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对接种于I型和II型胶原蛋白植入物中的犬软骨细胞进行了体外研究。

Canine chondrocytes seeded in type I and type II collagen implants investigated in vitro.

作者信息

Nehrer S, Breinan H A, Ramappa A, Shortkroff S, Young G, Minas T, Sledge C B, Yannas I V, Spector M

机构信息

Department of Orthopedic Surgery, Brigham & Women's Hospital, Harvard Medical School, Boston, Massachusetts, USA.

出版信息

J Biomed Mater Res. 1997 Summer;38(2):95-104. doi: 10.1002/(sici)1097-4636(199722)38:2<95::aid-jbm3>3.0.co;2-b.

Abstract

Synthetic and natural absorbable polymers have been used as vehicles for implantation of cells into cartilage defects to promote regeneration of the articular joint surface. Implants should provide a pore structure that allows cell adhesion and growth, and not provoke inflammation or toxicity when implanted in vivo. The scaffold should be absorbable and the degradation should match the rate of tissue regeneration. To facilitate cartilage repair the chemical structure and pore architecture of the matrix should allow the seeded cells to maintain the chondrocytic phenotype, characterized by synthesis of cartilage-specific proteins. We investigated the behavior of canine chondrocytes in two spongelike matrices in vitro: a collagen-glycosaminoglycan (GAG) copolymer produced from bovine hide consisting of type I collagen and a porous scaffold made of type II collagen by extraction of porcine cartilage. Canine chondrocytes were seeded on both types of matrices and cultured for 3 h, 7 days, and 14 days. The histology of chondrocyte-seeded implants showed a significantly higher percentage of cells with spherical morphology, consistent with chondrocytic morphology, in the type II sponge at each time point. Pericellular matrix stained for proteoglycans and for type II collagen after 14 days. Biochemical analysis of the cell seeded sponges for GAG and DNA content showed increases with time. At day 14 there was a significantly higher amount of DNA and GAG in the type II matrix. This is the first study that directly compares the behavior of chondrocytes in type I and type II collagen matrices. The type II matrix may be of value as a vehicle for chondrocyte implantation on the basis of the higher percentage of chondrocytes retaining spherical morphology and greater biosynthetic activity that was reflected in the greater increase of GAG content.

摘要

合成和天然可吸收聚合物已被用作将细胞植入软骨缺损以促进关节表面再生的载体。植入物应提供一种允许细胞粘附和生长的孔隙结构,并且在体内植入时不会引发炎症或毒性。支架应是可吸收的,其降解速率应与组织再生速率相匹配。为了促进软骨修复,基质的化学结构和孔隙结构应使接种的细胞能够维持软骨细胞表型,其特征是合成软骨特异性蛋白质。我们在体外研究了犬软骨细胞在两种海绵状基质中的行为:一种由牛皮制成的胶原蛋白 - 糖胺聚糖(GAG)共聚物,其包含I型胶原蛋白,以及一种通过提取猪软骨制成的II型胶原蛋白多孔支架。将犬软骨细胞接种在两种类型的基质上,并培养3小时、7天和14天。接种软骨细胞的植入物的组织学显示,在每个时间点,II型海绵中具有球形形态的细胞百分比显著更高,这与软骨细胞形态一致。14天后,细胞周围基质对蛋白聚糖和II型胶原蛋白进行了染色。对接种细胞的海绵进行的GAG和DNA含量的生化分析显示,其随时间增加。在第14天,II型基质中的DNA和GAG含量显著更高。这是第一项直接比较软骨细胞在I型和II型胶原蛋白基质中行为的研究。基于保留球形形态的软骨细胞百分比更高以及GAG含量的更大增加所反映的更高生物合成活性,II型基质可能作为软骨细胞植入的载体具有价值。

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