Influence of biomechanical and biochemical stimulation on the proliferation and differentiation of bone marrow stromal cells seeded on polyurethane scaffolds

Teng,Songsong; Liu,Chaoxu; Guenther,Daniel; Omar,Mohamed; Neunaber,Claudia; Krettek,Christian; Jagodzinski,Michael

doi:10.3892/etm.2016.3206

Influence of biomechanical and biochemical stimulation on the proliferation and differentiation of bone marrow stromal cells seeded on polyurethane scaffolds

Authors:
- Songsong Teng
- Chaoxu Liu
- Daniel Guenther
- Mohamed Omar
- Claudia Neunaber
- Christian Krettek
- Michael Jagodzinski
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Affiliations: Department of Orthopedic Trauma, Hannover Medical School, 30625 Hannover, Germany
Published online on: March 30, 2016 https://doi.org/10.3892/etm.2016.3206
Pages: 2086-2094
Copyright: © Teng et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

The aim of the present investigation was to compare the effects of cyclic compression, perfusion, dexamethasone (DEX) and bone morphogenetic protein‑7 (BMP‑7) on the proliferation and differentiation of human bone marrow stromal cells (hBMSCs) in polyurethane scaffolds in a perfusion bioreactor. Polyurethane scaffolds seeded with hBMSCs were cultured under six different conditions, as follows: 10% Cyclic compression at 0.5 and 5 Hz; 10 ml/min perfusion; 100 nM DEX; 100 ng/ml BMP‑7; and 1 ml/min perfusion without mechanical and biochemical stimulation (control). On days 7 and 14, samples were tested for the following data: Cell proliferation; mRNA expression of Runx2, COL1A1 and osteocalcin; osteocalcin content; calcium deposition; and the equilibrium modulus of the tissue specimen. The results indicated that BMP‑7 and 10 ml/min perfusion promoted cell proliferation, which was inhibited by 5 Hz cyclic compression and DEX. On day 7, the 5 Hz cyclic compression inhibited Runx2 expression, whereas the 0.5 Hz cyclic compression and BMP‑7 upregulated the COL1A1 mRNA levels on day 7 and enhanced the osteocalcin expression on day 14. The DEX‑treated hBMSCs exhibited downregulated osteocalcin expression. After 14 days, the BMP‑7 group exhibited the highest calcium deposition, followed by the 0.5 Hz cyclic compression and the DEX groups. The equilibrium modulus of the engineered constructs significantly increased in the BMP‑7, 0.5 Hz cyclic compression and DEX groups. In conclusion, the present results suggest that BMP‑7 and perfusion enhance cell proliferation, whereas high frequency cyclic compression inhibits the proliferation and osteogenic differentiation of hBMSCs. Low frequency cyclic compression is more effective than DEX, but less effective compared with BMP‑7 on the osteogenic differentiation of hBMSCs seeded on polyurethane scaffolds.

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1	Murphy CM, O'Brien FJ, Little DG and Schindeler A: Cell-scaffold interactions in the bone tissue engineering triad. Eur Cell Mater. 26:120–132. 2013.PubMed/NCBI
2	Colnot C: Cell sources for bone tissue engineering: Insights from basic science. Tissue Eng Part B Rev. 17:449–457. 2011. View Article : Google Scholar : PubMed/NCBI
3	Wang YK and Chen CS: Cell adhesion and mechanical stimulation in the regulation of mesenchymal stem cell differentiation. J Cell Mol Med. 17:823–832. 2013. View Article : Google Scholar : PubMed/NCBI
4	Langenbach F and Handschel J: Effects of dexamethasone, ascorbic acid and beta-glycerophosphate on the osteogenic differentiation of stem cells in vitro. Stem Cell Res Ther. 4:1172013. View Article : Google Scholar : PubMed/NCBI
5	Sittichokechaiwut A, Edwards JH, Scutt AM and Reilly GC: Short bouts of mechanical loading are as effective as dexamethasone at inducing matrix production by human bone marrow mesenchymal stem cell. Eur Cell Mater. 20:45–57. 2010.PubMed/NCBI
