从少量起始材料生成无支架的类软骨植入物。
Generation of a scaffold free cartilage-like implant from a small amount of starting material.
作者信息
Stoddart M J, Ettinger L, Häuselmann H J
机构信息
Laboratory for Experimental Cartilage Research, Centre for Rheumatology and Bone Disease, Klinik Im Park, Zürich, Switzerland.
出版信息
J Cell Mol Med. 2006 Apr-Jun;10(2):480-92. doi: 10.1111/j.1582-4934.2006.tb00413.x.
INTRODUCTION
An autologous cellular based treatment of a traumatic cartilage injury requires a procedure whereby a biopsy of healthy cartilage is removed from the patient and the cells isolated and expanded by monolayer passage. This increases the cell number to required levels but also leads to a de-differentiation of the cells. We aim to produce a scaffold-free, de-novo implant from a biopsy of cartilage.
METHODS
Bovine chondrocytes were isolated from a small biopsy and expanded. The chondrocytic phenotype of the monolayer expanded cells was recovered during a period of culture in alginate and the effect of factors such as IGF1, TFGbeta1 and dexamethasone was investigated.
RESULTS
During the alginate culture period a pre-treatment with IGF1 and dexamethasone was shown to have little effect. IGF1 however increased the glycosaminoglycan/DNA (GAG/DNA) content on day 14 to 84.95+/-5 ng/ng compared with 37.3+/-1.8 ng/ng in the controls (P<0.001). 35S labeling demonstrated an increased GAG synthesis in the presence of IGF1 (P<0.001). IGF1 also induced a increase of DNA content 1383+/-314 ng/bead compared to 512+/-19 ng/bead in the controls (P<0.001). The cells were released from the alginate and cultured in a silicon mould for a further 14 days to obtain a three dimensional implant. Releasing the cells from the alginate and casting in a mould produced an implant of defined shape which contained no foreign material. After 31 days of culture the implants contained 152.4+/-13.14 ng/ng GAG/DNA and 42.93+/-10.23 ng/ng collagen II.
DISCUSSION
We believe alginate released chondrocytes provide a real alternative to artificial scaffolds.
引言
基于自体细胞的创伤性软骨损伤治疗需要一个程序，即从患者身上取出健康软骨的活检样本，分离细胞并通过单层传代进行扩增。这增加了细胞数量至所需水平，但也导致细胞去分化。我们旨在从软骨活检样本中制备无支架的全新植入物。
方法
从一个小活检样本中分离牛软骨细胞并进行扩增。在藻酸盐中培养一段时间期间恢复单层扩增细胞的软骨细胞表型，并研究诸如IGF1、TFGβ1和地塞米松等因素的作用。
结果
在藻酸盐培养期间，显示IGF1和地塞米松预处理几乎没有效果。然而，IGF1在第14天将糖胺聚糖/DNA（GAG/DNA）含量增加到84.95±5 ng/ng，而对照组为37.3±1.8 ng/ng（P<0.001）。35S标记表明在IGF1存在下GAG合成增加（P<0.001）。与对照组的512±19 ng/珠相比，IGF1还诱导DNA含量增加至1383±314 ng/珠（P<0.001）。将细胞从藻酸盐中释放并在硅模具中进一步培养14天以获得三维植入物。将细胞从藻酸盐中释放并浇铸到模具中产生了形状明确且不含异物的植入物。培养31天后，植入物含有152.4±13.14 ng/ng GAG/DNA和42.93±10.23 ng/ng II型胶原蛋白。
讨论
我们认为藻酸盐释放的软骨细胞为人工支架提供了一个真正的替代方案。
Zotero 插件