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Bone & Joint Research
Vol. 8, Issue 2 | Pages 101 - 106
1 Feb 2019
Filardo G Petretta M Cavallo C Roseti L Durante S Albisinni U Grigolo B

Objectives

Meniscal injuries are often associated with an active lifestyle. The damage of meniscal tissue puts young patients at higher risk of undergoing meniscal surgery and, therefore, at higher risk of osteoarthritis. In this study, we undertook proof-of-concept research to develop a cellularized human meniscus by using 3D bioprinting technology.

Methods

A 3D model of bioengineered medial meniscus tissue was created, based on MRI scans of a human volunteer. The Digital Imaging and Communications in Medicine (DICOM) data from these MRI scans were processed using dedicated software, in order to obtain an STL model of the structure. The chosen 3D Discovery printing tool was a microvalve-based inkjet printhead. Primary mesenchymal stem cells (MSCs) were isolated from bone marrow and embedded in a collagen-based bio-ink before printing. LIVE/DEAD assay was performed on realized cell-laden constructs carrying MSCs in order to evaluate cell distribution and viability.


Orthopaedic Proceedings
Vol. 87-B, Issue SUPP_I | Pages 67 - 67
1 Mar 2005
Grigolo B Roseti L Fiorini M De Franceschi L Piacentini A Facchini A
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Aims: The maintenance of the original phenotype by isolated chondrocytes grown in vitro is an important requisite for their use in repairing damaged articular cartilage. The methods to verify the expression of cartilage specific molecules usually involve destructive procedures to recover the cells from the scaffolds for tests. The aim of this study was to find a soluble marker able to attest the occurrence of a differentiation process by chondrocytes grown onto a biomaterial used for cell transplantation. We turned our attention to cathepsin B which is known to be abnormally synthesized in de-differentiated chondrocytes and scarcely produced in the differentiated ones.

Methods: The production of cathepsin B by human articular chondrocytes expanded in vitro and then grown onto a hyaluronan-based polymer derivative (Hyaff“-11) three-dimensional scaffold was evaluated with a specific ELISA and by immunohistochemical analysis at different experimental times (1hour, 1 day, 7, 14, 21 days) together with the expression of mRNA by Real Time PCR.

Results: Cathepsin B is always secreted by the cells grown onto the biomaterial but the protein levels increased from the first day after seeding up to 7 days (p< 0.01), then decreased progressively and significantly until day 21 (p< 0.01). The immunohistological data confirmed those obtained by the ELISA test. Cathepsin B staining was particularly evident at day 7 after cells were seeded onto the biomaterial, and then progressively decreased up to 21 days; at this experimental time point, the totality of cells were negative. Real-time PCR monitoring with the LightCycler using fluorescent dye allowed rapid and sensitive detection of cathepsin B mRNAs from the patient samples. The mRNA levels increased for up to 7 days of culture and slightly decreased until day 21. However, no significant differences were observed.

Conclusions: We can identify in cathepsin B a soluble marker of differentiated chondrocytes phenotype useful in the monitoring of autologous chondrocyte transplantation performed by means of different carriers. Its low concentration in the constructs culture medium could be indicative of a phenotypic stability. The introduction of mature cells inside the chondral defects could help to regenerate damaged hyaline articular cartilage better and faster.