A new method using insert-based systems (IBS) to improve cell behavior study on flexible and rigid biomaterials

Abstract

In vitro studies about biomaterials biological properties are essential screening tests. Yet cell cultures encounter difficulties related to cell retention on material surface or to the observation of both faces of permeable materials. The objective of the present study was to develop a reliable in vitro method to study cell behavior on rigid and flexible/permeable biomaterials elaborating two specific insert-based systems (IBS-R and IBS-F respectively). IBS-R was designed as a specific cylindrical polytetrafluoroethylene (PTFE) system to evaluate attachment, proliferation and morphology of human gingival fibroblasts (HGFs) on grade V titanium and lithium disilicate glassceramic discs characteristics of dental prostheses. The number of cells, their covering on discs and their morphology were determined from MTS assays and microscopic fluorescent images after 24, 48 and 72 h. IBS-F was developed as a two components system to study HGFs behavior on guided bone regeneration polyester membranes. The viability and the membrane barrier effect were evaluated by metabolic MTS assays and by scanning electron microscopy. IBS-R and IBS-F were shown to promote (1) easy and rapid handling; (2) cell retention on biomaterial surface; (3) accurate evaluation of the cellular proliferation, spreading and viability; (4) use of non-toxic material. Moreover IBS-F allowed the study of the cell migration through degradable membranes, with an access to both faces of the biomaterial and to the bottom of culture wells for medium changing.

In vitro and in vivo biocompatibility of calcium‐phosphate scaffolds three‐dimensional printed by stereolithography for bone regeneration

“…Although cell loading was low, the use of a PTFE insert allowed optimal cell seeding and attachment on the scaffolds by preventing cell loss at the bottom of the culture well without affecting cell behavior (Grenade et al, 2016). Although cell loading was low, the use of a PTFE insert allowed optimal cell seeding and attachment on the scaffolds by preventing cell loss at the bottom of the culture well without affecting cell behavior (Grenade et al, 2016).…”

“…Both biomaterials induced similar cell behavior as the number of cells per pellet, cell coverage, and viability were comparable after 3 days of culture. Although cell loading was low, the use of a PTFE insert allowed optimal cell seeding and attachment on the scaffolds by preventing cell loss at the bottom of the culture well without affecting cell behavior (Grenade et al, 2016). The human tumor cell line MG-63 was employed in this study given their osteoblastic features like alkaline phosphatase activity and osteoblast-like markers as well as their use in dental biomaterial biocompatibility experimentation (Aisa et al, 1996;Bächle & Kohal, 2004;Boyan, Schwartz, Bonewald, & Swain, 1989;Clover & Gowen, 1994;Czuba, Quintana, De Pauw-Gillet, et al, 2018;Wu et al, 2014).…”

“…Arrow, periosteum. B, cranial bone; CaP, calciumphosphate; P, HA-TCP pellet; PTFE, polytetrafluoroethylene; S, skin proliferation at the periphery of the pellet as previously described (Grenade et al, 2016) (Figure 1b).…”

“…Each experiment was performed in triplicate ( n = 3) for each group and repeated three times. Pellets were maintained during culture by a polytetrafluoroethylene (PTFE) insert in order to inhibit cell proliferation at the periphery of the pellet as previously described (Grenade et al, ) (Figure b).…”