title: Comparing effects of perfusion and hydrostatic pressure on gene profiles of human chondrocyte.
abstract: Hydrostatic pressure and perfusion have been shown to regulate the chondrogenic potential of articular chondrocytes. In order to compare the effects of hydrostatic pressure plus perfusion (HPP) and perfusion (P) we investigated the complete gene expression profiles of human chondrocytes under HPP and P. A simplified bioreactor was constructed to apply loading (0.1 MPa for 2 h) and perfusion (2 ml) through the same piping by pressurizing the medium directly. High-density monolayer cultures of human chondrocytes were exposed to HPP or P for 4 days. Controls (C) were maintained in static cultures. Gene expression was evaluated by sequencing (RNAseq) and quantitative real-time PCR analysis. Both treatments changed gene expression levels of human chondrocytes significantly. Specifically, HPP and P increased COL2A1 expression and decreased COL1A1 and MMP-13 expression. Despite of these similarities, RNAseq revealed a list of cartilage genes including ACAN, ITGA10 and TNC, which were differentially expressed by HPP and P. Of these candidates, adhesion related molecules were found to be upregulated in HPP. Both HPP and P treatment had beneficial effects on chondrocyte differentiation and decreased catabolic enzyme expression. The study provides new insight into how hydrostatic pressure and perfusion enhance cartilage differentiation and inhibit catabolic effects.
authors.affiliation: Department of Orthopaedic Surgery, Physical Medicine and Rehabilitation, Campus Großhadern, Ludwig-Maximilians-University, Marchioninistr. 15, 81377 Munich, Germany.
authors.forename: Ge
authors.initials: G
authors.lastname: Zhu
authors.affiliation: Department of Orthopaedic Surgery, Physical Medicine and Rehabilitation, Campus Großhadern, Ludwig-Maximilians-University, Marchioninistr. 15, 81377 Munich, Germany. Electronic address: susanne.mayer@med.uni-muenchen.de.
authors.forename: Susanne
authors.initials: S
authors.lastname: Mayer-Wagner
authors.affiliation: Department of Orthopaedic Surgery, Physical Medicine and Rehabilitation, Campus Großhadern, Ludwig-Maximilians-University, Marchioninistr. 15, 81377 Munich, Germany.
authors.forename: Christian
authors.initials: C
authors.lastname: Schröder
authors.affiliation: Department of Orthopaedic Surgery, Physical Medicine and Rehabilitation, Campus Großhadern, Ludwig-Maximilians-University, Marchioninistr. 15, 81377 Munich, Germany.
authors.forename: Matthias
authors.initials: M
authors.lastname: Woiczinski
authors.affiliation: LAFUGA Genomics, Gene Center Munich, Ludwig-Maximilians-University, Feodor-Lynen-Str. 25, 81377 Munich, Germany.
authors.forename: Helmut
authors.initials: H
authors.lastname: Blum
authors.affiliation: LAFUGA Genomics, Gene Center Munich, Ludwig-Maximilians-University, Feodor-Lynen-Str. 25, 81377 Munich, Germany.
authors.forename: Ilaria
authors.initials: I
authors.lastname: Lavagi
authors.affiliation: LAFUGA Genomics, Gene Center Munich, Ludwig-Maximilians-University, Feodor-Lynen-Str. 25, 81377 Munich, Germany.
authors.forename: Stefan
authors.initials: S
authors.lastname: Krebs
authors.affiliation: Department of Orthopaedic Surgery, Physical Medicine and Rehabilitation, Campus Großhadern, Ludwig-Maximilians-University, Marchioninistr. 15, 81377 Munich, Germany.
authors.forename: Julia I
authors.initials: JI
authors.lastname: Redeker
authors.affiliation: Department of Orthopaedic Surgery, Physical Medicine and Rehabilitation, Campus Großhadern, Ludwig-Maximilians-University, Marchioninistr. 15, 81377 Munich, Germany.
authors.forename: Andreas
authors.initials: A
authors.lastname: Hölzer
authors.affiliation: Department of Orthopaedic Surgery, Physical Medicine and Rehabilitation, Campus Großhadern, Ludwig-Maximilians-University, Marchioninistr. 15, 81377 Munich, Germany.
authors.forename: Volkmar
authors.initials: V
authors.lastname: Jansson
authors.affiliation: Department of Orthopaedic Surgery, Physical Medicine and Rehabilitation, Campus Großhadern, Ludwig-Maximilians-University, Marchioninistr. 15, 81377 Munich, Germany.
authors.forename: Oliver
authors.initials: O
authors.lastname: Betz
authors.affiliation: Department of Orthopaedic Surgery, Physical Medicine and Rehabilitation, Campus Großhadern, Ludwig-Maximilians-University, Marchioninistr. 15, 81377 Munich, Germany.
authors.forename: Peter E
authors.initials: PE
authors.lastname: Müller
mesh_terms: D000293:Adolescent; D019149:Bioreactors; D002454:Cell Differentiation; D002478:Cells, Cultured; D019902:Chondrocytes / Q000166:cytology*; D015870:Gene Expression; D020869:Gene Expression Profiling / Q000379:methods*; D005786:Gene Expression Regulation; D006801:Humans; D006874:Hydrostatic Pressure; D008297:Male; D010477:Perfusion / Q000295:instrumentation; D017423:Sequence Analysis, RNA / Q000379:methods*
keywords: Bioreactor; Human chondrocytes; Hydrostatic pressure; Perfusion; Sequencing