Abstract
Identifying collagen produced de novo by cells in a background of purified collagenous biomaterials poses a major problem in for example the evaluation of tissue-engineered constructs and cell biological studies to tumor dissemination. We have developed a universal strategy to detect and localize newly deposited collagen based on its inherent association with dermatan sulfate. The method is applicable irrespective of host species and collagen source.
Highlights
Identifying collagen produced de novo by cells in a background of purified collagenous biomaterials poses a major problem in for example the evaluation of tissue-engineered constructs and cell biological studies to tumor dissemination
Dermatan sulfate is the glycosaminoglycan part of the proteoglycans decorin and biglycan, which are both collagen fibril-associated molecules that play a role in the regulation of collagen fibril diameter
These proteoglycans remain present on the mature collagen fibril (Fig. 1a, cartoon), and dermatan sulfate is associated with collagen fibrils[5,6]
Summary
Identifying collagen produced de novo by cells in a background of purified collagenous biomaterials poses a major problem in for example the evaluation of tissue-engineered constructs and cell biological studies to tumor dissemination. We have developed a universal strategy to detect and localize newly deposited collagen based on its inherent association with dermatan sulfate. In this study we evaluated newly synthesized fibrillar collagen (e.g. type I collagen), by making use of the inherent and intrinsic association of the glycosaminoglycan dermatan sulfate with collagen fibrils. Dermatan sulfate is the glycosaminoglycan part of the proteoglycans decorin and biglycan, which are both collagen fibril-associated molecules that play a role in the regulation of collagen fibril diameter. These proteoglycans remain present on the mature collagen fibril (Fig. 1a, cartoon), and dermatan sulfate is associated with collagen fibrils[5,6]. We tested the technique both in vivo and in vitro using a number of collagenous biomaterials including gels cultured with human fibroblasts with or without keratinocytes (denovoSkin and denovoDerm respectively)[8], experimental and commercially available scaffolds, and glycerol preserved acellular human dermis
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