Hyaluronan influence on the onset of chondrogenic differentiation of mesenchymal stem cells.
OBJECTIVE
Hyaluronan (HA) is an abundant component of chondrogenic tissue hence it is often used as a fundamental constituent in cartilage tissue substitutes. However, effects of different molecular weight HA on chondrogenic differentiation are not clear. The aim was to evaluate modulation of mesenchymal stem cell (MSC) early chondrogenesis by HA of molecular weights 100, 600 and 1 500 kDa.


METHODS
HA was applied on MSCs cultured in a pellet system for one, two and three weeks. Chondrogenesis was evaluated by determinations of gene expression of transcription factor Sox-9 and extracellular matrix proteins collagen type II and XI, aggrecan, and COMP by Real-Time PCR and completed with histological analysis.


RESULTS
Upon chondrogenic induction, the respective pellets revealed active transcription of the chondrogenic genes together with proceeding accumulation of glycosaminoglycan (GAG) rich extracellular matrix. Sox-9 was also expressed in non-chondrogenic MSC controls. HA treated pellets were not significantly influenced on day 7 of culture. However, on day 14, lowered expression in some of the extracellular matrix proteins appeared together with a moderately smaller amount of GAG content in pellet sections. Nevertheless, the analysis on day 21 has demonstrated that HA did not affect the outcome of the differentiation by the end of the culture. Any difference regarding the molecular weight of the HA was not found.


CONCLUSIONS
It could be speculated that HA induced a time shift in the phase of the dominant matrix protein onset which was in full compensated by the end of the evaluated time period. Thus, data suggest that HA of any tested molecular weight does not significantly modulate chondrogenesis of MSCs in pellet system.
Dvorakova
Jana
J
CPN s.r.o, Dolní Dobrouc, Czech Republic. dvorakova@contipro.cz
Velebny
Vladimir
V
Kubala
Lukas
L
D002356:Cartilage / Q000166:cytology / Q000378:metabolism; D002454:Cell Differentiation / Q000187:drug effects*; D019902:Chondrocytes / Q000187:drug effects*; D024043:Collagen Type II / Q000096:biosynthesis / Q000235:genetics; D024161:Collagen Type XI / Q000096:biosynthesis / Q000235:genetics; D005109:Extracellular Matrix / Q000187:drug effects / Q000378:metabolism; D015870:Gene Expression / Q000187:drug effects; D006025:Glycosaminoglycans / Q000378:metabolism; D006801:Humans; D006820:Hyaluronic Acid / Q000737:chemistry / Q000494:pharmacology*; D059630:Mesenchymal Stem Cells / Q000187:drug effects*; D008970:Molecular Weight; D012333:RNA, Messenger / Q000096:biosynthesis / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction; D055758:SOX9 Transcription Factor / Q000096:biosynthesis / Q000235:genetics
D024043:Collagen Type II; D024161:Collagen Type XI; D006025:Glycosaminoglycans; D012333:RNA, Messenger; D055758:SOX9 Transcription Factor; D006820:Hyaluronic Acid