title: Poly(γ-Glutamic Acid) as an Exogenous Promoter of Chondrogenic Differentiation of Human Mesenchymal Stem/Stromal Cells.
abstract: Cartilage damage and/or aging effects can cause constant pain, which limits the patient's quality of life. Although different strategies have been proposed to enhance the limited regenerative capacity of cartilage tissue, the full production of native and functional cartilaginous extracellular matrix (ECM) has not yet been achieved. Poly(γ-glutamic acid) (γ-PGA), a naturally occurring polyamino acid, biodegradable into glutamate residues, has been explored for tissue regeneration. In this work, γ-PGA's ability to support the production of cartilaginous ECM by human bone marrow mesenchymal stem/stromal cells (MSCs) and nasal chondrocytes (NCs) was investigated. MSC and NC pellets were cultured in basal medium (BM), chondrogenic medium (CM), and CM-γ-PGA-supplemented medium (CM+γ-PGA) over a period of 21 days. Pellet size/shape was monitored with time. At 14 and 21 days of culture, the presence of sulfated glycosaminoglycans (sGAGs), type II collagen (Col II), Sox-9, aggrecan, type XI collagen (Col XI), type X collagen (Col X), calcium deposits, and type I collagen (Col I) was analyzed. After excluding γ-PGA's cytotoxicity, earlier cell condensation, higher sGAG content, Col II, Sox-9 (day 14), aggrecan, and Col X (day 14) production was observed in γ-PGA-supplemented MSC cultures, with no signs of mineralization or Col I. These effects were not evident with NCs. However, Sox-9 (at day 14) and Col X (at days 14 and 21) were increased, decreased, or absent, respectively. Overall, γ-PGA improved chondrogenic differentiation of MSCs, increasing ECM production earlier in culture. It is proposed that γ-PGA incorporation in novel biomaterials has a beneficial impact on future approaches for cartilage regeneration.
authors.affiliation: 1Instituto de Investigação e Inovação em Saúde, Universidade do Porto, Porto, Portugal.|2INEB-Instituto de Engenharia Biomédica, Universidade do Porto, Porto, Portugal.|3Faculdade de Engenharia, Universidade do Porto, Porto, Portugal.
authors.forename: Joana C
authors.initials: JC
authors.lastname: Antunes
authors.affiliation: 3Faculdade de Engenharia, Universidade do Porto, Porto, Portugal.|4Max Planck Institute for Molecular Biomedicine, Münster, Germany.
authors.forename: Roman
authors.initials: R
authors.lastname: Tsaryk
authors.affiliation: 1Instituto de Investigação e Inovação em Saúde, Universidade do Porto, Porto, Portugal.|2INEB-Instituto de Engenharia Biomédica, Universidade do Porto, Porto, Portugal.
authors.forename: Raquel M
authors.initials: RM
authors.lastname: Gonçalves
authors.affiliation: 1Instituto de Investigação e Inovação em Saúde, Universidade do Porto, Porto, Portugal.|2INEB-Instituto de Engenharia Biomédica, Universidade do Porto, Porto, Portugal.|5ICBAS-Instituto de Ciências Biomédicas Abel Salazar, Universidade do Porto, Porto, Portugal.
authors.forename: Catarina Leite
authors.initials: CL
authors.lastname: Pereira
authors.affiliation: 6Department of Oral, Cranio-Maxillofacial and Facial Plastic Surgery, University Medical Center of the Goethe University, Frankfurt am Main, Germany.|7Sana Hospital Offenbach, Offenbach, Germany.
authors.forename: Constantin
authors.initials: C
authors.lastname: Landes
authors.affiliation: 8REPAIR Lab, Institute of Pathology, University Medical Center of the Johannes Gutenberg University, Mainz, Germany.
authors.forename: Christoph
authors.initials: C
authors.lastname: Brochhausen
authors.affiliation: 6Department of Oral, Cranio-Maxillofacial and Facial Plastic Surgery, University Medical Center of the Goethe University, Frankfurt am Main, Germany.|7Sana Hospital Offenbach, Offenbach, Germany.|8REPAIR Lab, Institute of Pathology, University Medical Center of the Johannes Gutenberg University, Mainz, Germany.
authors.forename: Shahram
authors.initials: S
authors.lastname: Ghanaati
authors.affiliation: 1Instituto de Investigação e Inovação em Saúde, Universidade do Porto, Porto, Portugal.|2INEB-Instituto de Engenharia Biomédica, Universidade do Porto, Porto, Portugal.|5ICBAS-Instituto de Ciências Biomédicas Abel Salazar, Universidade do Porto, Porto, Portugal.
authors.forename: Mário A
authors.initials: MA
authors.lastname: Barbosa
authors.affiliation: 8REPAIR Lab, Institute of Pathology, University Medical Center of the Johannes Gutenberg University, Mainz, Germany.
authors.forename: C James
authors.initials: CJ
authors.lastname: Kirkpatrick
mesh_terms: D053674:Aggrecans / Q000378:metabolism; D002118:Calcium / Q000378:metabolism; D018929:Cell Culture Techniques; D002478:Cells, Cultured; D019902:Chondrocytes / Q000166:cytology / Q000187:drug effects*; D020219:Chondrogenesis / Q000187:drug effects*; D003094:Collagen / Q000378:metabolism; D003470:Culture Media / Q000494:pharmacology; D017077:Culture Media, Conditioned / Q000737:chemistry; D016326:Extracellular Matrix Proteins / Q000378:metabolism; D006025:Glycosaminoglycans / Q000378:metabolism; D006801:Humans; D059630:Mesenchymal Stem Cells / Q000166:cytology / Q000187:drug effects*; D055171:Nasal Cartilages / Q000166:cytology; D011099:Polyglutamic Acid / Q000031:analogs & derivatives* / Q000494:pharmacology / Q000633:toxicity; D055758:SOX9 Transcription Factor / Q000378:metabolism; D017154:Stromal Cells / Q000166:cytology / Q000187:drug effects*; D053773:Transforming Growth Factor beta1 / Q000378:metabolism
chemical_list: C508319:ACAN protein, human; D053674:Aggrecans; D003470:Culture Media; D017077:Culture Media, Conditioned; D016326:Extracellular Matrix Proteins; D006025:Glycosaminoglycans; D055758:SOX9 Transcription Factor; C527932:SOX9 protein, human; D053773:Transforming Growth Factor beta1; C511775:poly(gamma-glutamic acid); D011099:Polyglutamic Acid; C013786:A73025; D003094:Collagen; D002118:Calcium