Abstract
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 (p < 0.0001) and GAG production (p = 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.
Highlights
adipose tissue-derived stem cells (ASCs) were selected as a stem cell source to investigate the influence of the cartilage extracellular matrix (ECM) of the Decellularized porcine NC (DPNC) scaffold on the stem cell fate
These results demonstrate that the attachment of ASCs was comparable on both DPNC surfaces and suggest that further experiments were not affected by the seeding per se
Our results indicate that the stem cell fate differs significantly between ECM zones in the DPNC, and an interaction with a specialized ECM can be used to guide ASCs towards chondrogenesis
Summary
The destruction of nasal septal cartilage due to trauma, skin cancer surgery, autoimmune disease or infection can lead to various problems, including impaired breathing, smelling disorders and obvious cosmetic deformity [1,2,3,4]. The surgical repair of those defects often requires costal or auricular cartilage to reconstruct nasal septal cartilage (NC). The harvesting of autologous cartilage bears the risk of donor site morbidity [1,5,6]. Those complications could be avoided if a viable, biocompatible and biomimetic equivalent to NC were available [7]. Tissue engineering approaches aim to produce cartilage tissue satisfying surgical needs [8,9]
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