Fuchs Julie R, Terada Shinichi, Hannouche Didier, Ochoa Erin R, Vacanti Joseph P, Fauza Dario O
Harvard Center for Minimally Invasive Surgery, Harvard Medical School, Massachusetts General Hospital, Boston, MA, USA.
J Pediatr Surg. 2002 Dec;37(12):1720-5. doi: 10.1053/jpsu.2002.36705.
BACKGROUND/PURPOSE: This study was aimed at characterizing the structure and function of engineered fetal cartilage in vitro.
Chondrocytes from ovine specimens of fetal elastic, fetal hyaline, and adult elastic cartilage were expanded in culture and their growth rates determined. Cells were seeded onto synthetic scaffolds, which were then maintained in a bioreactor. Matrix deposition was determined by specific staining and quantitative assays for glycosaminoglycans (GAG), type II collagen (CII), and elastin, as well as compared with native tissue. Statistical analysis was by analysis of variance (ANOVA) and Students' t test, with significance set at P less than.01.
Fetal elastic chondrocytes grew significantly faster than all other cell types. All fetal constructs resembled hyaline cartilage, regardless of the cell source. There were significantly higher levels of GAG and CII in fetal versus adult constructs, but no significant difference between fetal constructs from different sources. Unlike their adult counterparts, fetal constructs had GAG and CII levels similar to native tissues.
Fetal chondrocytes can be rapidly expanded in culture. Compared with adult constructs, matrix deposition is enhanced in engineered fetal cartilage, which closely resembles native tissue, regardless of the cell source. Engineered fetal cartilage may be a preferable option during surgical reconstruction of select congenital anomalies.
背景/目的:本研究旨在体外表征工程化胎儿软骨的结构与功能。
从绵羊胎儿弹性软骨、胎儿透明软骨和成年弹性软骨标本中分离软骨细胞,在培养中进行扩增并测定其生长速率。将细胞接种到合成支架上,然后置于生物反应器中培养。通过特异性染色以及对糖胺聚糖(GAG)、II型胶原蛋白(CII)和弹性蛋白的定量测定来确定基质沉积情况,并与天然组织进行比较。采用方差分析(ANOVA)和学生t检验进行统计分析,显著性设定为P小于0.01。
胎儿弹性软骨细胞的生长速度显著快于所有其他细胞类型。所有胎儿构建体均类似于透明软骨,与细胞来源无关。与成年构建体相比,胎儿构建体中的GAG和CII水平显著更高,但不同来源的胎儿构建体之间无显著差异。与成年构建体不同,胎儿构建体中的GAG和CII水平与天然组织相似。
胎儿软骨细胞可在培养中快速扩增。与成年构建体相比,工程化胎儿软骨中的基质沉积增强,无论细胞来源如何,其都与天然组织非常相似。在某些先天性畸形的手术重建中,工程化胎儿软骨可能是一个更优选择。