Wong Chin-Chean, Sheu Shi-Da, Chung Pei-Chun, Yeh Yi-Yen, Chen Chih-Hwa, Chang Yen-Wei, Kuo Tzong-Fu
Department of Orthopedics, Shuang Ho Hospital, Taipei Medical University, New Taipei City 23561, Taiwan.
Department of Orthopedics, School of Medicine, College of Medicine, Taipei Medical University, Taipei 11031, Taiwan.
Pharmaceutics. 2021 Mar 23;13(3):432. doi: 10.3390/pharmaceutics13030432.
The main aim of this study is to investigate the therapeutic efficacy of direct intra-articular injection of bone-marrow-derived stem/stromal cells (BMSCs) and the adjuvant role of hyaluronic acid (HA) in facilitating rabbit articular cartilage repair. First, rabbit BMSCs were treated with a medium containing different concentrations of HA. Later, HA's influence on BMSCs' CD44 expression, cell viability, extracellular glycosaminoglycan (GAG) synthesis, and chondrogenic gene expression was evaluated during seven-day cultivation. For the in vivo experiment, 24 rabbits were used for animal experiments and 6 rabbits were randomly allocated to each group. Briefly, chondral defects were created at the medial femoral condyle; group 1 was left untreated, group 2 was injected with HA, group 3 was transplanted with 3 × 10 BMSCs, and group 4 was transplanted with 3 × 10 BMSCs suspended in HA. Twelve weeks post-treatment, the repair outcome in each group was assessed and compared both macroscopically and microscopically. Results showed that HA treatment can promote cellular CD44 expression. However, the proliferation rate of BMSCs was downregulated when treated with 1 mg/mL (3.26 ± 0.03, = 0.0002) and 2 mg/mL (2.61 ± 0.04, = 0.0001) of HA compared to the control group (3.49 ± 0.05). In contrast, 2 mg/mL (2.86 ± 0.3) of HA treatment successfully promoted normalized GAG expression compared to the control group (1.88 ± 0.06) ( = 0.0009). The type II collagen gene expression of cultured BMSCs was significantly higher in BMSCs treated with 2 mg/mL of HA ( = 0.0077). In the in vivo experiment, chondral defects treated with combined BMSC and HA injection demonstrated better healing outcomes than BMSC or HA treatment alone in terms of gross grading and histological scores. In conclusion, this study helps delineate the role of HA as a chondrogenic adjuvant in augmenting the effectiveness of stem-cell-based injection therapy for in vivo cartilage repair. From a translational perspective, the combination of HA and BMSCs is a convenient, ready-to-use, and effective formulation that can improve the therapeutic efficacy of stem-cell-based therapies.
本研究的主要目的是探讨直接关节腔内注射骨髓源性干/基质细胞（BMSCs）的治疗效果以及透明质酸（HA）在促进兔关节软骨修复中的辅助作用。首先，用含有不同浓度HA的培养基处理兔BMSCs。随后，在为期七天的培养过程中评估HA对BMSCs的CD44表达、细胞活力、细胞外糖胺聚糖（GAG）合成以及软骨生成基因表达的影响。对于体内实验，使用24只兔子进行动物实验，每组随机分配6只兔子。简要地说，在股骨内侧髁制造软骨缺损；第1组不进行处理，第2组注射HA，第3组移植3×10个BMSCs，第4组移植悬浮于HA中的3×10个BMSCs。治疗后十二周，对每组的修复结果进行宏观和微观评估及比较。结果显示，HA处理可促进细胞CD44表达。然而，与对照组（3.49±0.05）相比，用1mg/mL（3.26±0.03，P = 0.0002）和2mg/mL（2.61±0.04，P = 0.0001）的HA处理时，BMSCs的增殖率下调。相比之下，与对照组（1.88±0.06）（P = 0.0009）相比，2mg/mL（2.86±0.3）的HA处理成功促进了正常化的GAG表达。在用2mg/mL的HA处理的BMSCs中，培养的BMSCs的II型胶原基因表达显著更高（P = 0.0077）。在体内实验中，就大体分级和组织学评分而言，联合注射BMSC和HA治疗的软骨缺损显示出比单独使用BMSC或HA治疗更好的愈合结果。总之，本研究有助于阐明HA作为软骨生成辅助剂在增强基于干细胞的注射疗法对体内软骨修复有效性方面的作用。从转化医学的角度来看，HA和BMSCs的组合是一种方便、即用型且有效的制剂，可提高基于干细胞疗法的治疗效果。
