Cartilage tissue engineering of nasal septal chondrocyte-macroaggregates in human demineralized bone matrix
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Tissue Engineering is an important method for generating cartilage tissue with isolated autologous cells and the support of biomaterials. In contrast to various gel-like biomaterials, human demineralized bone matrix (DBM) guarantees some biomechanical stability for an application in biomechanically loaded regions. The present study combined for the first time the method of seeding chondrocyte-macroaggregates in DBM for the purpose of cartilage tissue engineering. After isolating human nasal chondrocytes and creating a three-dimensional macroaggregate arrangement, the DBM was cultivated in vitro with the macroaggregates. The interaction of the cells within the DBM was analyzed with respect to cell differentiation and the inhibitory effects of chondrocyte proliferation. In contrast to chondrocyte-macroaggregates in the cell-DBM constructs, morphologically modified cells expressing type I collagen dominated. The redifferentiation of chondrocytes, characterized by the expression of type II collagen, was only found in low amounts in the cell-DBM constructs. Furthermore, caspase 3, a marker for apoptosis, was detected in the chondrocyte-DBM constructs. In another experimental setting, the vitality of chondrocytes as related to culture time and the amount of DBM was analyzed with the BrdU assay. Higher amounts of DBM tended to result in significantly higher proliferation rates of the cells within the first 48 h. After 96 h, the vitality decreased in a dose-dependent fashion. In conclusion, this study provides the proof of concept of chondrocyte-macroaggregates with DBM as an interesting method for the tissue engineering of cartilage. The as-yet insufficient redifferentiation of the chondrocytes and the sporadic initiation of apoptosis will require further investigations.
KeywordsTissue engineering Human demineralized bone matrix Cartilage Chondrocyte-macroaggregates
The authors thank Mr. Schurig and Mr. Schweiger for their technical support and for making the DBM available.
Conflict of interest
No competing financial interests exist.
- Alexander TH, Sage AB, Chen AC, Schumacher BL, Shelton E, Masuda K, Sah RL, Watson D (2010) Insulin-like growth factor-I and growth differentiation factor-5 promote the formation of tissue-engineered human nasal septal cartilage. Tissue Eng Part C Methods 16(5):1213–1221. doi: 10.1089/ten.TEC.2009.0396 PubMedCrossRefGoogle Scholar
- Bruns J (2003) Tissue engineering—neues zum gewebeersatz im muskel-skelett-system. Steinkopff Verlag, DarmstadtGoogle Scholar
- Jäckel M (1998) Die genetische Kontrolle des programmierten Zelltods (Apoptose), vol 46. Springer, HNO, pp 614–625Google Scholar
- Leatherman BD, Dornhoffer JL (2004) The use of demineralized bone matrix for mastoid cavity obliteration. Otol Neurotol 25 (1):22–25; discussion 25–26Google Scholar
- Maor G, Hochberg Z, Silbermann M (1993) Insulin-like growth factor I accelerates proliferation and differentiation of cartilage progenitor cells in cultures of neonatal mandibular condyles. Acta Endocrinol (Copenh) 128(1):56–64Google Scholar
- Pirsig W, Bean JK, Lenders H, Verwoerd CD, Verwoerd-Verhoef HL (1995) Cartilage transformation in a composite graft of demineralized bovine bone matrix and ear perichondrium used in a child for the reconstruction of the nasal septum. Int J Pediatr Otorhinolaryngol 32(2):171–181PubMedCrossRefGoogle Scholar
- ten Koppel PG, van Osch GJ, Verwoerd CD, Verwoerd-Verhoef HL (1998) Efficacy of perichondrium and a trabecular demineralized bone matrix for generating cartilage. Plast Reconstr Surg 102(6):2012–2020; discussion 2021Google Scholar
- Wang JC, Kanim LE, Nagakawa IS, Yamane BH, Vinters HV, Dawson EG (2001) Dose-dependent toxicity of a commercially available demineralized bone matrix material. Spine 26(13):1429–1435; discussion 1435–1426Google Scholar