There is a current need for a small diameter vascular graft due to the limited supply of autogenous grafts and the failure of synthetic grafts due to thrombosis and/or intimal hyperplasia. The use of living cells and tissues to fabricate a small diameter graft (i.e., tissue engineered blood vessel, TEBV) could be useful given the endothelialization potential and biocompatibility benefits of such a graft. However, while sufficient strength has been attained in a TEBV, coordinate compliance has yet to be fine-tuned. In this study we investigate the effects of biological response modifiers, retinoic acid (RA) and ascorbic acid (AA) on TEBV biomechanics as a function of time and subsequently correlate observed RA/AA induced changes in TEBV mechanics with alterations in smooth muscle cell (SMC) biochemistry. TEBVs were constructed using a fibrillar type I collagen network populated by human aortic smooth muscle cells (AoSMC). Following construction this TEBV was treated with 0.3 mM AA and 0.1 mM RA (concentrations found to induce changes in VSMC phenotype). Ultimate tensile stress (UTS), rate of relaxation (RR) and elastic efficiency (EE) of RA/AA treated and untreated TEBVs were measured following 1, 7, 15, 30, 45, and 60 days of treatment. At corresponding time points, the effect of these treatments on collagen and elastin protein synthesis and mRNA expression was examined. RA/AA treated TEBV strength increased and stiffness decreased compared to controls as a function of time. Relative collagen synthesis in treated TEBVs exceeded control levels by nearly two-fold at 15 and 30 days of incubation. RA/AA treated collagen gene expression followed a similar trend. Relative elastin synthesis was also greater in treated TEBVs as compared to untreated TEBVs at 15 and 30 days of incubation and correspondingly elastin mRNA expression was significantly elevated at 15 days of incubation. These data provide evidence that RA/AA treated TEBVs exhibit mechanical properties which more closely mimic those of a native vessel than their untreated counterparts and that changes in extracellular matrix composition and matrix gene expression in the presence of RA/AA treatment may play an important role in the development of said mechanical properties.

1.
Stoney, R. J., 1977, “The Arterial Autograft,” In Vascular Surgery, R. Rutherford, editor. WB Saunders, Philadelphia, pp. 363.
2.
Weinberg
,
C. B.
, and
Bell
,
E.
,
1986
, “
A Blood Vessel Model Constructed from Collagen and Cultured Vascular Cells
,”
Science
,
231
, pp.
397
400
.
3.
Tranquillo
,
R. T.
,
Girton
,
T. S.
,
Bromberek
,
B. A.
,
Tribes
,
T. G.
, and
Mooradian
,
D. L.
,
1996
, “
Magnetically Orientated Tissue-equivalent Tubes: Application to a Circumferentially Orientated Media-equivalent
,”
Biomaterials
,
17
, pp.
349
357
.
4.
Kanda
,
K. T.
,
Matsuda
,
T.
, and
Oka
,
T.
,
1993
, “
Mechanical Stress Induced Cellular Orientation and Phenotypic Modulation of 3-D Cultured Smooth Muscle Cells
,”
ASAIO J.
,
39
, pp.
M278–M282
M278–M282
.
5.
L’Heureux
,
N. L.
,
Germain
,
L.
,
Labbe
,
R.
, and
Auger
,
F. A.
,
1993
, “
In vitro Construction of a Human Blood Vessel from Cultured Vascular Cells: A Morphologic Study
,”
Journal of Vascular Surgery
17
, pp.
499
509
.
6.
Niklason
,
L. E.
,
Gao
,
J.
,
Abbott
,
W. M.
,
Hirschi
,
K. K.
,
Houser
,
S.
,
Marini
,
R.
,
Langer
,
R.
,
1999
, “
Functional Arteries Grown in Vitro
,”
Science
,
284
, pp.
489
493
.
7.
Kaushal
,
S.
,
Amiel
,
G. E.
,
Guleserian
,
K. J.
,
Shapira
,
O. M.
,
Perry
,
T.
,
Sutherland
,
F. W.
,
Rabkin
,
E.
,
Moran
,
A. M.
,
Schoen
,
F. J.
,
Atala
,
A.
,
Soker
,
S.
,
Bischoff
,
J.
,
Mayer
,
J. E.
, “
Functional Small-diameter Neovessels Created Using Endothelial Progenitor Cells Expanded Ex Vivo
,”
Nature Medicine
,
7
, pp.
1035
1040
.
8.