6	Li R, Liang L, Dou Y, Huang Z, Mo H, Wang Y and Yu B: Mechanical strain regulates osteogenic and adipogenic differentiation of bone marrow mesenchymal stem cells. Biomed Res Int. 2015:8732512015.PubMed/NCBI
7	Cardwell RD, Kluge JA, Thayer PS, Guelcher SA, Dahlgren LA, Kaplan DL and Goldstein AS: Static and cyclic mechanical loading of mesenchymal stem cells on elastomeric, electrospun polyurethane meshes. J Biomech Eng. 137:2015. View Article : Google Scholar : PubMed/NCBI
8	Kim TJ, Joo C, Seong J, Vafabakhsh R, Botvinick EL, Berns MW, Palmer AE, Wang N, Ha T, Jakobsson E, et al: Distinct mechanisms regulating mechanical force-induced Ca²⁺ signals at the plasma membrane and the ER in human MSCs. eLife. 4:e048762015. View Article : Google Scholar : PubMed/NCBI
9	Delaine-Smith RM, MacNeil S and Reilly GC: Matrix production and collagen structure are enhanced in two types of osteogenic progenitor cells by a simple fluid shear stress stimulus. Eur Cell Mater. 24:162–174. 2012.PubMed/NCBI
10	Yourek G, McCormick SM, Mao JJ and Reilly GC: Shear stress induces osteogenic differentiation of human mesenchymal stem cells. Regen Med. 5:713–724. 2010. View Article : Google Scholar : PubMed/NCBI
11	Liu Y, Wu J, Zhu Y and Han J: Therapeutic application of mesenchymal stem cells in bone and joint diseases. Clin Exp Med. 14:13–24. 2014. View Article : Google Scholar : PubMed/NCBI
12	Gautschi OP, Frey SP and Zellweger R: Bone morphogenetic proteins in clinical applications. ANZ J Surg. 77:626–631. 2007. View Article : Google Scholar : PubMed/NCBI
13	Lee KB, Taghavi CE, Murray SS, Song KJ, Keorochana G and Wang JC: BMP induced inflammation: A comparison of rhBMP-7 and rhBMP-2. J Orthop Res. 30:1985–1994. 2012. View Article : Google Scholar : PubMed/NCBI
14	Weiskirchen R and Meurer SK: BMP-7 counteracting TGF-beta1 activities in organ fibrosis. Front Biosci (Landmark Ed). 18:1407–1434. 2013. View Article : Google Scholar : PubMed/NCBI
15	Black CR, Goriainov V, Gibbs D, Kanczler J, Tare RS and Oreffo RO: Bone Tissue Engineering. Curr Mol Biol Rep. 1:132–140. 2015. View Article : Google Scholar : PubMed/NCBI
16	Gong T, Xie J, Liao J, Zhang T, Lin S and Lin Y: Nanomaterials and bone regeneration. Bone Res. 3:150292015. View Article : Google Scholar : PubMed/NCBI
17	Liu C, Abedian R, Meister R, Haasper C, Hurschler C, Krettek C, von Lewinski G and Jagodzinski M: Influence of perfusion and compression on the proliferation and differentiation of bone mesenchymal stromal cells seeded on polyurethane scaffolds. Biomaterials. 33:1052–1064. 2012. View Article : Google Scholar : PubMed/NCBI
18	Verdonk P, Beaufils P, Bellemans J, Djian P, Heinrichs EL, Huysse W, Laprell H, Siebold R and Verdonk R: Actifit Study Group: Successful treatment of painful irreparable partial meniscal defects with a polyurethane scaffold: Two-year safety and clinical outcomes. Am J Sports Med. 40:844–853. 2012. View Article : Google Scholar : PubMed/NCBI
19	Hannink G, de Mulder EL, van Tienen TG and Buma P: Effect of load on the repair of osteochondral defects using a porous polymer scaffold. J Biomed Mater Res B Appl Biomater. 100:2082–2089. 2012. View Article : Google Scholar : PubMed/NCBI
20	Steinert AF, Rackwitz L, Gilbert F, Noth U and Tuan RS: Concise review: the clinical application of mesenchymal stem cells for musculoskeletal regeneration: current status and perspectives. Stem Cells Transl Med. 1:237–247. 2012. View Article : Google Scholar : PubMed/NCBI
21	Jagodzinski M, Drescher M, Zeichen J, Hankemeier S, Krettek C, Bosch U and van Griensven M: Effects of cyclic longitudinal mechanical strain and dexamethasone on osteogenic differentiation of human bone marrow stromal cells. Eur Cell Mater. 7:35–41, Discussion 41. 2004.PubMed/NCBI
