Institute of Anatomy and Cell Biology, Paracelsus Medical University, Nuremberg and Salzburg, Prof. Ernst Nathan Str. 1, 90419 Nuremberg, Germany.
Department Materials Engineering, Institute of Polymers Materials, Leibniz-Institut für Polymerforschung Dresden e.V. (IPF), Hohe Straße 6, 01069 Dresden, Germany.
Int J Mol Sci. 2023 Apr 4;24(7):6714. doi: 10.3390/ijms24076714.
Successful anterior cruciate ligament (ACL) reconstructions strive for a firm bone-ligament integration. With the aim to establish an enthesis-like construct, embroidered functionalized scaffolds were colonized with spheroids of osteogenically differentiated human mesenchymal stem cells (hMSCs) and lapine (l) ACL fibroblasts in this study. These triphasic poly(L-lactide-co-ε-caprolactone) and polylactic acid (P(LA-CL)/PLA) scaffolds with a bone-, a fibrocartilage transition- and a ligament zone were colonized with spheroids directly after assembly (DC) or with 14-day pre-cultured lACL fibroblast and 14-day osteogenically differentiated hMSCs spheroids (=longer pre-cultivation, LC). The scaffolds with co-cultures were cultured for 14 days. Cell vitality, DNA and sulfated glycosaminoglycan (sGAG) contents were determined. The relative gene expressions of and , , and were analyzed. Compared to the lACL spheroids, those with hMSCs adhered more rapidly. Vimentin and collagen type I immunoreactivity were mainly detected in the hMSCs colonizing the bone zone. The DNA content was higher in the DC than in LC whereas the sGAG content was higher in LC. The gene expression of ECM components and transcription factors depended on cell type and pre-culturing condition. Zonal colonization of triphasic scaffolds using spheroids is possible, offering a novel approach for enthesis tissue engineering.
成功的前交叉韧带（ACL）重建力求实现牢固的骨-韧带整合。本研究旨在建立类似附着点的结构，使用具有功能化绣花的支架来接种骨诱导分化的人骨髓间充质干细胞（hMSCs）和成纤维细胞球体和兔（l）ACL 成纤维细胞球体。这些三相聚（L-丙交酯-co-ε-己内酯）和聚乳酸（P（LA-CL）/PLA）支架具有骨、纤维软骨过渡和韧带区，在组装后直接接种（DC）或经过 14 天预培养的 lACL 成纤维细胞和 14 天骨诱导分化的 hMSCs 球体（=更长的预培养，LC）。将接种了细胞的支架培养 14 天。测定细胞活力、DNA 和硫酸化糖胺聚糖（sGAG）含量。分析 和 、 、 和 的相对基因表达。与 lACL 球体相比，hMSCs 球体的附着速度更快。在骨区中主要检测到 hMSCs 接种的波形蛋白和 I 型胶原免疫反应。DC 中的 DNA 含量高于 LC，而 LC 中的 sGAG 含量较高。细胞外基质成分和转录因子的基因表达取决于细胞类型和预培养条件。使用球体对三相支架进行分区接种是可行的，为附着点组织工程提供了一种新方法。
