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Research Articles
Effects of CTHRC1 on odontogenic differentiation and angiogenesis in human dental pulp stem cells
Jong-soon Kim, Bin-Na Lee, Hoon-Sang Chang, In-Nam Hwang, Won-Mann Oh, Yun-Chan Hwang
Restor Dent Endod 2023;48(2):e18.   Published online April 28, 2023
DOI: https://doi.org/10.5395/rde.2023.48.e18
AbstractAbstract PDFPubReaderePub
Objectives

This study aimed to determine whether collagen triple helix repeat containing-1 (CTHRC1), which is involved in vascular remodeling and bone formation, can stimulate odontogenic differentiation and angiogenesis when administered to human dental pulp stem cells (hDPSCs).

Materials and Methods

The viability of hDPSCs upon exposure to CTHRC1 was assessed with the WST-1 assay. CTHRC1 doses of 5, 10, and 20 µg/mL were administered to hDPSCs. Reverse-transcription polymerase reaction was used to detect dentin sialophosphoprotein, dentin matrix protein 1, vascular endothelial growth factor, and fibroblast growth factor 2. The formation of mineralization nodules was evaluated using Alizarin red. A scratch wound assay was conducted to evaluate the effect of CTHRC1 on cell migration. Data were analyzed using 1-way analysis of variance followed by the Tukey post hoc test. The threshold for statistical significance was set at p < 0.05.

Results

CTHRC1 doses of 5, 10, and 20 µg/mL had no significant effect on the viability of hDPSCs. Mineralized nodules were formed and odontogenic markers were upregulated, indicating that CTHRC1 promoted odontogenic differentiation. Scratch wound assays demonstrated that CTHRC1 significantly enhanced the migration of hDPSCs.

Conclusions

CTHRC1 promoted odontogenic differentiation and mineralization in hDPSCs.

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Is dentin biomodification with collagen cross-linking agents effective for improving dentin adhesion? A systematic review and meta-analysis
Julianne Coelho Silva, Edson Luiz Cetira Filho, Paulo Goberlânio de Barros Silva, Fábio Wildson Gurgel Costa, Vicente de Paulo Aragão Saboia
Restor Dent Endod 2022;47(2):e23.   Published online May 6, 2022
DOI: https://doi.org/10.5395/rde.2022.47.e23
AbstractAbstract PDFSupplementary MaterialPubReaderePub
Objectives

The aim of this investigation was to evaluate the effectiveness of collagen cross-linking agents (CCLAs) used in combination with the adhesive technique in restorative procedures.

Materials and Methods

In this systematic review, the authors followed the Preferred Reporting Items for Systematic Reviews and Meta-Analyses checklist. An electronic search was performed using PubMed, Scopus, Web of Science, Cochrane Library, LILACS, and DOSS, up to October 2020. The gray literature was also researched. Only randomized clinical trials were selected.

Results

The selection process yielded 3 studies from the 838 retrieved. The addition of CCLAs in the retention of restorations increased the number of events. The postoperative sensitivity scores and marginal adaptation scores showed no significant difference between the CCLA and control groups, and the marginal pigmentation scores showed a significant increase in the CCLA group. There were no caries events in any group throughout the evaluation period.

Conclusions

This systematic review showed that there is no clinical efficacy to justify the use of CCLAs in the protocols performed.

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Interplay of collagen and mast cells in periapical granulomas and periapical cysts: a comparative polarizing microscopic and immunohistochemical study
Deepty Bansal, Mala Kamboj, Anjali Narwal, Anju Devi, Nisha Marwah
Restor Dent Endod 2022;47(1):e12.   Published online February 14, 2022
DOI: https://doi.org/10.5395/rde.2022.47.e12
AbstractAbstract PDFPubReaderePub
Objectives

This pilot study aimed to establish the interrelationship between collagen and mast cells in periapical granulomas and periapical cysts.

Materials and Methods

An observational cross-sectional study was conducted on the paraffin-embedded tissue sections of 68 specimens (34 periapical granulomas and 34 periapical cysts). The specimens were stained with picrosirius to observe collagen fiber birefringence and anti-tryptase antibody to evaluate the mast cell count immunohistochemically. The mean number and birefringence of collagen fibers, as well as the mean number of mast cells (total, granulated, and degranulated), and the mean inflammatory cell density were calculated. The data obtained were analyzed using the Kruskal Wallis test, Mann Whitney U test, and Spearman correlation test (p < 0.05).

Results

The mean number of thick collagen fibers was higher in periapical cysts, while that of thin fibers was higher in granulomas (p = 0.00). Cysts emitted orange-yellow to red birefringence, whereas periapical granulomas had predominantly green fibers (p = 0.00). The mean inflammatory cell density was comparable in all groups (p = 0.129). The number of total, degranulated, and granulated mast cells exhibited significant results (p = 0.00) in both groups. Thick cyst fibers showed significant inverse correlations with inflammation and degranulated mast cells (p = 0.041, 0.04 respectively).

