Chinese Journal of Tissue Engineering Research ›› 2019, Vol. 23 ›› Issue (34): 5436-5441.doi: 10.3969/j.issn.2095-4344.1439

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Preparation and evaluation of human articular cartilage-derived extracellular matrix tissue engineering scaffold

Peng Liqing1, Luo Xujiang1, Zhang Bin1, Shen Shi1, Huang Bo1, Gao Chao2, Lu Xiaobo1, Liu Shuyun2, Guo Quanyi1, 2
  

  1. 1Department of Bone and Joint Surgery, Affiliated Hospital of Southwest Medical University, Luzhou 646000, Sichuan Province, China; 2Institute of Orthopedics, First Medical Center, Chinese PLA General Hospital, Beijing Key Lab of Regenerative Medicine in Orthopedics, Key Laboratory of Musculoskeletal Trauma & War Injuries, PLA, Beijing 100853, China
  • Received:2019-06-10 Online:2019-12-08 Published:2019-12-08
  • Contact: Guo Quanyi, MD, Chief physician, Professor, Doctoral supervisor, Department of Bone and Joint Surgery, Affiliated Hospital of Southwest Medical University, Luzhou 646000, Sichuan Province, China; Institute of Orthopedics, First Medical Center, Chinese PLA General Hospital, Beijing Key Lab of Regenerative Medicine in Orthopedics, Key Laboratory of Musculoskeletal Trauma & War Injuries, PLA, Beijing 100853, China
  • About author:Peng Liqing, Master candidate, Department of Bone and Joint Surgery, Affiliated Hospital of Southwest Medical University, Luzhou 646000, Sichuan Province, China
  • Supported by:

    National R & D Program of China, No. 2018YFC1105900 (project participant: LSY); the National Natural Science Foundation of China, No. 81772319 (to GQY); the Natural Science Foundation of Beijing, No. 7172203 (to GQY)

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Abstract:

BACKGROUND: Cartilage-derived extracellular matrix as a natural material provides an ideal microenvironment for cells that depend on tissue growth and has become an ideal scaffold material for cartilage repair.
OBJECTIVE: To prepare human articular cartilage-derived extracellular matrix as a cartilage tissue engineering scaffold and to evaluate its physicochemical properties and biological properties.
METHODS: The human articular cartilage tissue was collected, pulverized into homogenate, decellularized, and freeze-dried to prepare a cartilage tissue engineering scaffold. Scaffold morphology was observed under scanning electron microscope. The porosity and water absorption and mechanical properties of the cartilage tissue engineering scaffold were determined. The composition of tissue engineering scaffold was observed by histochemistry and immunohistochemistry. Rabbit chondrocytes were cultured with the leaching liquor of human cartilage-derived extracellular matrix tissue engineering scaffold for different time periods. Routine cell culture fluid was used as control. The cytotoxicity of tissue engineering scaffold was detected by MTT colorimetry. Human chondrocytes and tissue engineering scaffold were co-cultured and cell adhesion and proliferation were observed under the scanning electron microscope and confocal microscope. The cell-scaffold complex sections were stained by histochemistry and immunohistochemistry.
RESULTS AND CONCLUSION: The prepared scaffold had three-dimensional porous sponge-like longitudinally oriented structure. There were cartilage fibers around the scaffold pores. After hematoxylin-eosin staining, no nucleus was observed. Both safranin O and Sirius red staining confirmed that cartilage tissue engineering scaffold contained collagen and cartilage matrix. The porosity and water absorption of the scaffold was (91.8±2.9)% and (93.5±1.4)%, respectively. MTT results showed that the leaching liquor of human cartilage-derived extracellular matrix was non-toxic to chondrocytes. After co-culture, human chondrocytes adhered, proliferated and evenly distributed on the peripheral wall of the scaffold pores. The results showed that human articular cartilage-derived extracellular matrix had similar composition to natural cartilage, provided the structure suitable for cell adhesion and proliferation, and exhibited good histocompatibility. Therefore, human articular cartilage-derived extracellular matrix can be used as a scaffold material for repairing cartilage defects by tissue engineering technique.

Key words: tissue engineering, extracellular matrix, scaffold, cartilage, allogenic

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