Regeneration of elastic fibers by three-dimensional culture on a collagen scaffold and the addition of latent TGF-β binding protein 4 to improve elastic matrix deposition

• Published in: Biomaterials 2015-12, Vol.72, p.29-37
• Main Authors: Aya, Rino, Ishiko, Toshihiro, Noda, Kazuo, Yamawaki, Satoko, Sakamoto, Yuki, Tomihata, Kenji, Katayama, Yasuhiro, Yoshikawa, Katsuhiro, Kubota, Hiroshi, Nakamura, Tomoyuki, Naitoh, Motoko, Suzuki, Shigehiko
• Format: Article
• Language: English
• Subjects: Advanced Basic Science; Animals; Cell Culture Techniques - methods; Cells, Cultured; Collagen - pharmacology; Collagen scaffold; Collagens; Crosslinking; Culture; Dentistry; Dermis - cytology; Elastic fibers; Elastic Tissue - drug effects; Elastic Tissue - metabolism; Extracellular Matrix - drug effects; Extracellular Matrix - metabolism; Fibers; Fibroblasts - cytology; Fibroblasts - drug effects; Fibroblasts - metabolism; Fluorescent Antibody Technique; Humans; Latent TGF-beta Binding Proteins - pharmacology; LTBP-4; Male; Mice, Inbred BALB C; Mice, Nude; Recombinant Proteins - pharmacology; Regeneration; Scaffolds; Sponges; Sus scrofa; Three dimensional; Tissue Scaffolds - chemistry
• ISSN: 0142-9612; 1878-5905
• DOI: 10.1016/j.biomaterials.2015.08.036

Abstract The objective of this study was to investigate the effects of latent TGF-β binding protein 4 (LTBP-4) on elastic fiber regeneration in three-dimensional cultures of human dermal fibroblasts (HDFs). Appropriate collagen scaffold for elastic fiber regeneration was also examined. Collagen sponges cross-linked at 120 °C and composed of small pores (25 μm on average) was favorable for elastic fiber regeneration by HDFs. Addition of LTBP-4, followed by culture for 21 days, accelerated elastic fiber accumulation within the scaffolds. Conditioned scaffolds containing either HDFs or LTBP-4-built mature elastic fibers were implanted between the dermis and the cutaneous muscle of mice. The combined use of HDFs and LTBP-4 resulted in thicker tissues containing elastic fibers. These results indicate that weakly cross-linked collagen sponges can be used as scaffolds for regenerating elastic fibers both in vitro and in vivo , and that the addition of LTBP-4 accelerates the deposition of both elastin and fibrillin-1, and increases cell proliferation. These techniques may be useful for generating cutaneous or cardiovascular tissue equivalents; furthermore, they may serve as a useful method for the three-dimensional analyses of drugs used to treat skin diseases or to examine the microstructure of elastin networks.