Improving the immunosuppressive potential of articular chondroprogenitors in a three-dimensional culture setting

Abstract

Cartilage repair in osteoarthritic patients remains a challenge. Identifying resident or donor stem/progenitor cell populations is crucial for augmenting the low intrinsic repair potential of hyaline cartilage. Furthermore, mediating the interaction between these cells and the local immunogenic environment is thought to be critical for long term repair and regeneration. In this study we propose articular cartilage progenitor/stem cells (CPSC) as a valid alternative to bone marrow-derived mesenchymal stem cells (BMMSC) for cartilage repair strategies after trauma. Similar to BMMSC, CPSC isolated from osteoarthritic patients express stem cell markers and have chondrogenic, osteogenic, and adipogenic differentiation ability. In an in vitro 2D setting, CPSC show higher expression of SPP1 and LEP, markers of osteogenic and adipogenic differentiation, respectively. CPSC also display a higher commitment toward chondrogenesis as demonstrated by a higher expression of ACAN. BMMSC and CPSC were cultured in vitro using a previously established collagen-chondroitin sulfate 3D scaffold. The scaffold mimics the cartilage niche, allowing both cell populations to maintain their stem cell features and improve their immunosuppressive potential, demonstrated by the inhibition of activated PBMC proliferation in a co-culture setting. As a result, this study suggests articular cartilage derived-CPSC can be used as a novel tool for cellular and acellular regenerative medicine approaches for osteoarthritis (OA). In addition, the benefit of utilizing a biomimetic acellular scaffold as an advanced 3D culture system to more accurately mimic the physiological environment is demonstrated.

Strategies for Articular Cartilage Repair and Regeneration

“…A new population of cells termed as cartilage-derived stem/progenitor cells (CPSCs) have been identified in the cartilage, which unlike the chondrocytes, may have the ability to self-renew (Jiang and Tuan, 2015). The application potential of CPSCs is still unclear, but research is ongoing to better understand this cell phenotype and its therapeutic prospects for AC repair (Jiang et al, 2016;Bauza et al, 2020).…”

3D Printed Scaffold Based on Type I Collagen/PLGA_TGF-β1 Nanoparticles Mimicking the Growth Factor Footprint of Human Bone Tissue

“…In addition, to avoid the premature release of incorporated biomolecules and GFs while providing a more targeted and sustained release to support bone regeneration, these NPs can be further integrated into biocompatible and biodegradable polymeric-based scaffolds [ 45 , 46 ]. These structures may be designed using either natural polymers such as alginate [ 47 , 48 , 49 , 50 ], collagen [ 17 , 51 , 52 , 53 , 54 , 55 , 56 ], gelatin [ 57 , 58 , 59 , 60 ], and fibrin [ 61 , 62 , 63 ], or synthetic polymers like polycaprolactone (PCL) [ 64 , 65 ] and polyurethanes (PU) [ 66 , 67 , 68 ]. Compared to synthetic materials, naturally derived polymers generally show superior biocompatibility and bioactivity although they lack mechanical strength and appropriate degradation rates.…”

Immunosuppressive potential evaluation of synovial fluid mesenchymal stem cells grown on 3D scaffolds as an alternative source of MSCs for osteoarthritis cartilage studies

“…4 ml of synovial fluid were washed three times with a solution of phosphate-buffered saline (PBS) 1:1 and then the whole extract was seeded in culture media (α-MEM) supplemented with 20% fetal bovine serum (FBS) and 1% penicillin (100 UI/ml)-streptomycin (100 mg/ml). Human bone marrow aspirate from two patients undergoing a total knee replacement surgery, was also obtained from the orthopedic biorepository at Houston Methodist Hospital Orthopedics and Sports Medicine Department (IRB CR00006624) and processed as previously reported (Bauza et al, 2020). Cells obtained after this process (p0) once at confluence were characterized for the expression of MSC-associated markers by Flow analysis (as describe bellow), expanded and cultured in α-MEM (Sigma-Aldrich, St. Louis, MO, United States) containing 20% (vol/ vol) FBS supplemented with 1% penicillin (100 UI/ml)streptomycin (100 mg/ml) (Gibco) and incubated at 37 °C in low oxygen conditions Media was changed every 2 days until the cells were at 80% confluency, at which point they were passaged.…”

“…To ameliorate in vitro studies relevance in studying both degeneration and inflammation mechanisms, threedimensional (3D) culture systems were explored to investigate chondrogenesis and other processes related to OA progression in vitro, involving multiple cell types, including stem cells, stromal cells, macrophages, and other chondrogenic progenitors (Khurshid et al, 2018;Santos et al, 2018;Evans and Fiederlein 2020). To this aim, implementation of 3D culture systems with biomimetic acellular scaffold to accurately simulate cartilage physiological environment led to significant improvements in cartilage progenitor/stem cells immunomodulatory effects (Bauza et al, 2020). Combining cell-based therapy with 3D scaffolds chondrogenic and osteogenic potential those hybrid systems offer great potential as in vitro systems to model OA (Samvelyan et al, 2021).…”