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.
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.
Department of Plastic and Reconstructive Surgery, Graduate School of Medicine, Kyoto University, 54 Kawaharacho, Shogoin, Sakyo-ku, Kyoto 606-8507, Japan.
Rino
R
Aya
Department of Plastic and Reconstructive Surgery, Otsu Red Cross Hospital, 1-1-35 Nagara Otsu, Shiga 520-8511, Japan.
Toshihiro
T
Ishiko
Department of Pediatrics, The University of Texas Medical School at Houston, TX 77030, USA.
Kazuo
K
Noda
Department of Plastic and Reconstructive Surgery, Graduate School of Medicine, Kyoto University, 54 Kawaharacho, Shogoin, Sakyo-ku, Kyoto 606-8507, Japan.
Satoko
S
Yamawaki
GUNZE LIMITED QOL Research Center Laboratory, 1 Aonocho Zeze Ayabe, Kyoto 623-8512, Japan.
Yuki
Y
Sakamoto
GUNZE LIMITED Research and Development Department, 1 Aonocho Zeze Ayabe, Kyoto 623-8512, Japan.
Kenji
K
Tomihata
Department of Plastic and Reconstructive Surgery, Graduate School of Medicine, Kyoto University, 54 Kawaharacho, Shogoin, Sakyo-ku, Kyoto 606-8507, Japan.
Yasuhiro
Y
Katayama
Department of Plastic and Reconstructive Surgery, Graduate School of Medicine, Kyoto University, 54 Kawaharacho, Shogoin, Sakyo-ku, Kyoto 606-8507, Japan.
Katsuhiro
K
Yoshikawa
Department of Life Science, Faculty of Engineering and Resource Science, Akita University, 1-1 Tegata Gakuenmachi, Akita 010-8502, Japan.
Hiroshi
H
Kubota
Department of Pharmacology, Kansai Medical University, 2-5-1 Shinmachi Hirakata, Osaka 573-1191, Japan.
Tomoyuki
T
Nakamura
Department of Plastic and Reconstructive Surgery, Graduate School of Medicine, Kyoto University, 54 Kawaharacho, Shogoin, Sakyo-ku, Kyoto 606-8507, Japan. Electronic address: mnaitoh@kuhp.kyoto-u.ac.jp.
Motoko
M
Naitoh
Department of Plastic and Reconstructive Surgery, Graduate School of Medicine, Kyoto University, 54 Kawaharacho, Shogoin, Sakyo-ku, Kyoto 606-8507, Japan.
Shigehiko
S
Suzuki
D000818:Animals; D018929:Cell Culture Techniques / Q000379:methods*; D002478:Cells, Cultured; D003094:Collagen / Q000494:pharmacology*; D020405:Dermis / Q000166:cytology; D004547:Elastic Tissue / Q000187:drug effects / Q000378:metabolism*; D005109:Extracellular Matrix / Q000187:drug effects / Q000378:metabolism*; D005347:Fibroblasts / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D005455:Fluorescent Antibody Technique; D006801:Humans; D053779:Latent TGF-beta Binding Proteins / Q000494:pharmacology*; D008297:Male; D008807:Mice, Inbred BALB C; D008819:Mice, Nude; D011994:Recombinant Proteins / Q000494:pharmacology; D012038:Regeneration; D034421:Sus scrofa; D054457:Tissue Scaffolds / Q000737:chemistry*
C509040:LTBP4 protein, human; D053779:Latent TGF-beta Binding Proteins; D011994:Recombinant Proteins; D003094:Collagen
Collagen scaffold; Elastic fibers; LTBP-4; Regeneration