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Published in: Clinical Oral Investigations 1/2013

01-01-2013 | Review

Scaffold-free microtissues: differences from monolayer cultures and their potential in bone tissue engineering

Authors: Fabian Langenbach, Christian Naujoks, Ralf Smeets, Karin Berr, Rita Depprich, Norbert Kübler, Jörg Handschel

Published in: Clinical Oral Investigations | Issue 1/2013

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Abstract

Objectives

Cell-based therapies for bone augmentation after tooth loss and for the treatment of periodontal defects improve healing defects. Usually, osteogenic cells or stem cells are cultivated in 2D primary cultures, before they are combined with scaffold materials, even though this means a loss of the endogenous 3D microenvironment for the cells. Moreover, the use of single-cell suspensions for the inoculation of scaffolds or for the direct application into an area of interest has the disadvantages of low initial cell numbers and susceptibility to unwanted cellular distribution, respectively.

Materials and methods

We addressed the question whether an alternative to monolayer cultures, namely 3D microtissues, has the potential to improve osteogenic tissue engineering and its clinical outcome.

Results

By contrast, to monolayer cultures, osteogenic differentiation of 3D microtissues is enhanced by mimicking in vivo conditions. It seems that the osteogenic differentiation in microtissues is enhanced by strong integrin–extracellular matrix interaction and by stronger autocrine BMP2 signaling. Moreover, microtissues are less prone to wash out by body fluids and allow the precise administration of large cell numbers.

Conclusion

Microtissue cultures have closer characteristics with cells in vivo and their enhanced osteogenic differentiation makes scaffold-free microtissues a promising concept in osteogenic tissue engineering.

