Bollmann Anne, Sons Hans Christian, Schiefer Jennifer Lynn, Fuchs Paul C, Windolf Joachim, Suschek Christoph Viktor
Department for Orthopedics and Trauma Surgery, Medical Faculty and University Hospital Duesseldorf, Heinrich-Heine-University Düsseldorf, Moorenstraße 5, 40225 Düsseldorf, Germany.
Department of Plastic Surgery, Hand Surgery, Burn Center, Merheim Hospital Cologne, University of Witten/Herdecke, Ostmerheimer Straße 200, 51109 Köln, Germany.
Biomedicines. 2022 Nov 29;10(12):3071. doi: 10.3390/biomedicines10123071.
Adipose tissue-derived stromal cells (ASCs) play an important role in various therapeutic approaches to bone regeneration. However, such applications become challenging when the obtained cells show a functional disorder, e.g., an impaired osteogenic differentiation potential (ODP). In addition to ASCs, human adipose tissue is also a source for another cell type with therapeutic potential, the dedifferentiated fat cells (DFATs), which can be obtained from mature adipocytes. Here, we for the first time compared the ODPs of each donors ASC and DFAT obtained from the same adipose tissue sample as well as the role of oxidative stress or antioxidative catalase on their osteogenic outcome. Osteogenic potential of ASC and DFAT from nine human donors were compared in vitro. Flow cytometry, staining for calcium accumulation with alizarin red, alkaline phosphatase assay and Western blots were used over an osteogenic induction period of up to 14 days. HO was used to induce oxidative stress and catalase was used as an antioxidative measure. We have found that ASC and DFAT cultures' ODPs are nearly identical. If ASCs from an adipose tissue sample showed good or bad ODP, so did the corresponding DFAT cultures. The inter-individual variability of the donor ODPs was immense with a maximum factor of about 20 and correlated neither with the age nor the sex of the donors of the adipose tissue. Oxidative stress in the form of exogenously added HO led to a significant ODP decrease in both cell types, with this ODP decrease being significantly lower in DFAT cultures than in the corresponding ASC cultures. Regardless of the individual cell culture-specific ODP, however, exogenously applied catalase led to an approx. 2.5-fold increase in osteogenesis in the ASC and DFAT cultures. Catalase appears to be a potent pro-osteogenic factor, at least in vitro. A new finding that points to innovative strategies and therapeutic approaches in bone regeneration. Furthermore, our results show that DFATs behave similarly to ASCs of the same adipose tissue sample with respect to ODPs and could therefore be a very attractive and readily available source of multipotent stem cells in bone regenerative therapies.
脂肪组织来源的基质细胞（ASC）在骨再生的各种治疗方法中发挥着重要作用。然而，当获得的细胞出现功能障碍，例如成骨分化潜能（ODP）受损时，此类应用就会变得具有挑战性。除了ASC，人类脂肪组织还是另一种具有治疗潜力的细胞类型——去分化脂肪细胞（DFAT）的来源，DFAT可从成熟脂肪细胞中获得。在此，我们首次比较了从同一脂肪组织样本中获取的每个供体的ASC和DFAT的ODP，以及氧化应激或抗氧化过氧化氢酶对它们成骨结果的作用。对来自9名人类供体的ASC和DFAT的成骨潜能进行了体外比较。在长达14天的成骨诱导期内，使用了流式细胞术、茜素红染色检测钙积累、碱性磷酸酶测定和蛋白质免疫印迹法。使用过氧化氢（HO）诱导氧化应激，并使用过氧化氢酶作为抗氧化措施。我们发现ASC和DFAT培养物的ODP几乎相同。如果来自脂肪组织样本的ASC表现出良好或不良的ODP，相应的DFAT培养物也是如此。供体ODP的个体间变异性极大，最大差异约为20倍，且与脂肪组织供体的年龄和性别均无相关性。外源性添加HO形式的氧化应激导致两种细胞类型的ODP均显著降低，DFAT培养物中的ODP降低幅度明显低于相应的ASC培养物。然而，无论个体细胞培养特异性的ODP如何，外源性应用过氧化氢酶都会使ASC和DFAT培养物中的成骨增加约2.5倍。过氧化氢酶似乎是一种有效的促骨生成因子，至少在体外是这样。这一新发现为骨再生的创新策略和治疗方法指明了方向。此外，我们的结果表明，就ODP而言，DFAT的行为与同一脂肪组织样本的ASC相似，因此在骨再生治疗中可能是一种非常有吸引力且易于获得的多能干细胞来源。
