Effects of auricular chondrocyte expansion on neocartilage formation in photocrosslinked hyaluronic acid networks
Effects of auricular chondrocyte expansion on neocartilage formation in photocrosslinked hyaluronic acid networks
复制标题
DOI：
10.1089/ten.2006.12.2665
复制
发表时间：
2006-09-01
影响因子：
--
通讯作者：
Burdick, Jason A.
中科院分区：
文献类型：
--
作者：
Chung, Cindy;Mesa, John;Burdick, Jason A.
本研究的总体目标是检查体外扩增对由透明质酸（HA）水凝胶中光包封的耳廓软骨细胞形成新软骨的影响，作为组织工程疗法治疗受损软骨的临床应用的下一步。将猪耳廓软骨细胞在分离后（p = 0）或进一步体外扩增后（p = 1和p = 2）直接包封在2wt%、50-kDa HA水凝胶中，并皮下植入裸鼠背部。12周后，将构建体进行机械测试、生物化学和免疫组织化学分析，并与单独的HA凝胶和天然软骨的对照进行比较。p = 0和p = 1结构的压缩平衡模量（51.2 +/- 8.0和72.5 +/- 35.2 kPa，分别）大于p = 2构建体（26.8 +/- 14.9 kPa）和单独的对照HA凝胶（12.3 +/- 1.3 kPa），并且与耳软骨（35.1 +/- 12.2 kPa）相当。生化分析显示，软骨细胞传代后糖胺聚糖（GAG）、胶原蛋白和弹性蛋白含量普遍降低，但在任何分析中，p = 0和p = 1构建体之间均未发现显著差异。组织学染色显示聚集蛋白聚糖染色强烈且均匀，并且所有构建体中II型胶原蛋白染色均高于I型胶原蛋白染色。总体而言，本研究表明，p = 0和p = 1的耳廓软骨细胞构建体在体内12周后产生了类似于天然耳廓软骨的新软骨组织。然而，这些结果表明软骨细胞的进一步扩增（p = 2）可导致受损的组织性质。
The overall objective of this study was to examine the effects of in vitro expansion on neocartilage formation by auricular chondrocytes photoencapsulated in a hyaluronic acid (HA) hydrogel as a next step toward the clinical application of tissue engineering therapies for treatment of damaged cartilage. Swine auricular chondrocytes were encapsulated either directly after isolation (p = 0), or after further in vitro expansion (p = 1 and p = 2) in a 2 wt%, 50-kDa HA hydrogel and implanted subcutaneously in the dorsum of nude mice. After 12 weeks, constructs were explanted for mechanical testing and biochemical and immunohistochemical analysis and compared to controls of HA gels alone and native cartilage. The compressive equilibrium moduli of the p = 0 and p = 1 constructs (51.2 +/- 8.0 and 72.5 +/- 35.2 kPa, respectively) were greater than the p = 2 constructs (26.8 +/- 14.9 kPa) and the control HA gel alone (12.3 +/- 1.3 kPa) and comparable to auricular cartilage (35.1 +/- 12.2 kPa). Biochemical analysis showed a general decrease in glycosaminoglycan (GAG), collagen, and elastin content with chondrocyte passage, though no significant differences were found between the p = 0 and p = 1 constructs for any of the analyses. Histological staining showed intense and uniform staining for aggrecan, as well as greater type II collagen versus type I collagen staining in all constructs. Overall, this study illustrates that constructs with the p = 0 and p = 1 auricular chondrocytes produced neocartilage tissue that resembled native auricular cartilage after 12 weeks in vivo. However, these results indicate that further expansion of the chondrocytes (p = 2) can lead to compromised tissue properties.