L’Heureux
,
N. L.
,
Paquet
,
S.
,
Labbe
,
R.
,
Germain
,
L.
, and
Auger
,
F. A.
,
1998
, “
A Completely Biological Tissue-engineered Human Blood Vessel
,”
FASEB J.
,
12
, pp.
47
56
.
9.
Baird
,
R. N.
, and
Abbott
,
W. M.
,
1976
, “
Pulsatile Blood Flow in Arterial Grafts
,”
Lancet
,
2
, pp.
948
949
.
10.
Gayna
,
E.
,
Mason
,
W.
, and
Marble
,
A.
,
1974
, “
Necessity for Elastic Properties in Synthetic Arterial Grafts
,”
Can. J. Surg.
,
17
, pp.
176
176
.
11.
Clark
,
R. E.
,
Apostolou
,
S.
, and
Kardos
,
J. L.
,
1976
, “
Mismatch of Mechanical Properties as a Cause of Arterial Prostheses Thrombosis
,”
Surg. Forum
,
27
, pp.
208
208
.
12.
Echave
,
V.
,
Koornick
,
A. R.
, and
Harmov
,
M.
,
1979
, “
Intimal Hyperplasia as a Complication of the use of PTFE Graft for Femoral-popliteal Bypass
,”
Surgery (St. Louis)
,
86
, pp.
791
796
.
13.
Kidson
,
I. G.
, and
Abbott
,
W. M.
,
1978
, “
Low Compliance and Arterial Graft Occlusion
,”
Circulation
,
58
, pp.
1
4
.
14.
Lyman
,
D. J.
,
Albo
,
D.
, and
Jackson
,
R.
,
1977
, “
Development of Small Diameter Vascular Prostheses
,”
Trans. Am. Soc. Artif. Intern. Organs
,
22
, pp.
474
484
.
15.
Clark
,
J. H.
,
1933
, “
Elasticity of Veins
,”
Am. J. Physiol.
,
105
, pp.
418
427
.
16.
Reuterwall
,
O. P.
,
1921
, “
Die Elasticitat der Gerfasswade und die Methoden Ihrer Naheren Prufung
,”
Acta Med. Scand.
,
Suppl. 2
, pp.
175
175
.
17.
Strandness, E., and Summer, D. S., 1975, Hemodynamics for Surgeons, Grune & Stratton, New York, pp. 150–175.
18.
Orlandi
,
A.
,
Ehrlich
,
H. P.
,
Ropraz
,
P.
,
Spagnoli
,
L. G.
, and
Gabbiani
,
G.
,
1994
, “
Rat Aortic Smooth Muscle Cells Isolated from Different Layers and at Different Times after Endothelial Denudation Show Distinct Biological Features In vitro
,”
Arterioscler. Thromb.
,
14
, pp.
982
989
.
19.
Barone
,
L. M.
,
Faris
,
B.
,
Chipman
,
S.
,
Toselli
,
P.
,
Oakes
,
B. W.
, and
Franzblau
,
C.
,
1985
, “
Alteration of the Extracellular Matrix of Smooth Muscle Cells by Ascorbate Treatment
,”
Biochim. Biophys. Acta
,
840
, pp.
245
254
.
20.
Ortwerth, B. J., and Monnier, V. M., 1996, “Protein Glycation by the Oxidation Products of Ascorbic Acid,” In Ascorbic Acid, New York, pp. 123–134.
21.
Murad, S., Grove, D., and Lindgerg, K. A., 1981, “Regulation of Collagen Synthesis by Ascorbic Acid,” Proceedings of the National Academy of Science USA, 78, pp. 2879–2882.
22.
Geesin
,
J. C.
,
Gordon
,
J. S.
, and
Berg
,
R. A.
,
1990
, “
Retinoids Affect Collagen Synthesis Through Inhibition of Ascorbate-induced Lipid Peroxidation in Cultured Human Dermal Fibroblasts
,”
Arch. Biochem. Biophys.
,
278
, pp.
350
55
.
23.
Oikarinen
,
H.
,
Oikarinen
,
A. I.
,
Tan
,
E. M.
,
Abergel
,
R. P.
,
Meeker
,
C. A.
,
Chu
,
M. L.
,
Prockop
,
D. J.
, and
Uitto
,
J.
,
1985
, “
Modulation of Procollagen Gene Expression by Retinoids. Inhibition of Collagen Production by Retinoic Acid Accompanied by Reduced Type I Procollagen Messenger Ribonucleic Acid Levels in Human Skin Fibroblast Cultures
,”
J. Clin. Invest.
,
75
, pp.
