Human mesenchymal stem cells (hMSCs) from bone marrow are considered a promising cell source for bone tissue engineering applications because of their ability to differentiate into cells of the osteoblastic lineage. Mechanical stimulation is able to promote osteogenic differentiation of hMSC; however, the use of hydrostatic pressure (HP) has not been well studied. Artificial extracellular matrices containing collagen and chondroitin sulfate (CS) have promoted the expression of an osteoblastic phenotype by hMSCs. However, there has been little research into the combined effects of biochemical stimulation by matrices and simultaneous mechanical stimulation. In this study, artificial extracellular matrices generated from collagen and/or CS were coated onto polycaprolactone-co-lactide substrates, seeded with hMSCs and subjected to cyclic HP at various time points during 21 days after cell seeding to investigate the effects of biochemical, mechanical, and combined biochemical and mechanical stimulations. Cell differentiation was assessed by analyzing the expression of alkaline phosphatase (ALP) at the protein- and mRNA levels, as well as for calcium accumulation. The timing of HP stimulation affected hMSC proliferation and expression of ALP activity. HP stimulation after 6 days was most effective at promoting ALP activity. CS-containing matrices promoted the osteogenic differentiation of hMSCs. A combination of both CS-containing matrices and cyclic HP yields optimal effects on osteogenic differentiation of hMSCs on scaffolds compared with individual responses.

1.
Alhadlaq
,
A.
, and
Mao
,
J. J.
, 2003, “
Tissue-Engineered Neogenesis of Human-Shaped Mandibular Condyle From Rat Mesenchymal Stem Cells
,”
J. Dent. Res.
0022-0345,
82
(
12
), pp.
951
956
.
2.
Pittenger
,
M. F.
,
Mackay
,
A. M.
,
Beck
,
S. C.
,
Jaiswal
,
R. K.
,
Douglas
,
R.
,
Mosca
,
J. D.
,
Moorman
,
M. A.
,
Simonetti
,
D. W.
,
Craig
,
S.
, and
Marshak
,
D. R.
, 1999, “
Multilineage Potential of Adult Human Mesenchymal Stem Cells
,”
Science
0036-8075,
284
(
5411
), pp.
143
147
.
3.
Jagodzinski
,
M.
,
Drescher
,
M.
,
Zeichen
,
J.
,
Hankemeier
,
S.
,
Krettek
,
C.
,
Bosch
,
U.
, and
van Griensven
,
M.
, 2004, “
Effects of Cyclic Longitudinal Mechanical Strain and Dexamethasone on Osteogenic Differentiation of Human Bone Marrow Stromal Cells
,”
Eur. Cells Mater
1473-2262,
7
, pp.
35
41
.
4.
Mauney
,
J. R.
,
Sjostorm
,
S.
,
Blumberg
,
J.
,
Horan
,
R.
,
O’Leary
,
J. P.
,
Vunjak-Novakovic
,
G.
,
Volloch
,
V.
, and
Kaplan
,
D. L.
, 2004, “
Mechanical Stimulation Promotes Osteogenic Differentiation of Human Bone Marrow Stromal Cells on 3-D Partially Demineralized Bone Scaffolds In Vitro
,”
Calcif. Tissue Int.
0171-967X,
74
(
5
), pp.
458
468
.
5.
Hildebrand
,
A.
,
Romaris
,
M.
,
Rasmussen
,
L. M.
,
Heinegard
,
D.
,
Twardzik
,
D. R.
,
Border
,
W. A.
, and
Ruoslahti
,
E.
, 1994, “
Interaction of the Small Interstitial Proteoglycans Biglycan, Decorin and Fibromodulin With Transforming Growth Factor Beta
,”
Biochem. J.
0264-6021,
302
, pp.
527
534
.
6.
Simmons
,
C. A.
,
Matlis
,
S.
,
Thornton
,
A. J.
,
Chen
,
S.
,
Wang
,
C. Y.
, and
Mooney
,
D. J.
, 2003, “
Cyclic Strain Enhances Matrix Mineralization by Adult Human Mesenchymal Stem Sells Via the Extracellular Signal-Regulated Kinase (ERK1/2) Signaling Pathway
,”
J. Biomech.
0021-9290,
36
(
8
), pp.
