Kutaish Halah, Tscholl Philippe Matthias, Cosset Erika, Bengtsson Laura, Braunersreuther Vincent, Mor Flavio Maurizio, Laedermann Jeremy, Furfaro Ivan, Stafylakis Dimitrios, Hannouche Didier, Gerstel Eric, Krause Karl-Heinz, Assal Mathieu, Menetrey Jacques, Tieng Vannary
Department of Pathology and Immunology, Medical School, University of Geneva, Geneva, Switzerland. University Medical Center, University of Geneva, Geneva, Switzerland. Foot and Ankle Surgery Centre, Centre Assal, Clinique La Colline, Hirslanden Geneva, Switzerland.
Investigation performed at the Faculty of Medicine, University of Geneva, in collaboration with Geneva University Hospitals, Geneva, Switzerland.
Am J Sports Med. 2023 Jan;51(1):237-249. doi: 10.1177/03635465221138099.
Chondrocyte-based cell therapy to repair cartilage has been used for >25 years despite current limitations. This work presents a new treatment option for cartilage lesions.
High-quality hyaline cartilage microtissues called Cartibeads are capable of treating focal chondral lesions once implanted in the defect, by complete fusion of Cartibeads among themselves and their integration with the surrounding native cartilage and subchondral bone.
Controlled laboratory study.
Cartibeads were first produced from human donors and characterized using histology (safranin O staining of glycosaminoglycan [GAG] and immunohistochemistry of collagen I and II) and GAG dosage. Cartibeads from 6 Göttingen minipigs were engineered and implanted in an autologous condition in the knee (4 or 5 lesions per knee). One group was followed up for 3 months and the other for 6 months. Feasibility and efficacy were measured using histological analysis and macroscopic and microscopic scores.
Cartibeads revealed hyaline features with strong staining of GAG and collagen II. High GAG content was obtained: 24.6-µg/mg tissue (wet weight), 15.52-µg/mg tissue (dry weight), and 35 ± 3-µg GAG/bead (mean ± SD). Histological analysis of Göttingen minipigs showed good integration of Cartibeads grafts at 3 and 6 months after implantation. The Bern Score of the histological assay comparing grafted versus empty lesions was significant at 3 months (grafted, n = 10; nongrafted, n = 4; score, 3.3 and 5.3, respectively) and 6 months (grafted, n = 11; nongrafted, n = 3; score, 1.6 and 5.1).
We developed an innovative 3-step method allowing, for the first time, the use of fully dedifferentiated adult chondrocytes with a high number of cell passage (owing to the extensive amplification in culture). Cartibeads engineered from chondrocytes hold potential as an advanced therapy medicinal product for treating cartilage lesions with established efficacy.
This successful preclinical study, combined with standardized manufacturing of Cartibeads according to good manufacturing practice guidelines, led to the approval of first-in-human clinical trial by the ethics committee and local medical authority. The generated data highlighted a promising therapy to treat cartilage lesions from a small amount of starting biopsy specimen. With our innovative cell amplification technology, very large lesions can be treated, and older active patients can benefit from it.
尽管存在当前的局限性，但基于软骨细胞的细胞疗法修复软骨已应用超过25年。这项工作提出了一种治疗软骨损伤的新选择。
一种称为软骨珠（Cartibeads）的高质量透明软骨微组织一旦植入缺损处，通过软骨珠之间的完全融合以及它们与周围天然软骨和软骨下骨的整合，能够治疗局灶性软骨损伤。
对照实验室研究。
首先从人类供体中制备软骨珠，并使用组织学（糖胺聚糖[GAG]的番红O染色以及I型和II型胶原的免疫组织化学）和GAG定量分析对其进行表征。对6只哥廷根小型猪的软骨珠进行工程化处理，并在自体条件下植入膝关节（每只膝关节植入4或5个损伤处）。一组随访3个月，另一组随访6个月。使用组织学分析以及宏观和微观评分来评估可行性和疗效。
软骨珠显示出透明软骨特征，GAG和II型胶原染色强烈。获得了高GAG含量：24.6μg/mg组织（湿重）、15.52μg/mg组织（干重）以及35±3μg GAG/珠（平均值±标准差）。对哥廷根小型猪的组织学分析表明，植入后3个月和6个月时软骨珠移植物整合良好。组织学检测中比较移植损伤与未移植损伤的伯尔尼评分在3个月时具有显著差异（移植组，n = 10；未移植组，n = 4；评分分别为3.3和5.3），在6个月时也具有显著差异（移植组，n = 11；未移植组，n = 3；评分分别为1.6和5.1）。
我们开发了一种创新的三步法，首次允许使用经过大量传代（由于在培养中大量扩增）的完全去分化的成年软骨细胞。由软骨细胞工程化制备的软骨珠作为一种治疗软骨损伤且疗效已确定的先进治疗用医药产品具有潜力。
这项成功的临床前研究，结合按照良好生产规范指南对软骨珠进行标准化生产，使得伦理委员会和当地医疗当局批准了首例人体临床试验。所产生的数据突出了一种从少量起始活检标本治疗软骨损伤的有前景的疗法。借助我们创新的细胞扩增技术，可以治疗非常大的损伤，并且老年活跃患者也能从中受益。
