Observation of Solute Transport between Articular Cartilage and Subchondral Bone in Live Mice

Author:

Huang Yang1ORCID,Chen Cheng1ORCID,Wang Fuyou1,Chen Guangxin1,Cheng Shidi2,Tang Zhexiong1,Li Zheng2,Gong Xiaoyuan1,Yang Liu1

Affiliation:

1. Center for Joint Surgery, Southwest Hospital, Third Military Medical University (Army Medical University), Chongqing, People’s Republic of China

2. Hematology Department, Army Medical Center of PLA, Chongqing, People’s Republic of China

Abstract

Objective To establish a method for investigating the permeability of calcified cartilage zone (CCZ) and to observe solute transport between articular cartilage (AC) and subchondral bone (SB) through intact CCZ in vivo. Design We developed a novel fixing device combined with un-decalcified fluorescence observation method to address the permeability of CCZ in live mice. Twenty-four Balb/c female mice aged 1 to 8 months were used to observe the development of CCZ. Eighty-four Balb/c female mice (aged 1 or 6 months) with mature or immature CCZ of distal femur were used to investigate the permeability of intact CCZ in vivo. Diffusivity of rhodamine B (476 Da) and tetramethyl-rhodamine isothicyanate-dextran (TRITC-Dextran, 20 kDa) was tested from AC to SB in 0 minutes, 1 minute, 15 minutes, 30 minutes, 1 hour, and 2 hours. None diffused knee joints (0 minutes) served as blank control, while in vitro immersion of distal femurs in rhodamine B or TRITC-Dextran for 72 hours served as positive control. Results CCZ was well developed in 6-month mice. Both tracers penetrated immature CCZ down to SB in less than 1 hour in live mice, while the diffusion of both tracers decreased rapidly at tidemark in all testing time points. Conclusion Current study provided direct evidence of blocking effect of CCZ in solute transportation during short diffusion period in live animal, indicating the important role of CCZ in joint development and microenvironment maintenance.

Funder

National Natural Science Foundation of China

southwest hospital

Chongqing Social Welfare Innovation Project

Publisher

SAGE Publications

Subject

Physical Therapy, Sports Therapy and Rehabilitation,Biomedical Engineering,Immunology and Allergy

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