Department of Plastic Surgery, Hand Surgery-Burn Center, University Hospital RWTH Aachen, Pauwelsstraße 30, Aachen 52074, Germany.
Tissue Cell. 2022 Oct;78:101886. doi: 10.1016/j.tice.2022.101886. Epub 2022 Aug 3.
Human adipose stem cells (ASCs) hold great potential for regenerative medicine approaches, including osteogenic regeneration of bone defects, that fail to heal autonomously. Osteogenic differentiation of stem cells is dependent on the stimulation of biophysical factors. In the present study, the effects of hypergravity, hypoxia, and hyperbaric treatment were investigated on adipose stem cell (ASC) metabolic activity, quantified by PrestoBlue conversion, and cell numbers, evaluated by crystal violet staining. Osteogenic differentiation was assessed by alkaline phosphatase (ALP) activity and cresolphthalein staining of calcium deposition. Differentiation was performed for 12 days, which was accompanied by periodical stimulation. Increasing gravity forces up to 50x g did not affect ASC viability, but it enhanced osteogenic markers with a strongest effect between 20 and 30x g. Hyperbaric stimulation at 3 bar decreased ASC cell numbers but increased ALP activity and calcium deposition. Hypoxia at 8 % atmospheric oxygen did not affect ASC proliferation, while cell numbers were reduced at 3 % oxygen. Furthermore, hypoxic conditions produced opposing results on osteogenic markers, as ALP activity increased whereas cresolphthalein staining decreased upon stimulation. These data demonstrated that intermittent short duration of basal physical or chemical impulses interfere with the osteogenic differentiation of ASCs. Our findings could be of specific relevance in ASC based therapies for regenerative medicine and bone tissue engineering approaches.
人类脂肪干细胞 (ASCs) 在再生医学方法中具有巨大的潜力，包括对无法自主愈合的骨缺损进行成骨再生。干细胞的成骨分化依赖于生物物理因素的刺激。在本研究中，研究了超重力、低氧和高压处理对脂肪干细胞 (ASC) 代谢活性的影响，通过 PrestoBlue 转化进行量化，并通过结晶紫染色评估细胞数量。通过碱性磷酸酶 (ALP) 活性和钙沉积的 cresolphthalein 染色评估成骨分化。分化进行了 12 天，并伴有周期性刺激。增加高达 50x g 的重力不会影响 ASC 的活力，但在 20 到 30x g 之间具有最强的增强成骨标志物的作用。3 巴的高压刺激会降低 ASC 细胞数量，但会增加 ALP 活性和钙沉积。8%大气氧的低氧不会影响 ASC 的增殖，而 3%氧会减少细胞数量。此外，低氧条件对成骨标志物产生相反的结果，因为 ALP 活性增加，而刺激时 cresolphthalein 染色减少。这些数据表明，基础物理或化学脉冲的间歇性短暂持续时间会干扰 ASCs 的成骨分化。我们的发现可能对基于 ASC 的再生医学和骨组织工程方法的治疗具有特定的相关性。