Clinical relevance

Microtissues are particularly suitable for tissue engineering because they improve seeding efficiency of biomaterials by increasing the cell load of a scaffold. This results in accelerated osteogenic tissue formation and could contribute to earlier implant stability in mandibular bone augmentation.
Literature
1.
go back to reference Sasso RC, LeHuec JC, Shaffrey C (2005) Iliac crest bone graft donor site pain after anterior lumbar interbody fusion: a prospective patient satisfaction outcome assessment. J Spinal Disord Tech 18(Suppl):S77–S81PubMedCrossRef Sasso RC, LeHuec JC, Shaffrey C (2005) Iliac crest bone graft donor site pain after anterior lumbar interbody fusion: a prospective patient satisfaction outcome assessment. J Spinal Disord Tech 18(Suppl):S77–S81PubMedCrossRef
2.
go back to reference Meyer U, Wiesmann HP, Berr K, Kubler NR, Handschel J (2006) Cell-based bone reconstruction therapies—principles of clinical approaches. Int J Oral Maxillofac Implants 21(6):899–906PubMed Meyer U, Wiesmann HP, Berr K, Kubler NR, Handschel J (2006) Cell-based bone reconstruction therapies—principles of clinical approaches. Int J Oral Maxillofac Implants 21(6):899–906PubMed
3.
go back to reference Petite H, Vandamme K, Monfoulet L, Logeart-Avramoglou D (2011) Strategies for improving the efficacy of bioengineered bone constructs: a perspective. Osteoporos Int 22(6):2017–2021PubMedCrossRef Petite H, Vandamme K, Monfoulet L, Logeart-Avramoglou D (2011) Strategies for improving the efficacy of bioengineered bone constructs: a perspective. Osteoporos Int 22(6):2017–2021PubMedCrossRef
4.
go back to reference Zhang ZY, Teoh SH, Hui JH, Fisk NM, Choolani M, Chan JK (2012) The potential of human fetal mesenchymal stem cells for off-the-shelf bone tissue engineering application. Biomaterials 33(9):2656–2672PubMedCrossRef Zhang ZY, Teoh SH, Hui JH, Fisk NM, Choolani M, Chan JK (2012) The potential of human fetal mesenchymal stem cells for off-the-shelf bone tissue engineering application. Biomaterials 33(9):2656–2672PubMedCrossRef
5.
go back to reference Zhang Z (2011) Bone regeneration by stem cell and tissue engineering in oral and maxillofacial region. Front Med 5(4):401–413PubMedCrossRef Zhang Z (2011) Bone regeneration by stem cell and tissue engineering in oral and maxillofacial region. Front Med 5(4):401–413PubMedCrossRef
6.
go back to reference Abbott A (2003) Cell culture: biology’s new dimension. Nat 424(6951):870–872CrossRef Abbott A (2003) Cell culture: biology’s new dimension. Nat 424(6951):870–872CrossRef
7.
go back to reference Burns JS, Rasmussen PL, Larsen KH, Schroder HD, Kassem M (2010) Parameters in three-dimensional osteospheroids of telomerized human mesenchymal (stromal) stem cells grown on osteoconductive scaffolds that predict in vivo bone-forming potential. Tissue Eng Part A 16(7):2331–2342PubMedCrossRef Burns JS, Rasmussen PL, Larsen KH, Schroder HD, Kassem M (2010) Parameters in three-dimensional osteospheroids of telomerized human mesenchymal (stromal) stem cells grown on osteoconductive scaffolds that predict in vivo bone-forming potential. Tissue Eng Part A 16(7):2331–2342PubMedCrossRef
8.
go back to reference Muraglia A, Corsi A, Riminucci M, Mastrogiacomo M, Cancedda R, Bianco P, Quarto R (2003) Formation of a chondro-osseous rudiment in micromass cultures of human bone-marrow stromal cells. J Cell Sci 116(Pt 14):2949–2955PubMedCrossRef Muraglia A, Corsi A, Riminucci M, Mastrogiacomo M, Cancedda R, Bianco P, Quarto R (2003) Formation of a chondro-osseous rudiment in micromass cultures of human bone-marrow stromal cells. J Cell Sci 116(Pt 14):2949–2955PubMedCrossRef
9.
go back to reference Trojani C, Weiss P, Michiels JF, Vinatier C, Guicheux J, Daculsi G, Gaudray P, Carle GF, Rochet N (2005) Three-dimensional culture and differentiation of human osteogenic cells in an injectable hydroxypropylmethylcellulose hydrogel. Biomaterials 26(27):5509–5517PubMedCrossRef Trojani C, Weiss P, Michiels JF, Vinatier C, Guicheux J, Daculsi G, Gaudray P, Carle GF, Rochet N (2005) Three-dimensional culture and differentiation of human osteogenic cells in an injectable hydroxypropylmethylcellulose hydrogel. Biomaterials 26(27):5509–5517PubMedCrossRef
10.
go back to reference Boehrs J, Zaharias RS, Laffoon J, Ko YJ, Schneider GB (2008) Three-dimensional culture environments enhance osteoblast differentiation. J Prosthodont 17(7):517–521PubMedCrossRef Boehrs J, Zaharias RS, Laffoon J, Ko YJ, Schneider GB (2008) Three-dimensional culture environments enhance osteoblast differentiation. J Prosthodont 17(7):517–521PubMedCrossRef
11.
go back to reference 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(9):954–958PubMedCrossRef 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(9):954–958PubMedCrossRef
12.
go back to reference 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 17:1–14PubMed 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 17:1–14PubMed
13.