22	van Tienen TG, Heijkants RG, Buma P, de Groot JH, Pennings AJ and Veth RP: Tissue ingrowth and degradation of two biodegradable porous polymers with different porosities and pore sizes. Biomaterials. 23:1731–1738. 2002. View Article : Google Scholar : PubMed/NCBI
23	Jagodzinski M, Breitbart A, Wehmeier M, Hesse E, Haasper C, Krettek C, Zeichen J and Hankemeier S: Influence of perfusion and cyclic compression on proliferation and differentiation of bone marrow stromal cells in 3-dimensional culture. J Biomech. 41:1885–1891. 2008. View Article : Google Scholar : PubMed/NCBI
24	Livak KJ and Schmittgen TD: Analysis of relative gene expression data using real-time quantitative PCR and the 2(−Delta Delta C(T)) Method. Methods. 25:402–408. 2001. View Article : Google Scholar : PubMed/NCBI
25	Liu J, Zhang B, Song S, Ma M, Si S, Wang Y, Xu B, Feng K, Wu J and Guo Y: Bovine collagen peptides compounds promote the proliferation and differentiation of MC3T3-E1 pre-osteoblasts. PloS One. 9:e999202014. View Article : Google Scholar : PubMed/NCBI
26	Kelly DJ, Crawford A, Dickinson SC, Sims TJ, Mundy J, Hollander AP, Prendergast PJ and Hatton PV: Biochemical markers of the mechanical quality of engineered hyaline cartilage. J Mater Sci Mater Med. 18:273–281. 2007. View Article : Google Scholar : PubMed/NCBI
27	Skerry TM: The response of bone to mechanical loading and disuse: fundamental principles and influences on osteoblast/osteocyte homeostasis. Arch Biochem Biophys. 473:117–123. 2008. View Article : Google Scholar : PubMed/NCBI
28	Turner CH, Owan I, Alvey T, Hulman J and Hock JM: Recruitment and proliferative responses of osteoblasts after mechanical loading in vivo determined using sustained-release bromodeoxyuridine. Bone. 22:463–469. 1998. View Article : Google Scholar : PubMed/NCBI
29	Jones LC, Yeoumans B, Hungerford DS and Frondoza CG: The response of osteoblast-like cells to dexamethasone and cyclic loading. Biomed Sci Instrum. 42:273–277. 2006.PubMed/NCBI
30	Makhdom AM and Hamdy RC: The role of growth factors on acceleration of bone regeneration during distraction osteogenesis. Tissue Eng Part B Rev. 19:442–453. 2013. View Article : Google Scholar : PubMed/NCBI
31	Huang C, Dai J and Zhang XA: Environmental physical cues determine the lineage specification of mesenchymal stem cells. Biochim Biophys Acta. 1850:1261–1266. 2015. View Article : Google Scholar : PubMed/NCBI
32	Michalopoulos E, Knight RL, Korossis S, Kearney JN, Fisher J and Ingham E: Development of methods for studying the differentiation of human mesenchymal stem cells under cyclic compressive strain. Tissue Eng Part C Methods. 18:252–262. 2012. View Article : Google Scholar : PubMed/NCBI
33	Delaine-Smith RM and Reilly GC: Mesenchymal stem cell responses to mechanical stimuli. Muscles Ligaments Tendons J. 2:169–180. 2012.PubMed/NCBI
34	Murphy CM, Haugh MG and O'Brien FJ: The effect of mean pore size on cell attachment, proliferation and migration in collagen-glycosaminoglycan scaffolds for bone tissue engineering. Biomaterials. 31:461–466. 2010. View Article : Google Scholar : PubMed/NCBI
35	Karageorgiou V and Kaplan D: Porosity of 3D biomaterial scaffolds and osteogenesis. Biomaterials. 26:5474–5491. 2005. View Article : Google Scholar : PubMed/NCBI
36	Roosa SM, Kemppainen JM, Moffitt EN, Krebsbach PH and Hollister SJ: The pore size of polycaprolactone scaffolds has limited influence on bone regeneration in an in vivo model. J Biomed Mater Res A. 92:359–368. 2010. View Article : Google Scholar : PubMed/NCBI
37	Kuboki Y, Jin Q and Takita H: Geometry of carriers controlling phenotypic expression in BMP-induced osteogenesis and chondrogenesis. J Bone Joint Surg Am. 83-A(Suppl 1): S105–S115. 2001.PubMed/NCBI
