title: In vitro Sirius Red collagen assay measures the pattern shift from soluble to deposited collagen.
abstract: In this study, we compared two in vitro collagen production assays ([(3)H]-proline incorporation and Sirius Red) for their ability to determine the pattern shift from soluble to deposited collagen. The effect of the antifibrotic agent, triptolide (TPL), on collagen production was also studied. The results showed that: (1) 48 h after NIH 3T3 (murine embryo fibroblast) and HFL-1(human fetal lung fibroblast) were exposed to transforming growth factor-beta 1 (TGF-β), there was an increase in soluble collagen in the culture medium; (2) on day 4, soluble collagen declined, whereas deposited collagen increased; (3) Sirius Red was easier to use than [(3)H]-proline incorporation and more consistently reflected the collagen pattern shift from soluble to deposited; (4) the in vitro Sirius Red assay took less time than the in vivo assay to determine the effect of TPL. Our results suggest that: (a) the newly synthesized soluble collagen can sensitively evaluate an agent's capacity for collagen production and (b) Sirius Red is more useful than [(3)H]-proline because it is easier to use, more convenient, less time consuming, and does not require radioactive material.
authors.affiliation: Department of Radiation Oncology, UF Shands Cancer Center, University of Florida, 2033 Mowry Road, Gainesville, 100385, FL, 32610, USA.
authors.forename: Chun
authors.initials: C
authors.lastname: Chen
authors.affiliation: Department of Radiation Oncology, UF Shands Cancer Center, University of Florida, 2033 Mowry Road, Gainesville, 100385, FL, 32610, USA.
authors.forename: Shanmin
authors.initials: S
authors.lastname: Yang
authors.affiliation: Department of Radiation Oncology, UF Shands Cancer Center, University of Florida, 2033 Mowry Road, Gainesville, 100385, FL, 32610, USA.
authors.forename: Mei
authors.initials: M
authors.lastname: Zhang
authors.affiliation: Department of Radiation Oncology, UF Shands Cancer Center, University of Florida, 2033 Mowry Road, Gainesville, 100385, FL, 32610, USA.
authors.forename: Zhenhuan
authors.initials: Z
authors.lastname: Zhang
authors.affiliation: Department of Radiation Oncology, UF Shands Cancer Center, University of Florida, 2033 Mowry Road, Gainesville, 100385, FL, 32610, USA.
authors.forename: Bingrong
authors.initials: B
authors.lastname: Zhang
authors.affiliation: Department of Radiation Oncology, UF Shands Cancer Center, University of Florida, 2033 Mowry Road, Gainesville, 100385, FL, 32610, USA.
authors.forename: Deping
authors.initials: D
authors.lastname: Han
authors.affiliation: Department of Radiation Oncology, UF Shands Cancer Center, University of Florida, 2033 Mowry Road, Gainesville, 100385, FL, 32610, USA.
authors.forename: Jun
authors.initials: J
authors.lastname: Ma
authors.affiliation: Department of Radiation Oncology, UF Shands Cancer Center, University of Florida, 2033 Mowry Road, Gainesville, 100385, FL, 32610, USA.
authors.forename: Xiaohui
authors.initials: X
authors.lastname: Wang
authors.affiliation: Department of Radiation Oncology, UF Shands Cancer Center, University of Florida, 2033 Mowry Road, Gainesville, 100385, FL, 32610, USA.
authors.forename: Jingshen
authors.initials: J
authors.lastname: Hong
authors.affiliation: Department of Radiation Oncology, UF Shands Cancer Center, University of Florida, 2033 Mowry Road, Gainesville, 100385, FL, 32610, USA.
authors.forename: Yansong
authors.initials: Y
authors.lastname: Guo
authors.affiliation: Department of Radiation Oncology, UF Shands Cancer Center, University of Florida, 2033 Mowry Road, Gainesville, 100385, FL, 32610, USA.
authors.forename: Paul
authors.initials: P
authors.lastname: Okunieff
authors.affiliation: Department of Radiation Oncology, UF Shands Cancer Center, University of Florida, 2033 Mowry Road, Gainesville, 100385, FL, 32610, USA. lurongzhang@ufl.edu.
authors.forename: Lurong
authors.initials: L
authors.lastname: Zhang
mesh_terms: D000818:Animals; D018906:Antineoplastic Agents, Alkylating / Q000494:pharmacology; D001391:Azo Compounds; D001681:Biological Assay; D002478:Cells, Cultured; D003094:Collagen / Q000378:metabolism*; D004396:Coloring Agents; D004224:Diterpenes / Q000494:pharmacology; D004622:Embryo, Mammalian / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D004852:Epoxy Compounds / Q000494:pharmacology; D005333:Fetus / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D005347:Fibroblasts / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D006801:Humans; D008168:Lung / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D051379:Mice; D041681:NIH 3T3 Cells; D010616:Phenanthrenes / Q000494:pharmacology; D011392:Proline / Q000378:metabolism; D016212:Transforming Growth Factor beta / Q000494:pharmacology
chemical_list: D018906:Antineoplastic Agents, Alkylating; D001391:Azo Compounds; D004396:Coloring Agents; D004224:Diterpenes; D004852:Epoxy Compounds; D010616:Phenanthrenes; D016212:Transforming Growth Factor beta; C009798:C.I. direct red 80; C001899:triptolide; D003094:Collagen; D011392:Proline