic system fibers consist of three types—oxytalan, elaunin, and elastic fibers—differing in their relative microfibril and elastin contents. All three types are found in human gingiva, but human periodontal ligaments contain only elastin-free fibers. We examined cultured human gingival fibroblasts (HGF) and cultured human periodontal ligament fibroblasts (HPLF) to determine the gene expression of fibrillin-1 and fibrillin-2 (the major components of microfibrils) and of tropoelastin. In addition, we assessed the degree of accumulation of these proteins in the extracellular matrix. Northern blot analysis revealed that the level of expression of fibrillin-1 and fibrillin-2 was higher in HGF than in HPLF. However, examination of matrix samples from HGF and HPLF cell layers showed that there was no difference in fibrillin-1 accumulation, although fibrillin-2 accumulated to a much greater extent in the HGF-derived matrix. Tropoelastin was expressed only in and around HGF. These results show a correlation between gene expression and the accumulation of tropoelastin and fibrillin-2 in HGF.

Identification of Genes Differentially Expressed in Cultured Human Periodontal Ligament Fibroblasts vs. Human Gingival Fibroblasts by DNA Microarray Analysis

2002 ◽  
Vol 81 (6) ◽  
pp. 399-405 ◽  
Author(s):  
X. Han ◽  
S. Amar
Keyword(s):  
Gene Expression ◽  
Dna Microarray ◽  
Periodontal Ligament ◽  
Expression Patterns ◽  
Differentially Expressed ◽  
Gingival Fibroblasts ◽  
Periodontal Ligament Fibroblasts ◽  
Genes Encoding ◽  
Different Characteristics ◽  
Related Proteins

Despite their similar spindle-shaped appearance, periodontal ligament fibroblasts (PDLF) and gingival fibroblasts (GF) appear to display distinct functional activities in the maintenance of tissue integrity and during inflammatory/immune responses. We postulated that different characteristics of PDLF and GF are defined by the differential expression of specific genes. To test this, we investigated the possible variance of gene expression profile between cultured PDLF and GF, using DNA microarray technology. One hundred sixty-three genes were found differentially expressed by at least three-fold between PDLF and GF. Genes encoding transmembrane proteins and cytoskeleton-related proteins tended to be up-regulated in PDLF, whereas genes encoding cell-cycle regulation proteins and metabolism-related proteins tended to be up-regulated in GF. We concluded that PDLF and GF appear to display different gene expression patterns that may reflect intrinsic functional differences of the two cell populations and may well coordinate with their tissue-specific activities.

Fibrillin-2 Degradation by Matrix Metalloproteinase-2 in Periodontium

2007 ◽  
Vol 86 (4) ◽  
pp. 352-356 ◽  
Author(s):  
E. Tsuruga ◽  
K. Irie ◽  
T. Yajima
Keyword(s):  
Extracellular Matrix ◽  
Cell Membrane ◽  
Periodontal Ligament ◽  
Elastic System ◽  
Fibroblast Cell ◽  
Matrix Metalloproteinase 2 ◽  
Periodontal Tissue ◽  
Gingival Fibroblasts ◽  
Mmp Inhibitor ◽  
Cell Layers

Elastic system fibers, comprised of microfibrils and tropoelastin, are extracellular components of periodontal tissue. During development, the microfibrils act as a template on which tropoelastin is deposited. However, the process of elastic system fiber remodeling is not fully understood. Therefore, we examined whether matrix metalloproteinases (MMPs) are involved in the remodeling of fibrillins (major components of microfibrils) by human gingival fibroblasts and periodontal ligament (PDL) fibroblasts. Gingival and PDL fibroblasts were cultured for 6 weeks. In some cultures, MMP inhibitor or tissue inhibitor of matrix metalloproteinsase-2 (TIMP-2) was added to the medium for an additional 2 weeks. Active MMP-2 (62 kDa) appeared as cell-membrane-associated or in extracellular matrix only in PDL fibroblast cell layers. The addition of MMP inhibitor or TIMP-2 significantly increased fibrillin-2 accumulation in PDL fibroblast cell layers, and decreased the amount of fibrillin-2 fragments, suggesting that active MMP-2 may degrade fibrillin-2, and that MMPs may play a role in the remodeling of elastic system fibers in PDL.

scholarly journals The role of 25-hydroxyvitamin-D3 and vitamin D receptor gene in human periodontal ligament fibroblasts as response to orthodontic compressive strain: an in vitro study

2021 ◽  
Vol 21 (1) ◽  
Author(s):  
Erika Calvano Küchler ◽  
Agnes Schröder ◽  
Vinicius Broska Teodoro ◽  
Ute Nazet ◽  
Rafaela Scariot ◽  
...  
Keyword(s):  
Gene Expression ◽  
Vitamin D ◽  
Single Nucleotide Polymorphisms ◽  
Periodontal Ligament ◽  
Compressive Strain ◽  
Nucleotide Polymorphisms ◽  
Periodontal Ligament Fibroblasts ◽  
Single Nucleotide ◽  
Cox 2

