Allen Kyle D, Erickson Kathryn, Athanasiou Kyriacos A
Department of Bioengineering, Rice University, Houston, TX, USA.
Arch Oral Biol. 2008 Jan;53(1):53-9. doi: 10.1016/j.archoralbio.2007.07.004. Epub 2007 Sep 7.
Implantation of synthetic temporomandibular joint (TMJ) disc replacements aimed to alleviate pain and restore functional losses caused by TMJ disorders. Unfortunately, these synthetic replacements have been largely unsuccessful and in some instances have incited severe immune responses. Tissue engineering, however, may provide viable TMJ disc replacements. Towards this end, we have studied TMJ disc gene expression as a measure of protein production potential. With passage, collagen type I and aggrecan gene expression decrease in TMJ disc cell cultures. We hypothesize that surfaces coated with TMJ disc proteins may rapidly recover the lost gene expression in passaged TMJ disc cells.
To study these effects, passages 0, 1, and 2 TMJ disc cells were plated in wells coated with aggrecan, collagen type I, collagen type II, or decorin. Safranin O staining was conducted to visualize cell aggregation.
At passage 0, cultures appeared similar on each surface; however, by passages 1 and 2, aggrecan-coated and decorin-coated surfaces appeared to have more cell aggregates. Gene expression data did not correspond to these visual changes. No treated surface offered a significant change in aggrecan, collagen type I, or decorin expression relative to untreated controls. Furthermore, aggrecan and collagen type I gene expression dropped relative to samples taken prior to plating.
These results indicate that, despite visual changes described by cell aggregates, protein coatings have limited effects for recovering TMJ disc gene expression in monolayer cultures.
植入人工颞下颌关节(TMJ)椎间盘置换物旨在缓解疼痛并恢复由TMJ紊乱引起的功能丧失。不幸的是,这些人工置换物在很大程度上并不成功,在某些情况下还引发了严重的免疫反应。然而,组织工程学可能提供可行的TMJ椎间盘置换物。为此,我们研究了TMJ椎间盘基因表达,以此作为蛋白质产生潜力的一种衡量指标。随着传代,TMJ椎间盘细胞培养物中I型胶原蛋白和聚集蛋白聚糖基因表达下降。我们推测,涂有TMJ椎间盘蛋白的表面可能会迅速恢复传代TMJ椎间盘细胞中丢失的基因表达。
为了研究这些影响,将第0代、第1代和第2代TMJ椎间盘细胞接种到涂有聚集蛋白聚糖、I型胶原蛋白、II型胶原蛋白或核心蛋白聚糖的孔中。进行番红O染色以观察细胞聚集情况。
在第0代时,各表面上的培养物看起来相似;然而,到第1代和第2代时,涂有聚集蛋白聚糖和核心蛋白聚糖的表面似乎有更多的细胞聚集体。基因表达数据与这些视觉变化并不相符。相对于未处理的对照,没有任何处理过的表面在聚集蛋白聚糖、I型胶原蛋白或核心蛋白聚糖表达上有显著变化。此外,相对于接种前采集的样本,聚集蛋白聚糖和I型胶原蛋白基因表达下降。
这些结果表明,尽管细胞聚集体描述了视觉变化,但蛋白质涂层对恢复单层培养中TMJ椎间盘基因表达的作用有限。