Conclusions

Mast cells and inflammatory cells influenced the nature of collagen fiber formation and its birefringence. This finding may assist in the prediction of the nature, pathogenesis, and biological behavior of periapical lesions.

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Review Article
Inhibition of matrix metalloproteinases: a troubleshooting for dentin adhesion
Izadora Quintela Souza de Moraes, Ticiano Gomes do Nascimento, Antonio Thomás da Silva, Lilian Maria Santos Silva de Lira, Abhishek Parolia, Isabel Cristina Celerino de Moraes Porto
Restor Dent Endod 2020;45(3):e31.   Published online May 22, 2020
DOI: https://doi.org/10.5395/rde.2020.45.e31
AbstractAbstract PDFPubReaderePub

Matrix metalloproteinases (MMPs) are enzymes that can degrade collagen in hybrid layer and reduce the longevity of adhesive restorations. As scientific understanding of the MMPs has advanced, useful strategies focusing on preventing these enzymes' actions by MMP inhibitors have quickly developed in many medical fields. However, in restorative dentistry, it is still not well established. This paper is an overview of the strategies to inhibit MMPs that can achieve a long-lasting material-tooth adhesion. Literature search was performed comprehensively using the electronic databases: PubMed, ScienceDirect and Scopus including articles from May 2007 to December 2019 and the main search terms were “matrix metalloproteinases”, “collagen”, and “dentin” and “hybrid layer”. MMPs typical structure consists of several distinct domains. MMP inhibitors can be divided into 2 main groups: synthetic (synthetic-peptides, non-peptide molecules and compounds, tetracyclines, metallic ions, and others) and natural bioactive inhibitors mainly flavonoids. Selective inhibitors of MMPs promise to be the future for specific targeting of preventing dentin proteolysis. The knowledge about MMPs functionality should be considered to synthesize drugs capable to efficiently and selectively block MMPs chemical routes targeting their inactivation in order to overcome the current limitations of the therapeutic use of MMPs inhibitors, i.e., easy clinical application and long-lasting effect.

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Research Articles
Differential diagnosis of periapical cyst using collagen birefringence pattern of the cyst wall
Hyo Jin Ji, Se-Hee Park, Kyung-Mo Cho, Suk Keun Lee, Jin Woo Kim
Restor Dent Endod 2017;42(2):111-117.   Published online February 9, 2017
DOI: https://doi.org/10.5395/rde.2017.42.2.111
AbstractAbstract PDFPubReaderePub
Objectives

Periapical lesions, including periapical cyst (PC), periapical granuloma (PG), and periapical abscess (PA), are frequently affected by chemical/physical damage during root canal treatment or severe bacterial infection, and thus, the differential diagnosis of periapical lesions may be difficult due to the presence of severe inflammatory reaction. The aim of this study was to make differential diagnosis among PC, PG, and PA under polarizing microscope.

Materials and Methods

The collagen birefringence patterns of 319 cases of PC (n = 122), PG (n = 158), and PA (n = 39) obtained using a polarizing microscope were compared. In addition, 6 cases of periodontal fibroma (PF) were used as positive controls.

Results

Collagen birefringence was condensed with a thick, linear band-like pattern in PC, but was short and irregularly scattered in PG, and scarce or absent in PA. PF showed intense collagen birefringence with a short, palisading pattern but no continuous band-like pattern. The linear band-like birefringence in PC was ascribed to pre-existing expansile tensile stress of the cyst wall.

Conclusions

In this study all PCs (n = 122) were distinguishable from PGs and PAs by their characteristic birefringence, despite the absence of lining epithelium (n = 20). Therefore, the authors suggest that the presence of linear band-like collagen birefringence of the cyst wall aids the diagnostic differentiation of PC from PG and PA.

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Effects of proanthocyanidin, a crosslinking agent, on physical and biological properties of collagen hydrogel scaffold
Yoorina Choi, Hee-Jin Kim, Kyung-San Min
Restor Dent Endod 2016;41(4):296-303.   Published online October 4, 2016
DOI: https://doi.org/10.5395/rde.2016.41.4.296
AbstractAbstract PDFPubReaderePub
Objectives

The purpose of the present study was to evaluate the effects of proanthocyanidin (PAC), a crosslinking agent, on the physical properties of a collagen hydrogel and the behavior of human periodontal ligament cells (hPDLCs) cultured in the scaffold.

Materials and Methods

Viability of hPDLCs treated with PAC was measured using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. The physical properties of PAC treated collagen hydrogel scaffold were evaluated by the measurement of setting time, surface roughness, and differential scanning calorimetry (DSC). The behavior of the hPDLCs in the collagen scaffold was evaluated by cell morphology observation and cell numbers counting.

