Summary

Current management of low-grade osteoarthritis (OA) has not been satisfying. Our study is the first study worldwide that evaluated the combined exosome derived adipose mesenchymal stem cell (Exos-AdMSC) and HA injection in large animal OA model with clinical, radiological, macroscopical, microscopical evaluation and its mechanism in the molecular level.

Abstract

Introduction

Cartilage has a poor regenerative potency hence current treatment of osteoarthritis (OA) mainly focused on treating and preventing inflammatory symptoms without the capability to restore the destroyed cartilage. Mesenchymal stem cell (MSC) which showed cartilage regenerative potency has many drawbacks including limited homing cells, long and intricate production process, difficulties in storage and distribution requiring very low temperature and several ethical issues. Exosome is cell-free nano-sized molecule that contain genetic materials that could enhance cartilage regeneration potency. Our study aimed to evaluate the combined effect of exosome-derived-adipose mesenchymal stem cell (Exos-AdMSC) in low grade OA model.

Methods

Eighteen skeletally mature Ovies aries sheep were performed total lateral meniscectomy on the right hind limb and performed conventional radiograph after 6 weeks. Low grade OA were confirmed by marginal osteophyte formation and the sheep were divided into 3 groups. Exos group received 3 dose of 1 ml Exos-AdMSC intra-articularly (IA), HA group received 2 dose of 1 ml hyaluronic acid (HA), and combi group receives the combination of exos and HA group. Clinical lameness (CLS) and radiological score were evaluated before, and 1st, 2nd, 3rd months after meniscectomy. After all sheep were euthanized, T2 map values by magnetic resonance imaging (MRI), macroscopic, microscopic and biomolecular evaluation using qualitative reverse transcriptase-polymerase chain reaction (qRT-PCR) along with micro-RNA (miRNA) analysis using microarray evaluation were performed. Bioinformatic analyses were done using KEGG-pathway application to evaluate cartilage regeneration pathway.

Results

Combi group showed lower CLS on the 2nd month after meniscectomy (p = 0.048), lower macroscopic OARSI on distal femur and tibia surface, and microscopic OARSI compared to HA group significantly while conventional radiographic evaluation showed no significant difference. Evaluation of T2 map values by MRI showed lowest on the combi group at the lateral compartment compared to all group albeit non-significant. Biomolecular evaluation by qRT-PCR showed significantly higher WNT-5A expression on the combi group vs exos group and microarray evaluation showed increased miR-140-3p, miR-27b-3p, miR-23a-3p and decreased miR-485-5p, miR-218-5p, miR-31-5p significantly in the combi vs exos group which affect increased USP34 gene expression, decreased ASAP1, EBF3, TAOK1, PPP1CB and CDK6 expression that influence the increased of cellular proliferation and apoptosis inhibition pathways.

Conclusion

Administration of IA combined exos-AdMSC and HA on low grade OA model could increase cartilage regeneration via upregulated miR-140-3p, miR-27b-3p, miR-23a-3p and downregulated miR-485-5p, miR-218-5p, miR-31-5p and increase cellular proliferation and apoptosis inhibition.


Keywords: exosome-adipose derived-mesenchymal stem cell (exos-AdMSC), hyaluronic acid (HA), low grade osteoarthritis (OA), sheep model, micro-RNA (miRNA), cartilage regeneration