Evaluation of in vitro human gingival fibroblast seeding on acellular dermal matrix

Abstract

The acellular dermal matrix (ADM) was introduced in periodontology as a substitute for the autogenous grafts, which became restricted because of the limited source of donor's tissue. The aim of this study was to investigate, in vitro, the distribution, proliferation and viability of human gingival fibroblasts seeded onto ADM. ADM was seeded with human gingival fibroblasts for up to 21 days. The following parameters were evaluated: cell distribution, proliferation and viability. Results revealed that, at day 7, fibroblasts were adherent and spread on ADM surface, and were unevenly distributed, forming a discontinuous single cell layer; at day 14, a confluent fibroblastic monolayer lining ADM surface was noticed. At day 21, the cell monolayer exhibited a reduction in cell density. At 7 days, about to 90% of adherent cells on ADM surface were cycling while at 14 and 21 days this proportion was significantly reduced. A high proportion of viable cell was detected on AMD surface both on 14 and 21 days. The results suggest that fibroblast seeding onto ADM for 14 days can allow good conditions for cell adhesion and spreading on the matrix; however, migration inside the matrix was limited.

Atomic force microscopy in the production of a biovital skin graft based on human acellular dermal matrix produced in‐house and in vitro cultured human fibroblasts

“…Similarly to observations made by Ng et al, Rodrigues et al, and Sorrel and Caplan, fibroblast inhabiting the surface of the studied ADMs did not migrate inwards toward the ECM structure. The reasonable explanation of this phenomenon may be attributed to dense, strict, and highly cross‐linked collagen structure of ADM .…”

“…The histological image of ADM‐2 depicts cell‐free ECM. The decellularization with the use of 0.05% trypsin and 0.02% EDTA solution (as for ADM‐2) was explored by other authors . They examined the efficiency of this procedure for decellularization of porcine heart valves.…”

“…A production of biovital skin grafts based on human ADMs and in vitro cultured human fibroblasts is a widely discussed issue. However, the vast majority of protocols involve the use of the commercially available cell‐free skin substitute‐ AlloDerm . In this study, because of the widespread availability of allogeneic human skin grafts stored in tissue bank, the research for an optimal, in‐house method of biovital skin substitute production was done.…”

A new approach to the production of a biovital skin graft based on human acellular dermal matrix produced in‐house, in vitro revitalized internally by human fibroblasts and keratinocytes on the surface

“…This phenomenon has also been observed by other authors, for example, observations made by Ng et al (), Rodrigues et al (), or Sorrel and Caplan (). They showed that fibroblasts colonizing the surface of the study ADMs cannot migrate inward towards the ECM structure (Ng et al, ; Rodrigues et al, ; Sorrel & Caplan, ). In order to facilitate fibroblast migration to the interior of the matrices, enzymatic and mechanical methods have been proposed.…”

Morphological, Functional and Biochemical Characterization of Canine Gingival Fibroblasts

“…Some authors have been evaluating non-enzymatic methods of gingival fibroblasts isolation in vitro for the use in gingival augmentation (6,(10)(11)(12)(13)(14). Gingival fibroblast culture in different matrices has been studied and has shown promising results in soft tissues regeneration (12)(13)(14). Fibroblasts are responsible for the normal growth of the organism and are essential in cases where tissue repairs are necessary, being one of the first cells to appear at injured sites.…”