Journal of Developmental Biology and Tissue Engineering
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Article Number - B82C1FA40378

Vol.5(1), pp. 7-12 , April 2013
ISSN: 2141-2251

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Full Length Research Paper

3-D osteoblast culture for biomaterials testing

Jörg Neunzehn*, Sascha Heinemann and Hans Peter Wiesmann1

Max Bergmann Center of Biomaterials and Institute of Material Science, Technische Universität Dresden, Budapester Str. 27, D-01069 Dresden, Germany.              

Email: [email protected]

 Accepted: 19 September 2012  Published: 30 April 2013

Copyright © 2013 Author(s) retain the copyright of this article.
This article is published under the terms of the Creative Commons Attribution License 4.0

Micromass cell cultures, osteomicrospheres, were formed by aggregation of primary osteoblasts. Cell differentiation during sphere formation and the integrity of the osteomicrospheres was evaluated by analyzing the immunohistochemical expression of osteonectin, osteocalcin, and collagen type I. Transmission electron microscopy facilitated the proof of the tissue-like microstructure inside the osteomicrospheres and the arrangement of collagen fibers similar to early stages of natural bone formation. The in vivo situation of osteomicrospheres transferred to host tissue was simulated by embedding mature osteomicrospheres in a fibrin matrix. Additionally, cell spreading out of the compartments was investigated by electron microscopy of microspheres cultivated on the surface of three-dimensional fiber-like scaffolds. Concluding, the properties of the osteomicrospheres represented in this work demonstrate the potential as a tissue-specific in vitro test method to replace early small animal tests in the future.

Key words: Biomaterials testing, micromass culture, osteoblast, osteomicrospheres, ultrastructure.

