Three-dimensional visualization and quantitative analysis of microvessels in rat cervical spinal cord using barium sulfate perfusion combined with micro-CT scanning

doi:10.3969/j.issn.2095-4344.1999

Abstract

Abstract:

BACKGROUND: Vascular structure is an important basis for maintaining cervical spinal cord function. To evaluate the involvement of microvessels in cervical spinal cord injury and treatment, it is necessary to carry out a good three-dimensional visualization and accurate quantitative analysis of the normal cervical spinal cord microvascular structure, but there are few reports about it.

OBJECTIVE: To investigate the three-dimensional visualization and quantitative analysis of normal cervical spinal cord microvascular structure in rats by barium sulfate perfusion combined with micro-CT scanning, and to lay a foundation for the later evaluation of the involvement degree of microvessels in cervical spinal cord injury and the measurement of therapeutic effect.

METHODS: Five adult male SD rats weighing 280-320 g were included. All animal operation procedures were implemented in accordance with the relevant national guidelines for the use of laboratory animals, and were approved by the Animal Use Ethics Committee of Southern Medical University. After rats were deeply anesthetized, vascularization was performed with warm heparin normal saline, polyformaldehyde and barium sulfate perfusion through cardiac puncture. Cervical spinal cord tissue was taken and placed in micro-CT for scanning and three-dimensional reconstruction. Quantitative analysis of the ratio of microvascular volume to tissue volume, ratio of vascular surface area to tissue volume, number of blood vessels, diameter of blood vessels, separation degree of blood vessels, connectivity density, structural model index and the number, percentage and volume of microvessels with different diameters were quantitatively analyzed.

RESULTS AND CONCLUSION: (1) After three-dimensional reconstruction and visualization, the microvascular structure in the cervical spinal cord was clearly visible, showing the anterior and posterior arteries of the cervical spinal cord as well as several branch arteries. (2) Quantitative analysis results of microvascular index showed that ratio of microvascular volume to tissue volume was 0.022±0.002; ratio of vascular surface area to tissue volume was (52.370±6.271) mm^-1; number of blood vessels was (0.604±0.076) mm^-1; diameter of blood vessels was (0.040±0.005) μm; separation degree of blood vessels was (1.762±0.255) μm; connectivity density was (2.661±0.757) mm^-3; structural model index was (4.400±0.129). The number, percentage and volume of microvessels in cervical spinal cord with different diameters were quantified successfully. (3) Barium sulfate perfusion combined with micro-CT scanning can achieve three-dimensional visualization and accurate quantitative analysis of normal cervical spinal cord microvascular structure in rats, laying a good foundation for the later evaluation of the involvement of microvessels during cervical spinal cord injury and the measurement of therapeutic effect.

Key words: microvessels, cervical spinal cord, three-dimensional visualization, quantitative analysis, rats, the National Natural Science Foundation of China

CLC Number:

R318|R445|R744

Liu Yapu, Lin Junyu, Yang Zhou, Wu Xiuhua, Wu Xiaoliang, Zhu Qing’an. Three-dimensional visualization and quantitative analysis of microvessels in rat cervical spinal cord using barium sulfate perfusion combined with micro-CT scanning[J]. Chinese Journal of Tissue Engineering Research, 2019, 23(36): 5836-5840.

Figures/Tables 2

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