Gelatin methacryloyl (GelMA) loaded with concentrated hypoxic pretreated adipose-derived mesenchymal stem cells(ADSCs) conditioned medium promotes wound healing and vascular regeneration in aged skin.

IF 11.3 1区 医学 Q1 Medicine
Shiyi Li, Jiachen Sun, Jinxiu Yang, Yi Yang, Hongfan Ding, Boya Yu, Kui Ma, Minliang Chen
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引用次数: 7

Abstract

Background: Aging skin is characterized by a disturbed structure and lack of blood supply, which makes it difficult to heal once injured. ADSCs secrete large amounts of cytokines, which promote wound healing and vascular regeneration through paracrine secretion, and the number of cytokines can be elevated by hypoxic pretreating. However, the components of ADSCs are difficult to retain in wounds. Gelatin methacrylate (GelMA) is a photopolymerizable hydrogel synthesized from gelatin and has recently emerged as a potentially attractive material for tissue engineering applications. GelMA loaded with concentrated hypoxic pretreated ADSCs conditioned medium could provide a new method of treating wounds in aged skin.

Methods: Primary ADSCs were isolated from human adipose tissue and characterized by flow cytometry and differentiation test. ADSCs in passages 4-6 were pretreated in the hypoxic and normoxic environments to collect conditioned medium, the conditioned medium was then concentrated to prepare concentrated ADSCs conditioned medium(cADSC-CM)(the one collected from ADSCs under hypoxia was called hypo-CM ,and the one from normoxia was called nor-CM). The concentration of cytokines was detected. After treated with cADSC-CM, the abilities of proliferation, migration, and tube formation of human umbilical vascular endothelial cells (HUVECs) were assayed, and Akt/mTOR and MAPK signal pathway was detected using western blotting. GelMA+hypo-CM hydrogel was prepared, and a comprehensive evaluation of morphology, protein release efficiency, degradation rate, mechanical properties, and rheology properties were performed. Full-thickness skin wounds were created on the backs of 20-month-old mice. After surgery, GelMA, GelMA+F12, GelMA+hypo-CM, and GelMA+nor-CM were applied to the wound surface respectively. H&E, Masson, and immunohistochemistry staining were performed, and a laser Doppler perfusion imager was used to evaluate the blood perfusion. The student's t-test was used for analysis between two groups and a one-way analysis of variance (ANOVA) was used for analysis among multi groups.

Results: Our results revealed that 1) wounds in aged skin healed more slowly than that in young skin and exhibited poorer perfusion; 2) hypoxic pretreated ADSCs secreted more cytokines including VEGF by activating HIF1α; 3) hypo-CM promoted proliferation and migration of HUVECs through VEGF/Akt/mTOR and MAPK signal pathway; 4) GelMA-hypoCM accelerated wound healing and angiogenesis in aged skin in vivo.

Conclusion: GelMA loaded with concentrated hypoxic pretreated adipose-derived mesenchymal stem cells conditioned medium could accelerate wound healing in aged skin by promoting angiogenesis.

明胶甲基丙烯酰(GelMA)装载浓缩缺氧预处理脂肪源性间充质干细胞(ADSCs)条件培养基,促进老化皮肤的伤口愈合和血管再生。
背景:皮肤老化的特点是结构紊乱,血液供应不足,一旦受伤就很难愈合。ADSCs分泌大量细胞因子,通过旁分泌促进创面愈合和血管再生,低氧预处理可提高细胞因子的数量。然而,ADSCs的成分很难在伤口中保留。明胶甲基丙烯酸酯(GelMA)是一种由明胶合成的光聚合水凝胶,近年来作为一种潜在的有吸引力的材料应用于组织工程。GelMA负载高浓度缺氧预处理的ADSCs条件培养基,为治疗老化皮肤创面提供了一种新的方法。方法:从人脂肪组织中分离原代ADSCs,采用流式细胞术和分化实验对其进行鉴定。将4 ~ 6代ADSCs分别在缺氧和常氧环境下进行预处理,收集条件培养基,将条件培养基浓缩制备ADSCs浓缩条件培养基(cADSC-CM)(缺氧条件下收集的ADSCs称为hypoo - cm,常氧条件下收集的ADSCs称为normo - cm)。检测细胞因子浓度。经cADSC-CM处理后,检测人脐血管内皮细胞(HUVECs)的增殖、迁移和成管能力,并采用western blotting检测Akt/mTOR和MAPK信号通路。制备GelMA+低- cm水凝胶,对其形态、蛋白质释放效率、降解率、力学性能、流变性能进行综合评价。在20个月大的小鼠背部创造了全层皮肤伤口。术后分别将GelMA、GelMA+F12、GelMA+低cm、GelMA+无cm涂抹于创面。行H&E、Masson、免疫组化染色,激光多普勒灌注成像仪评估血流灌注。两组间采用学生t检验,多组间采用单因素方差分析(ANOVA)。结果:1)老龄皮肤创面愈合速度慢于年轻皮肤创面,血流灌注较差;2)缺氧预处理的ADSCs通过激活HIF1α分泌更多的VEGF等细胞因子;3)低浓度cm通过VEGF/Akt/mTOR和MAPK信号通路促进HUVECs的增殖和迁移;4) GelMA-hypoCM在体内加速老化皮肤的伤口愈合和血管生成。结论:装载浓缩缺氧预处理脂肪间充质干细胞条件培养基的GelMA可通过促进血管生成来加速老化皮肤创面愈合。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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来源期刊
Biomaterials Research
Biomaterials Research Medicine-Medicine (miscellaneous)
CiteScore
10.20
自引率
3.50%
发文量
63
审稿时长
30 days
期刊介绍: Biomaterials Research, the official journal of the Korean Society for Biomaterials, is an open-access interdisciplinary publication that focuses on all aspects of biomaterials research. The journal covers a wide range of topics including novel biomaterials, advanced techniques for biomaterial synthesis and fabrication, and their application in biomedical fields. Specific areas of interest include functional biomaterials, drug and gene delivery systems, tissue engineering, nanomedicine, nano/micro-biotechnology, bio-imaging, regenerative medicine, medical devices, 3D printing, and stem cell research. By exploring these research areas, Biomaterials Research aims to provide valuable insights and promote advancements in the biomaterials field.
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