title: Phenol red inhibits chondrogenic differentiation and affects osteogenic differentiation of human mesenchymal stem cells in vitro.
abstract: The purpose with this study was to investigate the effect of phenol red (PR) on chondrogenic and osteogenic differentiation of human mesenchymal stem cells (hMSCs). hMSCs were differentiated into chondrogenic and osteogenic directions in DMEM with and without PR for 2, 7, 14, 21, and 28 days. Gene expression of chondrogenic and osteogenic markers were analyzed by RT-qPCR. The presence of proteoglycans was visualized histologically. Osteogenic matrix deposition and mineralization were examined measuring the alkaline phophatase activity and calcium deposition. During chondrogenic differentiation PR decreased sox9, collagen type 2, aggrecan on day 14 and 21 (P < 0.05), and proteoglycan synthesis on day 21 and 28. Collagen type 10 was decreased on day 21 (P < 0.05). During osteogenic differentiation PR increased alkaline phosphatase on day 7 while decreased on day 21 (P < 0.05). PR increased collagen type 1 on day 7, 14, and day 21 (P < 0.05). The alkaline phosphatase activity was increased after 2, 7, and 14 days (P < 0.05). The deposition of calcium was decreased on day 21 (P < 0.05). Our results indicate that PR should be removed from the culture media when differentiating hMSCs into chondrogenic and osteogenic directions due to the effects on these differentiation pathways.
authors.affiliation: Orthopaedic Research Laboratory, Aarhus University Hospital, Nørrebrogade 44, building 1A, 8000, Aarhus C, Denmark. helle.lysdahl@ki.au.dk
authors.forename: Helle
authors.initials: H
authors.lastname: Lysdahl
authors.forename: Anette
authors.initials: A
authors.lastname: Baatrup
authors.forename: Anna Bay
authors.initials: AB
authors.lastname: Nielsen
authors.forename: Casper Bindzus
authors.initials: CB
authors.lastname: Foldager
authors.forename: Cody
authors.initials: C
authors.lastname: Bünger
mesh_terms: D053674:Aggrecans / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism; D000469:Alkaline Phosphatase / Q000378:metabolism; D002118:Calcium / Q000378:metabolism; D002454:Cell Differentiation; D002478:Cells, Cultured; D019902:Chondrocytes / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D024043:Collagen Type II / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism; D024121:Collagen Type X / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism; D003470:Culture Media / Q000737:chemistry / Q000494:pharmacology*; D015870:Gene Expression / Q000187:drug effects; D006801:Humans; D008297:Male; D059630:Mesenchymal Stem Cells / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D010011:Osteocytes / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D010637:Phenolsulfonphthalein / Q000494:pharmacology*; D011509:Proteoglycans / Q000096:biosynthesis; D055758:SOX9 Transcription Factor / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism; D055815:Young Adult
chemical_list: D053674:Aggrecans; D024043:Collagen Type II; D024121:Collagen Type X; D003470:Culture Media; D011509:Proteoglycans; D055758:SOX9 Transcription Factor; C527932:SOX9 protein, human; D000469:Alkaline Phosphatase; D010637:Phenolsulfonphthalein; D002118:Calcium