Metabolism of collagen and proteoglycan in tissue-engineered artificial skin / 中国组织工程研究

ABSTRACT

BACKGROUND: The extracellular matrix (ECM) in demis consists of collagen, elastin and other matrix components, and as an in vitro skin model, the tissue-engineered artificial skin should have the synthesis, excretion and decomposition functions. OBJECTIVE: To observe the collagen and proteoglycan metabolism of tissue-engineered artificial skin. DESIGN: Single sampling observation SETTING: Department of Dermatology, Peking University Shenzhen Hospital. MATERIALS Siderophilin, trypsin, 3H-proline and sirius red, etc. METHODS: The experiment was conducted at Department of Dermatology, Peking University Shenzhen Hospital and Institute of Frontier Medical Science, Jilin University from June 2000 to December 2004.①The tissue-engineered artificial skin was prepared in accordance with literatures.②The collagen synthesis of tissue-engineered artificial skin was examined by 3H proline incorporation. The culture plate was added with 3H-proline in each hole and cultured for 4 hours respectively in the 1st, 2nd and 3rd weeks after tissue engineering culture, and compared with natural skin.③Picric acid sirius red staining was applied to observe collagen synthesis and excretion functions of tissue-engineered artificial skin, while proteoglycan synthesis and excretion were examined by AB-PAS staining respectively after 1, 2, 3, 4 and 6 weeks of culture. MAIN OUTCOME MEASURES: ①The detections of collagen synthesis and excretion; ②The detections of proteoglycan synthesis and excretion; ③ The results of 3H-proline incorporation experiment. RESULTS: ①The quantity of 3H-proline incorporation of tissue-engineered artificial skin was similar with that of normal skin after 7 and 14 days of culture, but obviously higher than that of normal skin at the 21st day after culture. ②Under polarization microscope, the tissue-engineered artificial skin presented fasciculate red double refraction and thin green double refraction, whereas simple matrix lattice only displayed red double refraction.③AB-PAS staining was negative in tissue-engineered artificial skin after 1 and 2 weeks culture together with simple matrix lattice, while positive in artificial skin after 3-6 weeks of culture. CONCLUSION: The tissue-engineered artificial skin has the capacity of synthesizing and excreting ECM collagen and proteoglycan.