1087
1096
.
7.
Wiesmann
,
A.
,
Buhring
,
H. J.
,
Mentrup
,
C.
, and
Wiesmann
,
H. P.
, 2006, “
Decreased CD90 Expression in Human Mesenchymal Stem Cells by Applying Mechanical Stimulation
,”
Head Face Med.
,
2
,
8
.
8.
Angele
,
P.
,
Yoo
,
J. U.
,
Smith
,
C.
,
Mansour
,
J.
,
Jepsen
,
K. J.
,
Nerlich
,
M.
, and
Johnstone
,
B.
, 2003, “
Cyclic Hydrostatic Pressure Enhances the Chondrogenic Phenotype of Human Mesenchymal Progenitor Cells Differentiated In Vitro
,”
J. Orthop. Res.
0736-0266,
21
(
3
), pp.
451
457
.
9.
Angele
,
P.
,
Abke
,
J.
,
Kujat
,
R.
,
Faltermeier
,
H.
,
Schumann
,
D.
,
Nerlich
,
M.
,
Kinner
,
B.
,
Englert
,
C.
,
Ruszczak
,
Z.
,
Mehrl
,
R.
, and
Mueller
,
R.
, 2004, “
Influence of Different Collagen Species on Physico-Chemical Properties of Crosslinked Collagen Matrices
,”
Biomaterials
0142-9612,
25
(
14
), pp.
2831
2841
.
10.
Lee
,
C. R.
,
Grodzinsky
,
A. J.
, and
Spector
,
M.
, 2003, “
Biosynthetic Response of Passaged Chondrocytes in a Type II Collagen Scaffold to Mechanical Compression
,”
J. Biomed. Mater. Res. Part A
1549-3296,
64
(
3
), pp.
560
569
.
11.
Miyanishi
,
K.
,
Trindade
,
M. C.
,
Lindsey
,
D. P.
,
Beaupre
,
G. S.
,
Carter
,
D. R.
,
Goodman
,
S. B.
,
Schurman
,
D. J.
, and
Smith
,
R. L.
, 2006, “
Effects of Hydrostatic Pressure and Transforming Growth Factor-Beta 3 on Adult Human Mesenchymal Stem Cell Chondrogenesis In Vitro
,”
Tissue Eng.
1076-3279,
12
(
6
), pp.
1419
1428
.
12.
Klein-Nulend
,
J.
,
Veldhuijzen
,
J. P.
, and
Burger
,
E. H.
, 1986, “
Increased Calcification of Growth Plate Cartilage as a Result of Compressive Force In Vitro
,”
Arthritis Rheum.
0004-3591,
29
(
8
), pp.
1002
1009
.
13.
Kim
,
S. H.
,
Choi
,
Y. R.
,
Park
,
M. S.
,
Shin
,
J. W.
,
Park
,
K. D.
,
Kim
,
S. J.
, and
Lee
,
J. W.
, 2007, “
ERK 1/2 Activation in Enhanced Osteogenesis of Human Mesenchymal Stem Cells in Poly(Lactic-Glycolic Acid) by Cyclic Hydrostatic Pressure
,”
J. Biomed. Mater. Res. Part A
1549-3296,
80
(
4
), pp.
826
836
.
14.
Roelofsen
,
J.
,
Klein-Nulend
,
J.
, and
Burger
,
E. H.
, 1995, “
Mechanical Stimulation by Intermittent Hydrostatic Compression Promotes Bone-Specific Gene Expression In Vitro
,”
J. Biomech.
0021-9290,
28
(
12
), pp.
1493
1503
.
15.
Jaiswal
,
N.
,
Haynesworth
,
S. E.
,
Caplan
,
A. I.
, and
Bruder
,
S. P.
, 1997, “
Osteogenic Differentiation of Purified, Culture-Expanded Human Mesenchymal Stem Cells In Vitro
,”
J. Cell. Biochem.
0730-2312,
64
(
2
), pp.
295
312
.
16.
Byers
,
R. J.
,
Brown
,
J.
,
Brandwood
,
C.
