In vitro and in vivo potentialities for cartilage repair from human advanced knee osteoarthritis synovial fluid-derived mesenchymal stem cells

Abstract

BackgroundMesenchymal stem cells (MSCs) are found in synovial fluid (SF) and can easily be harvested during arthrocentesis or arthroscopy. However, SF-MSC characterization and chondrogenicity in collagen sponges have been poorly documented as well as their hypothetical in vivo chondroprotective properties with intra-articular injections during experimental osteoarthritis (OA).MethodsSF-MSCs were isolated from human SF aspirates in patients suffering from advanced OA undergoing total knee joint replacements. SF-MSCs at passage 2 (P2) were characterized by flow cytometry for epitope profiling. SF-MSCs at P2 were subsequently cultured in vitro to assess their multilineage potentials. To assess their chondrogenicity, SF-MSCs at P4 were seeded in collagen sponges for 4 weeks under various oxygen tensions and growth factors combinations to estimate their gene profile and matrix production. Also, SF-MSCs were injected into the joints in a nude rat anterior cruciate ligament transection (ACLT) to macroscopically and histologically assess their possible chondroprotective properties,.ResultsWe characterized the stemness (CD73+, CD90+, CD105+, CD34−, CD45−) and demonstrated the multilineage potency of SF-MSCs in vitro. Furthermore, the chondrogenic induction (TGF-ß1 ± BMP-2) of these SF-MSCs in collagen sponges demonstrated a good capacity of chondrogenic gene induction and extracellular matrix synthesis. Surprisingly, hypoxia did not enhance matrix synthesis, although it boosted chondrogenic gene expression (ACAN, SOX9, COL2A1). Besides, intra-articular injections of xenogenic SF-MSCs did exert neither chondroprotection nor inflammation in ACLT-induced OA in the rat knee.ConclusionsAdvanced OA SF-MSCs seem better candidates for cell-based constructs conceived for cartilage defects rather than intra-articular injections for diffuse OA.

Combining Innovative Bioink and Low Cell Density for the Production of 3D-Bioprinted Cartilage Substitutes: A Pilot Study

“…In contrast, and as expected, ECM synthesis was more pronounced on D56 and the densitometry measurements after ECM staining confirmed the benefit of combining TGF-β1 and BMP-2 [75,[84][85][86][87]. As we previously reported in a study of synovial fluid MSCs, we observed no influence of BMP-2 alone on both gene and protein levels [37]. Unfortunately, the phenotype of MSCs in cartilage repair is unstable so that differentiation continues along the endochondral ossification pathway.…”

“…Five-micrometer sections were cut and stained using Immunohistochemistry analyses were performed with the LSAB®+ kit (HRP, Dako) using anti-type II collagen monoclonal antibodies (Labvision, France). Paraffin-embedded tissues (5 μm) were deparaffinized, treated with pepsin (0.4% w/v, Sigma) for 30 min at room temperature, and incubated with a hydrogen peroxide blocking solution for 5 min to block the endogenous peroxidases, as precisely described in our previous work [37]. The sections were counterstained with hematoxylin and mounted with resin.…”

“…For histology and immunohistochemistry, the tissues were imaged by using a DMD 108 optical microscope (Leica®, France) at a 4x magnification and evaluated with ImageJ software (U. S. National Institutes of Health, Bethesda, Maryland, USA). Transmission light images of Alcian blue staining and of collagen type II immunohistochemistry were recorded and evaluated by a semiquantitative method using the image analysis software ImageJ as we previously described [37]. Briefly, the transmitted light images were recorded and evaluated by a semiquantitative custom method to calculate the stained percentage area (Alcian blue stain and immunohistochemical markers of collagen type II).…”

Synovial Fluid Mesenchymal Stem Cells for Knee Arthritis and Cartilage Defects: A Review of the Literature

“…Despite many positive findings, results have not been consistent in the literature. Neybecker et al 48 used an anterior cruciate ligament transection (ACLT) model in a rat, followed by injection of human SF-MSC into the rat's knee joint. The results showed no significant differences in the articular lesions in the ACLT-induced OA in the nude rat knee following a single SF-MSC injection.…”

The Effect of Single Versus Multiple Intra-Articular Injection of Synovial Fluid Mesenchymal Stem Cells on Rat Temporomandibular Joint With Induced Arthritis. Biochemical and Histologic Analysis

“…Synovial fluid samples were diluted in expansion culture medium 1:6, then plated in Petri dishes. (21) MSCs were expanded in proliferation medium containing Dulbecco's modified Eagle's medium with low glucose (DMEM-LG, Gibco) with addition of 10% fetal bovine serum (FBS, Sigma), 1 ng/ml basic fibroblast growth factor (bFGF, Miltenyi Biotec), 1% glutamine (Gibco), and 1% penicillin-streptomycin (Gibco). Then they were cultured at 37°C with 5% humidified CO 2 .…”