Toothbrushing promotes gingival fibroblast proliferation more effectively than removal of dental plaque

Horiuchi M et. al. Journal of Clinical Peridontology 2002: 29: 791-795

Introduction
The major aim of periodontal treatment was a plaque removal. (Loe et al. 1965) The effective of toothbrush promoted of
Keratinization of oral epithelium(Orban, 1930, Mackenzie 1972) Sulcus epithelium (Caffesse et al. 1982) Enhance gingiva circulation (Brill & Krasse 1959 ) Formation of dense bundles of collagenous conective tissue(Tanaka et al. 1998) Thickening of alveola bone (Miake et al. 1988)

Objective
Assessing the effective of toothbrushing on dogs gingiva over time, compare with effective plaque removal of dental plaque.

Materials and Methods

6 mongrel dogs (7.6-9.8kg) Daily removing supragingival plaque with currete 2 weeks later sup & sub gingival plaque removing (baseline) Split mouth design into 4 quadrants
2 daily tooth brushing 2 daily supra plaque removal

Materials and Methods conts

Tooth brush was maintained 20s each tooth with 200g force. Tooth brush characteristic
2 rows of nylon bristles 3 tufts per row 50 filaments (diameter 0.2mm, length 11mm)

Histological & Immunohistochemical

Killing dog by injection of IV sodium thiamylal Teeth and gingivae were fixed wit 4 % paraformaldehyde in 0.1 M phospate buffer pH=7.4 at 4 C overnight Decalcification with a 10% tetrasodium-EDTA aqueous solution (pH=7.4) 4-8weeks at 4 C Staining with hematoxylin and erosin or immunohistochemically stained for proliferating cell nuclear antigen (PCNA) or procollagen type I on sectioning tooth (4 micrometer)

Materials and Methods

PCNA was stained with VECTAS TAIN Elite ABC kit. (Takata et al 1997) Immunohistochemical staining of procollagen type I Monoclonal antibody to human procollagen type I Cpeptide was diluted 1/150 in PBS. D64 square lattice test system was used to calculate the basement membrane Observed under microscope 400 magnification. PCNA positive and procollagen type I were counted in 7 areas

Clinical measurement
Plaque index (Silness & Loe 1964) Gingiva index (Silness & Loe 1963) Mean of Probing depth

Results
Dense infiltration by inflammatory cells was reduced in brushing area at 3 and 5 weeks. Density of PCNA positive fibroblast
2 times brushing = plaque removal area.

Fibroblast was increased significantly in

3rd week Double in 5th week

PCNA positive epithelial cells at

2.5 times brushing = plaque removal area.

Results conts
No statistic significant of reducing plaque index, gingiva index and pocket depths between brushing and plaque removal.

Discussion
Tooth brush area showed
PCNA-positive fibroblasts and basal cell in the 1st week. Density of type I procollagen-positive fibroblasts in 5th week. Lower inflammatory cell infiltration in 3rd week.

Plaque removal didnt change. Proliferation of fibroblasts by a direct effect of mechanical stimulation. TB > PR

Discussion
Previously studied by Tanaka found that
Pocket oxygen tension and hemoglobin oxygen saturation in gingivae Inflammatory cell infiltration by tooth brushing.

Collagen synthesis in gingivae was observed after 2 weeks of cell proliferation. (Rekhter & Gordon 1994) In this present study TB promoted proliferation of basal cells in the junctional epithelium.

Discussion
Possible mechanisms of modulation by mechanical forces include
Stretch-induce plasma membrane (Vandenburgh 1992) Autocrine and/or paracrin action of growth factor (Zhauang et al. 1996)

May prevent periodontal bacteria to gingiva tissue by decreasing epithelium turnover (Listgarten 1986) Fibrobalst prevents pocket formation (Nemeth et al. 1993)

Conclusion
Mechanical toothbrush enhances
Fibroblast proliferation within 1 week Collagen synthesis 5th week