1 Tissue Bioengineering and Cell Therapy Unit (GBTTC-CHUAC), CIBER-BBN/ISCIII, Rheumatology Group, Institute of Biomedical Research of A Coruña (INIBIC), University Hospital Complex A Coruña (CHUAC) , Galician Health Service (SERGAS), A Coruña, Spain .
2 Cell Therapy and Regenerative Medicine Unit, CIBER-BBN/ISCIII, Rheumatology Group, Institute of Biomedical Research of A Coruña (INIBIC), University Hospital Complex A Coruña (CHUAC), Galician Health Service (SERGAS), Department of Medicine, Faculty of Health Sciences, University of A Coruña , A Coruña, Spain .
Tissue Eng Part A. 2017 Sep;23(17-18):901-912. doi: 10.1089/ten.TEA.2016.0422. Epub 2017 Feb 8.
Localized trauma-derived breakdown of the hyaline articular cartilage may progress toward osteoarthritis, a degenerative condition characterized by total loss of articular cartilage and joint function. Tissue engineering technologies encompass several promising approaches with high therapeutic potential for the treatment of these focal defects. However, most of the research in tissue engineering is focused on potential materials and structural cues, while little attention is directed to the most appropriate source of cells endowing these materials. In this study, using human amniotic membrane (HAM) as scaffold, we defined a novel static in vitro model for cartilage repair. In combination with HAM, four different cell types, human chondrocytes, human bone marrow-derived mesenchymal stromal cells (hBMSCs), human amniotic epithelial cells, and human amniotic mesenchymal stromal cells (hAMSCs) were assessed determining their therapeutic potential.
A chondral lesion was drilled in human cartilage biopsies simulating a focal defect. A pellet of different cell types was implanted inside the lesion and covered with HAM. The biopsies were maintained for 8 weeks in culture. Chondrogenic differentiation in the defect was analyzed by histology and immunohistochemistry.
HAM scaffold showed good integration and adhesion to the native cartilage in all groups. Although all cell types showed the capacity of filling the focal defect, hBMSCs and hAMSCs demonstrated higher levels of new matrix synthesis. However, only the hAMSCs-containing group presented a significant cytoplasmic content of type II collagen when compared with chondrocytes. More collagen type I was identified in the new synthesized tissue of hBMSCs. In accordance, hBMSCs and hAMSCs showed better International Cartilage Research Society scoring although without statistical significance.
HAM is a useful material for articular cartilage repair in vitro when used as scaffold. In combination with hAMSCs, HAM showed better potential for cartilage repair with similar reparation capacity than chondrocytes.
透明软骨关节内的局部创伤性破坏可能会发展为骨关节炎，这是一种退行性疾病，其特征是完全丧失关节软骨和关节功能。组织工程技术包含几种有前途的方法，具有治疗这些局灶性缺陷的高治疗潜力。然而，组织工程学的大部分研究都集中在潜在的材料和结构线索上，而很少关注赋予这些材料最合适的细胞来源。在这项研究中，我们使用人羊膜（HAM）作为支架，定义了一种新的软骨修复体外静态模型。结合 HAM，评估了四种不同的细胞类型，即人软骨细胞、人骨髓间充质基质细胞（hBMSCs）、人羊膜上皮细胞和人羊膜间充质基质细胞（hAMSCs），以确定它们的治疗潜力。
在人软骨活检中模拟局灶性缺损钻取软骨损伤。将不同细胞类型的小球体植入损伤部位并用 HAM 覆盖。活检标本在培养中维持 8 周。通过组织学和免疫组织化学分析评估缺损内的软骨分化。
HAM 支架在所有组中均与天然软骨良好整合和粘附。尽管所有细胞类型都显示出填充局灶性缺陷的能力，但 hBMSCs 和 hAMSCs 表现出更高水平的新基质合成。然而，只有 hAMSCs 组的 II 型胶原细胞质含量与软骨细胞相比具有显著差异。在新合成的 hBMSCs 组织中鉴定出更多的 I 型胶原。相应地，hBMSCs 和 hAMSCs 表现出更好的国际软骨研究学会评分，尽管没有统计学意义。
HAM 作为支架在体外关节软骨修复中是一种有用的材料。与 hAMSCs 结合使用时，HAM 显示出更好的软骨修复潜力，与软骨细胞具有相似的修复能力。
