Le Visage Catherine, Yang Shu-Hua, Kadakia Leena, Sieber Ann N, Kostuik John P, Leong Kam W
Department of Biomedical Engineering, Johns Hopkins School of Medicine, Baltimore, MD, USA.
Spine (Phila Pa 1976). 2006 Oct 1;31(21):2423-30; discussion 2431. doi: 10.1097/01.brs.0000238684.04792.eb.
To evaluate the capacity of porcine small intestine submucosa to support the in vitro proliferation of human disc cells and the synthesis of extracellular matrix that could restore the biochemical properties of the disc.
To evaluate if porcine small intestine submucosa is a potential bioactive scaffold for rescuing degenerative disc cells.
Discogenic back pain is associated with alterations of the disc and abnormal turnover of the disc extracellular matrix. We hypothesize that a biodegradable and biocompatible acellular scaffold such as small intestine submucosa, which contains entrapped growth factors, may stimulate disc cells to synthesize extracellular matrix, thereby arresting the degeneration, or even promoting the regeneration, of the disc.
Human degenerative anulus and nucleus cells were seeded onto small intestine submucosa scaffolds, and evaluated over a 3-month period for cell growth (proliferation assay, deoxyribonucleic acid content) and matrix composition (glycosaminoglycan and collagen contents).
As hematoxylin and eosin staining revealed, more than 70% of seeded cells attached to the small intestine submucosa surface and invaded throughout the scaffold. The macroscopic appearance of cell-seeded scaffolds was dramatically modified over time. Cell metabolic activity was confirmed for up to 3 months. Seeded scaffolds showed a higher glycosaminoglycan content as compared to control scaffolds. Toluidine blue staining detected large areas of proteoglycans. Positive gene expression for collagens I, II, and X, aggrecan, and Sox-9 confirmed deposition of new extracellular matrix components.
This pilot study shows that small intestine submucosa is a promising bioactive material that could potentially serve as a temporary scaffold for intervertebral disc regeneration.
评估猪小肠黏膜下层支持人椎间盘细胞体外增殖以及合成可恢复椎间盘生化特性的细胞外基质的能力。
评估猪小肠黏膜下层是否为拯救退变椎间盘细胞的潜在生物活性支架。
椎间盘源性背痛与椎间盘改变及椎间盘细胞外基质异常更新有关。我们假设一种可生物降解且生物相容的无细胞支架，如含有包裹生长因子的小肠黏膜下层，可能刺激椎间盘细胞合成细胞外基质，从而阻止椎间盘退变，甚至促进其再生。
将人退变的纤维环和髓核细胞接种到小肠黏膜下层支架上，在3个月期间评估细胞生长（增殖测定、脱氧核糖核酸含量）和基质组成（糖胺聚糖和胶原蛋白含量）。
苏木精-伊红染色显示，超过70%的接种细胞附着于小肠黏膜下层表面并侵入整个支架。随着时间推移，接种细胞的支架宏观外观发生显著改变。细胞代谢活性在长达3个月的时间内得到证实。与对照支架相比，接种细胞的支架显示出更高的糖胺聚糖含量。甲苯胺蓝染色检测到大面积的蛋白聚糖。I、II和X型胶原蛋白、聚集蛋白聚糖和Sox-9的阳性基因表达证实了新细胞外基质成分的沉积。
这项初步研究表明，小肠黏膜下层是一种有前景的生物活性材料，有可能作为椎间盘再生的临时支架。