1545
1553
.
24.
Davidson
,
J. M.
,
LuValle
,
P. A.
,
Zoia
,
O.
,
Quaglino
,
D.
, Jr.
, and
Giro
,
M.
,
1997
, “
Ascorbate Differentially Regulates Elastin and Collagen Biosynthesis in Vascular Smooth Muscle Cells and Skin Fibroblasts by Pretranslational Mechanisms
,”
J. Biol. Chem.
,
272
, pp.
345
352
.
25.
Tajima
,
S.
, and
Pinnell
,
S. R.
,
1983
, “
Collagen Synthesis in Cultured Human Skin Fibroblasts: Effect of Ascorbic Acid and its Analogs
,”
J. Invest. Dermatol.
,
81
, pp.
158
162
.
26.
Geesin
,
J. C.
,
Darr
,
D.
,
Kaufman
,
R.
,
Murad
,
S.
, and
Pinnell
,
S. R.
,
1988
, “
Ascorbic Acid Specifically Increases Type I and Type III Procollagen Messenger RNA Levels in Human Skin Fibroblast
,”
J. Invest. Dermatol.
,
90
, pp.
420
424
.
27.
Hayashi
,
A.
,
Suzuki
,
T.
, and
Tajima
,
S.
,
1995
, “
Modulations of Elastin Expression and Cell Proliferation by Retinoids in Cultured Vascular Smooth Muscle Cells
,”
J. Biochem. (Tokyo)
,
117
, pp.
312
136
.
28.
Ogle
,
B. M.
, and
Mooradian
,
D. L.
,
1999
, “
The Role of Vascular Smooth Muscle Cell Integrins in the Compaction and Mechanical Strengthening of a Tissue Engineered Blood Vessel
,”
Tissue Eng.
,
5
, pp.
387
401
.
29.
Association for the Advancement of Medical Instrumentation, 1986, “American National Standard for Vascular Graft Prostheses,” In AAMI Standards and Recommended Practices, AAMI press, Philadelphia, pp. 567–572.
30.
ASTM, 1992, Designation: D 2290-92, “Standard Test Method for Apparent Tensile Strength of Ring or Tubular Plastics and Reinforced Plastics by Split Disk Method,” In Annual Book of ASTM Standard, Philadelphia, pp. 91–96.
31.
Gosline
,
J. M.
, and
Shadwick
,
R. E.
,
1996
, “
The Mechanical Properties of Fin Whale Arteries are Explained by Novel Connective Tissue Designs
,”
J. Exp. Biol.
,
199
, pp.
985
985
.
32.
Trojan, P., and Flinn, T., 1991, Engineering Materials and Their Applications, Houghton Mifflin Press, Boston.
33.
Atkinson
,
T. S.
,
Ewers
,
B. J.
, and
Haut
,
R. C.
,
1999
, “
The Tensile and Stress Relaxation Responses of Human Patellar Tendon Varies with Specimen Cross-sectional Area
,”
J. Biomech.
,
32
, pp.
907
914
.
34.
Goto
,
M.
, and
Kimoto
,
Y.
,
1966
, “
Hysteresis and Stress-relaxation of the Blood Vessels Studies by a Universal Tensile Testing Instrument
,”
Jpn. J. Physiol.
,
15
, pp.
169
184
.
35.
Oyen-Tiesma, M., Lewis, J. L., and Cook, R. F., 2000, “Viscoelastic Stress-relaxation of Cultured Neocartilage,” SEM IX International Congress and Exposition on Experimental Mechanics Society for Experimental Mechanics, 4, pp. 1–3.
36.
Hasegawa
,
M.
,
1983
, “
Rheological Properties and Wall Structures of Large Veins
,”
Biorheology
,
20
, pp.
531
545
.
37.
Grinnell
,
F.
,
Fukamizu
,
H.
,
Pawelek
,
P.
, and
Nakagawa
,
S.
,
1989
, “
Collagen Processing, Crosslinking and Fibril Bundle Assembly in Matrix Produced by Fibroblasts in Long-term Cultures Supplemented with Ascorbic Acid
,”
Exp. Cell Res.
,
181
, pp.
483
491
.
38.
Layman, D. L., McGoodwin, E. B., and Martin, G. R., 1971, “The Nature of Collagen Synthesis by Cultured Human Fibroblasts,” Proceedings of the National Academy of Science USA, 68, pp. 454–458.
