Lago Manuela E L, da Silva Lucília P, Henriques Catarina, Carvalho Andreia F, Reis Rui L, Marques Alexandra P
3B's Research Group-Biomaterials, Biodegradables and Biomimetics, Headquarters of the European Institute of Excellence on Tissue Engineering and Regenerative Medicine, University of Minho, Avepark, Barco, 4805-017 Guimarães, Portugal.
ICVS/3B's-PT Government Associate Laboratory, 4806-909 Braga/Guimarães, Portugal.
Bioengineering (Basel). 2018 Jun 27;5(3):52. doi: 10.3390/bioengineering5030052.
Adipose tissue is involved in many physiological processes. Therefore, the need for adipose tissue-like analogues either for soft tissue reconstruction or as in vitro testing platforms is undeniable. In this work, we explored the natural features of gellan gum (GG) to recreate injectable stable adipose-like microtissues. GG hydrogel particles with different percentages of polymer (0.5%, 0.75%, 1.25%) were developed and the effect of obtained mechanical properties over the ability of hASCs to differentiate towards the adipogenic lineage was evaluated based on the expression of the early (PPARγ) and late (FABP4) adipogenic markers, and on lipids formation and accumulation. Constructs were cultured in adipogenic induction medium up to 21 days or for six days in induction plus nine days in maintenance media. Overall, no significant differences were observed in terms of hASCs adipogenic differentiation within the range of Young’s moduli between 2.7 and 12.9 kPa. The long-term (up to six weeks) stability of the developed constructs supported its application in soft tissue reconstruction. Moreover, their ability to function as adipose-like microtissue models for drug screening was demonstrated by confirming its sensitivity to TNFα and ROCK inhibitor, respectively involved in the repression and induction of the adipogenic differentiation.
脂肪组织参与许多生理过程。因此,无论是用于软组织重建还是作为体外测试平台,对类似脂肪组织的类似物的需求都是不可否认的。在这项工作中,我们探索了结冷胶(GG)的天然特性,以创建可注射的稳定脂肪样微组织。我们制备了具有不同聚合物百分比(0.5%、0.75%、1.25%)的GG水凝胶颗粒,并基于早期(PPARγ)和晚期(FABP4)成脂标志物的表达以及脂质的形成和积累,评估了所获得的力学性能对人脂肪干细胞向脂肪生成谱系分化能力的影响。构建体在成脂诱导培养基中培养21天,或在诱导培养基中培养6天,然后在维持培养基中培养9天。总体而言,在2.7至12.9 kPa的杨氏模量范围内,人脂肪干细胞的成脂分化没有观察到显著差异。所开发构建体的长期(长达六周)稳定性支持其在软组织重建中的应用。此外,通过证实其对分别参与脂肪生成分化抑制和诱导的TNFα和ROCK抑制剂的敏感性,证明了它们作为药物筛选的脂肪样微组织模型的功能。