Carbodiimide Conjugation of Fibronectin on Collagen Basal Lamina Analogs Enhances Cellular Binding Domains and Epithelialization

Abstract

To improve the regenerative potential of biomaterials used as bioengineered scaffolds, it is necessary to strategically incorporate biologically active molecules that promote in vivo cellular processes that lead to a fully functional tissue. This work evaluates the effects of strategically binding fibronectin (FN) to collagen basal lamina analogs to enhance keratinocyte functions necessary for complete skin regeneration. We found that FN that was passively adsorbed to collagen-glycosaminoglycan basal lamina analogs enhanced epithelial thickness and keratinocyte proliferation compared with nontreated basal lamina analogs at 3 days of air/liquid (A/L) interface culture. Additionally, we evaluated the availability of FN cellular binding site domains when FN was either passively adsorbed or [1-ethyl-3-(3-dimethylaminopropyl) carbodiimide hydrochloride] conjugated to basal lamina analogs fabricated from collagen-glycosaminoglycan coprecipitate or self-assembled type I collagen. It was found that 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide hydrochloride conjugation of FN significantly enhanced FN binding site presentation as well as epithelial thickness. Overall, the results gained from this study will be used to improve the regenerative capacity of basal lamina analogs for bioengineered skin substitutes as well as the development of bioengineered scaffolds for other tissue engineering applications.