Preparation of cartilage decellularized extracellular matrix-loaded composite nanofiber scaffolds based on two-nozzle electrospinning

doi:10.12307/2023.839

Abstract

Abstract: BACKGROUND: Cartilage decellularized extracellular matrix (dECM) is an ideal biomaterial for the preparation of cartilage tissue engineering scaffolds, which can be used to repair cartilage defects.
OBJECTIVE: To use polyvinyl alcohol (PVA) as the loading material of dECM and poly3-hydroxybutyrate 4-hydroxybutyrate (P34HB) as the frame material to prepare P34HB-PVA-dECM composite nanofiber scaffolds by two-needle electrospinning, and preliminarily explore the bioactivity of the scaffolds in vitro.
METHODS: The cellular components in the cartilage tissue were removed by a combination of enzyme, chemical, and ultrasonic concussion cleaning methods to prepare cartilage dECM. P34HB, P34HB-PVA, and P34HB-PVA-dECM scaffolds were prepared by two-neezle electrospinning. The fiber morphology, composition, hydrophilicity, and mechanical properties of the scaffolds were characterized. Human bone marrow mesenchymal stem cells were co-cultured with the three kinds of scaffolds. The viability of cells on scaffolds was evaluated by the Live/Dead staining. The adhesion morphology of the cells on the scaffolds was observed by scanning electron microscopy. The proliferation performance of the cells on the scaffolds was detected by the alamar blue kit. The chondrogenic differentiation of human bone marrow stem cells on the scaffolds was evaluated by type II collagen immunofluorescence.

RESULTS AND CONCLUSION: (1) Scanning electron microscopy results showed that the scaffold fibers in each group were randomly distributed and the interconnections between the fibers showed a porous structure. The fiber diameter of P34HB-PVA-dECM scaffolds was the smallest. The P34HB-PVA-dECM scaffolds had the smallest water contact angle (P < 0.05) and the highest water absorption rate compared with the P34HB and P34HB-PVA scaffolds (P < 0.05). The elastic modulus of the P34HB-PVA-dECM scaffolds was higher than that of the P34HB and P34HB-PVA scaffolds (P < 0.05). (2) Live/Dead staining showed that most cells survived well on the three groups of scaffolds. The scanning electron microscope observation showed that the bone marrow mesenchymal stem cells on P34HB-PVA-dECM scaffolds extended pseudopod more fully and fused better with the scaffolds. Alamar blue staining exhibited that the proliferation rate of bone marrow mesenchymal stem cells on the P34HB-PVA-dECM scaffolds was faster than that on the P34HB and P34HB-PVA scaffolds (P < 0.05). The immunofluorescence staining results showed that more type II collagen produced after chondrogenic induction on the P34HB-PVA-dECM scaffolds was more than that on the P34HB and P34HB-PVA scaffolds (P < 0.05). (3) To sum up, the P34HB-PVA-dECM composite nanofiber scaffolds have smaller fiber structure, more optimized hydrophilicity and mechanical properties, which is more favorable for the adhesion, proliferation, and differentiation into chondrocytes of bone marrow mesenchymal stem cells.

Key words: two-nozzle electrospinning, decellularized extracellular matrix, polyvinyl alcohol, cartilage, tissue engineering, biomaterial, nanofiber scaffold

CLC Number:

Teng Jianxiang, Zhu Jisheng, Yuan Daizhu, Wang Zhen, Zhou Yuhu, Tian Xiaobin. Preparation of cartilage decellularized extracellular matrix-loaded composite nanofiber scaffolds based on two-nozzle electrospinning[J]. Chinese Journal of Tissue Engineering Research, 2023, 27(34): 5448-5454.

Figures/Tables 11

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