Department of Biomedicine, University Hospital Basel, Switzerland.
Osteoarthritis Cartilage. 2010 Aug;18(8):1067-76. doi: 10.1016/j.joca.2010.04.010. Epub 2010 Apr 29.
To characterize the post-expansion cartilage-forming capacity of chondrocytes harvested from detached fragments of osteochondral lesions (OCLs) of ankle joints (Damaged Ankle Cartilage Fragments, DACF), with normal ankle cartilage (NAC) as control.
DACF were obtained from six patients (mean age: 35 years) with symptomatic OCLs of the talus, while NAC were from 10 autopsies (mean age: 55 years). Isolated chondrocytes were expanded for two passages and then cultured in pellets for 14 days or onto HYAFF-11 meshes (FAB, Italy) for up to 28 days. Resulting tissues were assessed histologically, biochemically [glycosaminoglycan (GAG), DNA and type II collagen (CII)] and biomechanically.
As compared to NAC, DACF contained significantly lower amounts of DNA (3.0-fold), GAG (5.3-fold) and CII (1.5-fold) and higher amounts of type I collagen (6.2-fold). Following 14 days of culture in pellets, DACF-chondrocytes generated tissues less intensely stained for Safranin-O and CII, with significantly lower GAG contents (2.8-fold). After 28 days of culture onto HYAFF((R))-11, tissues generated by DACF-chondrocytes were less intensely stained for Safranin-O and CII, contained significantly lower amounts of GAG (1.9-fold) and CII (1.4-fold) and had lower equilibrium (1.7-fold) and dynamic pulsatile modulus (3.3-fold) than NAC-chondrocytes.
We demonstrated that DACF-chondrocytes have inferior cartilage-forming capacity as compared to NAC-chondrocytes, possibly resulting from environmental changes associated with trauma/disease. The study opens some reservations on the use of DACF-derived cells for the repair of ankle cartilage defects, especially in the context of tissue engineering-based approaches.
以正常踝关节软骨(NAC)为对照,研究踝关节骨软骨病变(OCL)游离碎块来源的软骨细胞(DACF)在扩张后的软骨形成能力。
从 6 名(平均年龄 35 岁)有症状的距骨 OCL 患者中获得 DACF,从 10 例尸检(平均年龄 55 岁)中获得 NAC。分离的软骨细胞经过两传代培养后,在微载体中培养 14 天,或接种到 HYAFF-11 网片(FAB,意大利)上培养 28 天。对组织进行组织学、生物化学[糖胺聚糖(GAG)、DNA 和 II 型胶原(CII)]和生物力学评估。
与 NAC 相比,DACF 中的 DNA(3.0 倍)、GAG(5.3 倍)和 CII(1.5 倍)含量明显较低,而 I 型胶原含量(6.2 倍)明显较高。在微载体中培养 14 天后,DACF 软骨细胞生成的组织经番红 O 和 CII 染色强度较弱,GAG 含量明显较低(2.8 倍)。在 HYAFF((R))-11 上培养 28 天后,DACF 软骨细胞生成的组织经番红 O 和 CII 染色强度较弱,GAG(1.9 倍)和 CII(1.4 倍)含量明显较低,平衡(1.7 倍)和动态脉冲模量(3.3 倍)也较低。
与 NAC 软骨细胞相比,DACF 软骨细胞的软骨形成能力较差,这可能是由于创伤/疾病相关的环境变化所致。该研究对使用 DACF 衍生细胞修复踝关节软骨缺损提出了一些保留意见,尤其是在基于组织工程的方法背景下。