go back to reference Kunz-Schughart LA, Kreutz M, Knuechel R (1998) Multicellular spheroids: a three-dimensional in vitro culture system to study tumour biology. Int J Exp Pathol 79(1):1–23PubMedCrossRef Kunz-Schughart LA, Kreutz M, Knuechel R (1998) Multicellular spheroids: a three-dimensional in vitro culture system to study tumour biology. Int J Exp Pathol 79(1):1–23PubMedCrossRef
14.
go back to reference Meyer U, Wiesmann HP, Libera J, Depprich R, Naujoks C, Handschel J (2012) Cartilage defect regeneration by ex vivo engineered autologous microtissue—preliminary results. Vivo 26(2):251–257 Meyer U, Wiesmann HP, Libera J, Depprich R, Naujoks C, Handschel J (2012) Cartilage defect regeneration by ex vivo engineered autologous microtissue—preliminary results. Vivo 26(2):251–257
15.
go back to reference Arufe MC, De la Fuente A, Mateos J, Fuentes I, De Toro FJ, Blanco FJ (2011) Analysis of the chondrogenic potential and secretome of mesenchymal stem cells derived from human umbilical cord stroma. Stem Cells Dev 20(7):1199–1212PubMedCrossRef Arufe MC, De la Fuente A, Mateos J, Fuentes I, De Toro FJ, Blanco FJ (2011) Analysis of the chondrogenic potential and secretome of mesenchymal stem cells derived from human umbilical cord stroma. Stem Cells Dev 20(7):1199–1212PubMedCrossRef
16.
go back to reference Estes BT, Diekman BO, Gimble JM, Guilak F (2010) Isolation of adipose-derived stem cells and their induction to a chondrogenic phenotype. Nat Protoc 5(7):1294–1311PubMedCrossRef Estes BT, Diekman BO, Gimble JM, Guilak F (2010) Isolation of adipose-derived stem cells and their induction to a chondrogenic phenotype. Nat Protoc 5(7):1294–1311PubMedCrossRef
17.
go back to reference Kafienah W, Al-Fayez F, Hollander AP, Barker MD (2003) Inhibition of cartilage degradation: a combined tissue engineering and gene therapy approach. Arthritis Rheum 48(3):709–718PubMedCrossRef Kafienah W, Al-Fayez F, Hollander AP, Barker MD (2003) Inhibition of cartilage degradation: a combined tissue engineering and gene therapy approach. Arthritis Rheum 48(3):709–718PubMedCrossRef
18.
go back to reference Keller GM (1995) In vitro differentiation of embryonic stem cells. Curr Opin Cell Biol 7(6):862–869PubMedCrossRef Keller GM (1995) In vitro differentiation of embryonic stem cells. Curr Opin Cell Biol 7(6):862–869PubMedCrossRef
19.
go back to reference Potter SW, Morris JE (1985) Development of mouse embryos in hanging drop culture. Anat Rec 211(1):48–56PubMedCrossRef Potter SW, Morris JE (1985) Development of mouse embryos in hanging drop culture. Anat Rec 211(1):48–56PubMedCrossRef
20.
go back to reference Wobus AM, Wolf E, Beier HM (2000) Embryonic stem cells and nuclear transfer strategies. Present state and future prospects. Cells Tissues Organs 166(1):1–5PubMedCrossRef Wobus AM, Wolf E, Beier HM (2000) Embryonic stem cells and nuclear transfer strategies. Present state and future prospects. Cells Tissues Organs 166(1):1–5PubMedCrossRef
21.
go back to reference Kelm JM, Timmins NE, Brown CJ, Fussenegger M, Nielsen LK (2003) Method for generation of homogeneous multicellular tumor spheroids applicable to a wide variety of cell types. Biotechnol Bioeng 83(2):173–180PubMedCrossRef Kelm JM, Timmins NE, Brown CJ, Fussenegger M, Nielsen LK (2003) Method for generation of homogeneous multicellular tumor spheroids applicable to a wide variety of cell types. Biotechnol Bioeng 83(2):173–180PubMedCrossRef
22.
go back to reference Handschel J, Naujoks C, Langenbach F, Berr K, Depprich RA, Ommerborn MA, Kubler NR, Brinkmann M, Kogler G, Meyer U (2010) Comparison of ectopic bone formation of embryonic stem cells and cord blood stem cells in vivo. Tissue Eng Part A 16(8):2475–2483PubMedCrossRef Handschel J, Naujoks C, Langenbach F, Berr K, Depprich RA, Ommerborn MA, Kubler NR, Brinkmann M, Kogler G, Meyer U (2010) Comparison of ectopic bone formation of embryonic stem cells and cord blood stem cells in vivo. Tissue Eng Part A 16(8):2475–2483PubMedCrossRef
23.
go back to reference Langenbach F, Berr K, Naujoks C, Hassel A, Hentschel M, Depprich R, Kubler NR, Meyer U, Wiesmann H-P, Kogler G, Handschel J (2011) Generation and differentiation of microtissues from multipotent precursor cells for use in tissue engineering. Nat Protocols 6(11):1726–1735CrossRef Langenbach F, Berr K, Naujoks C, Hassel A, Hentschel M, Depprich R, Kubler NR, Meyer U, Wiesmann H-P, Kogler G, Handschel J (2011) Generation and differentiation of microtissues from multipotent precursor cells for use in tissue engineering. Nat Protocols 6(11):1726–1735CrossRef
24.
go back to reference Hildebrandt C, Buth H, Thielecke H (2011) A scaffold-free in vitro model for osteogenesis of human mesenchymal stem cells. Tissue Cell 43(2):91–100PubMedCrossRef Hildebrandt C, Buth H, Thielecke H (2011) A scaffold-free in vitro model for osteogenesis of human mesenchymal stem cells. Tissue Cell 43(2):91–100PubMedCrossRef
25.
