Pourentezari Majid, Sharifian Zeinolabedin, Mardani Mohammad, Valiani Ali, Zamani Rarani Mohammad, Setayeshmehr Mohsen, Eini Fatemeh, Hashemibeni Batool
Department of Biology and Anatomical Sciences, Shahid Sadoughi University of Medical Sciences, Yazd, Iran.
Department of Anatomy, Faculty of Medicine, Hormozgan University of Medical Sciences, Bandar Abbas, Iran.
Iran J Basic Med Sci. 2021 Jan;24(1):24-29. doi: 10.22038/ijbms.2020.44409.10391.
Avocado/soybean unsaponifible (ASU) possesses properties including chondroprotective, anticatabolic, and anabolic. The goal behind this research was to detect the effect of ASU and TGF-β3 on the chondrogenesis of human adipose-derived stem cells (hADSCs) on poly (lactic-co-glycolic) acid (PLGA)/ hyaluronic acid (PLGA/HA) hybrid scaffold.
First hADSCs were seeded in PLGA/Hyaluronic acid scaffold and cultured in chondrogenic media. These cells were assigned into 4 groups: control, TGFβ-3, ASU, and TGFβ-3+ASU. The viability was assessed separately by MTT. Real-time PCR was used to quantify the expression of chondrogenic specific genes [] and . Moreover, Western blotting was employed to evaluate protein expression levels of collagens type II and X.
These findings indicated a significant increase in the proliferation and survival of hADSCs differentiated cells by ASU compared with the control group (=0.008). Real-time PCR results revealed significant differences in the expression of , , , and genes in the control group when compared with other groups (ASU, TGF-β3, and TGF-β3+ASU). ColII protein production significantly dropped in the TGF-β3 group in comparison with the TGF-β3+ASU group (0.000). The ColII (=0.002) and ColX (=0.002) protein production proved significantly higher in the TGF-β3+ASU group compared with the ASU group.
Using the synergist form TGFβ-3, ASU induces chondrogenesis in hADSCs in PLGA/HA composite scaffold. This can be deduced with reduction of special markers of hyaline cartilage in comparison with ASU and decreased hypertrophic marker compared with TGF-β3.
鳄梨/大豆不皂化物(ASU)具有软骨保护、抗分解代谢和合成代谢等特性。本研究的目的是检测ASU和转化生长因子-β3(TGF-β3)对人脂肪来源干细胞(hADSCs)在聚乳酸-乙醇酸共聚物(PLGA)/透明质酸(PLGA/HA)混合支架上软骨形成的影响。
首先将hADSCs接种于PLGA/透明质酸支架上,并在软骨形成培养基中培养。这些细胞被分为4组:对照组、TGFβ-3组、ASU组和TGFβ-3+ASU组。通过MTT法分别评估细胞活力。采用实时定量PCR法检测软骨形成特异性基因[ ]和[ ]的表达。此外,采用蛋白质免疫印迹法评估Ⅱ型和X型胶原蛋白的蛋白表达水平。
这些结果表明,与对照组相比,ASU可显著提高hADSCs分化细胞的增殖和存活率(P=0.008)。实时定量PCR结果显示,与其他组(ASU组、TGF-β3组和TGF-β3+ASU组)相比,对照组中[ ]、[ ]、[ ]和[ ]基因的表达存在显著差异。与TGF-β3+ASU组相比,TGF-β3组中Ⅱ型胶原蛋白的蛋白产量显著下降(P=0.000)。与ASU组相比,TGF-β3+ASU组中Ⅱ型胶原蛋白(P=0.002)和X型胶原蛋白(P=0.002)的蛋白产量显著更高。
联合使用TGFβ-3,ASU可诱导PLGA/HA复合支架上的hADSCs发生软骨形成。与单独使用ASU相比,透明软骨特异性标志物减少,与单独使用TGF-β3相比,肥大标志物减少,由此可推断上述结论。