Sodium alginate sponges with or without sodium hyaluronate: in vitro engineering of cartilage.
Studies are underway to design biosystems containing embedded chondrocytes to fill osteochondral defects and to produce a tissue close to native cartilage. In the present report, a new alginate three-dimensional support for chondrocyte culture is described. A sodium alginate solution, with or without hyaluronic acid (HA), was freeze-dried to obtain large-porosity sponges. This formulation was compared with a hydrogel of the same composition. In the sponge formulation, macroscopic and microscopic studies demonstrated the formation of a macroporous network (average pore size, 174 microm) associated with a microporous one (average pore size, 250 nm). Histological and biochemical studies showed that, when loaded with HA, the sponge provides an adapted environment for proteoglycan and collagen synthesis by chondrocytes. Cytoskeleton organization was studied by three-dimensional fluorescence microscopy (CellScan EPR). Chondrocytes exhibit a marked spherical shape with a nonoriented and sparse actin microfilament network. Type II collagen was detected in both types of sponges (with or without HA) using immunohistochemistry. In conclusion, the sponge formulation affords new perspectives with respect to the in vitro production of "artificial" cartilage. Furthermore, the presence of hyaluronate within the alginate sponge mimics a functional environment, suitable for the production by embedded chondrocytes of an extracellular matrix.
Miralles
G
G
Laboratoire de Physiopathologie et Pharmacologie Articulaires, UMR 7561 CNRS-UHP, Faculté de Médecine, BP 184, 54505, Vandoeuvre lès Nancy, France.
Baudoin
R
R
Dumas
D
D
Baptiste
D
D
Hubert
P
P
Stoltz
J F
JF
Dellacherie
E
E
Mainard
D
D
Netter
P
P
Payan
E
E
D000464:Alginates / Q000737:chemistry*; D000818:Animals; D001672:Biocompatible Materials; D002356:Cartilage; D002478:Cells, Cultured; D019902:Chondrocytes / Q000166:cytology / Q000502:physiology* / Q000648:ultrastructure; D020219:Chondrogenesis; D003094:Collagen / Q000378:metabolism; D003599:Cytoskeleton / Q000378:metabolism; D020723:Glucuronic Acid; D006603:Hexuronic Acids; D006820:Hyaluronic Acid / Q000737:chemistry*; D008297:Male; D008854:Microscopy, Electron; D011509:Proteoglycans / Q000096:biosynthesis; D051381:Rats; D017208:Rats, Wistar; D023822:Tissue Engineering / Q000379:methods*
D000464:Alginates; D001672:Biocompatible Materials; D006603:Hexuronic Acids; D011509:Proteoglycans; D020723:Glucuronic Acid; D006820:Hyaluronic Acid; D003094:Collagen