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人脂肪干细胞与石榴果实水醇提取物修复骨软骨缺损的动物模型研究

An animal model study of osteochondral defect repair by human adipose stem cells and pomegranate fruit hydroalchoholic extract.

作者信息

Teimourinejad Ahmad, Hashemibeni Batool, Salehi Hossein, Mostafavi Fateme Sadat, Kazemi Mohammad, Bahramian Hamid

机构信息

Department of Anatomical Sciences, Medical School, Isfahan University of Medical Sciences, Isfahan, Iran.

Department of Genetic and Molecular Biology, Medical School, Isfahan University of Medical Sciences, Isfahan, Iran.

出版信息

Avicenna J Phytomed. 2023 Mar-Apr;13(2):177-188. doi: 10.22038/AJP.2022.21243.

Abstract

OBJECTIVE

Articular cartilage damages do not repair spontaneously. Tissue engineering is a promising approach to repair cartilage damage. Transforming growth factor-beta (TGF-β) members are the known induction factors in chondrogenic differentiation. However, hypertrophy of the chondrocytes resulting from mesenchymal stem cells (MSCs) induction by TGF-β is inevitable. Pomegranate fruit contains many ingredients which are useful in ensuring the health of organs. This study was designed to investigate the Pomegranate Fruit hydroalchoholic Extract (PFE) capability in human adipose derived stem cells (hASCs) differentiation into the chondrocytes on fibrin scaffold.

MATERIALS AND METHODS

Pomegranate fruit hydroalchoholic extract (PFE) was prepared. hASCs were isolated, expanded, labeled, and seeded on the fibrin scaffold. The constructs were divided into three groups including TGF-β3, PFE, and control. The constructs were induced for 14 days, then, the MTT assay, Real-Time Polymerase Chain Reaction (PCR), and histochemistry assessments were run, and finally, the constructs were transplanted into the knee defect of rats. The gross and histological assessments of the transplants were done after 8 weeks.

RESULTS

The viability rate, and genes expression levels, and histological criterion of the PFE samples were significantly higher than that of the control. The macroscopic grades and histological results of the PFE samples were close to that of the TGF-β3. The number of positive cells for COLІI protein were higher significantly in the PFE group than the control.

CONCLUSION

PFE was effective in the chondrogenic induction of hASCs. Further studies are needed to find out the events of the chondrogenic induction using PFE.

摘要

目的

关节软骨损伤无法自发修复。组织工程是修复软骨损伤的一种有前景的方法。转化生长因子-β(TGF-β)成员是已知的软骨形成分化诱导因子。然而,由TGF-β诱导间充质干细胞(MSCs)导致的软骨细胞肥大是不可避免的。石榴果实含有许多对确保器官健康有益的成分。本研究旨在调查石榴果实水醇提取物(PFE)在人脂肪来源干细胞(hASCs)于纤维蛋白支架上分化为软骨细胞方面的能力。

材料与方法

制备石榴果实水醇提取物(PFE)。分离、扩增、标记hASCs,并将其接种在纤维蛋白支架上。构建体分为三组,包括TGF-β3、PFE和对照组。对构建体诱导14天,然后进行MTT试验、实时聚合酶链反应(PCR)和组织化学评估,最后将构建体移植到大鼠膝关节缺损处。8周后对移植组织进行大体和组织学评估。

结果

PFE样本的存活率、基因表达水平和组织学标准均显著高于对照组。PFE样本的宏观分级和组织学结果与TGF-β3组相近。PFE组中COLІI蛋白阳性细胞数量显著高于对照组。

结论

PFE在hASCs的软骨形成诱导中有效。需要进一步研究以查明使用PFE进行软骨形成诱导的相关情况。

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