go back to reference Langenbach F, Naujoks C, Laser A, Kelz M, Kersten-Thiele P, Berr K, Depprich R, Kubler N, Kogler G, Handschel J (2012) Improvement of the cell-loading efficiency of biomaterials by inoculation with stem cell-based microspheres, in osteogenesis. J Biomater Appl 26(5):549–564PubMedCrossRef Langenbach F, Naujoks C, Laser A, Kelz M, Kersten-Thiele P, Berr K, Depprich R, Kubler N, Kogler G, Handschel J (2012) Improvement of the cell-loading efficiency of biomaterials by inoculation with stem cell-based microspheres, in osteogenesis. J Biomater Appl 26(5):549–564PubMedCrossRef
26.
go back to reference Handschel JG, Depprich RA, Kubler NR, Wiesmann HP, Ommerborn M, Meyer U (2007) Prospects of micromass culture technology in tissue engineering. Head Face Med 3:4PubMedCrossRef Handschel JG, Depprich RA, Kubler NR, Wiesmann HP, Ommerborn M, Meyer U (2007) Prospects of micromass culture technology in tissue engineering. Head Face Med 3:4PubMedCrossRef
27.
go back to reference Cukierman E, Pankov R, Stevens DR, Yamada KM (2001) Taking cell–matrix adhesions to the third dimension. Sci 294(5547):1708–1712CrossRef Cukierman E, Pankov R, Stevens DR, Yamada KM (2001) Taking cell–matrix adhesions to the third dimension. Sci 294(5547):1708–1712CrossRef
28.
go back to reference 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(1):231–245PubMedCrossRef 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(1):231–245PubMedCrossRef
29.
go back to reference 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(6):569–591PubMedCrossRef 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(6):569–591PubMedCrossRef
30.
go back to reference Stevens MM, George JH (2005) Exploring and engineering the cell surface interface. Sci 310(5751):1135–1138CrossRef Stevens MM, George JH (2005) Exploring and engineering the cell surface interface. Sci 310(5751):1135–1138CrossRef
31.
go back to reference Kelm JM, Fussenegger M (2004) Microscale tissue engineering using gravity-enforced cell assembly. Trends Biotechnol 22(4):195–202PubMedCrossRef Kelm JM, Fussenegger M (2004) Microscale tissue engineering using gravity-enforced cell assembly. Trends Biotechnol 22(4):195–202PubMedCrossRef
32.
go back to reference Garcia AJ (2005) Get a grip: integrins in cell–biomaterial interactions. Biomaterials 26(36):7525–7529PubMedCrossRef Garcia AJ (2005) Get a grip: integrins in cell–biomaterial interactions. Biomaterials 26(36):7525–7529PubMedCrossRef
33.
go back to reference Heino J, Huhtala M, Kapyla J, Johnson MS (2009) Evolution of collagen-based adhesion systems. Int J Biochem Cell Biol 41(2):341–348PubMedCrossRef Heino J, Huhtala M, Kapyla J, Johnson MS (2009) Evolution of collagen-based adhesion systems. Int J Biochem Cell Biol 41(2):341–348PubMedCrossRef
35.
go back to reference Biggs MJ, Dalby MJ (2010) Focal adhesions in osteoneogenesis. Proc Inst Mech Eng H 224(12):1441–1453PubMedCrossRef Biggs MJ, Dalby MJ (2010) Focal adhesions in osteoneogenesis. Proc Inst Mech Eng H 224(12):1441–1453PubMedCrossRef
36.
go back to reference Lim JY, Dreiss AD, Zhou Z, Hansen JC, Siedlecki CA, Hengstebeck RW, Cheng J, Winograd N, Donahue HJ (2007) The regulation of integrin-mediated osteoblast focal adhesion and focal adhesion kinase expression by nanoscale topography. Biomaterials 28(10):1787–1797PubMedCrossRef Lim JY, Dreiss AD, Zhou Z, Hansen JC, Siedlecki CA, Hengstebeck RW, Cheng J, Winograd N, Donahue HJ (2007) The regulation of integrin-mediated osteoblast focal adhesion and focal adhesion kinase expression by nanoscale topography. Biomaterials 28(10):1787–1797PubMedCrossRef
37.
go back to reference Gaur T, Lengner CJ, Hovhannisyan H, Bhat RA, Bodine PV, Komm BS, Javed A, van Wijnen AJ, Stein JL, Stein GS, Lian JB (2005) Canonical WNT signaling promotes osteogenesis by directly stimulating Runx2 gene expression. J Biol Chem 280(39):33132–33140PubMedCrossRef Gaur T, Lengner CJ, Hovhannisyan H, Bhat RA, Bodine PV, Komm BS, Javed A, van Wijnen AJ, Stein JL, Stein GS, Lian JB (2005) Canonical WNT signaling promotes osteogenesis by directly stimulating Runx2 gene expression. J Biol Chem 280(39):33132–33140PubMedCrossRef
38.
go back to reference Hong JH, Hwang ES, McManus MT, Amsterdam A, Tian Y, Kalmukova R, Mueller E, Benjamin T, Spiegelman BM, Sharp PA, Hopkins N, Yaffe MB (2005) TAZ, a transcriptional modulator of mesenchymal stem cell differentiation. Science 309(5737):1074–1078PubMedCrossRef Hong JH, Hwang ES, McManus MT, Amsterdam A, Tian Y, Kalmukova R, Mueller E, Benjamin T, Spiegelman BM, Sharp PA, Hopkins N, Yaffe MB (2005) TAZ, a transcriptional modulator of mesenchymal stem cell differentiation. Science 309(5737):1074–1078PubMedCrossRef
39.
go back to reference Hong D, Chen HX, Xue Y, Li DM, Wan XC, Ge R, Li JC (2009) Osteoblastogenic effects of dexamethasone through upregulation of TAZ expression in rat mesenchymal stem cells. J Steroid Biochem Mol Biol 116(1–2):86–92PubMedCrossRef Hong D, Chen HX, Xue Y, Li DM, Wan XC, Ge R, Li JC (2009) Osteoblastogenic effects of dexamethasone through upregulation of TAZ expression in rat mesenchymal stem cells. J Steroid Biochem Mol Biol 116(1–2):86–92PubMedCrossRef
40.
go back to reference Phillips JE, Gersbach CA, Wojtowicz AM, Garcia AJ (2006) Glucocorticoid-induced osteogenesis is negatively regulated by Runx2/Cbfa1 serine phosphorylation. J Cell Sci 119(Pt 3):581–591PubMedCrossRef Phillips JE, Gersbach CA, Wojtowicz AM, Garcia AJ (2006) Glucocorticoid-induced osteogenesis is negatively regulated by Runx2/Cbfa1 serine phosphorylation. J Cell Sci 119(Pt 3):581–591PubMedCrossRef