39.
Peterkofsky
,
B.
, and
Diegelmann
,
R.
,
1971
, “
Use of a Mixture of Proteinase-free Collagenases for the Specific Assay of Radioactive Collagen in the Presence of other Proteins
,”
Biochemistry
,
10
,
988
994
.
40.
Johnson
,
D. J.
,
LaBourene
,
J.
,
Rabinoviatch
,
M.
, and
Keeley
,
F. W.
,
1993
, “
Relative Efficiency of Incorporation of Newly Synthesized Elastin and Collagen into Aorta, Pulmonary Artery and Pulmonary Vein of Growing Pigs
,”
Connect. Tissue Res.
,
29
, pp.
213
221
.
41.
Strauss
,
B. H.
,
Chisholm
,
R. J.
,
Keely
,
F. W.
,
Gotlieb
,
A. J.
,
Logan
,
R. A.
, and
Armstrong
,
P. W.
,
1994
, “
Extracellular Matrix Remodeling after Balloon Angioplasty Injury in a Rabbit Model of Restenosis
,”
Circ. Res.
,
75
, pp.
650
658
.
42.
Martora
,
A. M.
,
Sheng
,
G.
,
Springall
,
F.
,
Iland
,
H. J.
,
O’Grady
,
R. L.
, and
Lyons
,
J. G.
,
1999
, “
Absolute Quantitation of Specific mRNAs in Cell and Tissue Samples by Comparative PCR
,”
BioTechniques
,
27
, pp.
136
144
.
43.
Clark, G., 1981, “Methods for Connective Tissue,” In Staining Procedures, Fourth Edition, Clark, G., editor, pp. 113–130.
44.
Burton
,
A. C.
,
1954
, “
Relation of Structure to Function of Tissues of the Wall of Blood Vessels
,”
Physiol. Rev.
,
34
, pp.
619
642
.
45.
Burton, A. C., 1965, Physiology and Biophysics of the Circulation, Yearbook Medical Publishers, Chicago, pp. 67.
46.
Baird
,
R. N.
,
Kidson
,
I. G.
, and
L’Italien
,
G.
,
1977
, “
Dynamic Compliance of Arterial Grafts
,”
Am. J. Physiol.
,
233
, pp.
H568–H572
H568–H572
.
47.
Takehara
,
K.
,
Grotendorst
,
G. R.
,
Trojanowska
,
M.
, and
Leroy
,
E. C.
,
1986
, “
Ascorbate Effects on Type I Procollagen Synthesis by Human Adult Skin Fibroblasts: Different Migration Positions of Type I Procollagen Chains on SDS Polyacrylamide Gel after Incubation with Ascorbate
,”
Collagen Relevant Research
,
6
, pp.
455
466
.
48.
Barone
,
L. M.
,
Faris
,
B.
,
Chipman
,
S. D.
,
Toselli
,
P.
,
Oakes
,
B. W.
, and
Franzblau
,
C.
,
1985
, “
Alteration of the Extracellular Matrix of Smooth Muscle Cells by Ascorbate Treatment
,”
Biochim. Biophys. Acta
,
840
, pp.
245
254
.
49.
De Clerck
,
Y. A.
, and
Jones
,
P. A.
,
1980
, “
Effects of Ascorbic Acid on the Resistance of the Extracellular Matrix to Hydrolysis by Tumor Cells
,”
Cancer Res.
,
40
, pp.
3228
3231
.
50.
Scott-Burden
,
T.
,
Davies
,
P. J.
, and
Gevers
,
W.
,
1979
, “
Elastin Biosynthesis by Smooth Muscle Cells Cultured under Scorbutic Conditions
,”
Biochem. Biophys. Res. Commun.
,
91
, pp.
739
746
.
51.
Houglum
,
K. P.
,
Brenner
,
D. A.
, and
Chojkier
,
M.
,
1991
, “
Ascorbic Acid Stimulation of Collagen Biosynthesis Independent of Hydroxylation
,”
Am. J. Clin. Nutr.
,
54
, pp.
1141S–1143S
1141S–1143S
.
52.
Geesin
,
J. C.
,
Hendrick
,
J. F.
,
Gordon
,
J. S.
, and
Berg
,
R. A.
,
1991
, “
Modulation of Collagen Synthesis by Growth Factors: the Role of Ascorbate-stimulated Lipid Peroxidation
,”
Arch. Biochem. Biophys.
