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BY-NC-ND 3.0 license Open Access Published by De Gruyter Open Access May 28, 2010

Evaluation of different culture techniques of osteoblasts on 3D scaffolds

  • Ying-ying Wu EMAIL logo , Yu Ban , Ning Geng , Yong-yue Wang , Xiao-guang Liu , Tao Yu and Ping Gong
From the journal Open Life Sciences

Abstract

Bones adjust their structure to withstand the mechanical demands they experience. It is suggested that flow-derived shear stress may be the most significant and primary mediator of mechanical stimulation. In this study, we designed and fabricated a fluid flow cell culture system that can load shear stress onto cells cultured on 3D scaffolds. We evaluated the effect of different culture techniques, namely, (1) continuous perfusion fluid flow, (2) intermittent perfusion fluid flow, and (3) static condition, on the proliferation of osteoblasts seeded on partially deproteinized bones. The flow rate was set at 1 ml/min for all the cells cultured using flow perfusion and the experiment was conducted for 12 days. Scanning electron microscopy analysis indicated an increase in cell proliferation for scaffolds subjected to fluid shear stress. In addition, the long axes of these cells lengthened along the flowing fluid direction. Continuous perfusion significantly enhanced cell proliferation compared to either intermittent perfusion or static condition. All the results demonstrated that fluid shear stress is able to enhance the proliferation of cells and change the form of cells.

[1] Castellon P., Blatz M.B., Block M.S., Finger I.M., Rogers B., Immediate loading of dental implants in the edentulous mandible, J. Am. Dent. Assoc., 2004, 135, 1543–1549 10.14219/jada.archive.2004.0080Search in Google Scholar

[2] Esposito M., Grusovin M.G., Achille H., Coulthard P., Worthington H.V., Interventions for replacing missing teeth: different times for loading dental implants, Cochrane Database Syst. Rev., 2009, CD003878 10.1002/14651858.CD003878.pub4Search in Google Scholar

[3] Ibañez J.C., Jalbout Z.N., Immediate Loading of Osseotite Implants: Two-Year Results Clinical Science and Techniques, Implant. Dent., 2002, 11, 128–136 http://dx.doi.org/10.1097/00008505-200204000-0001310.1097/00008505-200204000-00013Search in Google Scholar

[4] Petropoulos V.C., Balshi T.J., Balshi S.F., Extractions, Implant Placement, and Immediate Loading of Mandibular Implants: A Case Report of a Functional Fixed Prosthesis in 5 Hours, Implant. Dent., 2003, 12, 283–290 http://dx.doi.org/10.1097/01.ID.0000091123.29073.5710.1097/01.ID.0000091123.29073.57Search in Google Scholar

[5] You L., Temiyasathit S., Lee P., Padmaja C.H.K., Yao W., Kingery W., et al., Osteocytes as mechanosensors in the inhibition of bone resorption due to mechanical loading, Bone, 2008, 42,172–179 http://dx.doi.org/10.1016/j.bone.2007.09.04710.1016/j.bone.2007.09.047Search in Google Scholar

[6] Bancroft G.N., Sikavitsas V.I., van den Dolder J., Fluid flow increases mineralized matrix deposition in 3D perfusion culture of marrow stromal osteoblasts in a dose-dependent manner, Proc. Natl. Acad. Sci. USA, 2002, 99, 12600–12605 http://dx.doi.org/10.1073/pnas.20229659910.1073/pnas.202296599Search in Google Scholar

[7] Cao L., Ding Y., Effects of estrogen and wall-shear stress on rat osteoblats in vitro, J. Pract. Stomatol., 2003, 19, 475–478 Search in Google Scholar

[8] Klein N.J., van der Plas A., Semeins C.M., Sensitivity of osteocytes to biomechanical stress in vitro, FASEB J., 1995, 9, 441–445 10.1096/fasebj.9.5.7896017Search in Google Scholar

[9] Knothe Tate M.L., Niederer P., Knothe U., In vivo tracer transport through the lacunocanalicular system of rat bone in an environment devoid of mechanical loading, Bone, 1998, 22, 107–17 http://dx.doi.org/10.1016/S8756-3282(97)00234-210.1016/S8756-3282(97)00234-2Search in Google Scholar

[10] Geng T., Luo F.S., Sun H.Y., Desiging a three-dimensional perfusion bioreactor system for bone tissue enginnering, J. Clin. Rehab. Tissue. Eng. Res., 2007, 11, 3476–3479 Search in Google Scholar

[11] Jaasma M.J., Plunkett N.A., Design and validation of a dynamic flow perfusion bioreactor for use with compliant tissue engineering scaffolds, J. Biotechnol., 2008, 133, 490–496 http://dx.doi.org/10.1016/j.jbiotec.2007.11.01010.1016/j.jbiotec.2007.11.010Search in Google Scholar PubMed

