Cross-linked Collagen–Chondroitin Sulfate–Hyaluronic Acid Imitating Extracellular Matrix as Scaffold for Dermal Tissue Engineering

Abstract

The objective of this study was to develop a novel scaffold imitating the ingredients and their ratios of natural dermal matrix and to evaluate its biological activity. We applied different ratios and different synthetic methods to fabricate nine kinds of cross-linked (CL) collagen/chondroitin sulfate/hyaluronic acid (Co/CS/HA) scaffolds for dermal tissue engineering. On the basis of comparison among the morphology, mechanical properties, and biodegradation rates of scaffolds, we selected the novel scaffold that was fabricated under unique procedures. In the procedures, Co, CS, and HA were firstly synthesized together in the ratio of 9:1:1 to form a membrane that was then CL with 5 mM of 1-ethyl-3-3-dimethylaminopropylcarbodiimide hydrochloride (EDC) (Co-CS-HA/CL 9:1:1). From the results of comparison, we also found that the ratio of 9:1:1 was better than other ratios. So the scaffold of Co-CS-HA/CL 9:1:1 was used as experimental group with the scaffolds of Co-HA/CS CL 9:1:1 and Co-CS/HA CL 9:1:1 as control groups to evaluate their characteristics in vitro. A control group of an open wound without scaffold was supplemented to evaluate their effects on promoting wound healing in vivo. Morphological observation showed that the novel Co-CS-HA/CL 9:1:1 scaffold had uniform and widely interconnected pores with mean diameters of 109 +/- 11 microm and adequate porosity of about 94%. Mechanical property and biodegradation assessment indicated that it had more degradation-resistant and higher elastic modulus than other scaffolds. Metabolic activity assay showed that it could more strongly promote cellular attachment and proliferation. When scaffolds were seeded with allogenic skin fibroblasts and implanted on the dorsum of Sprague-Dawley rats for 6 weeks, the novel Co-CS-HA/CL 9:1:1 skin equivalent could more successfully repair full thickness skin defects in Sprague-Dawley rats. The histology was more approximate to normal skin than those of the controls within 6 weeks. These results demonstrated that the novel CoCS-HA/CL 9:1:1 tri-copolymer has the potential to be used as a scaffold for dermal tissue engineering.

An Overview of Methods for theIn VivoEvaluation of Tissue-Engineered Skin Constructs

“…Transplanted cell ''label'' 131,133,139,146,150,152,153,166,173,184 b-galactosidase Transplanted cell label 143,163 Dil cell label Transplanted cell label 135 Green fluorescent protein Transplanted cell label 142,156 PKH26 red fluorescent cell Transplanted cell label 130,147,189 BM, basement membrane; BV, blood vessel; DEJ, dermal-epidermal junction.…”

Multifunctional Biomimetic Scaffolds Tailored for Cardiac Regeneration

“…149,150 Collagen has also been co-crosslinked with other proteins and macromolecules, such as fibroin, chondroitin sulfate and hyaluronic acid. 151,152 To fabricate of hydrogels for tissue engineering in general, the cross-linking strategy used must not use or result in cytotoxic products. It is also important that the degree of cross-linking can be tuned to such an extent that the materials obtained possess tissue specific properties, which in turn depends on the intended applications.…”

Leveraging “Raw Materials” as Building Blocks and Bioactive Signals in Regenerative Medicine

“…73 In addition, HA interacts with specific protein receptors on the surface of cells, such as CD44 and RHAMM, to modulate cell adhesion, proliferation, motility, and other signaling cascades. 74 For these reasons, HA has been utilized in recent tissue engineering strategies for skin, 50 89,91 Crosslinking can function to increase mechanical strength, while also prolonging degradation of HA. 65 Zhang et al 67 engineered a hydrogel scaffold by thermal crosslinking for cartilage tissue engineering that comprised solely of components found in the ECM of cartilage tissue using bovine collagen type I, HA, and CS (Table 3).…”

“…76 Additionally, the scaffolds were loaded with immobilized TGF-b3 and implanted in fullthickness cartilage defects in New Zealand white rabbits (Table 4). 76 Wang et al 50 employed a strategy of using solely raw materials to mimic the ECM of the dermis for skin tissue engineering grafts. The scaffold matrix consisted of collagen, CS, and HA with different ratios of each component, and was tested for optimal construct properties ( Table 5).…”

“…The scaffold matrix consisted of collagen, CS, and HA with different ratios of each component, and was tested for optimal construct properties ( Table 5). 50 This study, along with several others, embodied the emerging raw material approach for tissue engineering scaffolds. Overall, CS can be used to enhance mechanical integrity of a scaffold while also helping to mimic native ECM in connective tissues as well as articular cartilage.…”