Evaluation of Acellular Dermal Matrix as a Three-dimensional Scaffold

Objectives: The aim of this investigation was to verify if the acellular dermal matrix (ADM, Alloderm) is a viable three-dimensional scaffold, through its in vitro response to gingival fibroblasts (canine gingival fibroblasts – CGF and human gingival fibroblasts - HGF) and cancerous cells lineage (murine melanoma - B16F10). Methods: CGF, HGF and B16F10 were seeded on AMD at a density of 4 x 104 cells/well for periods up to 14 days. The following parameters were assessed: morphology and distribution of CGF, HGF and B16F10 by direct fluorescence; CGF and HGF viability and the effect of ADM conditioned medium (CM) on CGF viability by MTT. Results: Epifluorescence revealed that CGF and HGF were adherent and round shaped at 12 h and at 7 and 14 days they were spread, exhibiting an elongated shape. CGF were unevenly distributed on ADM surface leaving segments devoid of cells and showing no increase in cells number over the experimental periods; HGF formed a monolayer on ADM surface, in a higher number at 14 days (p<0,05); and B16F10 exhibited an increase in cells number in 7 days (p<0,05), and were mainly arranged in cell aggregates on ADM, forming a continuous layer at 14 days. A higher percentage of cells on the surface (p <0.05) compared to inside the matrix was observed for all cells type in all periods. MTT indicated higher cell viability in samples cultured with HGF compared to CGF (p=0.024). A significant lower cell viability in CGF grown in MC compared to cells grown in non conditioned medium was observed at 48 and 72 h (p <0.05). Conclusion: Gingival fibroblasts and even highly proliferative cells as B16F10 can be only superficially located on ADMA and CGF are negatively affected by ADM end products.