title: Bioengineered cartilage in a scaffold-free method by human cartilage-derived progenitor cells: a comparison with human adipose-derived mesenchymal stromal cells.
abstract: The objective of our study was to investigate chondrogenesis potential of human adipose-derived mesenchymal stromal cells (MSCs), using as a positive control a human source of cartilage-derived progenitor cells (PCs). This source of PCs was recently described by our group and dwells on the surface of nasoseptal cartilage. Histological analysis using Safranin O staining and immunofluorescence for actin filaments and collagen type II was performed on three-dimensional (3D) pellet cultures. Cartilage PCs and adipose MSCs showed similarities in monolayer culture related to cell morphology and proliferation. Our 3D pellet cultures substantially reduced the actin stress and after 21 days under chondrogenic medium, we observed an increase in the pellet diameter for cartilage PCs (7.4%) and adipose MSCs (21.2%). Adipose-derived MSCs responded to chondrogenic stimulus, as seen by positive areas for collagen type II, but they were not able to recreate a mature extracellular matrix. Using semi-quantitative analysis, we observed a majority of Safranin O areas rising from blue (no stain) to orange (moderate staining) and no changes in fibroblastic morphology (P < 0.0001). For cartilage PCs, chondrogenic induction is responsible for morphological changes and a high percentage of matrix area/number of cells (P ≤ 0.0001), evaluated by computerized histomorphometry. Morphological analyses reveal that adipose-derived MSCs were not able to recreate a bioengineered cartilage. The cost of culture was reduced, as the cartilage PCs under growth-factor free medium exhibit a high score for cartilage formation compared with the induced adipose mesenchymal stromal cells (P = 0.0021). Using a pellet 3D culture, our cartilage PCs were able to produce a cartilage tissue in vitro, leading to the future development of bioengineered products.
authors.affiliation: Federal University of Rio de Janeiro/Xerém, Duque de Caxias, Brazil; Division of Bioengineering, National Institute of Metrology, Quality and Technology (Inmetro), Duque de Caxias, Brazil.
authors.forename: Leandra S
authors.initials: LS
authors.lastname: Baptista
authors.forename: Karina R
authors.initials: KR
authors.lastname: Silva
authors.forename: Carolina S G
authors.initials: CS
authors.lastname: Pedrosa
authors.forename: Ronaldo J F C
authors.initials: RJ
authors.lastname: Amaral
authors.forename: João Vitor
authors.initials: JV
authors.lastname: Belizário
authors.forename: Radovan
authors.initials: R
authors.lastname: Borojevic
authors.forename: José Mauro
authors.initials: JM
authors.lastname: Granjeiro
mesh_terms: D008841:Actin Cytoskeleton / Q000378:metabolism; D000273:Adipose Tissue / Q000166:cytology / Q000378:metabolism*; D000293:Adolescent; D000328:Adult; D002356:Cartilage / Q000166:cytology / Q000378:metabolism*; D049109:Cell Proliferation; D048430:Cell Shape; D002478:Cells, Cultured; D019902:Chondrocytes / Q000378:metabolism*; D020219:Chondrogenesis; D024043:Collagen Type II / Q000378:metabolism; D005260:Female; D006801:Humans; D059630:Mesenchymal Stem Cells / Q000378:metabolism*; D008875:Middle Aged; D010641:Phenotype; D013234:Stem Cells / Q000378:metabolism*; D013997:Time Factors; D023822:Tissue Engineering / Q000379:methods*; D055815:Young Adult
chemical_list: D024043:Collagen Type II
keywords: Bioengineered cartilage; Human adipose mesenchymal stromal cells; Human cartilage progenitor cells