Fu Qiwei, Zhou Rong, Cao Jia, Chen Yi, Zhu Jun, Zhou Yiqin, Shao Jiahua, Xin Wei, Yuan Shuai
Department of Orthopedics, Shanghai Changzheng Hospital, Naval Medical University, Shanghai, China.
Department of Orthopedics, 72nd Group Army Hospital of the PLA, Huzhou City, China.
Front Bioeng Biotechnol. 2022 Aug 17;10:889306. doi: 10.3389/fbioe.2022.889306. eCollection 2022.
The aim of the study was to evaluate the advantages of without enzyme isolating patellar fat pad-derived mesenchymal stem cells (IPFP-SCs) and the feasibility of cartilage repair. The IPFP-SCs were isolated using the without enzyme method and compared with the IPFP-SCs obtained by the traditional enzyme digestion method in terms of cell proliferation ability, characterization, and differentiation ability, and the differences in chondrogenic induction and differentiation between the two groups were compared. Twenty-four New Zealand rabbits were randomly divided into four groups ( = 6). After the articular cartilage defects were modeled, different preparations were injected into the joint cavity. The rabbits in the group A were injected with the mixture of IPFP-SCs and pure PRP (P-PRP), separated using the without enzyme method, while those in the group B were injected with the mixture of IPFP-SCs and P-PRP separated with the digestion method, while those in the group C were injected with SVF separated using the without enzyme method, and those in the group D were injected with normal saline. At 6 weeks and 12 weeks after operation, the cartilage repair of rabbit joint specimens was observed and evaluated by gross observation and histological staining, and the effects of different IPFP-SCs application forms in repairing cartilage defects were compared. The time required to obtain IPFP-SCs by enzyme-free isolation was significantly less than that by enzyme digestion, while the acquisition rate of primary cells was significantly lower than that by enzyme digestion. After culture and amplification, the two IPFP-SCs from different sources did not show significant differences in cell proliferation, cell phenotype, and differentiation ability. In animal experiments, groups A and B had the best effect on the repair of cartilage defects, and there was no significant difference between the two groups. The repair effect in group C was weaker than that in the former two groups, but it was relatively better than that in group D. It is more time-saving to obtain IPFP-SCs by the without enzyme method than by enzymatic digestion, and there is no significant difference in cell identification and differentiation potential between the two methods. However, the rate of obtaining primary cells was significantly lower than that with the enzyme digestion method. IPFP-SCs showed good repair effect in the rabbit animal cartilage defect model, providing ideas and reference for the clinical application of stem cells in repairing articular cartilage.
本研究旨在评估无酶法分离髌下脂肪垫来源间充质干细胞（IPFP-SCs）的优势及软骨修复的可行性。采用无酶法分离IPFP-SCs，并将其与传统酶消化法获得的IPFP-SCs在细胞增殖能力、特性及分化能力方面进行比较，同时比较两组在软骨诱导分化方面的差异。将24只新西兰兔随机分为4组（每组n = 6）。构建关节软骨缺损模型后，向关节腔内注入不同制剂。A组注射无酶法分离的IPFP-SCs与纯PRP（P-PRP）的混合物，B组注射酶消化法分离的IPFP-SCs与P-PRP的混合物，C组注射无酶法分离的SVF，D组注射生理盐水。术后6周和12周，通过大体观察和组织学染色观察并评估兔关节标本的软骨修复情况，比较不同IPFP-SCs应用形式对软骨缺损修复的效果。无酶分离法获取IPFP-SCs所需时间明显少于酶消化法，而原代细胞获取率明显低于酶消化法。培养扩增后，两种不同来源的IPFP-SCs在细胞增殖、细胞表型及分化能力方面无显著差异。在动物实验中，A组和B组对软骨缺损修复效果最佳，两组间无显著差异。C组修复效果弱于前两组，但相对D组较好。无酶法获取IPFP-SCs比酶消化法更省时，两种方法在细胞鉴定及分化潜能方面无显著差异。然而，原代细胞获取率明显低于酶消化法。IPFP-SCs在兔动物软骨缺损模型中显示出良好的修复效果，为干细胞临床应用修复关节软骨提供了思路和参考。
