Three-Dimensional Culture of Cartilage Tissue on Nanogel-Cross-Linked Porous Freeze-Dried Gel Scaffold for Regenerative Cartilage Therapy: A Vibrational Spectroscopy Evaluation

被引:7
作者
Adachi, Tetsuya [1 ,2 ]
Miyamoto, Nao [1 ,3 ]
Imamura, Hayata [1 ,4 ]
Yamamoto, Toshiro [1 ]
Marin, Elia [1 ,4 ]
Zhu, Wenliang [4 ]
Kobara, Miyuki [5 ]
Sowa, Yoshihiro [2 ,6 ,7 ]
Tahara, Yoshiro [8 ]
Kanamura, Narisato [1 ]
Akiyoshi, Kazunari [9 ]
Mazda, Osam [2 ]
Nishimura, Ichiro [10 ,11 ]
Pezzotti, Giuseppe [1 ,2 ,4 ,12 ]
机构
[1] Kyoto Prefectural Univ Med, Grad Sch Med Sci, Dept Dent Med, Kamigyo Ku, Kyoto 6028566, Japan
[2] Kyoto Prefectural Univ Med, Grad Sch Med Sci, Dept Immunol, Kamigyo Ku, 465 Kajii Cho, Kyoto 6028566, Japan
[3] Kyoto Prefectural Univ Med, Infect Dis, Kamigyo Ku, 465 Kajii Cho, Kyoto 6028566, Japan
[4] Kyoto Inst Technol, Ceram Phys Lab, Sakyo Ku, Matsugasaki, Kyoto 6068585, Japan
[5] Kyoto Pharmaceut Univ, Dept Clin Pharmacol, Div Pathol Sci, Yamashina Ku, Misasagi Nakauchi Cho, Kyoto 6078414, Japan
[6] Kyoto Prefectural Univ Med, Dept Plast & Reconstruct Surg, Kamigyo Ku, 465 Kajii Cho, Kyoto 6028566, Japan
[7] Kyoto Univ, Grad Sch Med, Dept Plast & Reconstruct Surg, Sakyo Ku, Yoshida Konoe Cho, Kyoto 6068501, Japan
[8] Doshisha Univ, Dept Chem Engn & Mat Sci, 1-3 Tatara Miyakodani, Kyotanabe, Kyoto 6100394, Japan
[9] Kyoto Univ, Grad Sch Engn, Dept Polymer Chem, Nishikyo Ku, Kyoto 6158510, Japan
[10] Univ Calif Los Angeles, Sch Dent, Jane & Jerry Weintraub Ctr Reconstruct Biotechnol, Div Oral Biol & Med, Los Angeles, CA 90095 USA
[11] Univ Calif Los Angeles, Sch Dent, Jane & Jerry Weintraub Ctr Reconstruct Biotechnol, Div Adv Prosthodont, Los Angeles, CA 90095 USA
[12] Kyoto Inst Technol, Biomed Res Ctr, Sakyo Ku, Matsugasaki, Kyoto 6068585, Japan
关键词
NanoCliP-FD gel scaffold; tissue engineering; regenerative therapy; human periodontal ligament-derived stem cell; in situ Raman spectroscopy; LIGAMENT STEM-CELLS; RAMAN-SPECTROSCOPY; COLLAGEN; PROTEOGLYCAN; PROTEIN; PEPTIDE; SPECTRA; BONE; COORDINATE; SCATTERING;
D O I
10.3390/ijms23158099
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
This study presents a set of vibrational characterizations on a nanogel-cross-linked porous freeze-dried gel (NanoCliP-FD gel) scaffold for tissue engineering and regenerative therapy. This scaffold is designed for the in vitro culture of high-quality cartilage tissue to be then transplanted in vivo to enable recovery from congenital malformations in the maxillofacial area or crippling jaw disease. The three-dimensional scaffold for in-plate culture is designed with interface chemistry capable of stimulating cartilage formation and maintaining its structure through counteracting the dedifferentiation of mesenchymal stem cells (MSCs) during the formation of cartilage tissue. The developed interface chemistry enabled high efficiency in both growth rate and tissue quality, thus satisfying the requirements of large volumes, high matrix quality, and superior mechanical properties needed in cartilage transplants. We characterized the cartilage tissue in vitro grown on a NanoCliP-FD gel scaffold by human periodontal ligament-derived stem cells (a type of MSC) with cartilage grown by the same cells and under the same conditions on a conventional (porous) atelocollagen scaffold. The cartilage tissues produced by the MSCs on different scaffolds were comparatively evaluated by immunohistochemical and spectroscopic analyses. Cartilage differentiation occurred at a higher rate when MSCs were cultured on the NanoCliP-FD gel scaffold compared to the atelocollagen scaffold, and produced a tissue richer in cartilage matrix. In situ spectroscopic analyses revealed the cell/scaffold interactive mechanisms by which the NanoCliP-FD gel scaffold stimulated such increased efficiency in cartilage matrix formation. In addition to demonstrating the high potential of human periodontal ligament-derived stem cell cultures on NanoCliP-FD gel scaffolds in regenerative cartilage therapy, the present study also highlights the novelty of Raman spectroscopy as a non-destructive method for the concurrent evaluation of matrix quality and cell metabolic response. In situ Raman analyses on living cells unveiled for the first time the underlying physiological mechanisms behind such improved chondrocyte performance.
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页数:21
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