Díaz-Prado Silvia, Rendal-Vázquez Maria Esther, Muiños-López Emma, Hermida-Gómez Tamara, Rodríguez-Cabarcos Margarita, Fuentes-Boquete Isaac, de Toro Francisco J, Blanco Francisco J
Department of Medicine, INIBIC-University of A Coruña, A Coruña, Spain.
Cell Tissue Bank. 2010 May;11(2):183-95. doi: 10.1007/s10561-009-9144-1. Epub 2010 Apr 13.
The human amniotic membrane (HAM) is an abundant and readily obtained tissue that may be an important source of scaffold for transplanted chondrocytes in cartilage regeneration in vivo. To evaluate the potential use of cryopreserved HAMs as a support system for human chondrocytes in human articular cartilage repair. Chondrocytes were isolated from human articular cartilage, cultured and grown on the chorionic basement membrane side of HAMs. HAMs with chondrocytes were then used in 44 in vitro human osteoarthritis cartilage repair trials. Repair was evaluated at 4, 8 and 16 weeks by histological analysis. Chondrocytes cultured on the HAM revealed that cells grew on the chorionic basement membrane layer, but not on the epithelial side. Chondrocytes grown on the chorionic side of the HAM express type II collagen but not type I, indicating that after being in culture for 3-4 weeks they had not de-differentiated into fibroblasts. In vitro repair experiments showed formation on OA cartilage of new tissue expressing type II collagen. Integration of the new tissue with OA cartilage was excellent. The results indicate that cryopreserved HAMs can be used to support chondrocyte proliferation for transplantation therapy to repair OA cartilage.
人羊膜（HAM）是一种丰富且易于获取的组织，可能是体内软骨再生中移植软骨细胞的重要支架来源。为评估冷冻保存的HAM作为人软骨细胞在人关节软骨修复中的支持系统的潜在用途。从人关节软骨中分离软骨细胞，在HAM的绒毛膜基底膜侧培养生长。然后将带有软骨细胞的HAM用于44项体外人骨关节炎软骨修复试验。通过组织学分析在4周、8周和16周时评估修复情况。在HAM上培养的软骨细胞显示，细胞在绒毛膜基底膜层生长，但不在上皮侧生长。在HAM绒毛膜侧生长的软骨细胞表达II型胶原蛋白而不表达I型，表明在培养3 - 4周后它们没有去分化为成纤维细胞。体外修复实验显示在骨关节炎软骨上形成了表达II型胶原蛋白的新组织。新组织与骨关节炎软骨的整合良好。结果表明，冷冻保存的HAM可用于支持软骨细胞增殖，用于移植治疗修复骨关节炎软骨。