41.
go back to reference Franceschi RT, Iyer BS (1992) Relationship between collagen synthesis and expression of the osteoblast phenotype in MC3T3-E1 cells. J Bone Miner Res 7(2):235–246PubMedCrossRef Franceschi RT, Iyer BS (1992) Relationship between collagen synthesis and expression of the osteoblast phenotype in MC3T3-E1 cells. J Bone Miner Res 7(2):235–246PubMedCrossRef
42.
go back to reference Xiao G, Gopalakrishnan R, Jiang D, Reith E, Benson MD, Franceschi RT (2002) Bone morphogenetic proteins, extracellular matrix, and mitogen-activated protein kinase signaling pathways are required for osteoblast-specific gene expression and differentiation in MC3T3-E1 cells. J Bone Miner Res 17(1):101–110PubMedCrossRef Xiao G, Gopalakrishnan R, Jiang D, Reith E, Benson MD, Franceschi RT (2002) Bone morphogenetic proteins, extracellular matrix, and mitogen-activated protein kinase signaling pathways are required for osteoblast-specific gene expression and differentiation in MC3T3-E1 cells. J Bone Miner Res 17(1):101–110PubMedCrossRef
43.
go back to reference Kundu AK, Khatiwala CB, Putnam AJ (2009) Extracellular matrix remodeling, integrin expression, and downstream signaling pathways influence the osteogenic differentiation of mesenchymal stem cells on poly(lactide-co-glycolide) substrates. Tissue Eng Part A 15(2):273–283PubMedCrossRef Kundu AK, Khatiwala CB, Putnam AJ (2009) Extracellular matrix remodeling, integrin expression, and downstream signaling pathways influence the osteogenic differentiation of mesenchymal stem cells on poly(lactide-co-glycolide) substrates. Tissue Eng Part A 15(2):273–283PubMedCrossRef
44.
go back to reference Fatherazi S, Matsa-Dunn D, Foster BL, Rutherford RB, Somerman MJ, Presland RB (2009) Phosphate regulates osteopontin gene transcription. J Dent Res 88(1):39–44PubMedCrossRef Fatherazi S, Matsa-Dunn D, Foster BL, Rutherford RB, Somerman MJ, Presland RB (2009) Phosphate regulates osteopontin gene transcription. J Dent Res 88(1):39–44PubMedCrossRef
45.
go back to reference Tada H, Nemoto E, Foster BL, Somerman MJ, Shimauchi H (2011) Phosphate increases bone morphogenetic protein-2 expression through cAMP-dependent protein kinase and ERK1/2 pathways in human dental pulp cells. Bone 48(6):1409–1416PubMedCrossRef Tada H, Nemoto E, Foster BL, Somerman MJ, Shimauchi H (2011) Phosphate increases bone morphogenetic protein-2 expression through cAMP-dependent protein kinase and ERK1/2 pathways in human dental pulp cells. Bone 48(6):1409–1416PubMedCrossRef
46.
go back to reference Afzal F, Pratap J, Ito K, Ito Y, Stein JL, van Wijnen AJ, Stein GS, Lian JB, Javed A (2005) Smad function and intranuclear targeting share a Runx2 motif required for osteogenic lineage induction and BMP2 responsive transcription. J Cell Physiol 204(1):63–72PubMedCrossRef Afzal F, Pratap J, Ito K, Ito Y, Stein JL, van Wijnen AJ, Stein GS, Lian JB, Javed A (2005) Smad function and intranuclear targeting share a Runx2 motif required for osteogenic lineage induction and BMP2 responsive transcription. J Cell Physiol 204(1):63–72PubMedCrossRef
47.
go back to reference Payne KA, Meszaros LB, Phillippi JA, Huard J (2010) Effect of phosphatidyl inositol 3-kinase, extracellular signal-regulated kinases 1/2, and p38 mitogen-activated protein kinase inhibition on osteogenic differentiation of muscle-derived stem cells. Tissue Eng Part A 16(12):3647–3655PubMedCrossRef Payne KA, Meszaros LB, Phillippi JA, Huard J (2010) Effect of phosphatidyl inositol 3-kinase, extracellular signal-regulated kinases 1/2, and p38 mitogen-activated protein kinase inhibition on osteogenic differentiation of muscle-derived stem cells. Tissue Eng Part A 16(12):3647–3655PubMedCrossRef
48.
go back to reference Ito T, Sawada R, Fujiwara Y, Tsuchiya T (2008) FGF-2 increases osteogenic and chondrogenic differentiation potentials of human mesenchymal stem cells by inactivation of TGF-beta signaling. Cytotechnology 56(1):1–7PubMedCrossRef Ito T, Sawada R, Fujiwara Y, Tsuchiya T (2008) FGF-2 increases osteogenic and chondrogenic differentiation potentials of human mesenchymal stem cells by inactivation of TGF-beta signaling. Cytotechnology 56(1):1–7PubMedCrossRef
49.
go back to reference Maehata Y, Takamizawa S, Ozawa S, Kato Y, Sato S, Kubota E, Hata R (2006) Both direct and collagen-mediated signals are required for active vitamin D3-elicited differentiation of human osteoblastic cells: roles of osterix, an osteoblast-related transcription factor. Matrix Biol 25(1):47–58PubMedCrossRef Maehata Y, Takamizawa S, Ozawa S, Kato Y, Sato S, Kubota E, Hata R (2006) Both direct and collagen-mediated signals are required for active vitamin D3-elicited differentiation of human osteoblastic cells: roles of osterix, an osteoblast-related transcription factor. Matrix Biol 25(1):47–58PubMedCrossRef
50.
