Liang Rui, Woo Savio L-Y, Nguyen Tan D, Liu Ping-Cheng, Almarza Alejandro
Musculoskeletal Research Center, Department of Bioengineering, University of Pittsburgh, 300 Technology Drive, Pittsburgh, Pennsylvania 15219, USA.
J Orthop Res. 2008 Aug;26(8):1098-104. doi: 10.1002/jor.20616.
Bioscaffolds have been successfully used to improve the healing of ligaments and tendons. In a rabbit model, the application of porcine small intestine submucosa (SIS) to the healing medial collateral ligament (MCL) resulted in improved mechanical properties with the formation of larger collagen fibrils. Thus, the objective of the study was to find out whether the SIS bioscaffold could improve the gene expressions of fibrillogenesis-related molecules, specifically, collagen types I, III, V, and small leucine-rich proteoglycans including decorin, biglycan, lumican, and fibromodulin, as well as collagen fibril morphology and organization, in the healing rabbit MCL at an early time point (6 weeks postinjury). Twenty skeletally mature rabbits were equally divided into two groups. In the SIS-treated group, a 6-mm gap was surgically created and a layer of SIS was sutured to cover the gap, whereas the gap was left open in the nontreated group. At 6 weeks postinjury, Masson's trichrome staining showed that the SIS-treated group had more regularly aligned collagen fibers and cells. Transmission electron microscopy revealed that the SIS-treated group had larger collagen fibrils with a diameter distribution from 24 to 120 nm, whereas the nontreated group had only small collagen fibrils (ranging from 26 to 87 nm, p < 0.05). Finally, the quantitative real-time PCR showed that the mRNAs of collagen type V, decorin, biglycan, and lumican in the SIS-treated group were 41, 58, 51, and 43% lower than those in the nontreated group, respectively (p < 0.05). Such significant reduction in the gene expressions are closely related to the improved morphological characteristics, which are known to be coupled with better mechanical properties, as previously reported in longer term studies.
生物支架已成功用于改善韧带和肌腱的愈合。在兔模型中,将猪小肠黏膜下层(SIS)应用于内侧副韧带(MCL)愈合部位,可改善力学性能,并形成更大的胶原纤维。因此,本研究的目的是确定SIS生物支架是否能在损伤后早期(损伤后6周)改善兔MCL愈合过程中与纤维形成相关分子的基因表达,特别是I型、III型、V型胶原蛋白以及富含亮氨酸的小分子蛋白聚糖,包括核心蛋白聚糖、双糖链蛋白聚糖、光蛋白聚糖和纤调蛋白聚糖,以及胶原纤维的形态和排列。20只骨骼成熟的兔子被平均分为两组。在SIS治疗组中,手术制造一个6毫米的间隙,并用一层SIS缝合覆盖该间隙,而未治疗组的间隙保持开放。损伤后6周,Masson三色染色显示SIS治疗组的胶原纤维和细胞排列更规则。透射电子显微镜显示,SIS治疗组有直径为24至120纳米的更大的胶原纤维,而未治疗组只有小的胶原纤维(直径范围为26至87纳米,p<0.05)。最后,定量实时PCR显示,SIS治疗组中V型胶原蛋白、核心蛋白聚糖、双糖链蛋白聚糖和光蛋白聚糖的mRNA分别比未治疗组低41%、58%、51%和43%(p<0.05)。如先前长期研究所报道,基因表达的这种显著降低与改善的形态特征密切相关,而形态特征的改善又与更好的力学性能相关。