J Korean Acad Conserv Dent.  2004 Jul;29(4):370-377. 10.5395/JKACD.2004.29.4.370.

Tissue engineering of dental pulp on type I collagen

• Lee GH
• Huh SY
• Park SH

• KMID: 2175631
• DOI: http://doi.org/10.5395/JKACD.2004.29.4.370

Abstract

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 x 10(5) cells/ml/well) were cultured at 12-well plate at 37degrees C 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.

Keyword

Pulp tissue engineering; Pulp cells; Fibroblast; Tissue regeneration; Type I collagen gel; Matrix contraction

MeSH Terms

Blood Vessels
Collagen
Collagen Type I*
Dental Pulp*
Eosine Yellowish-(YS)
Fibroblasts
Gingiva
Hematoxylin
Humans
NIH 3T3 Cells
Skin
Tissue Engineering*
Collagen
Collagen Type I
Eosine Yellowish-(YS)
Hematoxylin

Figure

• 

  Figure 1 Collagen Gel Contraction Assay. Changes in the diameter of the different cell-seeded collagen gel matrix. 1 × 105 cells of the primary cultured gingival fibroblasts (GFB), skin fibroblast (SFB), NIH 3T3 cells (3T3) were mixed with type I collagen gel solution. Only collagen gel solution (500 µl/well) were plated as control, and the cell-seeded collagen gel solution (500 µl/well) were plated in the 12-well-plates. The values of the diameter represent the mean ± standard deviation calculated from two-samples at each point for 14 days.

• 

  Figure 2 Photographs of collagen gel matrix 1 hour after the collagen gel solution without cells (A), 1 × 105 cells of the primary cultured gingival fibroblasts (B), skin fibroblasts (C), pulp cells (D), and NIH 3T3 cells (E) were plated in the 12-well plates.

• 

  Figure 3 Photographs of collagen gel matrix 1 day after the collagen gel solution without cells (A), 1 × 105 cells of the primary cultured gingival fibroblasts (B), skin fibroblasts (C), pulp cells (D), and NIH 3T3 cells (E) were plated in the 12-well plates.

• 

  Figure 4 Photographs of collagen gel matrix 3 days after the collagen gel solution without cells (A), 1 × 105 cells of the primary cultured gingival fibroblasts (B), skin fibroblasts (C), pulp cells (D), and NIH 3T3 cells (E) were plated in the 12-well plates.

• 

  Figure 5 Photographs of collagen gel matrix 5 days after the collagen gel solution without cells (A), 1 × 105 cells of the primary cultured gingival fibroblasts (B), skin fibroblasts (C), pulp cells (D), and NIH 3T3 cells (E) were plated in the 12-well plates.

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  Figure 6 Photographs of collagen gel matrix 7 days after the collagen gel solution without cells (A), 1 × 105 cells of the primary cultured gingival fibroblasts (B), skin fibroblasts (C), pulp cells (D), and NIH 3T3 cells (E) were plated in the 12-well plates.

• 

  Figure 7 Microphotographs of histological sections of the normal adult pulp (A) and engineered pulp tissue (B) stained with hematoxylin and eosin. The original magnifications of the photographs were × 400.

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