All-in-One Hydrogel Realizing Adipose Derived Stem Cell Spheroids Production and in Vivo Injection via “Gel-Sol” Transition for Angiogenesis in Hind Limb Ischemia
All-in-One Hydrogel Realizing Adipose Derived Stem Cell Spheroids Production and in Vivo Injection via “Gel-Sol” Transition for Angiogenesis in Hind Limb Ischemia
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多合一水凝胶通过“凝胶-溶胶”过渡实现脂肪干细胞球体生产和体内注射，促进后肢缺血的血管生成
DOI：
10.1021/acsami.9b23534
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发表时间：
2020
影响因子：
9.5
通讯作者：
Jingbo Yin
中科院分区：
文献类型：
--
作者：
Yuhao Hong;Jialin Chen;Haowei Fang;Guifei Li;Shifeng Yan;Kunxi Zhang;Chen Wang;Jingbo Yin
脂肪源性干细胞（ASC）球状体表现出增强的血管生成功效，用于缺血治疗。因此，有必要开发一种一体化平台，使高效的球状体生产，收集和体内注射植入成为可能。本研究制备了一种基于聚（L-谷氨酸）（PLGA）的多孔水凝胶，不仅可以产生ASC球体，而且可以方便地收集球体，通过微创注射植入体内治疗后肢缺血。PLGA与含有二硫键的胱胺（Cys）交联形成多孔水凝胶，该水凝胶可通过谷胱甘肽（GSH）的还原作用实现"凝胶-溶胶"转变。一方面，发现在PLGA水凝胶中引入二硫键通过结合纤连蛋白促进细胞粘附，防止球状体的形成，而引入聚乙二醇单甲醚（mPEG）可以干扰二硫键对细胞粘附的影响，支持多孔水凝胶内的球状体形成。另一方面，转移到注射器中的多孔水凝胶可以在约40分钟内变成液体聚合物溶液，用于收集所产生的球状体和体内注射。此外，由于聚合物溶液的润滑，在通过25G注射器针头注射球状体/聚合物混悬液期间保护球状体，避免剪切损坏。体内注射后，ASC球状体旁分泌增加，促进血管生成和肌肉再生，21 d后对小鼠肢体缺血有明显治疗作用。同时，PLGA基材料在体内表现出良好的生物相容性。
Adipose-derived stem cell (ASC) spheroids exhibit enhanced angiogenic efficacy toward ischemia treatment. Thus, it is necessary to develop an all-in-one platform that enables efficient spheroid production, collection, and injectable implantation in vivo. The present study fabricated a poly(l-glutamic acid) (PLGA)-based porous hydrogel that can not only produce ASC spheroids but also conveniently collect spheroids for in vivo implantation via minimally invasive injection to treat hind limb ischemia. PLGA was cross-linked with cystamine (Cys), which contains disulfide bonds, to form a porous hydrogel that could realize “gel–sol” transition by the reduction effect of glutathione (GSH). For one thing, it was found that the introduction of the disulfide bond in the PLGA hydrogel promoted cellular adhesion via combining fibronectin, preventing the formation of spheroids, while the introduction of polyethylene glycol monomethyl ether (mPEG) could disturb the effect of the disulfide bond on cellular adhesion, supporting spheroid formation inside the porous hydrogel. For another, the porous hydrogel transferred into a syringe could turn into liquid polymer solution within about 40 min for collection of the produced spheroids and in vivo injection. In addition, because of the lubrication of polymer solution, the spheroids were protected during the injection of the spheroids/polymer suspensoid through a 25G syringe needle, avoiding damages from shearing. After the in vivo injection, the enhanced paracrine secretion of ASC spheroids resulted in promoted angiogenesis and muscle regeneration, exhibiting obvious therapeutic effect on limb ischemia in mice after 21 days. At the same time, PLGA-based material exhibited well-performed biocompatibility in vivo.