To determine the effects of bone morphogenetic proteins (BMP)-2 and −7, and serum, on extracellular matrix production by human septal chondrocytes in alginate.
Study Design
Human nasal septal chondrocytes were expanded, suspended in alginate, and cultured in BMP-2 or 7, with and without serum. The optimal concentration of each growth factor was determined based on matrix production. Next, the synergistic effects of BMP-2 and −7 at optimal concentrations were determined on separate beads, based on matrix quantity and histology.
Results
Matrix content was highest with concentrations of BMP-2 and −7 of 100 ng/ml and 20 ng/ml, respectively, with serum. Adding both BMP-2 and −7, with serum, increased matrix content by factors of 5.1 versus serum-only cultures, 2.7 versus only BMP-2 with serum, and 2.4 versus only BMP-7 with serum. All comparisons were statistically significant.
Conclusion
BMP-2 and −7 significantly increase production of extracellular matrix by septal chondrocytes suspended in alginate. The presence of serum improves matrix production.
Significance
BMP-2 and −7 have great potential for use in cartilage tissue engineering.
Get full access to this article
View all access options for this article.
References
1.
HomiczMRSchumacherBLSahRLEffects of serial expansion of septal chondrocytes on tissue-engineered neocartilage composition. Otolaryngol Head Neck Surg2002; 127:398–408.
2.
BenyaPDShafferJ.Dedifferentiated chondrocytes reexpress the differentiated collagen phenotype when cultured in agarose gels. Cell1982; 30:215–24.
3.
BujiaJPitzkePKastenbauerEEffect of growth factors on matrix synthesis by human nasal chondrocytes cultured in monolayer and in agar. Eur Arch Otorhinolaryngol1996; 253:336–40.
4.
DunhamBPKochRJ.Basic fibroblast growth factor and insulinlike growth factor I support the growth of human septal chondrocytes in a serum-free environment. Arch Otolaryngol Head Neck Surg1998; 124:1325–30.
5.
van OschGJVMMarijnissenWJCMvan der VeenSWThe potency of culture-expanded nasal septum chondrocytes for tissue engineering of cartilage. Am J Rhinol2001; 15:187–92.
6.
SampathTKReddiAH.Dissociative extraction and reconstitution of extracellular matrix components involved in local bone differentiation. Proc Natl Acad Sci U S A1981; 78:7599–603.
7.
UristMRMikulskiALietzeA.Solubilized and insolubilized bone morphogenetic protein. Proc Natl Acad Sci U S A1979; 76:1828–32.
8.
ChubinskayaSKuettnerKE.Regulation of osteogenic proteins by chondrocytes. Int J Biochem Cell Biol2003; 35:1323–40.
9.
FlechtenmacherJHuchKThonarEJRecombinant human osteogenic protein 1 is a potent stimulator of the synthesis of cartilage proteoglycans and collagens by human articular chondrocytes. Arthritis Rheum1996; 39:1896–904.
10.
NishidaYKnudsonCBEgerWOsteogenic protein-1 stimulates cell-associated matrix assembly by normal human articular chondrocytes: upregulation of hyaluronan synthase. Arthritis Rheum2000; 43:206–14.
11.
SailorLZHewickRMMorrisEA.Recombinant human bone morphogenetic protein-2 maintains the articular chondrocyte phenotype in long-term culture. J Orthop Res1996; 14:937–45.
12.
ChiaSHHomiczMRSchumacherBLCharacterization of human nasal septal chondrocytes cultured in alginate. J Am Coll Surg2005; 200:691–704.
13.
KimYJSahRLDoongJYFluorometric assay of DNA in cartilage explants using Hoechst 33258. Anal Biochem1988; 174:168–76.
14.
RichmonJDSageABSheltonEEffect of growth factors on cell proliferation, matrix deposition, and morphology of human nasal septal chondrocytes cultured in monolayer. Laryngoscope2005; 115:1553–60.
15.
VukicevicSLuytenFPReddiAH.Stimulation of expression of osteogenic and chondrogenic phenotypes in vitro by osteogenin. Proc Natl Acad Sci U S A1989; 86:8793–7.
16.
LuytenFPYuUMYanagishitaMNatural bovine osteogenin and recombinant human bone morphogenetic protein-2B are equipotent in the maintenance of proteoglycans in bovine articular cartilage explant culture. J Biol Chem1992; 267:3691–5.
17.
LuytenFPChenPParalkarVRecombinant bone morphogenetic protein-4, transforming growth factor-β, and activin A enhance the cartilage phenotype of auricular chondrocytes in vitro. Exp Cell Res1994; 210:224–9.
18.
MorrisE.Differential effects of TGF-beta superfamily members on articular cartilage metabolism: stimulation by rhBMP-9 and rhBMP-2 and inhibition of TGF-beta. Trans Orthop Res Soc1996; 42:175.
19.
ChenPVukicevicSSampathTKOsteogenic protein-1 promotes growth and maturation of chick sternal chondrocytes in serum-free cultures. J Cell Sci1995; 108:105–14.
20.
ChuaKHAminuddinBSFuzinaNHHuman serum provided additional values in growth factors supplemented medium for human chondrocytes monolayer expansion and engineered cartilage construction.Med J Malaysia2004; 59(Suppl B):194–5.
21.
GlowackiJYatesKEMacLeanRIn vitro engineering of cartilage: effects of serum substitutes, TGF-β, and IL-1α. Orthod Craniofac Res2005; 8:200–8.
22.
KimBSYooSPParkHW.Tissue engineering of cartilage with chondrocytes cultured in a chemically-defined, serum-free medium. Biotechnol Lett2004; 26:709–12.
23.
MandlEWvan der VeenSWVerhaarJANSerum-free medium supplemented with high-concentration FGF2 for cell expansion culture of human ear chondrocytes promotes redifferentiation capacity. Tiss Eng2002; 8:573–80.
24.
NaumannARotterNBujiaJTissue engineering of autologous cartilage transplants for rhinology. Am J Rhinol1998; 12:59–63.
25.
KafienahWJakobMDemarteauOThree-dimensional tissue engineering of hyaline cartilage: comparison of adult nasal and articular chondrocytes. Tiss Eng2002; 8:817–26.