Results

The setting time of the collagen scaffold was shortened in the presence of PAC (p < 0.05). The surface roughness of the PAC-treated collagen was higher compared to the untreated control group (p < 0.05). The thermogram of the crosslinked collagen exhibited a higher endothermic peak compared to the uncrosslinked one. Cells in the PAC-treated collagen were observed to attach in closer proximity to one another with more cytoplasmic extensions compared to cells in the untreated control group. The number of cells cultured in the PAC-treated collagen scaffolds was significantly increased compared to the untreated control (p < 0.05).

Conclusions

Our results showed that PAC enhanced the physical properties of the collagen scaffold. Furthermore, the proliferation of hPDLCs cultured in the collagen scaffold crosslinked with PAC was facilitated. Conclusively, the application of PAC to the collagen scaffold may be beneficial for engineering-based periodontal ligament regeneration in delayed replantation.

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Case Report
Biodentine-a novel dentinal substitute for single visit apexification
Gurudutt Nayak, Mohammad Faiz Hasan
Restor Dent Endod 2014;39(2):120-125.   Published online March 21, 2014
DOI: https://doi.org/10.5395/rde.2014.39.2.120
AbstractAbstract PDFPubReaderePub

Use of an apical plug in management of cases with open apices has gained popularity in recent years. Biodentine, a new calcium silicate-based material has recently been introduced as a dentine substitute, whenever original dentine is damaged. This case report describes single visit apexification in a maxillary central incisor with necrotic pulp and open apex using Biodentine as an apical barrier, and a synthetic collagen material as an internal matrix. Following canal cleaning and shaping, calcium hydroxide was placed as an intracanal medicament for 1 mon. This was followed by placement of small piece of absorbable collagen membrane beyond the root apex to serve as matrix. An apical plug of Biodentine of 5 mm thickness was placed against the matrix using pre-fitted hand pluggers. The remainder of canal was back-filled with thermoplasticized gutta-percha and access cavity was restored with composite resin followed by all-ceramic crown. One year follow-up revealed restored aesthetics and function, absence of clinical signs and symptoms, resolution of periapical rarefaction, and a thin layer of calcific tissue formed apical to the Biodentine barrier. The positive clinical outcome in this case is encouraging for the use of Biodentine as an apical plug in single visit apexification procedures.

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Basic Research
The effect of the removal of chondroitin sulfate on bond strength of dentin adhesives and collagen architecture
Jong-Ryul Kim, Sang-Jin Park, Gi-Woon Choi, Kyoung-Kyu Choi
J Korean Acad Conserv Dent 2010;35(3):211-221.   Published online May 31, 2010
DOI: https://doi.org/10.5395/JKACD.2010.35.3.211
AbstractAbstract PDFPubReaderePub

Proteoglycan is highly hydrophilic and negatively charged which enable them attract the water. The objective of study was to investigate the effects of Proteoglycan on microtensile bond strength of dentin adhesives and on architecture of dentin collagen matrix of acid etched dentin by removing the chondroitin sulphate attached on Proteoglycan. A flat dentin surface in mid-coronal portion of tooth was prepared. After acid etching, half of the specimens were immersed in 0.1 U/mL chondroitinase ABC (C-ABC) for 48 h at 37℃, while the other half were stored in distilled water. Specimens were bonded with the dentin adhesive using three different bonding techniques (wet, dry and re-wet) followed by microtensile bond strength test. SEM examination was done with debonded specimen, resin-dentin interface and acid-etched dentin surface with/without C-ABC treatment.

For the subgroups using wet-bonding or dry-bonding technique, microtensile bond strength showed no significant difference after C-ABC treatment (p > 0.05). Nevertheless, the subgroup using rewetting technique after air dry in the Single Bond 2 group demonstrated a significant decrease of microtensile bond strength after C-ABC treatment. Collagen architecture is loosely packed and some fibrils are aggregated together and relatively collapsed compared with normal acid-etched wet dentin after C-ABC treatment. Further studies are necessary for the contribution to the collagen architecture of noncollagenous protein under the various clinical situations and several dentin conditioners and are also needed about long-term effect on bond strength of dentin adhesive.

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Original Articles
The comparison of gene expression from human dental pulp cells and periodontal ligament cells
Hyoun So, Sang-Hyuk Park, Gi-Woon Choi
J Korean Acad Conserv Dent 2009;34(5):430-441.   Published online September 30, 2009
DOI: https://doi.org/10.5395/JKACD.2009.34.5.430
AbstractAbstract PDFPubReaderePub

The purpose of this study was to characterize functional distinction between human dental pulp cells(PC) and periodontal ligament cells(PDLC) using cDNA microarray assay and to confirm the results of the microarray assay using RT-PCR. 3 genes out of 51 genes which were found to be more expressed(>2 fold) in PC were selected, and 3 genes out of 19 genes which were found to be more expressed(>2 fold) in PDLC were selected for RT-PCR as well.