Abbott A (2003). Cell culture: biology's new dimension. Nature. 424: 870-872.
Aubin JE (1998). Advances in the osteoblast lineage. Biochem. Cell Biol. 76: 899-910.
Bates RC, Edwards NS, Yates JD (2000). Spheroids and cell survival. Crit. Rev. Oncol. Hematol. 36:61-74.
Battistelli M, Borzi RM, Olivotto E, Vitellozzi R, Burattini S, Facchini A, Falcieri E (2005). Cell and matrix morpho-functional analysis in chondrocyte micromasses. Microsc. Res. Tech. 67:286-295.
Boudreau NJ, Jones PL (1999). Extracellular matrix and integrin signalling: the shape of things to come. Biochem. J. 339 (Pt 3):481-488.
PMid:10215583 PMCid:PMC1220180
Chen CS, Mrksich M, Huang S, Whitesides GM, Ingber DE (1997). Geometric control of cell life and death. Science. 276: 1425-1428.
Cukierman E, Pankov R, Stevens DR, Yamada KM (2001). Taking cell-matrix adhesions to the third dimension. Science. 294:1708-1712.
Edwards PC, Mason JM (2006). Gene-enhanced tissue engineering for dental hard tissue regeneration: (1) overview and practical considerations. Head Face Med. 2:12.
Gerber I, ap Gwynn I, Alini M, Wallimann T (2005). Stimulatory effects of creatine on metabolic activity, differentiation and mineralization of primary osteoblast-like cells in monolayer and micromass cell cultures. Eur. Cell Mater. 10:8-22.
Handschel J, Naujoks C, Depprich R, Lammers L, Kubler N, Meyer U, Wiesmann HP (2011). Embryonic stem cells in scaffold-free three-dimensional cell culture: osteogenic differentiation and bone generation. Head Face Med. 7:12.
PMid:21752302 PMCid:PMC3143924
Handschel JG, Depprich RA, Kubler NR, Wiesmann HP, Ommerborn M, Meyer U (2007). Prospects of micromass culture technology in tissue engineering. Head Face Med. 3:4.
PMid:17212823 PMCid:PMC1781066
Heinemann C, Heinemann S, Worch H, Hanke T (2011). Development of an osteoblast/osteoclast co-culture derived by human bone marrow stromal cells and human monocytes for biomaterials testing. Eur Cell Mater. 21:80-93.
Iwasaki A, Matsumoto T, Tazaki G, Tsuruta H, Egusa H, Miyajima H, Sohmura T (2009). Mass Fabrication of Small Cell Spheroids by Using Micro-patterned Tissue Culture Plate. Adv. Eng. Mater. 11: 801-804.
Kale S, Biermann S, Edwards C, Tarnowski C, Morris M, Long MW (2000). Three-dimensional cellular development is essential for ex vivo formation of human bone. Nat. Biotechnol. 18:954-958.
Langenbach F, Berr K, Naujoks C, Hassel A, Hentschel M, Depprich R, Kubler NR, Meyer U, Wiesmann HP, Kogler G, Handschel J (2011). Generation and differentiation of microtissues from multipotent precursor cells for use in tissue engineering. Nat. Protoc. 6:1726-1735.
Meyer U, Joos U, Wiesmann HP (2004). Biological and biophysical principles in extracorporal bone tissue engineering. Part I. Int. J. Oral Maxillofac Surg. 33:325-332.
Owen TA, Aronow M, Shalhoub V, Barone LM, Wilming L, Tassinari MS, Kennedy MB, Pockwinse S, Lian JB, Stein GS (1990). Progressive development of the rat osteoblast phenotype in vitro: reciprocal relationships in expression of genes associated with osteoblast proliferation and differentiation during formation of the bone extracellular matrix. J. Cell Physiol. 143:420-430.
Rossi MI, Barros AP, Baptista LS, Garzoni LR, Meirelles MN, Takiya CM, Pascarelli BM, Dutra HS, Borojevic R (2005). Multicellular spheroids of bone marrow stromal cells: a three-dimensional in vitro culture system for the study of hematopoietic cell migration. Braz. J. Med. Biol. Res. 38:1455-1462.
Sivaraman A, Leach JK, Townsend S, Iida T, Hogan BJ, Stolz DB, Fry R, Samson LD, Tannenbaum SR, Griffith LG (2005). A microscale in vitro physiological model of the liver: predictive screens for drug metabolism and enzyme induction. Curr. Drug Metab. 6: 569-591.
Tare RS, Howard D, Pound JC, Roach HI, Oreffo RO (2005). Tissue engineering strategies for cartilage generation--micromass and three dimensional cultures using human chondrocytes and a continuous cell line. Biochem. Biophys. Res. Commun. 333:609-621.
Tortelli F, Cancedda R (2009). Three-dimensional cultures of osteogenic and chondrogenic cells: a tissue engineering approach to mimic bone and cartilage in vitro. Eur. Cell Mater. 17:1-14.
Weaver VM, Petersen OW, Wang F, Larabell CA, Briand P, Damsky C, Bissell MJ (1997). Reversion of the malignant phenotype of human breast cells in three-dimensional culture and in vivo by integrin blocking antibodies. J. Cell Biol. 137:231-245.
PMid:9105051 PMCid:PMC2139858
Wiesmann HP, Joos U, Meyer U (2004). Biological and biophysical principles in extracorporal bone tissue engineering. Part II. Int. J. Oral


APA (2013). 3-D osteoblast culture for biomaterials testing. Journal of Developmental Biology and Tissue Engineering, 5(1), 7-12.
Chicago Jörg Neunzehn, Sascha Heinemann and Hans Peter Wiesmann. "3-D osteoblast culture for biomaterials testing." Journal of Developmental Biology and Tissue Engineering 5, no. 1 (2013): 7-12.
MLA Jörg Neunzehn, Sascha Heinemann and Hans Peter Wiesmann. "3-D osteoblast culture for biomaterials testing." Journal of Developmental Biology and Tissue Engineering 5.1 (2013): 7-12.

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