Richert, Jordan Chay (2016-04). A Comparison of Rat Gingival Fibroblast Attachment on Commercial Acellular Dermal Matrices: An In Vitro Study. Master's Thesis.
Thesis
Gingival recession is defined as a mucogingival deformity in which the marginal soft tissues are positioned apical to cemento-enamel junction, with concomitant loss of attached gingiva and exposure of root surfaces. Surgical correction to obtain root coverage can be done in several ways including, sliding flaps, free gingival grafts, connective tissue grafts, coronally advanced flaps, or allografts. The purpose of this study was to characterize in vitro, fibroblast viability, and morphological appearance, when grown on three commercial acellular dermal matrices used in gingival grafting. Acellular dermal matrices were tested AlloDerm(R) RTM (BioHorizons, Birmingham, AL), Puros Dermis (Zimmer Dental, Carlsbad, CA), and PerioDerm(TM) (Dentsply Implants, Watham, MA). Primary rat gingival fibroblasts were cultured and used between the second and fourth passage for all experiments. All dermal matrices were obtained as packaged for surgical use, cut into 4 mm x 4mm squares, rehydrated in 0.9% sterile saline, and seeded with 3.5 x 10^5 cells to 4.0 x 10^5 cells/well. Cells were cultured for 24 hours, 1 week, and 2 weeks in media containing DMEM, 10% fetal bovine serum, and 1% antibiotic/antimycotic. The samples for each dermal matrix per time point (n=3) were either harvested for viability/morphology using confocal, scanning electron, and light microscopy. The samples harvested for viability where stained with live/dead fluorescent dye immediately after culture. The samples harvested for morphology were fixed in 4% paraformaldehyde for 30 minutes, washed and processed for scanning electron microscopy (SEM) or hemotoxylin and eosin (H&E) staining. All three dermal matrix products appeared to facilitate cell growth. Viability determined via live/dead assay revealed no significant differences between groups. Histologic examination showed fibroblasts throughout various layers of the dermis. SEM data revealed that as time progressed, the matrices exhibited a smoother topography, with dead cells and cellular extensions visualized at higher magnification. All commercial acellular dermal matrices supported rat gingival fibroblast growth. There were no significant differences in cell viability or cell distribution based on live/dead assay and H&E sections. SEM data supported the theory that fibroblasts consumed and remodeled all acellular dermal matrices.
