Wu Kun-Chi, Chang Yu-Hsun, Liu Hwan-Wun, Ding Dah-Ching
Department of Orthopedics, Hualien Tzu Chi Hospital, Buddhist Tzu Chi Medical Foundation and Tzu Chi University, Hualien, Taiwan.
Department of Pediatrics, Hualien Tzu Chi Hospital, Buddhist Tzu Chi Medical Foundation and Tzu Chi University, Hualien, Taiwan.
Tzu Chi Med J. 2019 Jan-Mar;31(1):11-19. doi: 10.4103/tcmj.tcmj_87_18.
Osteoarthritis (OA) is a chronic disease of degenerative joints. Mesenchymal stem cells (MSCs) have been used for cartilage regeneration in OA. We investigated the therapeutic potential of human umbilical cord-derived MSCs (HUCMSCs) with hyaluronic acid (HA) hydrogel transplanted into a porcine OA preclinical model.
The HUCMSCs were characterized with respect to morphology, surface markers, and differentiation capabilities. Quantitative reverse-transcriptase polymerase chain reaction (qRT-PCR) was used to examine gene expressions in a HUCMSC-HA coculture. Two healthy female minipigs weighing 30-40 kg and aged approximately 4 months were used in this large animal study. A full-thickness chondral injury was created in the trochlear groove of each of the pig's rear knees. After 3 weeks, a second osteochondral defect was created. Then, 1.5 mL of a HUCMSC (5 × 10 cells) and HA composite (4%) was transplanted into the chondral-injured area in the right knee of each pig. Using the same surgical process, an osteochondral defect (untreated) was created in the left knee as a control. The pigs were sacrificed 12 weeks after transplantation. Macroscopic and microscopic histologies, qRT-PCR, and immunostaining evaluated the degree of chondral degradation.
The HUCMSCs exhibited typical MSC characteristics, including spindle morphology, expression of surface markers (positive for CD29, CD4, CD73, CD90, and human leukocyte antigen [HLA]-ABC; negative for CD34, CD45, and HLA-DR), and multipotent differentiation (adipogenesis, osteogenesis, and chondrogenesis). More extensive proliferation of HUCMSCs was noted with 4% and 25% of HA than without HA. Expression of and in the HUCMSC-derived chondrocytes was increased when HA was included. The treated knees showed significant gross and histological improvements in hyaline cartilage regeneration when compared to the control knees. The International Cartilage Repair Society histological score was higher for the treated knees than the control knees.
Our findings suggest that cartilage regeneration using a mixture of HUCMSCs and HA in a large animal model may be an effective treatment for OA, and this study is a stepping stone toward the future clinical trials.
骨关节炎（OA）是一种关节退行性慢性疾病。间充质干细胞（MSCs）已被用于OA的软骨再生。我们研究了将人脐带间充质干细胞（HUCMSCs）与透明质酸（HA）水凝胶移植到猪OA临床前模型中的治疗潜力。
对HUCMSCs的形态、表面标志物和分化能力进行了表征。使用定量逆转录聚合酶链反应（qRT-PCR）检测HUCMSC-HA共培养中的基因表达。本大型动物研究使用了两只体重30-40 kg、年龄约4个月的健康雌性小型猪。在每只猪后膝关节的滑车沟处制造全层软骨损伤。3周后，制造第二个骨软骨缺损。然后，将1.5 mL的HUCMSC（5×10个细胞）和HA复合物（4%）移植到每只猪右膝的软骨损伤区域。使用相同的手术过程，在左膝制造一个骨软骨缺损（未治疗）作为对照。移植12周后处死猪。通过宏观和微观组织学、qRT-PCR和免疫染色评估软骨降解程度。
HUCMSCs表现出典型的MSC特征，包括纺锤形形态、表面标志物表达（CD29、CD4、CD73、CD90和人类白细胞抗原[HLA]-ABC呈阳性；CD34、CD45和HLA-DR呈阴性）以及多能分化（脂肪生成、骨生成和成软骨）。与无HA相比，4%和25%的HA使HUCMSCs的增殖更广泛。当加入HA时，HUCMSC来源的软骨细胞中 和 的表达增加。与对照膝相比，治疗膝在透明软骨再生方面显示出明显的大体和组织学改善。治疗膝的国际软骨修复协会组织学评分高于对照膝。
我们的研究结果表明，在大型动物模型中使用HUCMSCs和HA混合物进行软骨再生可能是OA的一种有效治疗方法，本研究是未来临床试验的一块垫脚石。
