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An Off-the-Shelf Tissue Engineered Cartilage Composed of Optimally Sized Pellets of Cartilage Progenitor/Stem Cells
ACS Biomaterials Science & Engineering ( IF 5.4 ) Pub Date : 2020-09-15 , DOI: 10.1021/acsbiomaterials.9b01863 Jisheng Ran 1 , Yang Fei 1 , Canlong Wang 2 , Dengfeng Ruan 1 , Yejun Hu 1, 3 , Zefeng Zheng 4 , Xiao Chen 3 , Zi Yin 3 , Chenqi Tang 1 , Yangwu Chen 1 , Jiayun Huang 1 , Lingfang Shen 5 , Lidong Wu 1 , Boon Chin Heng 6 , Dominique Pioletti 7 , Weiliang Shen 1, 3 , Hongwei Ouyang 3
ACS Biomaterials Science & Engineering ( IF 5.4 ) Pub Date : 2020-09-15 , DOI: 10.1021/acsbiomaterials.9b01863 Jisheng Ran 1 , Yang Fei 1 , Canlong Wang 2 , Dengfeng Ruan 1 , Yejun Hu 1, 3 , Zefeng Zheng 4 , Xiao Chen 3 , Zi Yin 3 , Chenqi Tang 1 , Yangwu Chen 1 , Jiayun Huang 1 , Lingfang Shen 5 , Lidong Wu 1 , Boon Chin Heng 6 , Dominique Pioletti 7 , Weiliang Shen 1, 3 , Hongwei Ouyang 3
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Articular cartilage focal lesion remains an intractable challenge in sports medicine, and autologous chondrocytes’ implantation (ACI) is one of the most commonly utilized treatment modality for this ailment. However, the current ACI technique requires two surgical steps which increases patients’ morbidity and incurs additional medical costs. In the present study, we developed a one-step cryopreserved off-the-shelf ACI tissue-engineered (TE) cartilage by seeding pellets of spheroidal cartilage stem/progenitor cells (CSPCs) on a silk scaffold. The pellets were developed through a hanging-drop method, and the incubation time of 1 day could efficiently produce spheroidal pellets without any adverse influence on the cell activity. The pellet size was also optimized. Under chondrogenic induction, pellets consisting of 40 000 CSPCs were found to exhibit the most abundant cartilage matrix deposition and the highest mRNA expression levels of SOX9, aggrecan, and COL2A1, as compared with pellets consisting of 10 000, 100 000, or 200 000 CSPCs. Scaffolds seeded with CSPCs pellets containing 40 000 cells could be preserved in liquid nitrogen with the viability, migration, and chondrogenic ability remaining unaffected for as long as 3 months. When implanted in a rat trochlear cartilage defect model for 3 months, the ready-to-use, cryopreserved TE cartilage yielded fully cartilage reconstruction, which was comparable with the uncryopreserved control. Hence, our study provided preliminary data that our off-the-shell TE cartilage with optimally sized CSPCs pellets seeded within silk scaffolds exhibited strong cartilage repair capacity, which provided a convenient and promising one-step surgical approach to ACI.
中文翻译：
由软骨祖细胞/干细胞最佳大小的小球组成的现成的组织工程化软骨。
关节软骨局灶性病变在运动医学中仍然是一个棘手的挑战，自体软骨细胞的植入（ACI）是该疾病最常用的治疗方式之一。然而，当前的ACI技术需要两个手术步骤，这增加了患者的发病率并招致了额外的医疗费用。在本研究中，我们通过在球形支架上接种球形软骨干/祖细胞（CSPC）的颗粒，一步一步冷冻保存了现成的ACI组织工程（TE）软骨。沉淀物是通过悬滴法显影的，孵育1天的时间可以有效地生产球形沉淀物，而对细胞活性没有任何不利影响。颗粒尺寸也被优化。在软骨诱导下，与由1万，10万或20万个CSPC组成的团块相比，由40000个CSPC组成的团块表现出最丰富的软骨基质沉积和最高的SOX9，聚集蛋白聚糖和COL2A1 mRNA表达水平。用CSPC沉淀的含有40 000个细胞的颗粒的支架可以保存在液氮中，其存活力，迁移和软骨形成能力可以保持长达3个月的时间。当在大鼠滑车软骨缺损模型中植入3个月后，即用型，冷冻保存的TE软骨可完全重建软骨，这与未冷冻保存的对照组相当。因此，我们的研究提供了初步的数据，即我们的带有合适尺寸CSPC颗粒的丝状脚手架内的壳外TE软骨具有强大的软骨修复能力，
更新日期：2020-09-15
中文翻译：
由软骨祖细胞/干细胞最佳大小的小球组成的现成的组织工程化软骨。
关节软骨局灶性病变在运动医学中仍然是一个棘手的挑战，自体软骨细胞的植入（ACI）是该疾病最常用的治疗方式之一。然而，当前的ACI技术需要两个手术步骤，这增加了患者的发病率并招致了额外的医疗费用。在本研究中，我们通过在球形支架上接种球形软骨干/祖细胞（CSPC）的颗粒，一步一步冷冻保存了现成的ACI组织工程（TE）软骨。沉淀物是通过悬滴法显影的，孵育1天的时间可以有效地生产球形沉淀物，而对细胞活性没有任何不利影响。颗粒尺寸也被优化。在软骨诱导下，与由1万，10万或20万个CSPC组成的团块相比，由40000个CSPC组成的团块表现出最丰富的软骨基质沉积和最高的SOX9，聚集蛋白聚糖和COL2A1 mRNA表达水平。用CSPC沉淀的含有40 000个细胞的颗粒的支架可以保存在液氮中，其存活力，迁移和软骨形成能力可以保持长达3个月的时间。当在大鼠滑车软骨缺损模型中植入3个月后，即用型，冷冻保存的TE软骨可完全重建软骨，这与未冷冻保存的对照组相当。因此，我们的研究提供了初步的数据，即我们的带有合适尺寸CSPC颗粒的丝状脚手架内的壳外TE软骨具有强大的软骨修复能力，
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