Gualdi Thomas, Laurent Romain, Moutarlier Virginie, Fenelon Mathilde, Nallet Aurélie, Pouthier Fabienne, Obert Laurent, de Billy Benoit, Meyer Christophe, Gindraux Florelle
Orthopaedic, Traumatology and Plastic Surgery Service, University Hospital of Besançon, Besançon, France.
Nanomedicine Lab, Imagery and Therapeutics (EA 4662), SFR FED 4234, University of Franche-Comté, Besançon, France.
Cell Tissue Bank. 2019 Sep;20(3):435-446. doi: 10.1007/s10561-019-09778-3. Epub 2019 Jun 17.
The human amniotic membrane (hAM) is an attractive biomaterial for regenerative medicine, as it contains amniotic mesenchymal stromal cells (hAMSC), epithelial cells (hAEC) and growth factors. We examined the potential use of hAM in orthopaedic and maxillofacial bone surgery, integrating the requirements of current regulations regarding advanced therapy medicinal products (ATMP) in the European Union. Previous studies have described the potential osteodifferentiation of intact hAM during whole-tissue culture in osteogenic conditions. The present study aims to determine whether in vitro osteodifferentiation of hAM is needed in the context bone repair, and the influence of this process on tissue structure, cell phenotype and cell function. Different conditions (fresh or cultured hAM; intact or hAM-derived cells) were tested. Phenotypic and functional analyses were performed with standard approaches (cell culture and staining, histological and immunolabelling) as well as original approaches (tissue staining, energy dispersive X-ray and X-ray diffraction). In our study, non-osteodifferentiated hAM (i.e., fresh or native hAM) exhibited innate pre-osteoblastic potential. Osteodifferentiation of fresh hAM induced a change in tissue structure, cell phenotype and function. Therefore, we hypothesize that pre-osteodifferentiation may not be necessary, especially if it induces unwanted changes. To our surprise, in these osteogenic conditions, hAEC had a mesenchymal phenotype with osteocyte function, and even native synthesis of hydroxyapatite, focusing osteogenic potential mainly in this epithelial layer. In conclusion, in vitro osteodifferentiation by tissue culture does not appear to be necessary for hAM to be used as an innovative ATMP for bone repair.
人羊膜（hAM）是一种用于再生医学的有吸引力的生物材料，因为它含有羊膜间充质基质细胞（hAMSC）、上皮细胞（hAEC）和生长因子。我们结合欧盟当前关于高级治疗药品（ATMP）的法规要求，研究了hAM在骨科和颌面骨手术中的潜在用途。先前的研究描述了完整的hAM在成骨条件下全组织培养过程中的潜在骨分化情况。本研究旨在确定在骨修复背景下hAM的体外骨分化是否必要，以及这一过程对组织结构、细胞表型和细胞功能的影响。测试了不同条件（新鲜或培养的hAM；完整或hAM衍生细胞）。采用标准方法（细胞培养和染色、组织学和免疫标记）以及原始方法（组织染色、能量色散X射线和X射线衍射）进行表型和功能分析。在我们的研究中，未进行骨分化的hAM（即新鲜或天然hAM）表现出固有的前成骨细胞潜力。新鲜hAM的骨分化诱导了组织结构、细胞表型和功能的变化。因此，我们假设前骨分化可能没有必要，特别是如果它会引起不必要的变化。令我们惊讶的是，在这些成骨条件下，hAEC具有骨细胞功能的间充质表型，甚至能天然合成羟基磷灰石，主要将成骨潜力集中在这一上皮层。总之，对于hAM作为用于骨修复的创新型ATMP而言，通过组织培养进行体外骨分化似乎并非必要。
