Experience coming from many cell culture studies has opened new perspectives: the hypothesis that tissue and organ reconstruction should be performed in a three-dimensional environment as it normally occurs in vivo, is now not so far off. The base of every study is that all in vitro tissue substitutes need an important neoangiogenesis to be successfully transplanted. As far as endothelial cell culture is concerned, it has been shown that angiogenesis can be successfully achieved only when cells are cultured in the presence of collagen-based matrices or basal membrane substrates. The aim of the present investigation was to demonstrate that human umbilical vein endothelial cells (HUVEC) can be grown and differentiated on an artificial dermis obtained by human fibroblasts cultured on hyaluronic acid-based scaffolds. For this purpose we have cultured HUVEC, retrieved by collagenase digestion of perfused human umbilical veins either alone or with fibroblast at a 1/1 ratio into HYAFF-11. The product is an uncross-linked linear polymer with an undetermined molecular weight, insoluble in aqueous solution yet spontaneously hydrolyzes over time, releasing benzyl alcohol and hyaluronan. HYAFF-11 was used to create non-woven meshes. Cultures were maintained for up to three weeks. Samples were taken at different time-points within this period for the MTT proliferation test and for immunohistochemical analysis. One-way analysis of variance (Anova test) of the software package Excel was used for data analyses. Repeat measurement analysis of variance and paired t-tests were used to determine if there were significant changes (p < 0.05). Repeatability was calculated as the standard deviation of the difference between measurements of the MTT test performed. The proliferation rate of HUVEC cultured alone and co-cultured with fibroblasts was compared. Our results demonstrate that hyaluronan-based biomaterials (HYAFF-11 NW mesh) represent a suitable substrate for HUVEC adhesion, proliferation and reorganization in the microcapillary network.

In vitro reconstruction of a microcapillary network into a tissue-engineered human dermal equivalent

ZAVAN, BARBARA;CORTIVO, ROBERTA;BRUN, PAOLA;BASSETTO, FRANCO;ABATANGELO, GIOVANNI
2002

Abstract

Experience coming from many cell culture studies has opened new perspectives: the hypothesis that tissue and organ reconstruction should be performed in a three-dimensional environment as it normally occurs in vivo, is now not so far off. The base of every study is that all in vitro tissue substitutes need an important neoangiogenesis to be successfully transplanted. As far as endothelial cell culture is concerned, it has been shown that angiogenesis can be successfully achieved only when cells are cultured in the presence of collagen-based matrices or basal membrane substrates. The aim of the present investigation was to demonstrate that human umbilical vein endothelial cells (HUVEC) can be grown and differentiated on an artificial dermis obtained by human fibroblasts cultured on hyaluronic acid-based scaffolds. For this purpose we have cultured HUVEC, retrieved by collagenase digestion of perfused human umbilical veins either alone or with fibroblast at a 1/1 ratio into HYAFF-11. The product is an uncross-linked linear polymer with an undetermined molecular weight, insoluble in aqueous solution yet spontaneously hydrolyzes over time, releasing benzyl alcohol and hyaluronan. HYAFF-11 was used to create non-woven meshes. Cultures were maintained for up to three weeks. Samples were taken at different time-points within this period for the MTT proliferation test and for immunohistochemical analysis. One-way analysis of variance (Anova test) of the software package Excel was used for data analyses. Repeat measurement analysis of variance and paired t-tests were used to determine if there were significant changes (p < 0.05). Repeatability was calculated as the standard deviation of the difference between measurements of the MTT test performed. The proliferation rate of HUVEC cultured alone and co-cultured with fibroblasts was compared. Our results demonstrate that hyaluronan-based biomaterials (HYAFF-11 NW mesh) represent a suitable substrate for HUVEC adhesion, proliferation and reorganization in the microcapillary network.
2002
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Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/11577/2525532
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