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慢性外侧踝关节不稳患者滑膜间充质干细胞的分离与鉴定:踝关节滑液、脂肪滑膜和纤维滑膜的比较分析

Isolation and Characterization of Synovial Mesenchymal Stem Cells Derived From Patients With Chronic Lateral Ankle Instability: A Comparative Analysis of Synovial Fluid, Adipose Synovium, and Fibrous Synovium of the Ankle Joint.

作者信息

Nakashima Hirotaka, Uchida Soshi, Hatakeyama Akihisa, Murata Yoichi, Yamanaka Yoshiaki, Tsukamoto Manabu, Sekiya Ichiro, Sakai Akinori

机构信息

Department of Orthopaedic Surgery, University of Occupational and Environmental Health, Kitakyushu, Japan.

Department of Orthopaedic Surgery, Wakamatsu Hospital, University of Occupational and Environmental Health, Kitakyushu, Japan.

出版信息

Orthop J Sports Med. 2022 May 13;10(5):23259671221094615. doi: 10.1177/23259671221094615. eCollection 2022 May.

Abstract

BACKGROUND

Synovial mesenchymal stem cells (MSCs) have high proliferative potential and are considered an excellent source for stem cell therapy.

PURPOSES

To isolate MSCs from the synovium of ankle joints in patients with chronic lateral ankle instability (CLAI) and to compare the characteristics of MSCs derived from the synovium anterior to the talus with those from the surrounding anterior talofibular ligament (ATFL) synovium.

STUDY DESIGN

Controlled laboratory study.

METHODS

The synovium was harvested from 2 locations in the ankle, the synovium anterior to the talus and the surrounding ATFL synovium, of 14 patients who underwent arthroscopic ATFL repair for CLAI without osteochondral lesions of the talus (OLTs). Synovial fluid was also harvested. MSCs were isolated from both types of synovial tissue, as well as synovial fluid. The number of MSCs in the synovium and their viability, proliferation, colony-forming units, and potential to differentiate into adipose, bone, and cartilage tissues were determined and compared between groups. Additionally, real-time polymerase chain reaction was used to assess the differentiation capacity of adipose, bone, and cartilage tissues from both samples. The Wilcoxon signed rank test was used to compare the sample weight, number of colonies, number of nucleated cells per colony, yield obtained, and phenotypic characteristics of MSCs derived from different locations of the synovium.

RESULTS

No significant differences were observed in the sample weight ( = .051), number of nucleated cells per milligram ( = .272), number of colonies ( = .722), and yield obtained ( = .099) between the 2 groups. MSCs could not be isolated from synovial fluid. The frequency of oil red O-positive adipogenic colonies ( = .028) and the expression of the adipsin gene ( < .05) were significantly increased in the cells from the synovium anterior to the talus compared to those in the cells from the surrounding ATFL synovium. However, chondrogenic and osteogenic potentials were not significantly different between the 2 groups.

CONCLUSION

Synovial MSCs obtained from the ankle joint had self-renewal and multilineage differentiation potential, although the adipogenesis potential of MSCs from the synovium anterior to the talus was superior to that from the surrounding ATFL synovium.

CLINICAL RELEVANCE

Both the adipose synovium and fibrous synovium in the ankle joints of patients with CLAI may be a good source of MSCs for stem cell therapy applications, whereas synovial fluid appeared unsuitable.

摘要

背景

滑膜间充质干细胞(MSCs)具有高增殖潜能,被认为是干细胞治疗的优质来源。

目的

从慢性外侧踝关节不稳(CLAI)患者的踝关节滑膜中分离MSCs,并比较距骨前方滑膜来源的MSCs与周围距腓前韧带(ATFL)滑膜来源的MSCs的特性。

研究设计

对照实验室研究。

方法

从14例因CLAI接受关节镜下ATFL修复且无距骨骨软骨损伤(OLT)的患者踝关节的2个部位采集滑膜,即距骨前方滑膜和周围ATFL滑膜。同时采集滑液。从这两种滑膜组织以及滑液中分离MSCs。测定并比较两组滑膜中MSCs的数量及其活力、增殖能力、集落形成单位以及向脂肪、骨和软骨组织分化的潜能。此外,采用实时聚合酶链反应评估两个样本来源的脂肪、骨和软骨组织的分化能力。采用Wilcoxon符号秩检验比较滑膜不同部位来源的MSCs的样本重量、集落数量、每个集落的有核细胞数量、获得的产量以及表型特征。

结果

两组在样本重量(P = 0.051)、每毫克有核细胞数量(P = 0.272)、集落数量(P = 0.722)和获得的产量(P = 0.099)方面未观察到显著差异。无法从滑液中分离出MSCs。与周围ATFL滑膜来源的细胞相比,距骨前方滑膜来源的细胞中油红O阳性脂肪生成集落的频率(P = 0.028)和脂联素基因的表达(P < 0.05)显著增加。然而,两组间的软骨生成和成骨潜能无显著差异。

结论

踝关节来源的滑膜MSCs具有自我更新和多向分化潜能,尽管距骨前方滑膜来源的MSCs的脂肪生成潜能优于周围ATFL滑膜来源的MSCs。

临床意义

CLAI患者踝关节的脂肪滑膜和纤维滑膜均可能是干细胞治疗应用中MSCs的良好来源,而滑液似乎不适合。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7c45/9118449/5369675098f3/10.1177_23259671221094615-fig1.jpg

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