[12] Zhao F., Ma T., Perfusion bioreactor system for human mesenchymal stem cell tissue engineering using recirculation bioreactors, Biomaterials, 2005, 26, 7012–7014 http://dx.doi.org/10.1016/j.biomaterials.2005.04.06210.1016/j.biomaterials.2005.04.062Search in Google Scholar PubMed

[13] Bjerrea L., Büngera C.E., Kassemb M., Myginda T., Flow perfusion culture of human mesenchymal stem cells on silicate-substituted tricalcium phosphate scaffolds, Biomaterials, 2008, 29, 2616–2627 http://dx.doi.org/10.1016/j.biomaterials.2008.03.00310.1016/j.biomaterials.2008.03.003Search in Google Scholar

[14] van den Dolder J., Bancroft G.N., Sikavitasas V.I., Flow perfusion culture of marrow stromal osteoblasts in titanium fiber mesh, J. Biomed. Mater. Res. A., 2003, 64, 235–241 http://dx.doi.org/10.1002/jbm.a.1036510.1002/jbm.a.10365Search in Google Scholar

[15] Rieder M.J., Carmona R., Krieger JE., Pritchard K.A. Jr., Greene A.S., Suppression of angiotensin-converting enzyme expression and activity by shear stress, Circ. Res., 1997, 80, 312–319 10.1161/01.RES.80.3.312Search in Google Scholar

[16] Sikavitsas V.I., Bancroft G.N., Holtorf H.L., Jansen J.A., Mikos A.G., Mineralized matrix deposition by marrow stromal osteoblasts in 3D perfusion culture increases with increasing fluid shear forces, Proc. Natl. Acad. Sci. USA, 2003, 100,14683–14688 http://dx.doi.org/10.1073/pnas.243436710010.1073/pnas.2434367100Search in Google Scholar

[17] Sikavitsas V.I., Bancroft G.N., Lemoine J.J., Liebschner M.A., Dauner M., Mikos A.G., Flow perfusion enhances the calcified matrix deposition of marrow stromal cells in biodegradable nonwoven fiber mesh scaffolds, Ann. Biomed. Eng., 2005, 33, 63–70 http://dx.doi.org/10.1007/s10439-005-8963-x10.1007/s10439-005-8963-xSearch in Google Scholar

[18] Norvell S.M., Alvarez M., Bidwell J.P., Pavalko F.M., Fluid Shear Stress Induces b-Catenin Signaling in Osteoblasts, Calcif. Tissue. Int., 2004, 75, 396–404 http://dx.doi.org/10.1007/s00223-004-0213-y10.1007/s00223-004-0213-ySearch in Google Scholar

[19] Orr D.E., Burg K.J., Design of a modular bioreactor to incorporate both perfusion flow and hydrostatic compression for tissue engineering applications, Ann. Biomed. Eng., 2008, 36, 1228–1241 http://dx.doi.org/10.1007/s10439-008-9505-010.1007/s10439-008-9505-0Search in Google Scholar

[20] Wang S.L., Liu N., Yang S.Y., Culture and identification of SD rat osteoblasts by modified enzymatic digestion in vitro, J. Clin. Rehab. Tissue. Eng. Res., 2008, 12, 2983–2987 Search in Google Scholar

[21] Varghese S., Wyzga N., Griffiths A.M., Effects of Serum From Children with Newly Diagnosed Crohn Disease on Primary Cultures of Rat Osteoblasts, J. Pediatr. Gastroenterol. Nutr., 2002, 35, 641–648 http://dx.doi.org/10.1097/00005176-200211000-0001010.1097/00005176-200211000-00010Search in Google Scholar

[22] Goldstein A.S., Juarez T.M., Helmke C.D., Effect of convection on osteoblastic cell growth and function in biodegradable polymer foam scaffolds, Biomaterials, 2001, 22, 1279–1288 http://dx.doi.org/10.1016/S0142-9612(00)00280-510.1016/S0142-9612(00)00280-5Search in Google Scholar

[23] Myers K.A., Rattner J.B., Nigel G., Osteoblast-like cells and fluid flow: Cytoskeleton-dependent shear sensitivity, Biochem. Biophys. Res. Commun., 2007, 364, 214–219 http://dx.doi.org/10.1016/j.bbrc.2007.09.10910.1016/j.bbrc.2007.09.109Search in Google Scholar PubMed

[24] Wu D., Ding Y., Effects of fluid shear stress on the proliferation and cell cycle of rat osteoblasts in vitro, J. Clin. Stomatol., 2007, 23, 197–199 Search in Google Scholar

Published Online: 2010-5-28
Published in Print: 2010-8-1

© 2010 Versita Warsaw

This work is licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 3.0 License.

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