Successful Low-Cost Scaffold-Free Cartilage Tissue Engineering Using Human Cartilage Progenitor Cell Spheroids Formed by Micromolded Nonadhesive Hydrogel

The scaffold-free tissue engineering using spheroids is pointed out as an approach for optimizing the delivery system of cartilage construct. In this study, we aimed to evaluate the micromolded nonadhesive hydrogel (MicroTissues®) for spheroid compaction (2-day culture) and spontaneous chondrogenesi...

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Main Authors: Mellannie P. Stuart, Renata A. M. Matsui, Matheus F. S. Santos, Isis Côrtes, Mayra S. Azevedo, Karina R. Silva, Anderson Beatrici, Paulo Emílio C. Leite, Priscila Falagan-Lotsch, José M. Granjeiro, Vladimir Mironov, Leandra S. Baptista
Format: Article
Language:English
Published: Wiley 2017-01-01
Series:Stem Cells International
Online Access:http://dx.doi.org/10.1155/2017/7053465
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author Mellannie P. Stuart
Renata A. M. Matsui
Matheus F. S. Santos
Isis Côrtes
Mayra S. Azevedo
Karina R. Silva
Anderson Beatrici
Paulo Emílio C. Leite
Priscila Falagan-Lotsch
José M. Granjeiro
Vladimir Mironov
Leandra S. Baptista
author_facet Mellannie P. Stuart
Renata A. M. Matsui
Matheus F. S. Santos
Isis Côrtes
Mayra S. Azevedo
Karina R. Silva
Anderson Beatrici
Paulo Emílio C. Leite
Priscila Falagan-Lotsch
José M. Granjeiro
Vladimir Mironov
Leandra S. Baptista
author_sort Mellannie P. Stuart
collection DOAJ
description The scaffold-free tissue engineering using spheroids is pointed out as an approach for optimizing the delivery system of cartilage construct. In this study, we aimed to evaluate the micromolded nonadhesive hydrogel (MicroTissues®) for spheroid compaction (2-day culture) and spontaneous chondrogenesis (21-day culture) using cartilage progenitors cells (CPCs) from human nasal septum without chondrogenic stimulus. CPC spheroids showed diameter stability (486 μm ± 65), high percentage of viable cells (88.1 ± 2.1), and low percentage of apoptotic cells (2.3%). After spheroid compaction, the synthesis of TGF-β1, TGF-β2, and TGF-β3 was significantly higher compared to monolayer (p<0.005). Biomechanical assay revealed that the maximum forces applied to spheroids after chondrogenesis were 2.6 times higher than for those cultured for 2 days. After spontaneous chondrogenesis, CPC spheroids were entirely positive for N-cadherin, collagen type II and type VI, and aggrecan and chondroitin sulfate. Comparing to monolayer, the expression of SOX5 and SOX6 genes analyzed by qPCR was significantly upregulated (p<0.01). Finally, we observed the capacity of CPC spheroids starting to fuse. To the best of our knowledge, this is the first time in the scientific literature that human CPC spheroids were formed by micromolded nonadhesive hydrogel, achieving a successful scaffold-free cartilage engineering without chondrogenic stimulus (low cost).
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spelling doaj-art-4c0fe5efe55c4720b1ca9b1ee17e15b32025-08-20T03:36:31ZengWileyStem Cells International1687-966X1687-96782017-01-01201710.1155/2017/70534657053465Successful Low-Cost Scaffold-Free Cartilage Tissue Engineering Using Human Cartilage Progenitor Cell Spheroids Formed by Micromolded Nonadhesive HydrogelMellannie P. Stuart0Renata A. M. Matsui1Matheus F. S. Santos2Isis Côrtes3Mayra S. Azevedo4Karina R. Silva5Anderson Beatrici6Paulo Emílio C. Leite7Priscila Falagan-Lotsch8José M. Granjeiro9Vladimir Mironov10Leandra S. Baptista11Laboratório de Bioengenharia Tecidual, Instituto Nacional de Metrologia, Qualidade e Tecnologia (Inmetro), Duque de Caxias, RJ, BrazilLaboratório de Bioengenharia Tecidual, Instituto Nacional de Metrologia, Qualidade e Tecnologia (Inmetro), Duque de Caxias, RJ, BrazilLaboratório de Bioengenharia Tecidual, Instituto Nacional de Metrologia, Qualidade e Tecnologia (Inmetro), Duque de Caxias, RJ, BrazilLaboratório de Bioengenharia Tecidual, Instituto Nacional de Metrologia, Qualidade e Tecnologia (Inmetro), Duque de Caxias, RJ, BrazilLaboratório