The Use of Confocal Laser Scanning Microscopy to Assess the Potential Suitability of 3d Scaffolds for Tissue Regeneration, by Monitoring Extra‐cellular Matrix Deposition and by Quantifying Cellular Infiltration and Proliferation

An in vitro evaluation of fibrinogen and gelatin containing cryogels as dermal regeneration scaffolds

“…This was also noted when we used this method to evaluate cellular infiltration of a commercially available dermal replacement scaffold material, Integra. 18 These differences in cell distribution, with the majority of cells on the external surfaces of the scaffolds, represent a challenge in biomaterial design. In a clinical setting, it is important that dermal scaffolds provide a suitable environment for the proliferation and migration of the overlying epidermal cells that are ultimately responsible for wound closure.…”

“…At 80% confluence, cells were harvested with trypsin and seeded in 2 ml volumes at a concentration of 1 × 10 4 cells per ml into 12-well TC plates (Greiner Bio-one) and incubated as above for 5 days until confluent, and therefore contact inhibited. 18 To assess cellular proliferation and infiltration into the cryogel samples, cells were allowed to migrate against gravity from the TC surface into the cryogels by placing the bisected half-cylinder gels, flat surface down, onto the confluent cell layer ( Fig. 1) with incubation at 37°C, 100% RH, 5% CO 2 for Fig.…”

“…CLSM settings were as described previously, with the protein of interest shown as a green or blue colour, as indicated. 18 The autofluorescent properties of the cryogels were exploited to enable viewing of the scaffold structure; fluorescent output converted to white (Fig. 4) or magenta (Fig.…”