Comparison of two methods for preparing knee osteochondral injury models in mice_Chinese Journal of Reparative and Reconstructive Surgery

Authors：

LIU Huan ^1,2 , DING Qirui ¹ , MA Cheng ¹ , QIN Haonan ¹ , WEI Yifan ¹ ,  REN Yongxin ¹

1. Department of Orthopedics, the First Affiliated Hospital of Nanjing Medical University, Nanjing Jiangsu, 210000, P.R.China;
2. Department of Orthopedics, the Affiliated Huaian No.1 Hospital of Nanjing Medical University, Huaian Jiangsu, 223300, P.R.China;

Corresponding author：

REN Yongxin, Email: renyongxinjsph@126.com

Keywords：

Osteochondral injury; animal model; tungsten drill; mouse

DOI：

10.7507/1002-1892.202101098

Video：

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Objective To observe the effect of using tungsten drills to prepare mouse knee osteochondral injury model by comparing with the needle modeling method, in order to provide an appropriate animal modeling method for osteochondral injury research.Methods A total of 75 two-month-old male C57BL/6 mice were randomly divided into 3 groups (n=25). Mice in groups A and B were used to prepare the right knee osteochondral injury models by using needles and tungsten drills, respectively; group C was sham-operation group. The general condition of the mice was observed after operation. The samples were taken at 1 day and 1, 2, 4, and 8 weeks after modeling, and HE staining was performed. The depth, width, and cross-sectional area of the injury site at 1 day in groups A and B were measured, and the percentage of the injury depth to the thickness of the articular cartilage (depth/thickness) was calculated. Toluidine blue staining and immunohistochemical staining for collagen type Ⅱ were performed at 8 weeks, and the International Cartilage Research Society (ICRS) score was used to evaluate the osteochondral healing in groups A and B.Results All mice survived to the completion of the experiment. HE staining showed that group C had normal cartilage morphology. At 1 day after modeling, the injury in group A only broke through the cartilage layer and reached the subchondral bone without entering the bone marrow cavity; the injury in group B reached the bone marrow cavity. The depth, width, cross-sectional area, and depth/thickness of the injury in group A were significantly lower than those in group B (P<0.05). At 1, 2, 4, and 8 weeks after modeling, there was no obvious tissue filling in the injured part of group A, and no toluidine blue staining and expression of collagen type Ⅱ were observed at 8 weeks; while the injured part of group B was gradually filled with tissue, the toluidine blue staining and the expression of collagen type Ⅱ were seen at 8 weeks. At 8 weeks, the ICRS score of group A was 8.2±1.3, which was lower than that of group B (13.6±0.9), showing significant difference (t=−7.637, P=0.000).Conclusion The tungsten drills can break through the subchondral bone layer and enter the bone marrow cavity, and the injury can heal spontaneously. Compared with the needle modeling method, it is a better method for modeling knee osteochondral injury in mice.

Citation： LIU Huan, DING Qirui, MA Cheng, QIN Haonan, WEI Yifan, REN Yongxin. Comparison of two methods for preparing knee osteochondral injury models in mice. Chinese Journal of Reparative and Reconstructive Surgery, 2021, 35(7): 862-867. doi: 10.7507/1002-1892.202101098 Copy

1.	Azuma C, Yasuda K, Tanabe Y, et al. Biodegradation of high-toughness double network hydrogels as potential materials for artificial cartilage. J Biomed Mater Res A, 2007, 81(2): 373-380.
2.	Dias IR, Viegas CA, Carvalho PP. Large animal models for osteochondral regeneration. Adv Exp Med Biol, 2018, 1059: 441-501.
3.	Chevrier A, Kouao AS, Picard G, et al. Interspecies comparison of subchondral bone properties important for cartilage repair. J Orthop Res, 2015, 33(1): 63-70.
4.	Eltawil NM, De Bari C, Achan P, et al. A novel in vivo murine model of cartilage regeneration. Age and strain-dependent outcome after joint surface injury. Osteoarthritis Cartilage, 2009, 17(6): 695-704.
5.	Matsuoka M, Onodera T, Sasazawa F, et al. An articular cartilage repair model in common C57Bl/6 mice. Tissue Eng Part C Methods, 2015, 21(8): 767-772.
6.	Mainil-Varlet P, Aigner T, Brittberg M, et al. Histological assessment of cartilage repair: a report by the Histology Endpoint Committee of the International Cartilage Repair Society (ICRS). J Bone Joint Surg (Am), 2003, 85-A Suppl 2: 45-57.
7.	Serra CI, Soler C. Animal models of osteoarthritis in small mammals. Vet Clin North Am Exot Anim Pract, 2019, 22(2): 211-221.
8.	Chu CR, Szczodry M, Bruno S. Animal models for cartilage regeneration and repair. Tissue Eng Part B Rev, 2010, 16(1): 105-115.
9.	Meng X, Ziadlou R, Grad S, et al. Animal models of osteochondral defect for testing biomaterials. Biochem Res Int, 2020, 2020: 9659412. doi: 10.1155/2020/9659412.
10.	Mahmoud EE, Kamei N, Shimizu R, et al. Therapeutic potential of multilineage-differentiating stress-enduring cells for osteochondral repair in a rat model. Stem Cells Int, 2017, 2017: 8154569. doi: 10.1155/2017/8154569.
11.	Park KS, Kim BJ, Lih E, et al. Versatile effects of magnesium hydroxide nanoparticles in PLGA scaffold-mediated chondrogenesis. Acta Biomater, 2018, 73: 204-216.
12.	Chapelin F, Khurana A, Moneeb M, et al. Tumor formation of adult stem cell transplants in rodent arthritic joints. Mol Imaging Biol, 2019, 21(1): 95-104.
13.	Al-Ansari MM, Hendrayani SF, Shehata AI, et al. p16(INK4A) represses the paracrine tumor-promoting effects of breast stromal fibroblasts. Oncogene, 2013, 32(18): 2356-2364.
14.	Ye C, Chen J, Qu Y, et al. Naringin and bone marrow mesenchymal stem cells repair articular cartilage defects in rabbit knees through the transforming growth factor-β superfamily signaling pathway. Exp Ther Med, 2020, 20(5): 59. doi: 10.3892/etm.2020.9187.
15.	Li L, Duan X, Fan Z, et al. Mesenchymal stem cells in combination with hyaluronic acid for articular cartilage defects. Sci Rep, 2018, 8(1): 9900. doi: 10.1038/s41598-018-27737-y.
16.	Carballo CB, Nakagawa Y, Sekiya I, et al. Basic science of articular cartilage. Clin Sports Med, 2017, 36(3): 413-425.
17.	Arshi A, Petrigliano FA, Williams RJ, et al. Stem cell treatment for knee articular cartilage defects and osteoarthritis. Curr Rev Musculoskelet Med, 2020, 13(1): 20-27.
18.	Eldracher M, Orth P, Cucchiarini M, et al. Small subchondral drill holes improve marrow stimulation of articular cartilage defects. Am J Sports Med, 2014, 42(11): 2741-2750.
19.	Boyan B. Development of the ASTM standard guide for in vivo assessment of implantable devices intended to repair or regenerate articular cartilage. Tissue Eng Pt A, 2014, 20: S69. https://www.astm.org/Standards/F2451.htm.

