Comparison of Adhesion and Proliferation of Human Gingival Fibroblasts on Acellular Dermal Matrix with and without Low Level Diode Laser Irradiation, an in vitro Study

Abstract

Statement of the Problem: In recent years, regeneration of periodontal soft tissues in the reconstruction of periodontal defects and the finding of suitable membranes and graft materials for the placement of autogenous grafts have been of great interest in various studies. In this regard, the proliferation and adhesion of regenerative cells are two linchpins of the complete regenerative process. Purpose: This study aimed to evaluate the effects of low-level laser beams on the attachment and the proliferation of human gingival fibroblasts in the presence of acellular dermal matrix (ADM). Materials and Method: All the experiments were conducted compared to tissue culture plate in four groups as follows: (1) Fibroblast+ADM+laser, (2) Fibroblast+ADM+ no laser, (3) Fibroblast + laser radiation, and (4) Fibroblast+ no laser. In this experimental study, the primary attachment was evaluated by passing 8h from seeding of 5×105 gingival fibroblasts with or without a single dose (15.6 J/cm2) of laser radiation. Cell proliferation rate was also examined at 24, 48, and 72 hours after cell culture, following exposure to 5.2 J/cm2 of laser at each day of examination. Thereafter, fibroblasts were incubated under the normal culture condition (at 37°C, 5% CO2) in high glucose Dulbecco's Modified Eagle's medium (DMEM) medium supplemented with 10% fetal bovine serum, 1% glutamax, and 1% penicillin/streptomycin. Subsequently, the cellular viability was assessed on each time point using MTS calorimetric assay. The obtained data were statistically analyzed by applying ANOVA and Tukey tests. Results: There was a significant difference among the means of these four groups in terms of the proliferation of fibroblasts at 24, 48 and 72 hours (p< 0.001). Moreover, there was no significant difference among the means of two groups in terms of fibroblastic attachment in 8 hours (p< 0.2). The fibroblast group has shown the highest proliferation rate among all groups after laser radiation. Conclusion: It was indicated that the laser radiation increases the fibroblast cell proliferation. Accordingly, although this increase was higher in the fibroblast group alone compared to the fibroblasts cultured on acellular dermal matrix, the laser radiation did not significantly increase the attachment of fibroblast cells to acellular dermal matrix.

Keywords

• acellular dermal; fibroblasts; lasers; cell proliferation;