de Bioengenharia Tecidual, Instituto Nacional de Metrologia, Qualidade e Tecnologia (Inmetro), Duque de Caxias, RJ, BrazilLaboratório de Bioengenharia Tecidual, Instituto Nacional de Metrologia, Qualidade e Tecnologia (Inmetro), Duque de Caxias, RJ, BrazilPrograma de Pós-graduação em Biotecnologia, Instituto Nacional de Metrologia, Qualidade e Tecnologia (Inmetro), Duque de Caxias, RJ, BrazilLaboratório de Bioengenharia Tecidual, Instituto Nacional de Metrologia, Qualidade e Tecnologia (Inmetro), Duque de Caxias, RJ, BrazilLaboratório de Bioengenharia Tecidual, Instituto Nacional de Metrologia, Qualidade e Tecnologia (Inmetro), Duque de Caxias, RJ, BrazilLaboratório de Bioengenharia Tecidual, Instituto Nacional de Metrologia, Qualidade e Tecnologia (Inmetro), Duque de Caxias, RJ, BrazilLaboratório de Bioengenharia Tecidual, Instituto Nacional de Metrologia, Qualidade e Tecnologia (Inmetro), Duque de Caxias, RJ, BrazilLaboratório de Bioengenharia Tecidual, Instituto Nacional de Metrologia, Qualidade e Tecnologia (Inmetro), Duque de Caxias, RJ, BrazilThe scaffold-free tissue engineering using spheroids is pointed out as an approach for optimizing the delivery system of cartilage construct. In this study, we aimed to evaluate the micromolded nonadhesive hydrogel (MicroTissues®) for spheroid compaction (2-day culture) and spontaneous chondrogenesis (21-day culture) using cartilage progenitors cells (CPCs) from human nasal septum without chondrogenic stimulus. CPC spheroids showed diameter stability (486 μm ± 65), high percentage of viable cells (88.1 ± 2.1), and low percentage of apoptotic cells (2.3%). After spheroid compaction, the synthesis of TGF-β1, TGF-β2, and TGF-β3 was significantly higher compared to monolayer (p<0.005). Biomechanical assay revealed that the maximum forces applied to spheroids after chondrogenesis were 2.6 times higher than for those cultured for 2 days. After spontaneous chondrogenesis, CPC spheroids were entirely positive for N-cadherin, collagen type II and type VI, and aggrecan and chondroitin sulfate. Comparing to monolayer, the expression of SOX5 and SOX6 genes analyzed by qPCR was significantly upregulated (p<0.01). Finally, we observed the capacity of CPC spheroids starting to fuse. To the best of our knowledge, this is the first time in the scientific literature that human CPC spheroids were formed by micromolded nonadhesive hydrogel, achieving a successful scaffold-free cartilage engineering without chondrogenic stimulus (low cost).http://dx.doi.org/10.1155/2017/7053465
spellingShingle Mellannie P. Stuart
Renata A. M. Matsui
Matheus F. S. Santos
Isis Côrtes
Mayra S. Azevedo
Karina R. Silva
Anderson Beatrici
Paulo Emílio C. Leite
Priscila Falagan-Lotsch
José M. Granjeiro
Vladimir Mironov
Leandra S. Baptista
Successful Low-Cost Scaffold-Free Cartilage Tissue Engineering Using Human Cartilage Progenitor Cell Spheroids Formed by Micromolded Nonadhesive Hydrogel
Stem Cells International
title Successful Low-Cost Scaffold-Free Cartilage Tissue Engineering Using Human Cartilage Progenitor Cell Spheroids Formed by Micromolded Nonadhesive Hydrogel
title_full Successful Low-Cost Scaffold-Free Cartilage Tissue Engineering Using Human Cartilage Progenitor Cell Spheroids Formed by Micromolded Nonadhesive Hydrogel
title_fullStr Successful Low-Cost Scaffold-Free Cartilage Tissue Engineering Using Human Cartilage Progenitor Cell Spheroids Formed by Micromolded Nonadhesive Hydrogel
title_full_unstemmed Successful Low-Cost Scaffold-Free Cartilage Tissue Engineering Using Human Cartilage Progenitor Cell Spheroids Formed by Micromolded Nonadhesive Hydrogel
title_short Successful Low-Cost Scaffold-Free Cartilage Tissue Engineering Using Human Cartilage Progenitor Cell Spheroids Formed by Micromolded Nonadhesive Hydrogel
title_sort successful low cost scaffold free cartilage tissue engineering using human cartilage progenitor cell spheroids formed by micromolded nonadhesive hydrogel
url http://dx.doi.org/10.1155/2017/7053465
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