Chiba K, Andersson G B, Masuda K, Momohara S, Williams J M, Thonar E J
Department of Orthopedic Surgery, Rush Medical College at Rush-Presbyterian-St. Luke's Medical Center, Chicago, Ilinois, USA.
Spine (Phila Pa 1976). 1998 Sep 1;23(17):1821-7; discussion 1828. doi: 10.1097/00007632-199809010-00002.
This study determined whether entrapment of a rabbit intervertebral disc in alginate gel helped to promote the retention of normal metabolic activities by the nucleus pulposus and anulus fibrosus in tissue culture.
To establish an in vitro culture system to study the metabolism of the intervertebral disc as a whole integral organ.
In vitro studies of the metabolism of intervertebral discs have been scarce because of the difficulties involved in maintaining the integrity of the tissues, especially that of the nucleus pulposus, in culture medium.
Rabbit intervertebral discs were embedded in alginate gel and maintained in culture for as long as 1 month. At weekly intervals, experiments were performed to measure the rate of proteoglycan synthesis and to characterize proteoglycans newly synthesized by cells in the anulus fibrosus and nucleus pulposus. In addition, at these same time intervals, the contents of sulfated proteoglycans, antigenic keratan sulfate, hyaluronan, and collagen in these two intervertebral disc tissues were measured to evaluate tissue integrity. Intervertebral discs cultured in medium alone were used as controls and analyzed in parallel.
The anulus fibrosus and nucleus pulposus of intervertebral discs cultured in alginate gel sustained a higher rate of proteoglycan synthesis and maintained a higher content of extracellular matrix components than the respective controls at all times.
This new alginate tissue culture system should prove useful for studying the metabolism of whole intervertebral discs.
本研究确定将兔椎间盘包埋于藻酸盐凝胶中是否有助于促进髓核和纤维环在组织培养中保持正常代谢活动。
建立一种体外培养系统,以研究作为一个完整器官的椎间盘的代谢。
由于在培养基中维持椎间盘组织完整性存在困难,尤其是髓核的完整性,因此对椎间盘代谢的体外研究一直很少。
将兔椎间盘包埋于藻酸盐凝胶中,并在培养中维持长达1个月。每周进行实验,以测量蛋白聚糖合成速率,并对纤维环和髓核中的细胞新合成的蛋白聚糖进行表征。此外,在相同的时间间隔,测量这两种椎间盘组织中硫酸化蛋白聚糖、抗原性硫酸角质素、透明质酸和胶原蛋白的含量,以评估组织完整性。单独在培养基中培养的椎间盘用作对照并进行平行分析。
在藻酸盐凝胶中培养的椎间盘的纤维环和髓核在所有时间都比各自的对照维持更高的蛋白聚糖合成速率,并保持更高的细胞外基质成分含量。
这种新的藻酸盐组织培养系统应被证明对研究整个椎间盘的代谢有用。