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Bio-Design and Manufacturing  2021 Vol.4 No.3 P.441-451

http://doi.org/10.1007/s42242-021-00128-5


Adenosine-treated bioprinted muscle constructs prolong cell survival and improve tissue formation


Author(s):  Dongxu Ke, Adam M. Jorgensen, Sang J. Lee, James J. Yoo & Sean V. Murphy

Affiliation(s):  Wake Forest Institute for Regenerative Medicine, Wake Forest School of Medicine, Winston-Salem, NC, USA; more

Corresponding email(s):   semurphy@wakehealth.edu

Key Words:  Bioprinting, Hypoxia, Adenosine, Cell viability, In vivo


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Dongxu Ke, Adam M. Jorgensen, Sang J. Lee, James J. Yoo & Sean V. Murphy. Adenosine-treated bioprinted muscle constructs prolong cell survival and improve tissue formation[J]. Journal of Zhejiang University Science D, 2021, 4(3): 441-451.

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Abstract: 
It is crucial to maintaining the viability of biofabricated human-sized tissues to ensure their successful survival and function after transplantation. adenosine is a purine nucleoside that has the function to suppress cellular metabolism and has been previously proposed as a method to prolong cell viability under hypoxia. In this study, we optimized the dose concentration of adenosine for incorporation into bioprinted constructs to preserve long-term cell viability in vitro. Our results showed that muscle cells (C2C12) containing 6, 7, or 8 mM adenosine maintained high cell viability for 20 days under hypoxic conditions (0.1% O2), whereas cells without adenosine treatment showed 100% cell death after 11 days. After 20 days under hypoxic conditions, muscle cells treated with adenosine proliferated and differentiated when transferred to normoxic conditions. From these adenosine concentrations, 6 mM was picked as the optimized adenosine concentration for further investigations due to its most effective results on improving cell viability. The bioprinted muscle constructs containing adenosine (6 mM) maintained high cell viability for 11 days under hypoxic conditions, while the control constructs without adenosine had no live cells. For in vivo validation, the bioprinted constructs with adenosine implanted under the dorsal subcutaneous space in mice, showed the enhanced formation of muscle tissue with minimal central necrosis and apoptosis, when compared to the constructs without adenosine. These positive in vitro and in vivo results demonstrate that the use of adenosine is an effective approach to preventing the challenge of hypoxia-induced necrosis in bioprinted tissues for clinical translation.

维克森林大学医学院Sean V. Murphy等 | 腺苷处理的生物打印肌肉组织以延长细胞存活并改善组织形成

本研究论文聚焦腺苷的使用以提高3D打印肌肉组织的生物活性保持问题。维持细胞活性对于生物制造人体大小的组织和确保其在移植后成功存活和功能至关重要。腺苷是一种嘌呤核苷,具有抑制细胞代谢的功能,之前已被提出作为在缺氧下延长细胞活力的方法。在这项研究中,我们优化了腺苷的剂量浓度,并将其掺入生物打印的组织结构中,以保持体外的长期细胞活力。我们的结果表明,含有6、7或8 mM腺苷的肌肉细胞(C2C12)在缺氧条件下(0.1% O2)保持高细胞活力20天,而没有腺苷处理的细胞在11天后显示100%细胞死亡。在缺氧条件下20天后,用腺苷处理的肌肉细胞在转移到正常毒性条件下时增殖和分化。从这些腺苷浓度中,选择6 mM作为优化的腺苷浓度进行进一步研究,实验显示其在提高细胞活力方面具有最有效的结果。含有腺苷(6 mM)的生物打印肌肉构建体在缺氧条件下保持高细胞活力11天,而没有腺苷的对照构建体没有活细胞。对于体内验证,与没有腺苷的打印结果相比,将腺苷加入植入小鼠背皮下的打印组织显示出肌肉组织的增强形成,具有最小的中央坏死和凋亡。这些积极的体外和体内结果表明,使用腺苷是预防生物打印组织缺氧导致坏死的有效方法,从而使其具有进一步进行临床转化的可能性。

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