,
Wood
,
P.
,
Staley
,
W.
,
Hainey
,
L.
,
Freemont
,
A. J.
, and
Hoyland
,
J. A.
, 1999, “
Osteoblastic Differentiation and mRNA Analysis of STRO-1-Positive Human Bone Marrow Stromal Cells Using Primary In Vitro Culture and Poly (A) PCR
,”
J. Pathol.
0022-3417,
187
(
3
), pp.
374
381
.
17.
Salasznyk
,
R. M.
,
Williams
,
W. A.
,
Boskey
,
A.
,
Batorsky
,
A.
, and
Plopper
,
G. E.
, 2004, “
Adhesion to Vitronectin and Collagen I Promotes Osteogenic Differentiation of Human Mesenchymal Stem Cells
,”
J. Biomed. Biotechnol.
1110-7243,
2004
(
1
), pp.
24
34
.
18.
Bosnakovski
,
D.
,
Mizuno
,
M.
,
Kim
,
G.
,
Takagi
,
S.
,
Okumura
,
M.
, and
Fujinaga
,
T.
, 2006, “
Chondrogenic Differentiation of Bovine Bone Marrow Mesenchymal Stem Cells (MSCs) in Different Hydrogels: Influence of Collagen Type II Extracellular Matrix on MSC Chondrogenesis
,”
Biotechnol. Bioeng.
0006-3592,
93
(
6
), pp.
1152
1163
.
19.
Kinoshita
,
S.
,
Finnegan
,
M.
,
Bucholz
,
R. W.
, and
Mizuno
,
K.
, 1999, “
Three-Dimensional Collagen Gel Culture Promotes Osteoblastic Phenotype in Bone Marrow Derived Cells
,”
Kobe J. Med. Sci.
0023-2513,
45
(
5
), pp.
201
211
.
20.
Roehlecke
,
C.
,
Witt
,
M.
,
Kasper
,
M.
,
Schulze
,
E.
,
Wolf
,
C.
,
Hofer
,
A.
, and
Funk
,
R. W.
, 2001, “
Synergistic Effect of Titanium Alloy and Collagen Type I on Cell Adhesion, Proliferation and Differentiation of Osteoblast-Like Cells
,”
Cells Tissues Organs
1422-6405,
168
(
3
), pp.
178
187
.
21.
Geissler
,
U.
,
Hempel
,
U.
,
Wolf
,
C.
,
Scharnweber
,
D.
,
Worch
,
H.
, and
Wenzel
,
K.
, 2000, “
Collagen Type I-Coating of Ti6Al4V Promotes Adhesion of Osteoblasts
,”
J. Biomed. Mater. Res.
0021-9304,
51
(
4
), pp.
752
760
.
22.
Douglas
,
T.
,
Heinemann
,
S.
,
Mietrach
,
C.
,
Hempel
,
U.
,
Bierbaum
,
S.
,
Scharnweber
,
D.
, and
Worch
,
H.
, 2007, “
Interactions of Collagen Types I and II With Chondroitin Sulfates A-C and Their Effect on Osteoblast Adhesion
,”
Biomacromolecules
1525-7797,
8
(
4
), pp.
1085
1092
.
23.
Wollenweber
,
M.
,
Domaschke
,
H.
,
Hanke
,
T.
,
Boxberger
,
S.
,
Schmack
,
G.
,
Gliesche
,
K.
,
Scharnweber
,
D.
, and
Worch
,
H.
, 2006, “
Mimicked Bioartificial Matrix Containing Chondroitin Sulphate on a Textile Scaffold of Poly(3-hydroxybutyrate) Alters the Differentiation of Adult Human Mesenchymal Stem Cells
,”
Tissue Eng.
1076-3279,
12
(
2
), pp.
345
359
.
24.
Bierbaum
,
S.
,
Douglas
,
T.
,
Hanke
,
T.
,
Scharnweber
,
D.
,
Tippelt
,
S.
,
Monsees
,
T. K.
,
Funk
,
R. H.
, and
Worch
,
H.
