Department of Hand, Plastic and Aesthetic Surgery, LMU Klinikum, University Hospital, LMU Munich, Ziemsenstrasse 5, 80336 Munich, Germany.
Laboratory of Cartilage Development, Diseases and Regeneration, Department for Orthopaedics and Trauma Surgery, Musculoskeletal University Center Munich (MUM), University Hospital, LMU Munich, Fraunhoferstrasse 20, 82152 Planegg, Germany.
Int J Mol Sci. 2021 Sep 14;22(18):9917. doi: 10.3390/ijms22189917.
Previous anatomical studies have shown different functional zones in human nasal septal cartilage (NC). These zones differ in respect to histological architecture and biochemical composition. The aim of this study was to investigate the influence of these zones on the fate of stem cells from a regenerative perspective. Therefore, decellularized porcine septal cartilage was prepared and subjected to histological assessment to demonstrate its equivalence to human cartilage. Decellularized porcine NC (DPNC) exposed distinct surfaces depending on two different histological zones: the outer surface (OS), which is equivalent to the superficial zone, and the inner surface (IS), which is equivalent to the central zone. Human adipose tissue-derived stem cells (ASCs) were isolated from the abdominal fat tissue of five female patients and were seeded on the IS and OS of DPNC, respectively. Cell seeding efficiency (CSE), vitality, proliferation, migration, the production of sulfated glycosaminoglycans (sGAG) and chondrogenic differentiation capacity were evaluated by histological staining (DAPI, Phalloidin, Live-Dead), biochemical assays (alamarBlue, PicoGreen, DMMB) and the quantification of gene expression (qPCR). Results show that cell vitality and CSE were not influenced by DPNC zones. ASCs, however, showed a significantly higher proliferation and elevated expression of early chondrogenic differentiation, as well as fibrocartilage markers, on the OS. On the contrary, there was a significantly higher upregulation of hypertrophy marker MMP13 ( < 0.0001) and GAG production ( = 0.0105) on the IS, whereas cell invasion into the three-dimensional DPNC was higher in comparison to the OS. We conclude that the zonal-dependent distinct architecture and composition of NC modulates activities of ASCs seeded on DPNC. These findings might be used for engineering of cartilage substitutes needed in facial reconstructive surgery that yield an equivalent histological and functional structure, such as native NC.
先前的解剖学研究表明，人类鼻中隔软骨（NC）存在不同的功能区。这些区域在组织学结构和生化组成上存在差异。本研究旨在从再生的角度探讨这些区域对干细胞命运的影响。因此，制备了脱细胞猪鼻中隔软骨并进行组织学评估，以证明其与人软骨等效。脱细胞猪 NC（DPNC）根据两个不同的组织学区域呈现出不同的表面：外表面（OS），与浅层区域等效，内表面（IS），与中央区域等效。从五名女性患者的腹部脂肪组织中分离出人脂肪组织源性干细胞（ASCs），并分别接种在 DPNC 的 IS 和 OS 上。通过组织学染色（DAPI、鬼笔环肽、死活染色）、生化测定（alamarBlue、PicoGreen、DMMB）和基因表达定量（qPCR）评估细胞接种效率（CSE）、活力、增殖、迁移、硫酸化糖胺聚糖（sGAG）的产生和软骨分化能力。结果表明，DPNC 区域不会影响细胞活力和 CSE。然而，在 OS 上，ASCs 的增殖和早期软骨分化以及纤维软骨标志物的表达显著升高。相反，在 IS 上，肥大标志物 MMP13（<0.0001）和 GAG 产生（=0.0105）的上调显著更高，而与 OS 相比，细胞向三维 DPNC 的侵入更高。我们得出结论，NC 的区域依赖性不同结构和组成会调节接种在 DPNC 上的 ASCs 的活性。这些发现可用于工程化面部重建手术中所需的软骨替代物，这些替代物具有等效的组织学和功能结构，如天然 NC。
