Advances in Clinical and Experimental Medicine
2017, vol. 26, nr 2, March-April, p. 223–230
Publication type: original article
Collagenous scaffolds supplemented with hyaluronic acid and chondroitin sulfate used for wound fibroblast and embryonic nerve cell culture
1 Laboratory of Connective Tissue Metabolism, Department of Neuropeptides Research, Medical University of Lodz, Poland
2 Department of Material and Commodity Sciences and Textile Metrology, Lodz University of Technology, Poland
3 Department of Neuropeptides Research, Medical University of Lodz, Poland
4 Central Scientific Laboratory, Medical University of Lodz, Poland
Background. Tissue engineering is a strategy aimed at improving the regeneration of injured tissues.
Objectives. The aim of the present study was to determine whether a tri-copolymer composed of crosslinked collagen, chondroitin sulfate and hyaluronic acid (Col + CS + HA) provides a better environment for fibroblast and embryonic nerve cell culture than a collagenous scaffold (Col).
Material and Methods. The porosity of each of the matrices was characterized with a scanning electron microscope. Fibroblasts were isolated from rat wound granulation tissue (polypropylene net implanted subcutaneously). Embryonic nerve cells were obtained from the brains of rat embryos. The cells were applied to scaffolds and then stained with bisbenzimide to calculate cell entrapment within the material. The metabolic activity of the cells cultured within the scaffolds was tested using the 3-(4,5-dimethythiazol2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay.
Results. The Col scaffolds had a homogenously porous structure with a pore diameter of 50 μm for 70% of pores. The pore diameter in the tri-copolymer (Col + HA + CS) ranged from 24 to 160 μm (95% of total pore volume). Four times more cells (fibroblasts and embryonic nerve cells) were trapped within the superficial part of the collagenous scaffold than that of the tri-copolymer. On the third day of culture the metabolic activity of the fibroblasts within the 2 tested scaffolds was significantly higher than in the control conditions (cell culture on a laminin-coated surface). Also, the embryonic nerve cells demonstrated increased metabolic activity in Col + CS + HA scaffolds than the Col scaffolds.
Conclusion. Both fibroblasts and embryonic nerve cells could be seeded within the 2 tested scaffolds. Both the scaffolds provide good conditions for fibroblast culture. However, the Col + CS + HA tri-copolymer is preferable for embryonic nerve cell engineering.
collagen, fibroblasts in vitro, embryonic nerve cells
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