Chinese Journal of Tissue Engineering Research ›› 2024, Vol. 28 ›› Issue (22): 3451-3456.doi: 10.12307/2024.473

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Elastase quantitative analysis of elastin effect on mechanical response of anterior cruciate ligament

Zhang Wentian1, Deng Yuping2, 3, 4, Liu Xiaoyun2, 4, Li Caijuan5, Wang Mian2, 6, Liang Zeyu2, Xiong Liang3, Huang Gang3, Chen Guangxin7, Li Zitao8, 9, Huang Wenhua1, 2, 3, 6, 10   

  1. 1School of Basic Medical Sciences, Fujian Medical University, Fuzhou 350122, Fujian Province, China; 2Guangdong Engineering Research Center for Translation of Medical 3D Printing Application, Guangdong Provincial Key Laboratory of Medical Biomechanics, National Key Discipline of Human Anatomy, School of Basic Medical Sciences, Southern Medical University, Guangzhou 510515, Guangdong Province, China; 3Department of Orthopedics and Traumatology, Integrated Hospital of Traditional Chinese and Western Medicine, Southern Medical University, Guangzhou 510315, Guangdong Province, China; 4Shenzhen Bay Laboratory, Institute of Biomedical Engineering, Shenzhen 518132, Guangdong Province, China; 5Ultrasonography Lab, 9Department of Orthopedics, Hongqi Hospital Affiliated to Mudanjiang Medical University, Mudanjiang 157011, Heilongjiang Province, China; 6Pingshan District People’s Hospital of Shenzhen, Pingshan General Hospital of Southern Medical University, Shenzhen 518118, Guangdong Province, China; 7College of Medical Imaging, Mudanjiang Medical University, Mudanjiang 157011, Heilongjiang Province, China; 8Mudanjiang Beiyao Resources Development and Application Collaborative Innovation Center, Mudanjiang 157011, Heilongjiang Province, China; 10Guangdong Medical Innovation Platform for Translation of 3D Printing Application, Third Affiliated Hospital of Southern Medical University, Guangzhou 510000, Guangdong Province, China
  • Received:2023-08-05 Accepted:2023-08-26 Online:2024-08-08 Published:2024-01-19
  • Contact: Huang Wenhua, MD, Professor, Doctoral supervisor, School of Basic Medical Sciences, Fujian Medical University, Fuzhou 350122, Fujian Province, China; Guangdong Engineering Research Center for Translation of Medical 3D Printing Application, Guangdong Provincial Key Laboratory of Medical Biomechanics, National Key Discipline of Human Anatomy, School of Basic Medical Sciences, Southern Medical University, Guangzhou 510515, Guangdong Province, China; Department of Orthopedics and Traumatology, Integrated Hospital of Traditional Chinese and Western Medicine, Southern Medical University, Guangzhou 510315, Guangdong Province, China; Pingshan District People’s Hospital of Shenzhen, Pingshan General Hospital of Southern Medical University, Shenzhen 518118, Guangdong Province, China; Guangdong Medical Innovation Platform for Translation of 3D Printing Application, Third Affiliated Hospital of Southern Medical University, Guangzhou 510000, Guangdong Province, China Li Zitao, Master, Professor, Master’s supervisor, Mudanjiang Beiyao Resources Development and Application Collaborative Innovation Center, Mudanjiang 157011, Heilongjiang Province, China; Department of Orthopedics, Hongqi Hospital Affiliated to Mudanjiang Medical University, Mudanjiang 157011, Heilongjiang Province, China
  • About author:Zhang Wentian, Master candidate, School of Basic Medical Sciences, Fujian Medical University, Fuzhou 350122, Fujian Province, China
  • Supported by:
    National Key Research & Development Plan, No. 2022YFB4600600 (to HWH); National Natural Science Foundation of China, No. 31972915, 32271181 (to HWH); National Natural Science Foundation of China, No. 82171966 (to LCJ); Guangdong Basic and Applied Basic Research Fund, No. 2020B1515120001 (to HWH); China Postdoctoral Science Foundation, No. 2022M711533 (to DYP); Postdoctoral Project of the Guangdong Provincial Excellent Youth Talent International Training Program (to DYP); Guangdong Medical Science and Technology Research Fund Project, No. A2023129 (to XL); Heilongjiang Natural Science Foundation Project, No. SS2023H004 (to LZT); Science Foundation Torch Program of Mudanjiang Medical University, No. 2022-MYHJ-007 (to LCJ)

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Abstract: BACKGROUND: The anterior cruciate ligament has unique nonlinear mechanical properties under a complex physiological loading environment. Elastin is an important contributor to the mechanical properties of the anterior cruciate ligament, but its mechanical response to the anterior cruciate ligament under axial tension is not clear.
OBJECTIVE: To quantitatively analyze the effect of elastin on the tensile mechanical response of the anterior cruciate ligament.
METHODS: Elastase solution and control buffer were prepared. The porcine anterior cruciate ligament samples were prepared into small-size samples and randomly soaked in 0, 0.1, 1.0, 2.0, 5.0, and 10.0 U/mL elastase solution for 6 hours, and other small samples of the same size were soaked in 2 U/mL elastase solution for 0, 1, 3, 6, 9, and 12 hours. To determine suitable soaking conditions for elastin-targeted enzymes and verify the digestive effect, histological staining was used to compare the effects of enzyme treatment on tissue structure and composition. The ligament samples were randomly divided into elastase-treated group and PBS group, and immersed in 2 U/mL elastase solution and PBS buffer for 6 hours, respectively. A mechanical tensile test was performed before and after immersion.
RESULTS AND CONCLUSION: (1) The biochemical results showed that being treated in 2 U/mL elastase solution for 6 hours could reduce the elastin content by 31.1%, and had no significant effect on other mechanical-related components in the tissue. (2) The histological results showed that elastase was able to penetrate the tissue, and the loose degree of tissue increased after treatment. (3) In the mechanical results before and after treatment, the mechanical properties of the PBS group decreased significantly, only the low-tension elastic modulus increased significantly and the initial length increased significantly in the elastase-treated group. (4) The intergroup comparison results showed that there was no significant difference between the two groups in pre-treatment, but the low-tension elastic modulus, initial slopes, saturated slopes, and initial length of the elastase-treated group after treatment were significantly higher than those in the PBS group. (5) These results suggest that elastin degradation significantly affects the biomechanical properties of the anterior cruciate ligament and further complements our understanding of the structure-function relationship of the anterior cruciate ligament. 

Key words: anterior cruciate ligament, biomechanics, elastin, EVG staining, quasi-static tensile, viscoelasticity

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