Journal
BIOMATERIALS
Volume 24, Issue 18, Pages 3125-3132Publisher
ELSEVIER SCI LTD
DOI: 10.1016/S0142-9612(03)00156-X
Keywords
tissue engineering; human adipose tissue; preadipocytes; hyaluronic acid; scaffolds
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Background and aim of the study: Reconstruction of soft tissue defects is a challenge in plastic surgery and there is clinical need for adequate solutions. Aim of this study was to develop a biohybrid construct consisting of hyaluronic acid-based scaffolds and human adipocyte precursor cells as a soft tissue filler. Methods: Human adipocyte precursor cells were obtained by collagenase digestion of adipose tissue samples and seeded on hyaluronic acid-based spongy scaffolds of various degrees of esterification and pore size using different techniques. After cell attachment. adipose differentiation was induced by defined adipogenic factors under serum-free culture conditions. Results: Among the five different scaffold types under investigation the highest cell attachment rate was observed for the HYAFF(R) scaffold with 100% esterification and a mean pore size of 400 mum (HYAFF(R) 111p). For inoculation of human adipocyte precursor cells on hyaluronic acid-based scaffolds a drop-on technique and low-pressure centrifugation using a Speed Vac(R) airfuge were compared. With respect to efficacy, cell distribution and simpleness the drop-on method proved to be the method of choice. In a serum-free medium supplemented with 66 nm insulin, 100 nm cortisol and I mug/ml troglitazone a substantial proportion of cells underwent adipose differentiation as assessed by lipid accumulation and emergence of glycerol-3-phosphate dehydrogenase activity, a lipogenic marker enzyme. Conclusion: Hyaluronic acid-based scaffolds appear to be a suitable three-dimensional carrier for the culture and in vitro differentiation of human adipocyte precursor cells. (C) 2003 Elsevier Science Ltd. All rights reserved.
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