,
67
, pp.
952
956
.
53.
Kurata
,
S.
,
Senoo
,
H.
, and
Hata
,
R.
,
1993
, “
Transcriptional Activiation of Type I Collagen Genes by Ascorbic Acid 2-phosphate in Human Skin Fibroblasts and its Failure in Cells from a Patient with α2 (I) Chain Defective Ehlers-Danlos Syndrome
,”
Exp. Cell Res.
,
206
, pp.
63
71
.
54.
Lyons
,
B.
, and
Schwarz
,
R.
,
1984
, “
Ascorbate Stimulation of PAT Cells Causes an Increase in Transcription Rates and a Decrease in Degradation Rates of Procollagen mRNA
,”
Nucleic Acids Res.
,
5
, pp.
2569
2579
.
55.
Peterkofsky
,
B.
,
1991
, “
Ascorbate Requirement for Hydroxylation and Secretion of Procollagen: Relationship to Inhibition of Collagen Synthesis in Scurvy
,”
Am. J. Clin. Nutr.
,
54
, pp.
1135S–1140S
1135S–1140S
.
56.
Chojkier
,
M.
,
Houglum
,
K.
,
Solis-Herruzo
,
J.
,
Brenner
,
D.
,
1989
, “
Stimulation of Collagen Gene Expression by Ascorbic Acid in Cultured Human Fibroblasts
,”
J. Biol. Chem.
,
264
, pp.
16957
16962
.
57.
Lee
,
K. W.
,
Mossine
,
V.
, and
Ortwerth
,
B. J.
,
1998
, “
The Relative Ability of Glucose and Ascorbate to Glycate and Crosslink Lens Proteins In Vitro
,”
Exp. Eye Res.
,
67
, pp.
95
104
.
58.
Tsuchiya
,
H.
, and
Bates
,
C. J.
,
1997
, “
Vitamin C and Copper Interactions in Guinea-pigs and a Study of Collagen Cross-links
,”
Br. J. Nutrition
,
77
, pp.
315
325
.
59.
Broberg
,
A.
, and
Heino
,
J.
,
1996
, “
Integrin alpha2beta1-dependent Contraction of Floating Collagen Gels and Induction of Collagenase are Inhibited by Tyrosine Kinase Inhibitors
,”
Exp. Cell Res.
,
228
, pp.
29
35
.
60.
Langholz
,
O.
,
Rockel
,
D.
,
Mauch
,
C.
,
Kozlowska
,
E.
,
Bank
,
I.
,
Krieg
,
T.
, and
Eckes
,
B.
,
1995
, “
Collagen and Collagenase Gene Expression in Three-dimensional Collagen Lattices are Differentially Regulated by alpha1beta1 and alpha2beta1 Integrins
,”
J. Cell Biol.
,
131
, pp.
1903
1915
.
61.
Ewart
,
A. K.
,
Jin
,
W.
,
Atkinson
,
D.
,
Morris
,
D. A.
, and
Keating
,
M. T.
,
1994
, “
Supravalvular Aortic Stenosis Associated with a Deletion Disrupting the Elastin Gene
,”
J. Clin. Invest.
,
93
, pp.
1071
1077
.
62.
French
,
B. A.
,
1998
, “
Gene Therapy and Cardiovascular Disease
,”
Curr. Opin. Cardiol.
,
13
, pp.
205
213
.
63.
O’Brien
,
J. P.
,
1982
, “
Destruction of Elastic Tissue (elastolysis) as a Link Between Atherosclerosis and the Temporal Arteritis/Polymyalgia Rheumatica Syndrome
,”
Pathol. Biol. (Paris)
,
32
, pp.
123
138
.
64.
Fedewa
,
M. M.
,
Oegema
,
T. R.
,
Schwartz
,
M. H.
,
MacLeod
,
A.
, and
Lewis
,
J. L.
,
1998
, “
Chondrocytes in Culture Produce a Mechanically Functional Tissue
,”
J. Orthop. Res.
,
16
, pp.
227
236
.
65.
DeWitt
,
M. T.
,
Handley
,
C. J.
,
Oakes
,
B. W.
, and
Lowther
,
D. A.
,
1984
, “
In Vitro Response of Chondrocytes to Mechanical Loading: the Effect of Short Term Mechanical Tension
,”
Connective Tissue Research
12
, pp.
97
109
.
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