Chinese Journal of Tissue Engineering Research ›› 2020, Vol. 24 ›› Issue (16): 2544-2550.doi: 10.3969/j.issn.2095-4344.2256

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Good cell compatibility and permeability of silk fibroin/chitosan composite scaffolds 

Zhang Xiaoyun1, Chen Yueping1, Song Shilei2, Zhang Chi2, Zhuo Yinghong1, Yang Nan2, Zhan Huasong2, Xu Canhong2   

  1. 1Department of Orthopedics, Ruikang Hospital Affiliated to Guangxi University of Traditional Chinese Medicine, Nanning 530011, Guangxi Zhuang Autonomous Region, China; 2Graduate School of Guangxi University of Traditional Chinese Medicine, Nanning 530001, Guangxi Zhuang Autonomous Region, China
  • Received:2019-09-06 Revised:2019-09-07 Accepted:2019-10-09 Online:2020-06-08 Published:2020-03-26
  • Contact: Chen Yueping, Chief physician, Doctoral supervisor, Department of Orthopedics, Ruikang Hospital Affiliated to Guangxi University of Traditional Chinese Medicine, Nanning 530011, Guangxi Zhuang Autonomous Region, China
  • About author:Zhang Xiaoyun, Attending physician, Department of Orthopedics, Ruikang Hospital Affiliated to Guangxi University of Traditional Chinese Medicine, Nanning 530011, Guangxi Zhuang Autonomous Region, China
  • Supported by:
    the National Natural Science Foundation of China, No. 81760796, 81960803; the Basic Ability Improvement Project for Young Teachers in Guangxi Universities, No. 2019KY0352, KY2016YB204; the University-Level Scientific Research Project of Guangxi University of Traditional Chinese Medicine in 2019, No. 2019QN027 the First-Class Subject of Guangxi University of Traditional Chinese Medicine, No. 2019XK029; the Natural Science Foundation of Guangxi Zhuang Autonomous Region, No. 2015GXNSFAA139136; the Key Project of Health Department of Guangxi Zhuang Autonomous Region, No. S201419-05; the National Famous Traditional Chinese Medicine Inheritance Studio Construction Project in 2016

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Abstract:

BACKGROUND: Silk fibroin and chitosan are commonly used as scaffolds in tissue engineering, but there are some shortcomings in their separate application. When they are mixed, they can be modified each other. They give full play to each other’s advantages and become ideal composite scaffolds.

OBJECTIVE: To prepare Silk fibroin/chitosan composite scaffold and determineits properties.

METHODS: The silk fibroin/chitosan composite scaffolds were prepared by freeze-drying method. The morphology and structure of the composite scaffolds were examined by scanning electron microscopy. The properties of the composite scaffolds were tested by thermogravimetric analysis, mechanical properties test, and cytotoxicity test. The quaternion chitosan was prepared. The nuclear magnetic resonance spectrum was detected by nuclear magnetic resonance instrument. The potential and particle size distribution were detected by Zeta potentiometer. The protection of DNA was detected by gel electrophoresis. The binding with DNA was observed by transmission electron microscope.

RESULTS AND CONCLUSION: (1) The results of scanning electron microscopy showed that the silk fibroin/chitosan composite scaffolds had a good three-dimensional pore structure, with a pore size of 50-100 μm. (2) The results of thermogravimetric analysis showed that when the temperature was less than 200 °C, the mass loss rate of silk fibroin/chitosan composite scaffold was lower. When the temperature increased to 200-500 °C, the mass loss rate of the scaffold began to accelerate, and the loss amount increased. At 800 °C, the residual mass of the composite scaffold was 38%. (3) The maximum strain of silk fibroin/chitosan composite scaffold reached 94.94%, and the maximum stress was 7.01 MPa. (4) The results of CCK-8 experiment showed that silk fibroin/chitosan composite scaffolds had no cytotoxicity to rabbit bone marrow mesenchymal stem cells and had good cell compatibility. (5) The results of nuclear magnetic resonance spectra showed that the quaternion degree of quaternary ammonium chitosan was about 20%. (6) The particle size distribution of quaternized chitosan was (588.56±52.39) nm, and the surface of quaternized chitosan was positively charged with a potential of (16.3±3.92) mV, which was beneficial to the combination with DNA. (7) The results of gel electrophoresis experiments showed that the higher the proportion of quaternion chitosan, the better the encapsulation of DNA. When the ratio of chitosan/DNA was 1∶3, the encapsulation effect was achieved. (8) The results of transmission electron microscopy showed that most of the particles of quaternized chitosan/DNA were solid and round; the particle size difference was small; and the average particle size was about 200 nm. (9) The results showed that silk fibroin/chitosan composite scaffolds had a good biocompatibility and cell permeability, which was conducive to the growth of cells between scaffolds.

Key words: silk fibroin protein/chitosan composite scaffold, biodegradation rate, mechanical properties, cytotoxicity, quaternary ammonium chitosan, silk protein, bone marrow mesenchymal stem cells, biocompatibility

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