38	Gorna K and Gogolewski S: Biodegradable porous polyurethane scaffolds for tissue repair and regeneration. J Biomed Mater Res A. 79:128–138. 2006. View Article : Google Scholar : PubMed/NCBI
39	Gaspar DA, Gomide V and Monteiro FJ: The role of perfusion bioreactors in bone tissue engineering. Biomatter. 2:167–175. 2012. View Article : Google Scholar : PubMed/NCBI
40	Shea CM, Edgar CM, Einhorn TA and Gerstenfeld LC: BMP treatment of C3H10T1/2 mesenchymal stem cells induces both chondrogenesis and osteogenesis. J Cell Biochem. 90:1112–1127. 2003. View Article : Google Scholar : PubMed/NCBI
41	Cheng SL, Yang JW, Rifas L, Zhang SF and Avioli LV: Differentiation of human bone marrow osteogenic stromal cells in vitro: Induction of the osteoblast phenotype by dexamethasone. Endocrinology. 134:277–286. 1994. View Article : Google Scholar : PubMed/NCBI
42	Pelaez D, Huang CY and Cheung HS: Cyclic compression maintains viability and induces chondrogenesis of human mesenchymal stem cells in fibrin gel scaffolds. Stem Cells Dev. 18:93–102. 2009. View Article : Google Scholar : PubMed/NCBI
43	Mathews S, Bhonde R, Gupta PK and Totey S: Novel biomimetic tripolymer scaffolds consisting of chitosan, collagen type 1 and hyaluronic acid for bone marrow-derived human mesenchymal stem cells-based bone tissue engineering. J Biomed Mater Res B Appl Biomater. 102:1825–1834. 2014. View Article : Google Scholar : PubMed/NCBI
44	Kim K, Dean D, Mikos AG and Fisher JP: Effect of initial cell seeding density on early osteogenic signal expression of rat bone marrow stromal cells cultured on cross-linked poly(propylene fumarate) disks. Biomacromolecules. 10:1810–1817. 2009. View Article : Google Scholar : PubMed/NCBI
45	Frank O, Heim M, Jakob M, Barbero A, Schäfer D, Bendik I, Dick W, Heberer M and Martin I: Real-time quantitative RT-PCR analysis of human bone marrow stromal cells during osteogenic differentiation in vitro. J Cell Biochem. 85:737–746. 2002. View Article : Google Scholar : PubMed/NCBI
46	Xiao G, Wang D, Benson MD, Karsenty G and Franceschi RT: Role of the alpha2-integrin in osteoblast-specific gene expression and activation of the Osf2 transcription factor. J Biol Chem. 273:32988–32994. 1998. View Article : Google Scholar : PubMed/NCBI
47	Fiorentini E, Granchi D, Leonardi E, Baldini N and Ciapetti G: Effects of osteogenic differentiation inducers on in vitro expanded adult mesenchymal stromal cells. Int J Artif Organs. 34:998–1011. 2011. View Article : Google Scholar : PubMed/NCBI
48	Ito S, Suzuki N, Kato S, Takahashi T and Takagi M: Glucocorticoids induce the differentiation of a mesenchymal progenitor cell line, ROB-C26 into adipocytes and osteoblasts, but fail to induce terminal osteoblast differentiation. Bone. 40:84–92. 2007. View Article : Google Scholar : PubMed/NCBI
49	Kim HJ, Kim UJ, Vunjak-Novakovic G, Min BH and Kaplan DL: Influence of macroporous protein scaffolds on bone tissue engineering from bone marrow stem cells. Biomaterials. 26:4442–4452. 2005. View Article : Google Scholar : PubMed/NCBI
50	Dahlin RL, Gershovich JG, Kasper FK and Mikos AG: Flow perfusion co-culture of human mesenchymal stem cells and endothelial cells on biodegradable polymer scaffolds. Ann Biomed Eng. 42:1381–1390. 2014. View Article : Google Scholar : PubMed/NCBI
51	Kim J and Ma T: Bioreactor strategy in bone tissue engineering: Pre-culture and osteogenic differentiation under two flow configurations. Tissue Eng Part A. 18:2354–2364. 2012. View Article : Google Scholar : PubMed/NCBI
52	Duty AO, Oest ME and Guldberg RE: Cyclic mechanical compression increases mineralization of cell-seeded polymer scaffolds in vivo. J Biomech Eng. 129:531–539. 2007. View Article : Google Scholar : PubMed/NCBI