Flow perfusion culture of marrow stromal osteoblasts in titanium fiber mesh

Journal of Biomedical Materials Research - Part A - Tập 64A Số 2 - Trang 235-241 - 2003
Juliette van den Dolder1,2, Gregory N. Bancroft3,2, Vassilios I. Sikavitsas3,2, Paul H.M. Spauwen4, John A. Jansen1, Antonios G. Mikos3
1Department of Biomaterials, College of Dental Science, University Medical Center Nijmegen, Nijmegen, The Netherlands
2these authors contributed equally to this work
3Department of Bioengineering, Rice University, P.O. Box 1892, MS 142, Houston, Texas 77251-1892
4Department of Plastic and Reconstructive Surgery, University Medical Center Nijmegen, Nijmegen, The Netherlands

Tóm tắt

Abstract

The objective of this study was to evaluate the effect of two cell culture techniques, static and flow perfusion, on the osteogenic expression of rat bone marrow cells seeded into titanium fiber mesh for a period up to 16 days. A cell suspension of rat bone marrow stromal osteoblasts (5 × 105 cells/300 μL) was seeded into the mesh material. Thereafter, the constructs were cultured under static conditions or in a flow perfusion system for 4, 8, and 16 days. To evaluate cellular proliferation and differentiation, constructs were examined for DNA, calcium content, and alkaline phosphatase activity. Samples were also examined with scanning electron microscopy (SEM) and plastic‐embedded histological sections. Results showed an increase in DNA from day 4 to day 8 for the flow perfusion system. At day 8, a significant enhancement in DNA content was observed for flow perfusion culture compared with static culture conditions, but similar cell numbers were found for each culture system at 16 days. Calcium measurements showed a large increase in calcium content of the meshes subjected to flow perfusion at day 16. The SEM examination revealed that the 16‐day samples subjected to flow perfusion culture were completely covered with layers of cells and mineralized matrix. In addition, this matrix extended deep into the scaffolds. In contrast, meshes cultured under static conditions had only a thin sheet of matrix present on the upper surface of the meshes. Evaluation of the light microscopy sections confirmed the SEM observations. On the basis of our results, we conclude that a flow perfusion system can enhance the early proliferation, differentiation, and mineralized matrix production of bone marrow stromal osteoblasts seeded in titanium fiber mesh. © 2002 Wiley Periodicals, Inc. J Biomed Mater Res 64A: 235–241, 2003

Từ khóa


Tài liệu tham khảo

Burwell RG, 1963, Studies in the transplantation of bone. V. The capacity of fresh and treated homografts of bone to evoke transplantation immunity, Clin Orthop, 99, S5

Bancroft GN, 2001, Bone tissue engineering by cell transplantation, Tissue Eng Therapeutic Use, 5, 151

Owen M, 1985, Bone and mineral, 1

10.1002/(SICI)1097-4644(199702)64:2<278::AID-JCB11>3.0.CO;2-F

10.1016/8756-3282(92)90364-3

10.1016/S0963-6897(96)00279-5

10.1007/BF02724045

10.1007/BF00225804

10.1002/(SICI)1097-4644(19981001)71:1<55::AID-JCB6>3.0.CO;2-0

10.1002/jcp.1041430304

10.1002/(SICI)1097-0290(19980105)57:1<46::AID-BIT6>3.0.CO;2-V

10.1021/bp970120j

10.1002/(SICI)1097-0290(19990905)64:5<580::AID-BIT8>3.0.CO;2-X

10.1023/A:1008946832443

10.1007/BF02523224

10.1016/S0142-9612(00)00280-5

10.1089/10763270152436436

10.1034/j.1600-0501.2002.130112.x

VehofJWM van den DolderJ de RuijterJE SpauwenPHM JansenJA.The effect of seeding technique and incubation time on bone formation in titanium fiber mesh. Abstract. Society of Biomaterials Minneapolis2001.

van den DolderJ VehofJWM SpauwenPHM JansenJA.Bone formation by rat bone marrow cells cultured on titanium fiber mesh: effect of in vitro culture time. J Biomed Mater Res in press.

10.1002/1097-4636(200105)55:2<242::AID-JBM1011>3.0.CO;2-D

10.1089/107632702753725076

10.1006/bbrc.1996.1131

10.1016/S0021-9290(00)00231-1

10.1152/ajpendo.1996.271.1.E205

10.1359/jbmr.1997.12.1.45

10.1006/bbrc.1998.9270

10.1002/jcp.1041430113

10.1152/ajpcell.1991.261.3.C428