, 2006, “
Collageneous Matrix Coatings on Titanium Implants Modified With Decorin and Chondroitin Sulfate: Characterization and Influence on Osteoblastic Cells
,”
J. Biomed. Mater. Res. Part A
1549-3296,
77
(
3
), pp.
551
562
.
25.
Zhong
,
S.
,
Teo
,
W. E.
,
Zhu
,
X.
,
Beuerman
,
R.
,
Ramakrishna
,
S.
, and
Yung
,
L. Y.
, 2005, “
Formation of Collagen-Glycosaminoglycan Blended Nanofibrous Scaffolds and Their Biological Properties
,”
Biomacromolecules
1525-7797,
6
(
6
), pp.
2998
3004
.
26.
van Susante
,
J. L. C.
,
Pieper
,
J.
,
Buma
,
P.
,
van Kuppevelt
,
T. H.
,
van Beuningen
,
H.
,
van Der Kraan
,
P. M.
,
Veerkamp
,
J. H.
,
van den Berg
,
W. B.
, and
Veth
,
R. P. H.
, 2001, “
Linkage of Chondroitin-Sulfate to Type I Collagen Scaffolds Stimulates the Bioactivity of Seeded Chondrocytes In Vitro
,”
Biomaterials
0142-9612,
22
(
17
), pp.
2359
2369
.
27.
Coombes
,
A. G.
,
Rizzi
,
S. C.
,
Williamson
,
M.
,
Barralet
,
J. E.
,
Downes
,
S.
, and
Wallace
,
W. A.
, 2004, “
Precipitation Casting of Polycaprolactone for Applications in Tissue Engineering and Drug Delivery
,”
Biomaterials
0142-9612,
25
(
2
), pp.
315
325
.
28.
Matsuzaka
,
K.
,
Yoshinari
,
M.
,
Shimono
,
M.
, and
Inoue
,
T.
, 2004, “
Effects of Multigrooved Surfaces on Osteoblast-Like Cells In Vitro: Scanning Electron Microscopic Observation and mRNA Expression of Osteopontin and Osteocalcin
,”
J. Biomed. Mater. Res. Part A
1549-3296,
68
(
2
), pp.
227
234
.
29.
Zhou
,
Y.
,
Chen
,
F.
,
Ho
,
S. T.
,
Woodruff
,
M. A.
,
Lim
,
T. M.
, and
Hutmacher
,
D. W.
, 2007, “
Combined Marrow Stromal Cell-Sheet Techniques and High-Strength Biodegradable Composite Scaffolds for Engineered Functional Bone Grafts
,”
Biomaterials
0142-9612,
28
(
5
), pp.
814
824
.
30.
Myers
,
K. A.
,
Shrive
,
N. G.
, and
Hart
,
D. A.
, 2007, “
A Novel Apparatus Applying Long Term Intermittent Cyclic Hydrostatic Pressure to In Vitro Cell Cultures
,”
J. Biosci. Bioeng.
1389-1723,
103
(
6
), pp.
578
581
.
31.
Majima
,
T.
,
Marchuk
,
L. L.
,
Sciore
,
P.
,
Shrive
,
N. G.
,
Frank
,
C. B.
, and
Hart
,
D. A.
, 2000, “
Compressive Compared With Tensile Loading of Medial Collateral Ligament Scar In Vitro Uniquely Influences mRNA Levels for Aggrecan, Collagen Type II, and Collagenase
,”
J. Orthop. Res.
0736-0266,
18
(
4
), pp.
524
531
.
32.
Tasevski
,
V.
,
Sorbetti
,
J. M.
,
Chiu
,
S. S.
,
Shrive
,
N. G.
, and
Hart
,
D. A.
, 2005, “
Influence of Mechanical and Biological Signals on Gene Expression in Human MG-63 Cells: Evidence for a Complex Interplay Between Hydrostatic Compression and Vitamin D3 or TGF-beta1 on MMP-1 and MMP-3 mRNA Levels
,”
Biochem. Cell Biol.
0829-8211,
83
(
1
), pp.
96
107
.
33.
Myers
,
K. A.
,
Rattner
,
J. B.
,
Shrive
,
N. G.
, and
Hart
,
D. A.
, 2007, “
Osteoblast-Like Cells and Fluid Flow: Cytoskeleton-Dependent Shear Sensitivity
,”
Biochem. Biophys. Res. Commun.
