PMID- 22536910
OWN - NLM
STAT- MEDLINE
DCOM- 20130116
LR  - 20211021
IS  - 1937-335X (Electronic)
IS  - 1937-3341 (Print)
IS  - 1937-3341 (Linking)
VI  - 18
IP  - 17-18
DP  - 2012 Sep
TI  - Effects of perfusion and dynamic loading on human neocartilage formation in 
      alginate hydrogels.
PG  - 1784-92
LID - 10.1089/ten.TEA.2011.0506 [doi]
AB  - Dynamic loading and perfusion culture environments alone are known to enhance 
      cartilage extracellular matrix (ECM) production in dedifferentiated articular 
      chondrocytes. In this study, we explored whether a combination of these factors 
      would enhance these processes over a free-swelling (FS) condition using adult 
      human articular chondrocytes embedded in 2% alginate. The alginate constructs 
      were placed into a bioreactor for perfusion (P) only (100 muL/per minute) or 
      perfusion and dynamic compressive loading (PL) culture (20% for 1 h, at 0.5 Hz), 
      each day. Control FS alginate gels were maintained in six-well static culture. 
      Gene expression analysis was conducted on days 7 and 14, while cell viability, 
      immunostaining, and mechanical property testing were performed on day 14 only. 
      Total glycosaminoglycan (GAG) content and GAG synthesis were assessed after 14 
      days. Col2a1 mRNA expression levels were significantly higher (at least 
      threefold; p<0.05) in both bioreactor conditions compared with FS by days 7 and 
      14. For all gene studies, no significant differences were seen between P and PL 
      treatments. Aggrecan mRNA levels were not significantly altered in any condition 
      although both GAG/DNA and (35)S GAG incorporation studies indicated higher GAG 
      retention and synthesis in the FS treatment. Collagen type II protein deposition 
      was low in all samples, link protein distribution was more diffuse in FS 
      condition, and aggrecan deposition was located in the outer regions of the 
      alginate constructs in both bioreactor conditions, yet more uniformly in the FS 
      condition. Catabolic gene expression (matrix metalloproteinase 3 [MMP3] and 
      inducible nitric oxide synthase [iNOS]) was higher in bioreactor conditions 
      compared with FS, although iNOS expression levels decreased to approximately 
      fourfold less than the FS condition by day 14. Our data indicate that conditions 
      created in the bioreactor enhanced both anabolic and catabolic responses, similar 
      to other loading studies. Perfusion was sufficient alone to promote this dual 
      response. PL increased the deposition of aggrecan surrounding cells compared with 
      the other conditions; however, overall low GAG retention in the bioreactor system 
      was likely due to both perfusion and catabolic conditions created. Optimal 
      conditions, which permit appropriate anabolic and catabolic processes for 
      accumulation of ECM and tissue remodeling for neocartilage development, 
      specifically for humans, are needed.
FAU - Grogan, Shawn P
AU  - Grogan SP
AD  - Shiley Center for Orthopaedic Research and Education at Scripps Clinic, La Jolla, 
      CA 92037, USA.
FAU - Sovani, Sujata
AU  - Sovani S
FAU - Pauli, Chantal
AU  - Pauli C
FAU - Chen, Jianfen
AU  - Chen J
FAU - Hartmann, Andreas
AU  - Hartmann A
FAU - Colwell, Clifford W Jr
AU  - Colwell CW Jr
FAU - Lotz, Martin K
AU  - Lotz MK
FAU - D'Lima, Darryl D
AU  - D'Lima DD
LA  - eng
GR  - P01 AG007996/AG/NIA NIH HHS/United States
GR  - UL1 RR025774/RR/NCRR NIH HHS/United States
PT  - Journal Article
PT  - Research Support, N.I.H., Extramural
PT  - Research Support, Non-U.S. Gov't
DEP - 20120612
PL  - United States
TA  - Tissue Eng Part A
JT  - Tissue engineering. Part A
JID - 101466659
RN  - 0 (Alginates)
RN  - 0 (Glycosaminoglycans)
RN  - 0 (Hexuronic Acids)
RN  - 0 (Hydrogels)
RN  - 8A5D83Q4RW (Glucuronic Acid)
RN  - 9007-49-2 (DNA)
SB  - IM
MH  - Adolescent
MH  - Adult
MH  - Alginates/*pharmacology
MH  - Bioreactors
MH  - Cartilage, Articular/cytology/*physiology
MH  - Cell Survival/drug effects/genetics
MH  - Cells, Cultured
MH  - Chondrocytes/cytology/drug effects/metabolism
MH  - Chondrogenesis/*drug effects
MH  - DNA/metabolism
MH  - Female
MH  - Gene Expression Profiling
MH  - Gene Expression Regulation/drug effects
MH  - Glucuronic Acid/pharmacology
MH  - Glycosaminoglycans/metabolism
MH  - Hexuronic Acids/pharmacology
MH  - Humans
MH  - Hydrogels/*pharmacology
MH  - Immunohistochemistry
MH  - Male
MH  - Middle Aged
MH  - Perfusion/*methods
MH  - Weight-Bearing/physiology
MH  - Young Adult
PMC - PMC3432904
EDAT- 2012/04/28 06:00
MHDA- 2013/01/17 06:00
PMCR- 2013/09/01
CRDT- 2012/04/28 06:00
PHST- 2012/04/28 06:00 [entrez]
PHST- 2012/04/28 06:00 [pubmed]
PHST- 2013/01/17 06:00 [medline]
PHST- 2013/09/01 00:00 [pmc-release]
AID - 10.1089/ten.tea.2011.0506 [pii]
AID - 10.1089/ten.TEA.2011.0506 [doi]
PST - ppublish
SO  - Tissue Eng Part A. 2012 Sep;18(17-18):1784-92. doi: 10.1089/ten.TEA.2011.0506. 
      Epub 2012 Jun 12.