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A bioscaffold of decellularized whole osteochondral sheet improves proliferation and differentiation of loaded mesenchymal stem cells in a rabbit model

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Abstract

As a Natural decellularized extracellular matrix, osteochondral tissue is the best scaffold for the restoration of osteoarthritis defects. Bioscaffolds have the most similarly innate properties like biomechanical properties and the preserved connection of the bone-to-cartilage border. Although, their compacity and low porosity particularly, are proven to be difficulties of decellularization and cell penetration. This study aims to develop a new bioscaffold of decellularized osteochondral tissue (DOT) that is recellularized by bone marrow-derived mesenchymal stem cells (BM-MSCs), as a biphasic allograft, which preserved the interface between the cartilage section and subchondral bone of the joint. Whole osteochondral tissues of rabbit knee joints were sheeted in cartilaginous parts in 200–250 µm sections while connected to the subchondral bone and then fully decellularized. The BM-MSCs were seeded on the scaffolds in vitro; some constructs were subcutaneously implanted into the back of the rabbit. The cell penetration, differentiation to bone and cartilage, viability, and cell proliferation in vitro and in vivo were evaluated by qPCR, histological staining, MTT assay, and immunohistochemistry. DNA content analysis and SEM assessments confirmed the decellularization of the bioscaffold. Then, histological and SEM evaluations indicated that the cells could successfully penetrate the bone and cartilage lacunas in implanted grafts. MTT assay confirmed cell proliferation. Prominently, gene expression analysis showed that seeded cells differentiated into osteoblasts and chondrocytes in both bone and cartilage sections. More importantly, seeded cells on the bioscaffold started ECM secretion. Our results indicate that cartilage-to-bone border integrity was largely preserved. Additionally, ECM-sheeted DOT could be employed as a useful scaffold for promoting the regeneration of osteochondral defects.

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Acknowledgements

We would like to thank Dr. Mirnajafi (Tarbiat Modares University, Tehran, Iran) for his technical and scientific assistance.

Funding

This work was supported by Royan Institute, Tehran, Iran (Grant No: 91000291).

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Authors and Affiliations

  1. Department of Stem Cells and Developmental Biology, Cell Science Research Centre, Royan Institute for Stem Cell Biology and Technology, ACECR, Tehran, Iran

    Leila Taghiyar, Hamideh Asadi & Mohamadreza Baghaban Eslaminejad

  2. Department of Developmental Biology, University of Science and Culture, Tehran, Iran

    Hamideh Asadi

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  1. Leila Taghiyar
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  2. Hamideh Asadi
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Correspondence to Mohamadreza Baghaban Eslaminejad.

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Conflict of interest

All authors contributed to the study’s conception and design. Material preparation, ion, data collection, and analysis were performed by [leila taghiyar] and [Hamideh Asadi]. The first draft of the manuscript was written by [Mohamadreza baghaban Eslaminejad] and all authors commented on previous versions of the manuscript. All authors read and approved the final manuscript.

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The study protocol was approved by the institutional review board of the Royan Institute. The animal studies were performed after receiving approval from the Institutional Animal Care and Use Committee (IACUC) in the Royan Institutional (IACUC Approval No. J/90/1397).

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10561_2023_10084_MOESM1_ESM.tif

Supplementary Figure 1. Rabbit total knee used for osteochondral sample isolation (A), Macroscopic presentation of the osteochondral scaffold before cutting (B), CAMPDEN cutting system used for the preparation of cartilage sheets (C), Macroscopic presentation of osteochondral scaffold after producing the sheets in cartilage part (D), Constructs after seeding cells in culture media (E), Subcutaneous implantation of scaffolds onto rabbits back (F)

10561_2023_10084_MOESM2_ESM.docx

Table 1. Description of rabbit primers used in the quantitative reverse transcription polymerase chain reactions (qPCR).

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Taghiyar, L., Asadi, H. & Baghaban Eslaminejad, M. A bioscaffold of decellularized whole osteochondral sheet improves proliferation and differentiation of loaded mesenchymal stem cells in a rabbit model. Cell Tissue Bank 24, 711–724 (2023). https://doi.org/10.1007/s10561-023-10084-2

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