go back to reference Siddappa R, Martens A, Doorn J, Leusink A, Olivo C, Licht R, van Rijn L, Gaspar C, Fodde R, Janssen F, van Blitterswijk C, de Boer J (2008) cAMP/PKA pathway activation in human mesenchymal stem cells in vitro results in robust bone formation in vivo. Proc Natl Acad Sci U S A 105(20):7281–7286PubMedCrossRef Siddappa R, Martens A, Doorn J, Leusink A, Olivo C, Licht R, van Rijn L, Gaspar C, Fodde R, Janssen F, van Blitterswijk C, de Boer J (2008) cAMP/PKA pathway activation in human mesenchymal stem cells in vitro results in robust bone formation in vivo. Proc Natl Acad Sci U S A 105(20):7281–7286PubMedCrossRef
51.
go back to reference Salasznyk RM, Williams WA, Boskey A, Batorsky A, Plopper GE (2004) Adhesion to vitronectin and collagen I promotes osteogenic differentiation of human mesenchymal stem cells. J Biomed Biotechnol 2004(1):24–34PubMedCrossRef Salasznyk RM, Williams WA, Boskey A, Batorsky A, Plopper GE (2004) Adhesion to vitronectin and collagen I promotes osteogenic differentiation of human mesenchymal stem cells. J Biomed Biotechnol 2004(1):24–34PubMedCrossRef
52.
go back to reference Levi B, Nelson ER, Brown K, James AW, Xu D, Dunlevie R, Wu JC, Lee M, Wu B, Commons GW, Vistnes D, Longaker MT (2011) Differences in osteogenic differentiation of adipose-derived stromal cells from murine, canine, and human sources in vitro and in vivo. Plast Reconstr Surg 128(2):373–386PubMedCrossRef Levi B, Nelson ER, Brown K, James AW, Xu D, Dunlevie R, Wu JC, Lee M, Wu B, Commons GW, Vistnes D, Longaker MT (2011) Differences in osteogenic differentiation of adipose-derived stromal cells from murine, canine, and human sources in vitro and in vivo. Plast Reconstr Surg 128(2):373–386PubMedCrossRef
53.
go back to reference Zhang W, Deng ZL, Chen L, Zuo GW, Luo Q, Shi Q, Zhang BQ, Wagner ER, Rastegar F, Kim SH, Jiang W, Shen J, Huang E, Gao Y, Gao JL, Zhou JZ, Luo J, Huang J, Luo X, Bi Y, Su Y, Yang K, Liu H, Luu HH, Haydon RC, He TC, He BC (2010) Retinoic acids potentiate BMP9-induced osteogenic differentiation of mesenchymal progenitor cells. PLoS One 5(7):e11917PubMedCrossRef Zhang W, Deng ZL, Chen L, Zuo GW, Luo Q, Shi Q, Zhang BQ, Wagner ER, Rastegar F, Kim SH, Jiang W, Shen J, Huang E, Gao Y, Gao JL, Zhou JZ, Luo J, Huang J, Luo X, Bi Y, Su Y, Yang K, Liu H, Luu HH, Haydon RC, He TC, He BC (2010) Retinoic acids potentiate BMP9-induced osteogenic differentiation of mesenchymal progenitor cells. PLoS One 5(7):e11917PubMedCrossRef
54.
go back to reference Langenbach F, Naujoks C, Kersten-Thiele PV, Berr K, Depprich RA, Kubler NR, Kogler G, Handschel J (2010) Osteogenic differentiation influences stem cell migration out of scaffold-free microspheres. Tissue Eng Part A 16(2):759–766PubMedCrossRef Langenbach F, Naujoks C, Kersten-Thiele PV, Berr K, Depprich RA, Kubler NR, Kogler G, Handschel J (2010) Osteogenic differentiation influences stem cell migration out of scaffold-free microspheres. Tissue Eng Part A 16(2):759–766PubMedCrossRef
55.
go back to reference Lammers L, Naujoks C, Berr K, Depprich R, Kubler N, Meyer U, Langenbach F, Luttenberg B, Kogler G, Wiesmann HP, Handschel J (2012) Impact of DAG stimulation on mineral synthesis, mineral structure and osteogenic differentiation of human cord blood stem cells. Stem Cell Res 8(2):193–205PubMedCrossRef Lammers L, Naujoks C, Berr K, Depprich R, Kubler N, Meyer U, Langenbach F, Luttenberg B, Kogler G, Wiesmann HP, Handschel J (2012) Impact of DAG stimulation on mineral synthesis, mineral structure and osteogenic differentiation of human cord blood stem cells. Stem Cell Res 8(2):193–205PubMedCrossRef
56.
go back to reference Nagata T, Bellows CG, Kasugai S, Butler WT, Sodek J (1991) Biosynthesis of bone proteins [SPP-1 (secreted phosphoprotein-1, osteopontin), BSP (bone sialoprotein) and SPARC (osteonectin)] in association with mineralized-tissue formation by fetal-rat calvarial cells in culture. Biochem J 274(Pt 2):513–520PubMed Nagata T, Bellows CG, Kasugai S, Butler WT, Sodek J (1991) Biosynthesis of bone proteins [SPP-1 (secreted phosphoprotein-1, osteopontin), BSP (bone sialoprotein) and SPARC (osteonectin)] in association with mineralized-tissue formation by fetal-rat calvarial cells in culture. Biochem J 274(Pt 2):513–520PubMed
57.
go back to reference Wang W, Itaka K, Ohba S, Nishiyama N, Chung UI, Yamasaki Y, Kataoka K (2009) 3D spheroid culture system on micropatterned substrates for improved differentiation efficiency of multipotent mesenchymal stem cells. Biomaterials 30(14):2705–2715PubMedCrossRef Wang W, Itaka K, Ohba S, Nishiyama N, Chung UI, Yamasaki Y, Kataoka K (2009) 3D spheroid culture system on micropatterned substrates for improved differentiation efficiency of multipotent mesenchymal stem cells. Biomaterials 30(14):2705–2715PubMedCrossRef
58.
go back to reference Legros R, Balmain N, Bonel G (1987) Age-related changes in mineral of rat and bovine cortical bone. Calcif Tissue Int 41(3):137–144PubMedCrossRef Legros R, Balmain N, Bonel G (1987) Age-related changes in mineral of rat and bovine cortical bone. Calcif Tissue Int 41(3):137–144PubMedCrossRef
59.
