Allogeneic Mesenchymal Stem Cells in Combination with Hyaluronic Acid for the Treatment of Osteoarthritis in Rabbits

En-Rung Chiang, Hsiao-Li Ma, Jung-Pan Wang, Chien-Lin Liu, Tain-Hsiung Chen, Shih-Chieh Hung, En-Rung Chiang, Hsiao-Li Ma, Jung-Pan Wang, Chien-Lin Liu, Tain-Hsiung Chen, Shih-Chieh Hung

Abstract

Mesenchymal stem cell (MSC)-based therapies may aid in the repair of articular cartilage defects. The purpose of this study was to investigate the effects of intraarticular injection of allogeneic MSCs in an in vivo anterior cruciate ligament transection (ACLT) model of osteoarthritis in rabbits. Allogeneic bone marrow-derived MSCs were isolated and cultured under hypoxia (1% O2). After 8 weeks following ACLT, MSCs suspended in hyaluronic acid (HA) were injected into the knees, and the contralateral knees were injected with HA alone. Additional controls consisted of a sham operation group as well as an untreated osteoarthritis group. The tissues were analyzed by macroscopic examination as well as histologic and immunohistochemical methods at 6 and 12 weeks post-transplantation. At 6 and 12 weeks, the joint surface showed less cartilage loss and surface abrasion after MSC injection as compared to the tissues receiving HA injection alone. Significantly better histological scores and cartilage content were observed with the MSC transplantation. Furthermore, engraftment of allogenic MSCs were evident in surface cartilage. Thus, injection of the allogeneic MSCs reduced the progression of osteoarthritis in vivo.

Conflict of interest statement

Competing Interests: The authors have declared that no competing interests exist.

Figures

Fig 1. Study design.
Fig 1. Study design.
Evaluation of the effect of hypoxia-cultured MSCs for treatment of osteoarthritis in rabbits. The numbers in parentheses indicate number of rabbits receiving ACLT and analysis. CTR: control group; H&E: haematoxylin and eosin; S.O: safranin O; P.B: Prussian blue.
Fig 2. Macroscopic examination of the effects…
Fig 2. Macroscopic examination of the effects of hypoxia-cultured MSCs on osteoarthritis progression.
(A) Femur condyles 6 and 12 weeks post-treatment. Asterisks indicate osteophyte formation. (B) India ink staining for articular surface of tibial plateaus.
Fig 3. Histologic analysis.
Fig 3. Histologic analysis.
(A, B) Femur condyles of animals at 6 and 12 weeks post-treatment were assessed by (A) haematoxylin and eosin and (B) Safranin-O staining. (C) Quantitative analysis of safranin-O staining. Data are mean ± standard deviation. *P <0.05.
Fig 4. Histological grading of the effect…
Fig 4. Histological grading of the effect of hypoxia-cultured MSCs.
Mankin scores at different part of the joint at 6 and 12 weeks post-treatment. Data are mean ± standard deviation. *P

Fig 5. Immunohistochemistry analysis of type II…

Fig 5. Immunohistochemistry analysis of type II collagen.

(A) Femur condyles stained with type II…
Fig 5. Immunohistochemistry analysis of type II collagen.
(A) Femur condyles stained with type II collagen (Magnification, ×100). (B) Quantitative analysis of type II collagen. Data are mean ± standard deviation. *P <0.05.

Fig 6. Immunohistochemistry analysis of type X…

Fig 6. Immunohistochemistry analysis of type X collagen.

(A) Microscopic appearance of femur condyles stained…
Fig 6. Immunohistochemistry analysis of type X collagen.
(A) Microscopic appearance of femur condyles stained with type X collagen (Magnification, ×100; Upper box, ×400). (B) Quantitative analysis of type X collagen. Data are mean ± standard deviation. *P <0.05.

Fig 7. Identification and localization of SPIO-labeled…

Fig 7. Identification and localization of SPIO-labeled MSCs.

(A) Endocytosis of SPIO nanoparticles by MSCs…
Fig 7. Identification and localization of SPIO-labeled MSCs.
(A) Endocytosis of SPIO nanoparticles by MSCs visualized by Prussian blue staining (bar = 100 μm). (B-D) Engraftment of injected MSCs into cartilage of the (B) femoral condyle (Magnification, ×100), (C) tibial plateau (Magnification, ×100) and (D) meniscus (Magnification, ×50) by Prussian blue and Safranin-O staining. (Lower boxes, (B, C) ×200, (D) ×100).
All figures (7)
Fig 5. Immunohistochemistry analysis of type II…
Fig 5. Immunohistochemistry analysis of type II collagen.
(A) Femur condyles stained with type II collagen (Magnification, ×100). (B) Quantitative analysis of type II collagen. Data are mean ± standard deviation. *P <0.05.
Fig 6. Immunohistochemistry analysis of type X…
Fig 6. Immunohistochemistry analysis of type X collagen.
(A) Microscopic appearance of femur condyles stained with type X collagen (Magnification, ×100; Upper box, ×400). (B) Quantitative analysis of type X collagen. Data are mean ± standard deviation. *P <0.05.
Fig 7. Identification and localization of SPIO-labeled…
Fig 7. Identification and localization of SPIO-labeled MSCs.
(A) Endocytosis of SPIO nanoparticles by MSCs visualized by Prussian blue staining (bar = 100 μm). (B-D) Engraftment of injected MSCs into cartilage of the (B) femoral condyle (Magnification, ×100), (C) tibial plateau (Magnification, ×100) and (D) meniscus (Magnification, ×50) by Prussian blue and Safranin-O staining. (Lower boxes, (B, C) ×200, (D) ×100).

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