Abstract Background This study aimed to investigate, if different physiological concentrations of vitamin D (25(OH)D3) and single nucleotide polymorphisms in vitamin D receptor (VDR) gene have an impact on gene expression in human periodontal ligament (hPDL) fibroblasts induced by simulated orthodontic compressive strain. Methods A pool of hPDL fibroblasts was treated in absence or presence of 25(OH)D3 in 3 different concentrations (10, 40 and 60 ng/ml). In order to evaluate the role of single nucleotide polymorphisms in the VDR gene, hPDL fibroblasts from 9 patients were used and treated in absence or presence of 40 ng/ml 25(OH)D3. Each experiment was performed with and without simulated orthodontic compressive strain. Real-time PCR was used for gene expression and allelic discrimination analysis. Relative expression of dehydrocholesterol reductase (DHCR7), Sec23 homolog A, amidohydrolase domain containing 1 (AMDHD1), vitamin D 25-hydroxylase (CYP2R1), Hydroxyvitamin D-1-α hydroxylase, receptor activator of nuclear factor-κB ligand (RANKL), osteoprotegerin (OPG), cyclooxygenase-2 (COX-2) and interleukin-6 (IL6) was assessed. Three single nucleotide polymorphisms in VDR were genotyped. Parametric or non-parametric tests were used with an alpha of 5%. Results RANKL, RANKL:OPG ratio, COX-2, IL-6, DHCR7, CYP2R1 and AMDHD1 were differentially expressed during simulated orthodontic compressive strain (p < 0.05). The RANKL:OPG ratio was downregulated by all concentrations (10 ng/ml, 40 ng/ml and 60 ng/ml) of 25(OH)D3 (mean = 0.96 ± 0.68, mean = 1.61 ± 0.66 and mean = 1.86 ± 0.78, respectively) in comparison to the control (mean 2.58 ± 1.16) (p < 0.05). CYP2R1 gene expression was statistically modulated by the different 25(OH)D3 concentrations applied (p = 0.008). Samples from individuals carrying the GG genotype in rs739837 presented lower VDR mRNA expression and samples from individuals carrying the CC genotype in rs7975232 presented higher VDR mRNA expression (p < 0.05). Conclusions Simulated orthodontic compressive strain and physiological concentrations of 25(OH)D3 seem to regulate the expression of orthodontic tooth movement and vitamin-D-related genes in periodontal ligament fibroblasts in the context of orthodontic compressive strain. Our study also suggests that single nucleotide polymorphisms in the VDR gene regulate VDR expression in periodontal ligament fibroblasts in the context of orthodontic compressive strain.

Gingival fibroblasts are better at inhibiting osteoclast formation than periodontal ligament fibroblasts

2006 ◽  
Vol 98 (2) ◽  
pp. 370-382 ◽  
Author(s):  
Teun J. de Vries ◽  
Ton Schoenmaker ◽  
Nutthamon Wattanaroonwong ◽  
Marije van den Hoonaard ◽  
Arlies Nieuwenhuijse ◽  
...  
Keyword(s):  
Periodontal Ligament ◽  
Osteoclast Formation ◽  
Gingival Fibroblasts ◽  
Periodontal Ligament Fibroblasts

scholarly journals Gene Expression Profile in Immortalized Human Periodontal Ligament Fibroblasts Through hTERT Ectopic Expression: Transcriptome and Bioinformatic Analysis

2021 ◽  
Vol 8 ◽  
Author(s):  
Lygia S. Nogueira ◽  
Carolina P. Vasconcelos ◽  
Geovanni Pereira Mitre ◽  
Leonardo Oliveira Bittencourt ◽  
Jessica Rodrigues Plaça ◽  
...  
Keyword(s):  
Gene Expression ◽  
Life Span ◽  
Periodontal Ligament ◽  
Ectopic Expression ◽  
Global Gene Expression ◽  
Cell Types ◽  
Bioinformatic Analysis ◽  
Periodontal Ligament Fibroblasts ◽  
Genetic Changes ◽  
Htert Gene

Human periodontal ligament fibroblast (hPLF) cells play an important role in maintaining oral cavity homeostasis with special function in tissue regeneration and maintenance of dental alveoli. Although their primary cell cultures are considered a good experimental model with no genetic changes, the finite life span may limit some experimental designs. The immortalization process increases cell life span but may cause genetic changes and chromosomal instability, resulting in direct effects on physiological cell responses. In this way, we aimed to investigate the global gene expression of hPLFs after the immortalization process by the ectopic expression of the catalytic subunit of the enzyme telomerase reverse transcriptase (hTERT) through transcriptome analysis. The embryonic origin of the primary culture of hPLF cells and immortalized hPLF-hTERT was also tested by vimentin staining, hTERT synthesis evaluated by indirect immunocytochemistry, analysis of cell proliferation, and morphology. The results indicated that hPLFs and hPLF-hTERT were positive for vimentin. On the 20th cell passage, hPLFs were in senescence, while hPLF-hTERT maintained their proliferation and morphology characteristics. At the same passage, hPLF-hTERT presented a significant increase in hTERT synthesis, but transcriptome did not reveal overexpression of the hTERT gene. Fifty-eight genes had their expression altered (11 upregulated and 47 downregulated) with the absence of changes in the key genes related to these cell types and in the main cancer-associated genes. In addition, the increase in hTERT protein expression without the overexpression of its gene indicates posttranscriptional level regulation. Successful immortalization of hPLFs through the ectopic expression of hTERT encourages further studies to design experimental protocols to investigate clinical questions from a translational perspective.

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