C. Gentili (Genoa, IT)
University of Genoa Department of Experimental MedicinePresenter Of 2 Presentations
12.2.4 - Extracellular Vesicles Derived From Mesenchymal Stromal Cells Cultured in a Clinical-Grade Edium Promote Human Cartilage Recovery.
Abstract
Purpose
Osteoarthritis (OA) is a joint disorder causing articular cartilage degeneration. Currently, treatments are mainly pain- and symptom-modifying, rather than disease-modifying. Human bone marrow stromal cells (hBMSCs) have emerged as a promising paracrine mechanism-based approach for the treatment of OA. Many studies demonstrate that MSCs attend to tissue repair through the secretion of trophic factors or extracellular vesicles (EVs). We developed a “donor-to-patient” system for aseptic therapeutic cell manufacturing using a xeno-free medium. We validated the potential therapeutic benefits of secreted EVs isolated from BMSC culture in this innovative culture system, for cartilage repair.
Methods and Materials
We characterized (EVs) derived from hBMSCs, grown in a xeno-free culture system (XFS) compared to a conventional fetal bovine serum (FBS) culture system, in normoxic and hypoxic culture setting. We investigated also the therapeutic potential of EVs in an in vitro model of OA. We characterized the miRNA content of EVs in different culture setting to select putative miRNA that could be involved in a biological function.
Results
The biological effects of XFS- and FBS-cultured hBMSC-derived EVs were tested on IL-1α treated hACs in an experiment designed to mimic the OA environment. We observed that under inflammatory conditions hACs are able to recruit and internalize more MSC-derived EVs, especially those derived from cells cultured in our XFS system and in hypoxic conditions. XFS-EVs both in normoxia and hypoxia shows anti-inflammatory properties in an in vitro OA model. Analysis of miRNA content showed upregulation in XFS-hBMSC-derived EVs of miRNAs known to have a chondroprotective role, and also appear to be involved in cartilage homeostasis and influence TGF-beta signaling.
Conclusion
XFS medium was found to be suitable for isolation and expansion of hBMSCs with increased production of EVs. The application of these EVs overcomes the safety concerns associated with serum-containing media and makes ready-to-use clinical cartilage therapies more accessible.
P051 - Human articular chondrocyte re-differentiation in 3D-printed Platelet Rich Plasma (PRP)-alginate-based bioink
Abstract
Purpose
Tissue engineering strategies for the clinical treatment of cartilage defects are mainly based on the transplantation of expanded autologous chondrocytes in vitro into the patient's damaged tissue. However, the differentiation of chondrocytes during in vitro expansion often leads to suboptimal results in the therapeutic intervention. Three-dimensional (3D) culture systems can restore this de-differentiated state and re-establish the chondrogenic phenotype. In this scenario, 3D bioprinting is a suitable technology for creating patient-shaped grafts from cell-laden bio-ink by layer-by-layer fabrication. This allows for a more physiological environment for transplanted cells, promoting tissue regeneration and repair.
Methods and Materials
Human joint chondrocytes (hACs) were isolated from patient tissue biopsies and expanded in vitro in a monolayer (2D culture). 3D bioprinting was performed by incorporating expanded hACs in mixed bioinks of platelet-rich plasma (PRP) alginate or alginate. Cell morphology, viability, growth, and chondrogenic differentiation were studied in vitro in both types of printed constructs.
Results
3D culture in both inks significantly increased the expression of chondrogenic markers compared to the 2D condition. Furthermore, the addition of PRP to the alginate upregulated the expression of these markers. Functionalization of PRP ink also increased the vitality, growth, and metabolic activity of hACs compared to alginate alone.
Conclusion
3D bioprinting of hAC allowed them to preserve the chondrogenic phenotype, furthermore, the use of bioink integrated with PRP supported the maintenance of metabolically active articular cartilage collagen type II positive. In view of future clinical translation, the choice of cell source for the 3D bioprinting of patient-specific grafts will have to be oriented towards cell types with a higher potential, such as chondro-progenitors, than mature hACs in order to ensure a more efficient regeneration and repair of cartilage.
Presenter Of 1 Presentation
P051 - Human articular chondrocyte re-differentiation in 3D-printed Platelet Rich Plasma (PRP)-alginate-based bioink
Abstract
Purpose
Tissue engineering strategies for the clinical treatment of cartilage defects are mainly based on the transplantation of expanded autologous chondrocytes in vitro into the patient's damaged tissue. However, the differentiation of chondrocytes during in vitro expansion often leads to suboptimal results in the therapeutic intervention. Three-dimensional (3D) culture systems can restore this de-differentiated state and re-establish the chondrogenic phenotype. In this scenario, 3D bioprinting is a suitable technology for creating patient-shaped grafts from cell-laden bio-ink by layer-by-layer fabrication. This allows for a more physiological environment for transplanted cells, promoting tissue regeneration and repair.
Methods and Materials
Human joint chondrocytes (hACs) were isolated from patient tissue biopsies and expanded in vitro in a monolayer (2D culture). 3D bioprinting was performed by incorporating expanded hACs in mixed bioinks of platelet-rich plasma (PRP) alginate or alginate. Cell morphology, viability, growth, and chondrogenic differentiation were studied in vitro in both types of printed constructs.
Results
3D culture in both inks significantly increased the expression of chondrogenic markers compared to the 2D condition. Furthermore, the addition of PRP to the alginate upregulated the expression of these markers. Functionalization of PRP ink also increased the vitality, growth, and metabolic activity of hACs compared to alginate alone.
Conclusion
3D bioprinting of hAC allowed them to preserve the chondrogenic phenotype, furthermore, the use of bioink integrated with PRP supported the maintenance of metabolically active articular cartilage collagen type II positive. In view of future clinical translation, the choice of cell source for the 3D bioprinting of patient-specific grafts will have to be oriented towards cell types with a higher potential, such as chondro-progenitors, than mature hACs in order to ensure a more efficient regeneration and repair of cartilage.