Department of Orthopedics, University Medical Center Utrecht, Utrecht, The Netherlands.
Department of Pathology and Medical Biology, University Medical Center Groningen, Groningen, The Netherlands.
Spine (Phila Pa 1976). 2018 Mar 1;43(5):307-315. doi: 10.1097/BRS.0000000000000920.
An in vitro study using human degenerated nucleus pulposus cells.
To determine the effect of osmolality and different osmolytes on the regeneration by human nucleus pulposus cells through gene expression and extracellular matrix production.
Intervertebral disc (IVD) degeneration is a major problem in developed countries. Regeneration of the IVD can prevent pain and costs due to diminished work absence and health care, and improve quality of life. The osmotic value of a disc decreases during degeneration due to loss of proteoglycans and might increase degeneration. It is known that gene expression of matrix genes of nucleus pulposus (NP) cells increases when cultured in hyperosmotic medium. Thus, increasing the osmolality of the disc might be beneficial for disc regeneration.
In the current study, isolated degenerated human NP cells were used in regeneration culture with medium of different osmolalities, adjusted with different osmolytes. NaCl, urea and sucrose. The cells were cultured for 28 days and expression of matrix genes and production of glycosaminoglycans and collagen II were measured.
Gene expression for both collagen II and aggrecan increased with increasing osmolality using NaCl or sucrose, but not urea. Protein production however, was not affected by increasing osmolality and was decreased when using urea and sucrose. Expression of genes for Col1A1, MMP13, and MMP14 decreased with increasing osmolality, whereas expression of LOXL2 and LOXL3 increased. Transient expression of TonEBP was found 6 hours after the start of culture, but not at later time points.
Although expression of matrix genes is upregulated, hyperosmolality does not enhance matrix production by nucleus pulposus cells. Raising osmolality can potentially increase matrix production, but in itself is not sufficient to accomplish regeneration in the current in vitro culture system.
N /A.
一项使用人退变髓核细胞的体外研究。
通过基因表达和细胞外基质产生来确定渗透压和不同渗透物对人髓核细胞再生的影响。
椎间盘（IVD）退变是发达国家的一个主要问题。IVD 的再生可以预防因工作缺勤和医疗保健减少而导致的疼痛和成本，并提高生活质量。由于蛋白聚糖的丢失，椎间盘的渗透压值在退变过程中降低，并且可能会增加退变。已知在高渗培养基中培养时，髓核（NP）细胞的基质基因的基因表达会增加。因此，增加椎间盘的渗透压可能有利于椎间盘的再生。
在目前的研究中，使用不同渗透压的不同渗透物调节的培养基对分离的退变人 NP 细胞进行再生培养。使用 NaCl、尿素和蔗糖。将细胞培养 28 天，并测量基质基因的表达和糖胺聚糖和胶原 II 的产生。
使用 NaCl 或蔗糖增加渗透压时，胶原 II 和聚集蛋白聚糖的基因表达均增加，但尿素则不然。然而，蛋白产生不受渗透压增加的影响，并且当使用尿素和蔗糖时会减少。Col1A1、MMP13 和 MMP14 的基因表达随渗透压增加而降低，而 LOXL2 和 LOXL3 的表达增加。在培养开始后 6 小时发现瞬时表达 TonEBP，但在以后的时间点则没有。
尽管基质基因的表达上调，但高渗透压不会增强髓核细胞的基质产生。提高渗透压可能会增加基质的产生，但在当前的体外培养系统中，自身不足以完成再生。
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