0006-291X,
364
(
2
), pp.
214
219
.
34.
Williams
,
B. R.
,
Gelman
,
R. A.
,
Poppke
,
D. C.
, and
Piez
,
K. A.
, 1978, “
Collagen Fibril Formation
,”
J. Biol. Chem.
0021-9258,
253
(
18
), pp.
6578
6585
.
35.
Rentsch
,
C.
,
Rentsch
,
B.
,
Breier
,
A.
,
Hofmann
,
A.
,
Manthey
,
S.
,
Scharnweber
,
D.
,
Biewener
,
A.
, and
Zwipp
,
H.
, 2008, “
Evaluation of the Osteogenic Potential and Vascularisation of 3D Poly(3)hydroxybutyrate Scaffolds Subcutaneously Implanted in Nude Rats
,”
J. Biomed. Mater. Res. Part A
1549-3296, to be published.
36.
Benoit
,
D. S.
,
Durney
,
A. R.
, and
Anseth
,
K. S.
, 2007, “
The Effect of Heparin-Functionalized PEG Hydrogels on Three-Dimensional Human Mesenchymal Stem Cell Osteogenic Differentiation
,”
Biomaterials
0142-9612,
28
(
1
), pp.
66
77
.
37.
Bruder
,
S. P.
,
Jaiswal
,
N.
,
Ricalton
,
N. S.
,
Mosca
,
J. D.
,
Kraus
,
K. H.
, and
Kadiyala
,
S.
, 1998,
Mesenchymal Stem Cells in Osteobiology and Applied Bone Regeneration
,”
Clin. Orthop. Relat. Res.
0009-921X,
355
, pp.
S247
S256
.
38.
Ciapetti
,
G.
,
Ambrosio
,
L.
,
Marletta
,
G.
,
Baldini
,
N.
, and
Giunti
,
A.
, 2006, “
Human Bone Marrow Stromal Cells: In Vitro Expansion and Differentiation for Bone Engineering
,”
Biomaterials
0142-9612,
27
(
36
), pp.
6150
6160
.
39.
Tateishi
,
K.
,
Ando
,
W.
,
Higuchi
,
C.
,
Hart
,
D. A.
,
Hashimoto
,
J.
,
Nakata
,
K.
,
Yoshikawa
,
H.
, and
Nakamura
,
N.
, 2008, “
Comparison of Human Serum With Fetal Bovine Serum for Expansion and Differentiation of Human Synovial MSC: Potential Feasibility for Clinical Applications
,”
Cell Transplant
0963-6897,
17
(
5
), pp.
549
557
.
40.
De Bari
,
C.
,
Dell’Accio
,
F.
,
Karystinou
,
A.
,
Guillot
,
P. V.
,
Fisk
,
N. M.
,
Khan
,
I. M.
,
Archer
,
C. W.
,
Jones
,
E. A.
,
McGonagle
,
D.
,
Mitsiadis
,
T. A.
,
Donaldson
,
A. N.
,
Luyten
,
F. P.
, and
Pitzalis
,
C.
, 2008, “
A Biomarker-Based Mathematical Model to Predict Bone Forming Potency of Human Synovial and Periosteal Mesenchymal Stem Cells
,”
Arthritis Rheum.
0004-3591,
58
(
1
), pp.
240
250
.
41.
Ando
,
W.
,
Tateishi
,
K.
,
Katakai
,
D.
,
Hart
,
D. A.
,
Higuchi
,
C.
,
Nakata
,
K.
,
Hashimoto
,
J.
,
Fujie
,
H.
,
Shino
,
K.
,
Yoshikawa
,
H.
, and
Nakamura
,
N.
, 2008, “
In Vitro Generation of a Scaffold-Free Tissue-Engineered Construct (TEC) Derived From Human Synovial Mesenchymal Stem Cells: Biological and Mechanical Properties and Further Chondrogenic Potential
,”
Tissue Eng Part A
,
14
(
12
), pp.
2041
2049
.
You do not currently have access to this content.