Abstract
Elastic cartilage in the rabbit external ear is an important animal model with attractive potential value for researching the physiological and pathological states of cartilages especially during wound healing. In this work, nonlinear optical microscopy based on two-photon excited fluorescence and second harmonic generation were employed for imaging and quantifying the intact elastic cartilage. The morphology and distribution of main components in elastic cartilage including cartilage cells, collagen and elastic fibers were clearly observed from the high-resolution two-dimensional nonlinear optical images. The areas of cell nuclei, a parameter related to the pathological changes of normal or abnormal elastic cartilage, can be easily quantified. Moreover, the three-dimensional structure of chondrocytes and matrix were displayed by constructing three-dimensional image of cartilage tissue. At last, the emission spectra from cartilage were obtained and analyzed. We found that the different ratio of collagen over elastic fibers can be used to locate the observed position in the elastic cartilage. The redox ratio based on the ratio of nicotinamide adenine dinucleotide (NADH) over flavin adenine dinucleotide (FAD) fluorescence can also be calculated to analyze the metabolic state of chondrocytes in different regions. Our results demonstrated that this technique has the potential to provide more accurate and comprehensive information for the physiological states of elastic cartilage.
Highlights
Elastic cartilage is present in the ear pinna, external auditory meatus, eustachian tubes and epiglottis, etc
The major components of elastic cartilage are specialized cells called chondroblasts that produce a large amount of extracellular matrix (ECM) composed of Type II collagenbers, abundant ground substance rich in proteoglycan and elasticbers.[1,2]
An intact elastic cartilage is composed of large chondrocytes surrounded by an extensive ECM and covered with perichondrium
Summary
Elastic cartilage is present in the ear pinna, external auditory meatus, eustachian tubes and epiglottis, etc. The major components of elastic cartilage are specialized cells called chondroblasts that produce a large amount of extracellular matrix (ECM) composed of Type II collagenbers, abundant ground substance rich in proteoglycan and elasticbers.[1,2] The regulation of chondrocyte function by the ECM is critical to understanding normal physiological state of cartilage and its changes in disease states. Cartilage diseases include a wide variety of clinical phenotypes from common osteoarthrosis to several di®erent types of chondrodysplasias.[3,4] It is important to treat elastic cartilage defects or other cartilage diseases for improving the quality of life in patients.[5] some therapies such as reconstruction surgery that depend on development in tissue engineering technique have been developed for repairing cartilage lesions, visualizing these complex processes within cartilage tissues remains challenging.[6]
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