Cell Proliferation, Chondrogenic Differentiation, and Cartilaginous Tissue Formation in Recombinant Silk Fibroin with Basic Fibroblast Growth Factor Binding Peptide.

IF 5 3区 医学 Q1 ENGINEERING, BIOMEDICAL
Manabu Yamada, Arata Nakajima, Kayo Sakurai, Yasushi Tamada, Koichi Nakagawa
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Abstract

Regeneration of articular cartilage remains a challenge for patients who have undergone cartilage injury, osteochondritis dissecans and osteoarthritis. Here, we describe a new recombinant silk fibroin with basic fibroblast growth factor (bFGF) binding peptide, which has a genetically introduced sequence PLLQATLGGGS, named P7. In this study, we cultured a human mesenchymal cell line derived from bone marrow, UE6E7-16, in wild-type fibroin sponge (FS) and recombinant silk fibroin sponge with P7 peptide (P7 FS). We compared cell proliferation, chondrogenic differentiation and cartilaginous tissue formation between the two types of sponge. After stimulation with bFGF at 3 ng/mL, P7 FS showed significantly higher cell growth (1.2-fold) and higher cellular DNA content (5.6-fold) than did wild-type FS. To promote chondrogenic differentiation, cells were cultured in the presence of TGF-β at 10 ng/mL for 28 days. Immunostaining of P7 FS showed SOX9-positive cells comparable to wild-type FS. Alcian-Blue staining of P7 FS also showed cartilaginous tissue formation equivalent to wild-type FS. A significant increase in cell proliferation in P7 FS implies future clinical application of this transgenic fibroin for regeneration of articular cartilage. To produce cartilaginous tissue efficiently, transgenic fibroin sponges and culture conditions must be improved. Such changes should include the selection of growth factors involved in chondrogenic differentiation and cartilage formation.

含有碱性成纤维细胞生长因子结合肽的重组蚕丝纤维素的细胞增殖、软骨分化和软骨组织形成
对于软骨损伤、骨软骨炎和骨关节炎患者来说,关节软骨的再生仍然是一项挑战。在此,我们描述了一种含有碱性成纤维细胞生长因子(bFGF)结合肽的新型重组丝纤维蛋白,其基因序列为 PLLQATLGGGS,命名为 P7。在这项研究中,我们用野生型海绵纤维素(FS)和含有 P7 肽的重组海绵丝纤维素(P7 FS)培养了一种来源于骨髓的人类间充质细胞系 UE6E7-16。我们比较了两种海绵的细胞增殖、软骨分化和软骨组织形成情况。在 3 纳克/毫升的 bFGF 刺激下,P7 FS 的细胞生长速度(1.2 倍)和细胞 DNA 含量(5.6 倍)明显高于野生型 FS。为促进软骨分化,细胞在10 ng/mL的TGF-β存在下培养28天。P7 FS的免疫染色显示SOX9阳性细胞与野生型FS相当。P7 FS的Alcian-Blue染色也显示软骨组织的形成与野生型FS相当。P7 FS的细胞增殖明显增加,这意味着这种转基因纤维蛋白未来可用于关节软骨再生的临床应用。为了有效地生产软骨组织,必须改进转基因纤维蛋白海绵和培养条件。这些改变应包括选择参与软骨分化和软骨形成的生长因子。
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来源期刊
Journal of Functional Biomaterials
Journal of Functional Biomaterials Engineering-Biomedical Engineering
CiteScore
4.60
自引率
4.20%
发文量
226
审稿时长
11 weeks
期刊介绍: Journal of Functional Biomaterials (JFB, ISSN 2079-4983) is an international and interdisciplinary scientific journal that publishes regular research papers (articles), reviews and short communications about applications of materials for biomedical use. JFB covers subjects from chemistry, pharmacy, biology, physics over to engineering. The journal focuses on the preparation, performance and use of functional biomaterials in biomedical devices and their behaviour in physiological environments. Our aim is to encourage scientists to publish their results in as much detail as possible. Therefore, there is no restriction on the length of the papers. The full experimental details must be provided so that the results can be reproduced. Several topical special issues will be published. Scope: adhesion, adsorption, biocompatibility, biohybrid materials, bio-inert materials, biomaterials, biomedical devices, biomimetic materials, bone repair, cardiovascular devices, ceramics, composite materials, dental implants, dental materials, drug delivery systems, functional biopolymers, glasses, hyper branched polymers, molecularly imprinted polymers (MIPs), nanomedicine, nanoparticles, nanotechnology, natural materials, self-assembly smart materials, stimuli responsive materials, surface modification, tissue devices, tissue engineering, tissue-derived materials, urological devices.
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