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. 2019 Aug 28;9:12453. doi: 10.1038/s41598-019-47946-3

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© The Author(s) 2019

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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General experimental design and gross appearance, H&E and Alcian blue staining of chondrocyte encapsulated collagen microspheres, compared with pellet. Excised tissues from tibia plateau (A), lateral (B) and medial femoral condyle (C) were collected from total knee replacement surgery. Schematic diagram of the overall experimental design (D). In brief, chondrocytes in the cartilage were enzymatically isolated from the cartilage specimens and expand in monolayer culture. The cells (P3) were then cultured in 3D in order to resume its in vivo characteristics. Two kinds of 3D culture method, namely pellet culture and collagen microencapsulation were used for chondrocyte 3D culture and the effect in phenotypes restoration were compared. Collagen microspheres at different time points during a 3 week culture (E). H&E staining (F–I) and Alcian blue staining (J–M) of microsphere and pellet. Diameter of microsphere during a 3 week culture (O), from 5 independent experiments, each with diameter measurement of 30 microspheres at each time points.