Ascorbic Acid and Iron Supplement Treatment Improves Stem Cell–Mediated Cartilage Regeneration in a Minipig Model

Abstract

Background: The transplantation of mesenchymal stem cells (MSCs) into cartilage defects has led to variable cartilage repair outcomes. Previous in vitro studies have shown that ascorbic acid and reduced iron independently can improve the chondrogenic differentiation of MSCs. However, the combined effect of ascorbic acid and iron supplementation on MSC differentiation has not been investigated. Purpose: To investigate the combined in vivo effects of ascorbic acid and a US Food and Drug Administration (FDA)–approved iron supplement on MSC-mediated cartilage repair in mature Göttingen minipigs. Study Design: Controlled laboratory study. Methods: We pretreated bone marrow–derived MSCs with ascorbic acid and the FDA-approved iron supplement ferumoxytol and then transplanted the MSCs into full-thickness cartilage defects in the distal femurs of Göttingen minipigs. Untreated cartilage defects served as negative controls. We evaluated the cartilage repair site with magnetic resonance imaging at 4 and 12 weeks after MSC implantation, followed by histological examination and immunofluorescence staining at 12 weeks. Results: Ascorbic acid plus iron–pretreated MSCs demonstrated a significantly better MOCART (magnetic resonance observation of cartilage repair tissue) score (73.8 ± 15.5), better macroscopic cartilage regeneration score according to the International Cartilage Repair Society (8.6 ± 2.0), better Pineda score (2.9 ± 0.8), and larger amount of collagen type II (28,469 ± 21,313) compared with untreated controls (41.3 ± 2.5, 1.8 ± 2.9, 12.8 ± 1.9, and 905 ± 1326, respectively). The obtained scores were also better than scores previously reported in the same animal model for MSC implants without ascorbic acid. Conclusion: Pretreatment of MSCs with ascorbic acid and an FDA-approved iron supplement improved the chondrogenesis of MSCs and led to hyaline-like cartilage regeneration in the knee joints of minipigs. Clinical Relevance: Ascorbic acid and iron supplements are immediately clinically applicable. Thus, these results, in principle, could be translated into clinical applications.

Updates on mesenchymal stem cell therapies for articular cartilage regeneration in large animal models

“…Porcine aMSCs seeded onto decellularized cartilage extracellular matrix (DCECM) performed similarly to chondrocytes regarding histological chondral defect regeneration ( Lu L. et al, 2021 ). Scaffold-free implantation of porcine bMSCs and aMSCs at 5–30 million cells per defect, respectively, also improved cartilage histological and MRI score compared to nontreatment ( Yamasaki et al, 2019 ; Theruvath et al, 2021 ). Likewise, autologous porcine synovial tissue-derived MSCs (sMSCs) aggregates, at 400,000 cells/defect, bolstered histology and macroscopic scores in femoral condyle after 12 weeks ( Kondo et al, 2019 ).…”

Magnetic Nanoparticles and Magnetic Field Exposure Enhances Chondrogenesis of Human Adipose Derived Mesenchymal Stem Cells But Not of Wharton Jelly Mesenchymal Stem Cells

“…Iron availability and supplementation has been recently confirmed as a modality to increase chondrogenesis in vitro and in vivo in Guineea pigs. Ascorbic acid and iron supplementation in the form of commercially available ferumoxytol was found to significantly increase chondrogenesis in bone marrow ADSC in vitro as well as in vivo in Guinea minipig model of cartilage defect ( Theruvath et al, 2021 ). Ferrumoxytol, is used as iron supplement for intravenous administration and occasionally off label as a MRI contrast agent for vascular and nodal metastasis.…”

Magnetic Nanomaterials: Advanced Tools for Cartilage Extracellular Matrix Regeneration

“…Bone marrow-derived MSCs pretreated with ascorbic acid and iron supplement ferumoxytol were shown to accelerate the repair of full-thickness cartilage defects in minipigs. This could be produced by reducing metal ions to a biologically active ferrous form, preventing free-radical mediated tissue damage, promoting proline and lysine hydroxylation, and supporting the formation of mature ECM collagen molecules 76 . MNM-induced discrete mechanical stimulation could potentially be used to increase chondrogenesis by MSCs.…”