用GMP生产过程控制动力学指标预测ACI结局：初始软骨细胞产量和群体倍增时间作为预后临床生物标志物
简介
本前瞻性队列研究纳入67例接受第二代自体软骨细胞移植（ACI）治疗大面积膝关节软骨缺损的患者，评估GMP工艺中初始软骨细胞产量及群体倍增时间等动力学指标对预后预测价值。结果发现，较高的初始细胞产量和较短的倍增时间显著预测KOOS及IKDC评分的中期改善，而静态基因表达和融合度无预测作用。提示ACI质量控制应侧重细胞增殖动力学，以筛选适合的患者并优化再生治疗结…
英文摘要
PURPOSE: To determine whether specific in vitro biologic characteristics and manufacturing kinetics of cultured human articular chondrocytes (HACs) can serve as predictive biomarkers for patient-reported and structural outcomes following second-generation autologous chondrocyte implantation (ACI) in the knee. MATERIALS AND METHODS: This prospective cohort study evaluated 67 patients (mean age 25.1 ± 8.3 years) treated with second-generation ACI for large focal cartilage defects (mean size 5.4 ± 2.5 cm2) between 2017 and 2024. HACs were expanded under Good Manufacturing Practice (GMP) conditions using human platelet lysate-supplemented media. Four biological determinants were analyzed: (1) initial chondrocyte yield (ICY) isolated from the cartilage biopsy; (2) chondrogenic activity (relative ACAN and COL2A1 expression); (3) final culture confluence level; and (4) HAC population doubling time (PDT) pre- and post-cryopreservation. Clinical outcomes (KOOS and IKDC) and MRI outcomes (MOCART) were assessed at 2 years (T1) and at a mean final follow-up of 4.2 ± 1.7 years (T2) via univariate and multiple regression analyses. RESULTS: Static biosynthetic markers (ACAN/COL2A1 expression) and final culture confluence did not significantly correlate with longitudinal clinical (KOOS/IKDC) or structural (MOCART) outcomes. Conversely, the retained kinetic parameters served as strong prognostic indicators. Multiple regression revealed that higher ICY values significantly predicted improvements across most KOOS subscales from baseline to T2, including KOOS Symptoms (β = 63.56; p < 0.01), KOOS Pain (β = 88.83; p < 0.001), KOOS ADL (β = 107.84; p < 0.001), and KOOS Sport and Recreation Function (β = 92.3; p < 0.05). Furthermore, a prolonged PDT, indicative of diminished in vitro proliferative vigor, inversely correlated with functional recovery in IKDC (β = -26.12; p < 0.01), KOOS Pain (β = -23.09; p < 0.05), and KOOS ADL (β = -21.71; p < 0.05) scores. CONCLUSIONS: The intrinsic proliferative vigor of the HAC cellular payload (shorter PDT) and higher initial cell yields are robust kinetic biomarkers associated with markedly superior mid-term clinical outcomes following second-generation ACI for large focal chondral defects in the knee. In contrast, standard morphological and gene expression metrics failed to predict in vivo functional success. These findings advocate for a risk-based paradigm shift in GMP quality control methodologies, emphasizing dynamic HAC growth kinetics over static cellular features to optimize patient-specific regenerative potential.