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. 2020 Feb 7;10:2154. doi: 10.1038/s41598-020-59219-5

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© The Author(s) 2020

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Detection of local proteolytic damage by AFL measurement. (A) Representative photos of cartilage samples before and after treatment. Cartilage surface of porcine metacarpophalangeal joints were treated with bacterial collagenase (500 μg/ml), MMP-1 (500 μg/ml) and trypsin (200 μg/ml) by placing filter papers soaked with these proteinase solutions. Samples were incubated at 37 °C for 24 hours with enzymes or buffer prior to measurement in a humidified cell culture incubator to prevent drying of cartilage samples. (B) Mean autofluorescence lifetime of articular cartilage samples after treatment with bacterial collagenase, MMP-1 and trypsin. The data indicate that treatment of porcine cartilage with these enzymes decreased AFL in a statistically significant manner: bacterial collagenase, p < 0.0001; MMP-1, p < 0.0004; and trypsin, p < 0.0072 (n = 9 for each group). (C) Porcine joint cartilage was treated with increasing dose of Bacterial collagenase as indicated, and mean AFL was measured (n = 6). (D) Location of line profile measurements along the cartilage surface. (E) Mean autofluorescence lifetime along the cartilage surface treated with buffer (top), bacterial collagenase (middle) and MMP-1 (bottom).