1. Azuma C, Yasuda K, Tanabe Y, et al. Biodegradation of high-toughness double network hydrogels as potential materials for artificial cartilage. J Biomed Mater Res A, 2007, 81(2): 373-380.
2. Dias IR, Viegas CA, Carvalho PP. Large animal models for osteochondral regeneration. Adv Exp Med Biol, 2018, 1059: 441-501.
3. Chevrier A, Kouao AS, Picard G, et al. Interspecies comparison of subchondral bone properties important for cartilage repair. J Orthop Res, 2015, 33(1): 63-70.
4. Eltawil NM, De Bari C, Achan P, et al. A novel in vivo murine model of cartilage regeneration. Age and strain-dependent outcome after joint surface injury. Osteoarthritis Cartilage, 2009, 17(6): 695-704.
5. Matsuoka M, Onodera T, Sasazawa F, et al. An articular cartilage repair model in common C57Bl/6 mice. Tissue Eng Part C Methods, 2015, 21(8): 767-772.
6. Mainil-Varlet P, Aigner T, Brittberg M, et al. Histological assessment of cartilage repair: a report by the Histology Endpoint Committee of the International Cartilage Repair Society (ICRS). J Bone Joint Surg (Am), 2003, 85-A Suppl 2: 45-57.
7. Serra CI, Soler C. Animal models of osteoarthritis in small mammals. Vet Clin North Am Exot Anim Pract, 2019, 22(2): 211-221.
8. Chu CR, Szczodry M, Bruno S. Animal models for cartilage regeneration and repair. Tissue Eng Part B Rev, 2010, 16(1): 105-115.
9. Meng X, Ziadlou R, Grad S, et al. Animal models of osteochondral defect for testing biomaterials. Biochem Res Int, 2020, 2020: 9659412. doi: 10.1155/2020/9659412.
10. Mahmoud EE, Kamei N, Shimizu R, et al. Therapeutic potential of multilineage-differentiating stress-enduring cells for osteochondral repair in a rat model. Stem Cells Int, 2017, 2017: 8154569. doi: 10.1155/2017/8154569.
11. Park KS, Kim BJ, Lih E, et al. Versatile effects of magnesium hydroxide nanoparticles in PLGA scaffold-mediated chondrogenesis. Acta Biomater, 2018, 73: 204-216.
12. Chapelin F, Khurana A, Moneeb M, et al. Tumor formation of adult stem cell transplants in rodent arthritic joints. Mol Imaging Biol, 2019, 21(1): 95-104.
13. Al-Ansari MM, Hendrayani SF, Shehata AI, et al. p16(INK4A) represses the paracrine tumor-promoting effects of breast stromal fibroblasts. Oncogene, 2013, 32(18): 2356-2364.
14. Ye C, Chen J, Qu Y, et al. Naringin and bone marrow mesenchymal stem cells repair articular cartilage defects in rabbit knees through the transforming growth factor-β superfamily signaling pathway. Exp Ther Med, 2020, 20(5): 59. doi: 10.3892/etm.2020.9187.
15. Li L, Duan X, Fan Z, et al. Mesenchymal stem cells in combination with hyaluronic acid for articular cartilage defects. Sci Rep, 2018, 8(1): 9900. doi: 10.1038/s41598-018-27737-y.
16. Carballo CB, Nakagawa Y, Sekiya I, et al. Basic science of articular cartilage. Clin Sports Med, 2017, 36(3): 413-425.
17. Arshi A, Petrigliano FA, Williams RJ, et al. Stem cell treatment for knee articular cartilage defects and osteoarthritis. Curr Rev Musculoskelet Med, 2020, 13(1): 20-27.
18. Eldracher M, Orth P, Cucchiarini M, et al. Small subchondral drill holes improve marrow stimulation of articular cartilage defects. Am J Sports Med, 2014, 42(11): 2741-2750.
19. Boyan B. Development of the ASTM standard guide for in vivo assessment of implantable devices intended to repair or regenerate articular cartilage. Tissue Eng Pt A, 2014, 20: S69. https://www.astm.org/Standards/F2451.htm.

Chinese Journal of Reparative and Reconstructive Surgery

Comparison of two methods for preparing knee osteochondral injury models in mice

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