According to this study, the results were as follows:

1. From the microarray assay, 51 genes were more expressed (2 fold) from PC than PDLC.

2. RT-PCR confirmed that ITGA4 and TGF β2 were more expressed in PC than in PDLC.

3. From the microarray assay, 19 genes were more expressed (2 fold) from PDLC than PC.

4. RT-PCR confirmed that LUM, WISP1, and MMP1 were more expressed in PDLC than in PC.

From the present study, different expression of the genes between the PC and PDLC were characterized to show the genes which play an important role in dentinogenesis were more expressed from PC than PDLC, while the genes which were related with collagen synthesis were more expressed from PDLC than PC.

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The effect of cyanate methacrylate on the shear bond strengths to dentin
Hyang-Kyung Kim, Kyung-Kyu Choi, Gi-Woon Choi, Sang-Jin Park
J Korean Acad Conserv Dent 2007;32(3):236-247.   Published online May 31, 2007
DOI: https://doi.org/10.5395/JKACD.2007.32.3.236
AbstractAbstract PDFPubReaderePub

The purpose of this study was to evaluate the effects of cyanate methacylate on the shear bond strengths to bovine dentin surfaces as a dentin primers.

Seven experimental adhesives were made with different mass fraction of Isocyanatoetylmethacrylate (IEM), 40wt% HEMA (Wako Pure Chemical Industries Osaka, Japan), 0.6% camphoroquinone, 0.4% amine and ethanol as balance. dentin bonding agents (0, 2, 4, 6, 8, 10, 12%) were made and applied on the surface of bovine dentin specimens of 7 experimental groups.

Shear bond strengths were measured using a universal testing machine (Instro 4466).

To identify the ratio and modes of cohesive failures, microscopic examinationn was performed. The ultra-structure of resin tags were observed under scanning electron microscope.

The results were as follows ;

1) A higher shear bond strengths (33.62 MPa) in group 8% of Cyanate methacrylate to dentin were found, but there were no statistically significancy between Groups (p > 0.05).

2) The higher ratio of cohesive failures mode in group 2, 6, an 10% could be seen than that in any other groups.

3) A shorter resin tags were observed in all experimental groups.

This could be resulted that the preventing from the cyanate methacrylate penetrate into dentin owing to reacting it with dentin collagen.

Therefore the resin tags were shorter in lengths.

Whether the higher bonding strengths of dentin bonding agents can be affected was not been assured with statistic results.

The results indicated that the relation between tensile strengths of the dentin adhesives to bovine dentin and resin tags formed into the dentin could not affected.

The main reason of increasing the shear bond strength to bovine dentin in experimental groups could not be assured.

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Tissue engineering of dental pulp on type I collagen
Gwang-Hee Lee, Sung-Yoon Huh, Sang-Hyuk Park
J Korean Acad Conserv Dent 2004;29(4):370-377.   Published online July 31, 2004
DOI: https://doi.org/10.5395/JKACD.2004.29.4.370
AbstractAbstract PDFPubReaderePub

The purpose of this study was to regenerate human dental pulp tissues similar to native pulp tissues. Using the mixture of type I collagen solution, primary cells collected from the different tissues (pulp, gingiva, and skin) and NIH 3T3 (1 × 105 cells/ml/well) were cultured at 12-well plate at 37℃ for 14 days. Standardized photographs were taken with digital camera during 14 days and the diameter of the contracted collagen gel matrix was measured and statistically analyzed with student t-test. As one of the pulp tissue engineering, normal human dental pulp tissue and collagen gel matrix cultured with dental pulp cells for 14 days were fixed and stained with Hematoxyline & Eosin.

According to this study, the results were as follows:

1. The contraction of collagen gel matrix cultured with pulp cells for 14 days was significantly higher than other fibroblasts (gingiva, skin) (p < 0.05).

2. The diameter of collagen gel matrix cultured with pulp cells was reduced to 70.4% after 7 days, and 57.1% after 14 days.

3. The collagen gel without any cells did not contract, whereas the collagen gel cultured with gingiva and skin showed mild contraction after 14 days (88.1% and 87.6% respectively).

4. The contraction of the collagen gel cultured with NIH 3T3 cells after 14 days was higher than those cultured with gingival and skin fibroblasts, but it was not statistically significant (72.1%, p > 0.05).

5. The collagen gel matrix cultured with pulp cells for 14 days showed similar shape with native pulp tissue without blood vessels.

This approach may provide a means of engineering a variety of other oral tissue as well and these cell behaviors may provide information needed to establish pulp tissue engineering protocols.

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