Repair of Large Animal Partial-Thickness Cartilage Defects Through Intraarticular Injection of Matrix-Rejuvenated Synovium-Derived Stem Cells

Abstract

Cartilage defects have a limited ability to self-heal. Stem cell treatment is a promising approach; however, replicative senescence is a challenge to acquiring large-quantity and high-quality stem cells for cartilage regeneration. Synovium-derived stem cells (SDSCs) are a tissue-specific stem cell for cartilage regeneration. Our recent findings suggest that decellularized stem cell matrix (DSCM) can rejuvenate expanded SDSCs in cell proliferation and chondrogenic potential. In this study, we were investigating (1) whether transforming growth factor (TGF)-β1 and TGF-β3 played a similar role in chondrogenic induction of SDSCs after expansion on either DSCM or plastic flasks (plastic), and (2) whether DSCM-expanded SDSCs had an enhanced capacity in repairing partial-thickness cartilage defects in a minipig model. SDSCs were isolated from synovium in two 3-month-old pigs and DSCM was prepared using SDSCs. Passage 2 SDSCs were expanded on either DSCM or plastic for one passage. The expanded cells were evaluated for cell morphology, chondrogenic capacity, and related mechanisms. TGF-β1 and TGF-β3 were compared for their role in chondrogenesis of SDSCs after expansion on either DSCM or plastic. The chondrogenic induction medium without TGF-β served as a control. In 13 minipigs, we intraarticularly injected DSCM- or plastic-expanded SDSCs or saline into knee partial-thickness cartilage defects and assessed their repair using histology and immunohistochemistry. We found DSCM-expanded SDSCs were small, had a fibroblast-like shape, and grew quickly in a three-dimensional format with concomitant up-regulation of phosphocyclin D1 and TGF-β receptor II. Plastic-expanded SDSCs exhibited higher mRNA levels of chondrogenic markers when incubated with TGF-β3, while DSCM-expanded SDSCs displayed comparable chondrogenic potential when treated with either TGF-β isotype. In the minipig model, DSCM-expanded SDSCs were better than plastic-expanded SDSCs in enhancing collagen II and sulfated glycosaminoglycan expression in repair of partial-thickness cartilage defects, but both groups were superior to the saline control group. Our observations suggested that DSCM is a promising cell expansion system that can promote cell proliferation and enhance expanded cell chondrogenic potential in vitro and in vivo. Our approach could lead to a tissue-specific cell expansion system providing large-quantity and high-quality stem cells for the treatment of cartilage defects.

Anti-Inflammatory Strategies in Cartilage Repair

“…12 A study in mini pigs successfully proved its resurfacing effect on partial-thickness cartilage defects after intra-articular injection of DSCM-expanded allogeneic SDSCs. 13 In this review article, the strategies using pharmacologic drugs, biomechanical stimulation, tissue-specific stem cells, and ECMbased scaffolds are summarized. An emerging strategy using a DSCM preconditioning approach to rejuvenate tissuespecific stem cells in both cell amount and chondrogenic potential is emphasized for its vital role in cartilage repair and anti-inflammation (Fig.…”

Cartilage repair techniques in the knee: stem cell therapies

“…Jung showed that enhancement of cartilage repair with the use of collagen type I/III membrane significantly improves the repair tissue quality in minipigs [37]. In addition, use of allogenic MSC and xenoimplantation of human umbilical cord bloodderived MSC were tested in this animal model, showing induction of cartilage healing by those cell types [42,44]. Efficacy of MSC/scaffold constructs have been shown in other large animals such as sheep, horse, and goats [45][46][47][48][49].…”

“…With the intent of evaluation with models that is more relevant to clinical condition in humans, large animals such as pig and sheep have been used as experimental subjects [37][38][39][40][41][42][43][44][45][46][47][48][49][50][51][52]. Among those, a (mini)pig model has been most frequently employed in previous studies [37][38][39][40][41][42][43][44].…”

“…Among those, a (mini)pig model has been most frequently employed in previous studies [37][38][39][40][41][42][43][44]. Various cell types and materials were tested in the studies.…”

Current Applications of Mesenchymal Stem Cells for Cartilage Tissue Engineering

“…Pei et al found that in synovium-derived stem cells (SDSCs), TGF-β3 enhancing collagen II and sulfated glycosaminoglycan minimize the expression of collagen I in the repair of partial-thickness cartilage defects in porcine SDSC pellets compared with TGF-β1 [120].…”