Warnock Jennifer J, Spina Jason, Bobe Gerd, Duesterdieck-Zellmer Katja F, Ott Jesse, Baltzer Wendy I, Bay Brian K
Department of Clinical Sciences, College of Veterinary Medicine, Oregon State University, Corvallis, OR 97331, USA.
Department of Clinical Sciences, College of Veterinary Medicine, Oregon State University, Corvallis, OR 97331, USA.
Vet J. 2014 Jan;199(1):49-56. doi: 10.1016/j.tvjl.2013.10.038. Epub 2013 Nov 11.
Meniscal injury is a common cause of canine lameness. Tissue engineered bioscaffolds may be a treatment option for dogs suffering from meniscal damage. The aim of this study was to compare in vitro meniscal-like matrix formation and biomechanical properties of porcine intestinal submucosa sheets (SIS), used in canine meniscal regenerative medicine, to synoviocyte-seeded SIS bioscaffold (SSB), cultured with fetal bovine serum (SSBfbs) or chondrogenic growth factors (SSBgf). Synoviocytes from nine dogs were seeded on SIS and cultured for 30days with 17.7% fetal bovine serum or recombinant chondrogenic growth factors (IGF-1, TGFβ1 and bFGF). The effect on fibrochondrogenesis was determined by comparing mRNA expression of collagen types Iα and IIα, aggrecan, and Sry-type homeobox protein-9 (SOX9) as well as protein expression of collagens I and II, glycosaminoglycan (GAG), and hydroxyproline. The effect of synoviocyte seeding and culture conditions on biochemical properties was determined by measuring peak load, tensile stiffness, resilience, and toughness of bioscaffolds. Pre-culture SIS contained 13.6% collagen and 2.9% double-stranded DNA. Chondrogenic growth factor treatment significantly increased SOX9, collagens I and IIα, aggrecan gene expression (P<0.05), and histological deposition of fibrocartilage extracellular matrix (GAG and collagen II). Culture with synoviocytes increased SIS tensile peak load at failure, resilience, and toughness of bioscaffolds (P<0.05). In conclusion, culturing SIS with synoviocytes prior to implantation might provide biomechanical benefits, and chondrogenic growth factor treatment of cultured synoviocytes improves in vitro axial meniscal matrix formation.
半月板损伤是犬类跛行的常见原因。组织工程生物支架可能是治疗半月板损伤犬的一种选择。本研究的目的是比较用于犬半月板再生医学的猪小肠黏膜下层片(SIS)与接种滑膜细胞的SIS生物支架(SSB)(分别用胎牛血清培养的SSBfbs和用软骨生成生长因子培养的SSBgf)的体外半月板样基质形成和生物力学特性。将来自9只犬的滑膜细胞接种到SIS上,并用17.7%胎牛血清或重组软骨生成生长因子(胰岛素样生长因子-1、转化生长因子β1和成纤维细胞生长因子)培养30天。通过比较Iα型和IIα型胶原蛋白、聚集蛋白聚糖以及Sry型同源盒蛋白9(SOX9)的mRNA表达,以及I型和II型胶原蛋白、糖胺聚糖(GAG)和羟脯氨酸的蛋白表达,来确定对纤维软骨生成的影响。通过测量生物支架的峰值负荷、拉伸刚度、弹性和韧性,来确定滑膜细胞接种和培养条件对生化特性的影响。预培养的SIS含有13.6%的胶原蛋白和2.9%的双链DNA。软骨生成生长因子处理显著增加了SOX9、I型和IIα型胶原蛋白、聚集蛋白聚糖的基因表达(P<0.05),以及纤维软骨细胞外基质(GAG和胶原蛋白II)的组织学沉积。与滑膜细胞一起培养增加了生物支架在失效时的SIS拉伸峰值负荷、弹性和韧性(P<0.05)。总之,植入前用滑膜细胞培养SIS可能会带来生物力学益处,并且对培养的滑膜细胞进行软骨生成生长因子处理可改善体外轴向半月板基质形成。