go back to reference Leong L, Hughes PE, Schwartz MA, Ginsberg MH, Shattil SJ (1995) Integrin signaling: roles for the cytoplasmic tails of alpha IIb beta 3 in the tyrosine phosphorylation of pp 125FAK. J Cell Sci 108(Pt 12):3817–3825PubMed Leong L, Hughes PE, Schwartz MA, Ginsberg MH, Shattil SJ (1995) Integrin signaling: roles for the cytoplasmic tails of alpha IIb beta 3 in the tyrosine phosphorylation of pp 125FAK. J Cell Sci 108(Pt 12):3817–3825PubMed
60.
go back to reference Ducy P, Zhang R, Geoffroy V, Ridall AL, Karsenty G (1997) Osf2/Cbfa1: a transcriptional activator of osteoblast differentiation. Cell 89(5):747–754PubMedCrossRef Ducy P, Zhang R, Geoffroy V, Ridall AL, Karsenty G (1997) Osf2/Cbfa1: a transcriptional activator of osteoblast differentiation. Cell 89(5):747–754PubMedCrossRef
61.
go back to reference Xiao G, Wang D, Benson MD, Karsenty G, Franceschi RT (1998) Role of the alpha2-integrin in osteoblast-specific gene expression and activation of the Osf2 transcription factor. J Biol Chem 273(49):32988–32994PubMedCrossRef Xiao G, Wang D, Benson MD, Karsenty G, Franceschi RT (1998) Role of the alpha2-integrin in osteoblast-specific gene expression and activation of the Osf2 transcription factor. J Biol Chem 273(49):32988–32994PubMedCrossRef
62.
go back to reference Franceschi RT, Iyer BS, Cui Y (1994) Effects of ascorbic acid on collagen matrix formation and osteoblast differentiation in murine MC3T3-E1 cells. J Bone Miner Res Off J Am Soc Bone Miner Res 9(6):843–854CrossRef Franceschi RT, Iyer BS, Cui Y (1994) Effects of ascorbic acid on collagen matrix formation and osteoblast differentiation in murine MC3T3-E1 cells. J Bone Miner Res Off J Am Soc Bone Miner Res 9(6):843–854CrossRef
63.
go back to reference Naujoks C, Langenbach F, Berr K, Depprich R, Kubler N, Meyer U, Handschel J, Kogler G (2011) Biocompatibility of osteogenic predifferentiated human cord blood stem cells with biomaterials and the influence of the biomaterial on the process of differentiation. J Biomater Appl 25(5):497–512PubMedCrossRef Naujoks C, Langenbach F, Berr K, Depprich R, Kubler N, Meyer U, Handschel J, Kogler G (2011) Biocompatibility of osteogenic predifferentiated human cord blood stem cells with biomaterials and the influence of the biomaterial on the process of differentiation. J Biomater Appl 25(5):497–512PubMedCrossRef
64.
go back to reference Holy CE, Shoichet MS, Davies JE (2000) Engineering three-dimensional bone tissue in vitro using biodegradable scaffolds: investigating initial cell-seeding density and culture period. J Biomed Mater Res 51(3):376–382PubMedCrossRef Holy CE, Shoichet MS, Davies JE (2000) Engineering three-dimensional bone tissue in vitro using biodegradable scaffolds: investigating initial cell-seeding density and culture period. J Biomed Mater Res 51(3):376–382PubMedCrossRef
65.
go back to reference Bitar M, Brown RA, Salih V, Kidane AG, Knowles JC, Nazhat SN (2008) Effect of cell density on osteoblastic differentiation and matrix degradation of biomimetic dense collagen scaffolds. Biomacromolecules 9(1):129–135PubMedCrossRef Bitar M, Brown RA, Salih V, Kidane AG, Knowles JC, Nazhat SN (2008) Effect of cell density on osteoblastic differentiation and matrix degradation of biomimetic dense collagen scaffolds. Biomacromolecules 9(1):129–135PubMedCrossRef
66.
go back to reference Jahn K, Richards RG, Archer CW, Stoddart MJ (2010) Pellet culture model for human primary osteoblasts. Eur Cell Mater 20:149–161PubMed Jahn K, Richards RG, Archer CW, Stoddart MJ (2010) Pellet culture model for human primary osteoblasts. Eur Cell Mater 20:149–161PubMed
67.
go back to reference Kabiri M, Kul B, Lott WB, Futrega K, Ghanavi P, Upton Z, Doran MR (2012) 3D mesenchymal stem/stromal cell osteogenesis and autocrine signalling. Biochem Biophys Res Commun 419(2):142–147PubMedCrossRef Kabiri M, Kul B, Lott WB, Futrega K, Ghanavi P, Upton Z, Doran MR (2012) 3D mesenchymal stem/stromal cell osteogenesis and autocrine signalling. Biochem Biophys Res Commun 419(2):142–147PubMedCrossRef
68.
go back to reference Phimphilai M, Zhao Z, Boules H, Roca H, Franceschi RT (2006) BMP signaling is required for RUNX2-dependent induction of the osteoblast phenotype. J Bone Miner Res 21(4):637–646PubMedCrossRef Phimphilai M, Zhao Z, Boules H, Roca H, Franceschi RT (2006) BMP signaling is required for RUNX2-dependent induction of the osteoblast phenotype. J Bone Miner Res 21(4):637–646PubMedCrossRef
69.
go back to reference Abe E, Yamamoto M, Taguchi Y, Lecka-Czernik B, O'Brien CA, Economides AN, Stahl N, Jilka RL, Manolagas SC (2000) Essential requirement of BMPs-2/4 for both osteoblast and osteoclast formation in murine bone marrow cultures from adult mice: antagonism by noggin. J Bone Miner Res 15(4):663–673PubMedCrossRef Abe E, Yamamoto M, Taguchi Y, Lecka-Czernik B, O'Brien CA, Economides AN, Stahl N, Jilka RL, Manolagas SC (2000) Essential requirement of BMPs-2/4 for both osteoblast and osteoclast formation in murine bone marrow cultures from adult mice: antagonism by noggin. J Bone Miner Res 15(4):663–673PubMedCrossRef
70.
