A chondromimetic microsphere for in situ spatially controlled chondrogenic differentiation of human mesenchymal stem cells.
Human mesenchymal stem cells (hMSCs) have been identified as a viable cell source for cartilage tissue engineering. However, to undergo chondrogenic differentiation hMSCs require growth factors, in particular members of the transforming growth factor beta (TGF-β) family. While in vitro differentiation is feasible through continuous supplementation of TGF-β3, mechanisms to control and drive hMSCs down the chondrogenic lineage in their native microenvironment remain a significant challenge. The release of TGF-β3 from an injectable microsphere composed of the cartilage-associated extracellular matrix molecule hyaluronan represents a readily translatable approach for in situ differentiation of hMSCs for cartilage repair. In this study, chondromimetic hyaluronan microspheres were used as a growth factor delivery source for hMSC chondrogenesis. Cellular compatibility of the microspheres (1.2 and 14.1 μm) with hMSCs was shown and release of TGF-β3 from the most promising 14.1 μm microspheres to control differentiation of hMSCs was evaluated. Enhanced accumulation of cartilage-associated glycosaminoglycans by hMSCs incubated with TGF-β3-loaded microspheres was seen and positive staining for collagen type II and proteoglycan confirmed successful in vitro chondrogenesis. Gene expression analysis showed significantly increased expression of the chondrocyte-associated genes, collagen type II and aggrecan. This delivery platform resulted in significantly less collagen type X expression, suggesting the generation of a more stable cartilage phenotype. When evaluated in an ex vivo osteoarthritic cartilage model, implanted hMSCs with TGF-β3-loaded HA microspheres were detected within cartilage fibrillations and increased proteoglycan staining was seen in the tissue. In summary, data presented here demonstrate that TGF-β3-bound hyaluronan microspheres provide a suitable delivery system for induction of hMSC chondrogenesis and their use may represent a clinically feasible tissue engineering approach for the treatment of articular cartilage defects.
Regenerative Medicine Institute (REMEDI), National University of Ireland Galway, Galway, Ireland.
Sharon
S
Ansboro
Regenerative Medicine Institute (REMEDI), National University of Ireland Galway, Galway, Ireland.
Jessica S
JS
Hayes
Regenerative Medicine Institute (REMEDI), National University of Ireland Galway, Galway, Ireland.
Valerie
V
Barron
Network of Excellence for Functional Biomaterials (NFB), National University of Ireland Galway, Galway, Ireland.
Shane
S
Browne
Regenerative Medicine Institute (REMEDI), National University of Ireland Galway, Galway, Ireland.
Linda
L
Howard
Network of Excellence for Functional Biomaterials (NFB), National University of Ireland Galway, Galway, Ireland.
Udo
U
Greiser
The Centre for Microscopy and Imaging (CMI), National University of Ireland Galway, Galway, Ireland.
Pierce
P
Lalor
Discipline of Surgery, Clinical Science Institute, National University of Ireland Galway, Galway, Ireland.
Fintan
F
Shannon
Regenerative Medicine Institute (REMEDI), National University of Ireland Galway, Galway, Ireland.
Frank P
FP
Barry
Network of Excellence for Functional Biomaterials (NFB), National University of Ireland Galway, Galway, Ireland.
Abhay
A
Pandit
Regenerative Medicine Institute (REMEDI), National University of Ireland Galway, Galway, Ireland. Electronic address: mary.murphy@nuigalway.ie.
J Mary
JM
Murphy
D000293:Adolescent; D000328:Adult; D053674:Aggrecans / Q000235:genetics / Q000378:metabolism; D000818:Animals; D032701:Biomimetics; D002454:Cell Differentiation / Q000187:drug effects*; D002460:Cell Line; D019902:Chondrocytes / Q000187:drug effects* / Q000378:metabolism / Q000637:transplantation; D020219:Chondrogenesis / Q000187:drug effects*; D024043:Collagen Type II / Q000235:genetics / Q000378:metabolism; D024121:Collagen Type X / Q000378:metabolism; D004305:Dose-Response Relationship, Drug; D004337:Drug Carriers; D005786:Gene Expression Regulation; D006801:Humans; D006820:Hyaluronic Acid / Q000378:metabolism; D045164:Mesenchymal Stem Cell Transplantation; D059630:Mesenchymal Stem Cells / Q000187:drug effects* / Q000378:metabolism; D008863:Microspheres; D020370:Osteoarthritis, Knee / Q000378:metabolism / Q000473:pathology / Q000628:therapy; D010641:Phenotype; D013997:Time Factors; D023822:Tissue Engineering; D053782:Transforming Growth Factor beta3 / Q000494:pharmacology* / Q000633:toxicity; D055815:Young Adult
D053674:Aggrecans; D024043:Collagen Type II; D024121:Collagen Type X; D004337:Drug Carriers; C509044:TGFB3 protein, human; D053782:Transforming Growth Factor beta3; D006820:Hyaluronic Acid
Chondrogenesis; Delivery vehicle; Mesenchymal stem cell; Microspheres; Osteoarthritis; Tissue engineering