Taghavi Hossein, Soleimani Rad Jafar, Mehdipour Ahmad, Ferdosi Khosroshahi Ahad, Kheirjou Raziyeh, Hasanpour Milad, Roshangar Leila
Stem Cell Research Center, Tabriz University of Medical Sciences, Tabriz, Iran.
Department of Anatomical Sciences, Faculty of Medicine, Tabriz University of Medical Sciences, Tabriz, Iran.
Adv Pharm Bull. 2020 Sep;10(4):623-629. doi: 10.34172/apb.2020.075. Epub 2020 Aug 9.
Acellular scaffold extracted from extracellular matrix (ECM) have been used for constructive and regenerative medicine. Adipose derived stem cells (ADSCs) can enhance the vascularization capacity of scaffolds. High mobility group box 1 (HMGB1) and stromal derived factor1 (SDF1) are considered as two important factors in vascularization and immunologic system. In this study, the effect of mineral pitch on the proliferation of human ADSCs was evaluated. In addition to HMGB1 and SDF1, factors expression in acellular scaffold was also assessed. To determine acellular scaffold morphology and the degree of decellularization, hematoxylin & eosin (H&E), 6-diamidino-2-phenylindole (DAPI), and Masson's trichrome staining were applied. The scaffolds were treated with mineral pitch. Also, ADSCs were seeded on the scaffolds, and adhesion of the cells to the scaffolds were assessed using field emission scanning electron microscopy (FE-SEM). In addition, the efficiency of mineral pitch to induce the proliferation of ADSCs on the scaffolds was evaluated using 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyl tetrazolium bromide (MTT) assay. To measure HMGB1 and SDF1 mRNA expression, real-time polymerase chain reactions (RT-PCR) was used. FE-SEM showed that decellularized matrix possesses similar matrix morphology with a randomly oriented fibrillar structure and interconnecting pores. No toxicity was observed in all treatments, and cell proliferation were supported in scaffolds. The important point is that, the proliferation capacity of ADSCs on Mineral pitch loaded scaffolds significantly increased after 48 h incubation time compared to the unloaded scaffold (P<0.001). The results of this study suggest that mineral pitch has potentials to accelerate proliferation of ADSCs on the acellular scaffolds.
从细胞外基质（ECM）中提取的无细胞支架已用于组织工程和再生医学。脂肪来源干细胞（ADSCs）可增强支架的血管生成能力。高迁移率族蛋白B1（HMGB1）和基质细胞衍生因子1（SDF1）被认为是血管生成和免疫系统中的两个重要因子。在本研究中，评估了矿脂对人ADSCs增殖的影响。除了HMGB1和SDF1，还评估了无细胞支架中各因子的表达。为确定无细胞支架的形态和脱细胞程度，采用苏木精-伊红（H&E）染色、6-二脒基-2-苯基吲哚（DAPI）染色和Masson三色染色。对支架进行矿脂处理。此外，将ADSCs接种在支架上，用场发射扫描电子显微镜（FE-SEM）评估细胞与支架的黏附情况。另外，使用3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四氮唑溴盐（MTT）法评估矿脂诱导ADSCs在支架上增殖的效率。为检测HMGB1和SDF1 mRNA表达水平，采用实时聚合酶链反应（RT-PCR）。FE-SEM显示，脱细胞基质具有类似的基质形态，呈随机取向的纤维状结构和相互连通的孔隙。所有处理均未观察到毒性，且支架支持细胞增殖。重要的是，与未负载矿脂的支架相比，负载矿脂的支架在孵育48小时后ADSCs的增殖能力显著增强（P<0.001）。本研究结果表明，矿脂具有促进ADSCs在无细胞支架上增殖的潜力。