go back to reference Robey PG (2011) Cell sources for bone regeneration: the good, the bad, and the ugly (but promising). Tissue Eng Part B Rev 17(6):423–430PubMedCrossRef Robey PG (2011) Cell sources for bone regeneration: the good, the bad, and the ugly (but promising). Tissue Eng Part B Rev 17(6):423–430PubMedCrossRef
71.
go back to reference Sacchetti B, Funari A, Michienzi S, Di Cesare S, Piersanti S, Saggio I, Tagliafico E, Ferrari S, Robey PG, Riminucci M, Bianco P (2007) Self-renewing osteoprogenitors in bone marrow sinusoids can organize a hematopoietic microenvironment. Cell 131(2):324–336PubMedCrossRef Sacchetti B, Funari A, Michienzi S, Di Cesare S, Piersanti S, Saggio I, Tagliafico E, Ferrari S, Robey PG, Riminucci M, Bianco P (2007) Self-renewing osteoprogenitors in bone marrow sinusoids can organize a hematopoietic microenvironment. Cell 131(2):324–336PubMedCrossRef
72.
go back to reference Bianco P, Robey PG, Simmons PJ (2008) Mesenchymal stem cells: revisiting history, concepts, and assays. Cell Stem Cell 2(4):313–319PubMedCrossRef Bianco P, Robey PG, Simmons PJ (2008) Mesenchymal stem cells: revisiting history, concepts, and assays. Cell Stem Cell 2(4):313–319PubMedCrossRef
73.
go back to reference Jilka RL (2007) Molecular and cellular mechanisms of the anabolic effect of intermittent PTH. Bone 40(6):1434–1446PubMedCrossRef Jilka RL (2007) Molecular and cellular mechanisms of the anabolic effect of intermittent PTH. Bone 40(6):1434–1446PubMedCrossRef
74.
go back to reference Kelm JM, Breitbach M, Fischer G, Odermatt B, Agarkova I, Fleischmann BK, Hoerstrup SP (2012) 3D microtissue formation of undifferentiated bone marrow mesenchymal stem cells leads to elevated apoptosis. Tissue engineering Part A 18(7–8):692–702PubMedCrossRef Kelm JM, Breitbach M, Fischer G, Odermatt B, Agarkova I, Fleischmann BK, Hoerstrup SP (2012) 3D microtissue formation of undifferentiated bone marrow mesenchymal stem cells leads to elevated apoptosis. Tissue engineering Part A 18(7–8):692–702PubMedCrossRef
75.
go back to reference Altmann B, Steinberg T, Giselbrecht S, Gottwald E, Tomakidi P, Bächle-Haas M, Kohal RJ (2011) Promotion of osteoblast differentiation in 3D biomaterial micro-chip arrays comprising fibronectin-coated poly(methyl methacrylate) polycarbonate. Biomaterials 32(34):8947–8956PubMedCrossRef Altmann B, Steinberg T, Giselbrecht S, Gottwald E, Tomakidi P, Bächle-Haas M, Kohal RJ (2011) Promotion of osteoblast differentiation in 3D biomaterial micro-chip arrays comprising fibronectin-coated poly(methyl methacrylate) polycarbonate. Biomaterials 32(34):8947–8956PubMedCrossRef
76.
go back to reference Ferrera D, Poggi S, Biassoni C, Dickson GR, Astigiano S, Barbieri O, Favre A, Franzi AT, Strangio A, Federici A, Manduca P (2002) Three-dimensional cultures of normal human osteoblasts: proliferation and differentiation potential in vitro and upon ectopic implantation in nude mice. Bone 30(5):718–725PubMedCrossRef Ferrera D, Poggi S, Biassoni C, Dickson GR, Astigiano S, Barbieri O, Favre A, Franzi AT, Strangio A, Federici A, Manduca P (2002) Three-dimensional cultures of normal human osteoblasts: proliferation and differentiation potential in vitro and upon ectopic implantation in nude mice. Bone 30(5):718–725PubMedCrossRef
77.
go back to reference Kelm JM, Lorber V, Snedeker JG, Schmidt D, Broggini-Tenzer A, Weisstanner M, Odermatt B, Mol A, Zund G, Hoerstrup SP (2010) A novel concept for scaffold-free vessel tissue engineering: self-assembly of microtissue building blocks. J Biotechnol 148(1):46–55PubMedCrossRef Kelm JM, Lorber V, Snedeker JG, Schmidt D, Broggini-Tenzer A, Weisstanner M, Odermatt B, Mol A, Zund G, Hoerstrup SP (2010) A novel concept for scaffold-free vessel tissue engineering: self-assembly of microtissue building blocks. J Biotechnol 148(1):46–55PubMedCrossRef
78.
go back to reference Kelm JM, Fussenegger M (2010) Scaffold-free cell delivery for use in regenerative medicine. Adv Drug Deliv Rev 62(7–8):753–764PubMedCrossRef Kelm JM, Fussenegger M (2010) Scaffold-free cell delivery for use in regenerative medicine. Adv Drug Deliv Rev 62(7–8):753–764PubMedCrossRef
79.
go back to reference Berahim Z, Moharamzadeh K, Rawlinson A, Jowett AK (2011) Biologic interaction of three-dimensional periodontal fibroblast spheroids with collagen-based and synthetic membranes. J Periodontol 82(5):790–797PubMedCrossRef Berahim Z, Moharamzadeh K, Rawlinson A, Jowett AK (2011) Biologic interaction of three-dimensional periodontal fibroblast spheroids with collagen-based and synthetic membranes. J Periodontol 82(5):790–797PubMedCrossRef
Metadata
Title
Scaffold-free microtissues: differences from monolayer cultures and their potential in bone tissue engineering
Authors
Fabian Langenbach
Christian Naujoks
Ralf Smeets
Karin Berr
Rita Depprich
Norbert Kübler
Jörg Handschel
Publication date
01-01-2013
Publisher
Springer-Verlag
Published in
Clinical Oral Investigations / Issue 1/2013
Print ISSN: 1432-6981
Electronic ISSN: 1436-3771
DOI
https://doi.org/10.1007/s00784-012-0763-8

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