Objective The objective was to evaluate the proliferating, migratory and extracellular matrix (ECM) forming potential of annulus fibrosus cells derived from early (edAFC) or advanced (adAFC) degenerative tissue and their usability as a possible cell source for regenerative approaches for AF closure. Design EdAFC ( n = 5 Pfirrman score of 2-3) and adAFC (n = 5 Pfirrman score of 4-5) were isolated from tissue of patients undergoing spine stabilizing surgery. Cell migration on stimulation with human serum (HS), platelet-rich plasma (PRP), and transforming growth factor β-3 (TGFB3) was assessed by migration assay and proliferation was assessed on stimulation with HS. Induction of ECM synthesis was evaluated by gene expression analysis of AF-related genes in three-dimensional scaffold cultures that have been stimulated with 5% PRP or 10 ng/mL TGFB3 and histologically by collagen type I, type II, alcian blue, and safranin-O staining. Results EdAFC and adAFC were significantly attracted by 10% HS and 5% PRP. Additionally, both cell groups proliferated under stimulation with HS. Stimulation with 10 ng/mL TGFB3 showed significant induction of gene expression of collagen type II and aggrecan, while 5% PRP decreased the expression of collagen type I. Both cell groups showed formation of AF-like ECM after stimulation with TGFB3, whereas stimulation with PRP did not. Conclusions Our study demonstrated that AF cells retain their potential for proliferation, migration, and ECM formation independent of the degeneration status of the tissue. Proliferation, migration, and ECM synthesis of the endogenous AF cells can be supported by different supplements. Hence, endogenous AF cells might be a suitable cell source for a regenerative repair approaches.

中文翻译：

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人环纤维细胞的增殖，迁移和ECM形成潜能与变性状态无关。

目的目的是评估早期（edAFC）或晚期（adAFC）退化组织衍生的纤维环细胞的增殖，迁移和细胞外基质（ECM）形成潜力，以及它们作为房颤闭合再生方法的可能细胞来源的可用性。从进行脊柱稳定手术的患者组织中分离设计EdAFC（n = 5 Pfirrman评分为2-3）和adAFC（n = 5 Pfirrman评分为4-5）。用迁移测定法评估人血清（HS），富血小板血浆（PRP）和转化生长因子β-3（TGFB3）刺激下的细胞迁移，并评估HS刺激下的增殖。通过在5％的PRP或10 ng / mL TGFB3刺激下并在组织学上受到I型，II型，阿尔辛蓝和藏红素胶原蛋白刺激的三维支架培养物中AF相关基因的基因表达分析来评估ECM合成的诱导-O染色。结果10％的HS和5％的PRP显着吸引了EdAFC和adAFC。另外，两个细胞群在HS刺激下增殖。用10 ng / mL TGFB3刺激可显着诱导II型胶原蛋白和聚集蛋白聚糖的基因表达，而5％PRP则可降低I型胶原蛋白的表达。TGTG3刺激后，两个细胞组均显示出AF样ECM的形成，而TGFB3刺激PRP没有。结论我们的研究表明，AF细胞保留了其增殖，迁移，和ECM的形成与组织的退化状态无关。内源性AF细胞的增殖，迁移和ECM合成可以通过不同的补充剂来支持。因此，内源性AF细胞可能是再生修复方法的合适细胞来源。