Article
In vitro assessment of mesenchymal stromal cells in different scaffolds for dermal regeneration
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Published: | September 27, 2011 |
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Introduction: Seeding of mesenchymal stromal cells (MSC) into scaffolds for dermal regeneration (SDR) has been described as a promising approach to achieve wound healing in clinical settings. However, the interaction between MSC and different biomaterials has not yet been described. The work presented here compares the behavior of MSC after seeding in different commonly used biomaterials.
Materials and methods: First, scaffolds based on collagen (Integra matrix®), Fibrin (Smart Matrix), Chitosane (BioPiel®) and acellularised skin (Strattice®) were compared in terms of their general structure by SEM and hydrophilicity. After, MSC isolated from human fat tissue of healthy donors who gave informed consent were seeded on the scaffolds (1.8 x 105 cells / 6 mm diameter SDR). Seeding volume was chosen according to the maximum fluid uptake of the materials. Seeding efficiency was quantified by cell counting of the non adherent MSC after incubation and by measuring metabolic activity with a water-soluble tetrazolin Assay (WST) on Day 1. Metabolic activity and viability of the cells was quantified by WST and Lactate Dehydrogenase (LDH) measurements over a 14 day period.
Results: SEM Analysis revealed important differences in the microstructure of the SDR, including pore size, thickness and inner surface. Here we found that Chitosane has a film-like structure so it cannot be considered as a three dimensional SDR as the others. Fluid uptake of the SDR differed largely showing that Integra had the highest fluid uptake (p < 0.05). Seeding efficiency was significantly lower in Chitosane (p < 0.05) while in all other SDR, seeding led to around 90 % of efficiency. Afterwards, WST Assay performed 24 hours after seeding showed significantly lower metabolic activity in Strattice® (p < 0.05), while high metabolic rates were found in all the other scaffolds without significant differences among them, over a 14 day period of analysis. Similar results were obtained by LDH analysis.
Conclusions: Comparison of the four scaffolds shows that Integra®, Smart Matrix and BioPiel® are potential candidates for Application of MSC to wounds while seeding MSC to Strattice was unsuccessful due to low survival rates of the cells. This work represents the first step to optimize the use of MSC-seeded scaffolds for wound healing. However further studies have to be performed to evaluate their significance in vivo.