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生长因子处理的张紧滑膜细胞新组织:迈向半月板生物支架组织工程

Growth factor treated tensioned synoviocyte neotissues: towards meniscal bioscaffold tissue engineering.

作者信息

Warnock J J, Bobe G, Duesterdieck-Zellmer K F, Spina J, Ott J, Baltzer W I, Bay B K

机构信息

Department of Clinical Sciences, College of Veterinary Medicine, Oregon State University, Corvallis, OR 97331, USA.

Department of Animal and Rangeland Sciences, Oregon State University, Corvallis, OR 97331, USA; Linus Pauling Institute, Oregon State University, OR 97331, USA.

出版信息

Vet J. 2014 Apr;200(1):22-30. doi: 10.1016/j.tvjl.2014.01.010. Epub 2014 Jan 24.

Abstract

Meniscal injury is a common cause of osteoarthritis, pain, and disability in dogs and humans, but tissue-engineered bioscaffolds could be a treatment option for meniscal deficiency. The objective of this study was to compare meniscus-like matrix histology, composition, and biomechanical properties of autologous tensioned synoviocyte neotissues (TSN) treated with fetal bovine serum (TSNfbs) or three chondrogenic growth factors (TSNgf). Fourth passage canine synoviocytes from 10 dogs were grown in hyperconfluent monolayer culture, formed into TSN, and then cultured for 3 weeks with 17.7% FBS or three human recombinant TSNgf (bFGF, TGF-β1, and IGF-1). Cell viability was determined with laser microscopy. Histological architecture and the composition of fibrocartilage matrix were evaluated in TSN by staining tissues for glycosaminoglycan (GAG), α-smooth muscle actin, and collagen 1 and 2; quantifying the content of GAG, DNA, and hydroxyproline; and measuring the gene expression of collagens type 1α and 2α, the GAG aggrecan, and transcription factor Sry-type Homeobox Protein-9 (SOX9). Biomechanical properties were determined by materials testing force-deformation curves. The TSN contained components and histological features of mensical fibrocartilage extracellular matrix. Growth factor-treated TSN had higher DNA content but lower cell viability than TSNfbs. TSNgf had greater fibrocartilage-like matrix content (collagen 2 and GAG content with increased collagen 2α and SOX9 gene expression). Additionally, TSNgf collagen was more organized histologically and so had greater tensile biomechanical properties. The results indicate the potential of TSN when cultured with growth factors as implantable bioscaffolds for the treatment of canine meniscal deficiency.

摘要

半月板损伤是犬类和人类骨关节炎、疼痛及残疾的常见原因,但组织工程生物支架可能是半月板缺损的一种治疗选择。本研究的目的是比较用胎牛血清(TSNfbs)或三种软骨生成生长因子(TSNgf)处理的自体张紧滑膜细胞新组织(TSN)的半月板样基质组织学、组成和生物力学特性。从10只犬获取的第四代滑膜细胞在高汇合单层培养中生长,形成TSN,然后用17.7%胎牛血清或三种人重组TSNgf(碱性成纤维细胞生长因子、转化生长因子-β1和胰岛素样生长因子-1)培养3周。用激光显微镜测定细胞活力。通过对组织进行糖胺聚糖(GAG)、α-平滑肌肌动蛋白、胶原蛋白1和2染色;定量GAG、DNA和羟脯氨酸含量;以及测量1α型和2α型胶原蛋白、GAG聚集蛋白聚糖和转录因子Sry型同源盒蛋白9(SOX9)的基因表达,评估TSN中纤维软骨基质的组织结构和组成。通过材料测试力-变形曲线测定生物力学特性。TSN含有半月板纤维软骨细胞外基质的成分和组织学特征。生长因子处理的TSN比TSNfbs具有更高的DNA含量但更低的细胞活力。TSNgf具有更高的纤维软骨样基质含量(胶原蛋白2和GAG含量增加,同时胶原蛋白2α和SOX9基因表达增加)。此外,TSNgf胶原蛋白在组织学上更有序,因此具有更大的拉伸生物力学特性。结果表明,生长因子培养的TSN作为可植入生物支架治疗犬半月板缺损具有潜力。

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