Preclinical safety and efficacy of an allogeneic adipose-derived mesenchymal stem cell medicinal product for rotator...

Cell therapy with adipose-derived mesenchymal stem cells (ASCs) is a promising biological augmentation strategy for rotator cuff repair; however, before clinical translation, regulatory frameworks mandate rigorous preclinical characterization of safety. We report the complete preclinical program evaluating both the safety of an allogeneic ASC-based cell therapy medicinal product (CTMP) and its efficacy in a chronic rotator cuff tear model.

Phase I (safety, 164 immunodeficient nude BALB/c mice) assessed toxicity, biodistribution, and tumorigenicity after intra-articular and intravenous administration at doses ranging from 10× to 1,000× the projected clinical dose. Toxicity was evaluated by biochemical panels and full-body histopathology at 90 days. Biodistribution was quantified by fluorodeoxyglucose-positron emission tomography/computed tomography (days 45 and 90) and human-specific quantitative polymerase chain reaction (Alu-repeat) at 90 days. Tumorigenicity used an HT-29 positive-control model. Phase II (efficacy, 48 Sprague-Dawley rats) used a validated 4-week chronic supraspinatus tear-and-repair model; after repair, animals received ASCs in suspension (n = 24) or Hypothermosol vehicle (n = 24) and were assessed histologically (Åström-Rausing scale) and biomechanically (Instron 4,411) at 4 months.

In Phase I, no biochemical or histopathological toxicity attributable to the CTMP was detected at any dose or route. Three animals showed fluorodeoxyglucose-PET findings compatible with neoplasm but were histologically negative, confirming assay specificity. Quantitative polymerase chain reaction detected strong human DNA signals exclusively at the highest intravenous doses (≥500×), limited to the injection site (tail) and one heart sample; maximum retention was 1.75% at 90 days. After intra-articular administration at the clinical dose (10×), human ASCs persisted locally in 21% of animals at 90 days with no systemic dissemination. In Phase II, no statistically significant differences between groups were found in total histological score (median: 2 vs. 1; P = .811), any individual histological parameter, maximum load to failure (51.99 ± 9.1 N vs. 50.97 ± 9.1 N; P = .770), tendon displacement (5.33 ± 1.2 mm vs. 5.53 ± 1.4 mm; P = .852), or stiffness (17.28 ± 4.96 N/mm vs. 14.85 ± 4.01 N/mm; P = .669).

This allogeneic ASC-based CTMP demonstrated a favorable safety profile across all tested doses and administration routes, supporting its candidacy for clinical development. Efficacy in suspension delivery over a repaired chronic tear was not demonstrated; immune-mediated rejection and mechanical displacement represent additional explanatory factors alongside cell retention failure, and should guide the design of next-generation delivery strategies.