Calcific aortic valve disease (CAVD) is a significant cardiovascular disorder characterized by the formation of calcific nodules (CN) on the valve. In vitro assays studying the formation of these nodules were developed and have led to many significant mechanistic findings; however, the biophysical properties of CNs have not been clearly defined. A thorough analysis of dystrophic and osteogenic nodules utilizing scanning electron microscopy (SEM), energy dispersive spectrometry (EDS), and atomic force microscopy (AFM) was conducted to describe calcific nodule properties and provide a link between calcific nodule morphogenesis in vitro and in vivo. Unique nodule properties were observed for dystrophic and osteogenic nodules, highlighting the distinct mechanisms occurring in valvular calcification.

References

References
1.
Otto
,
C. M.
,
Kuusisto
,
J.
,
Reichenbach
,
D. D.
,
Gown
,
A. M.
, and
O'brien
,
K. D.
,
1994
, “
Characterization of the Early Lesion of ‘Degenerative' Valvular Aortic Stenosis. Histological and Immunohistochemical Studies
,”
Circulation
,
90
(
2
), pp.
844
853
.10.1161/01.CIR.90.2.844
2.
Freeman
,
R. V.
, and
Otto
,
C. M.
,
2005
, “
Spectrum of Calcific Aortic Valve Disease: Pathogenesis, Disease Progression, and Treatment Strategies
,”
Circulation
,
111
(
24
), pp.
3316
3326
.10.1161/CIRCULATIONAHA.104.486738
3.
Iung
,
B.
, and
Vahanian
,
A.
,
2011
, “
Epidemiology of Valvular Heart Disease in the Adult
,”
Nat. Rev. Cardiol.
,
8
(
3
), pp.
162
172
.10.1038/nrcardio.2010.202
4.
Dweck
,
M. R.
,
Boon
,
N. A.
, and
Newby
,
D. E.
,
2012
, “
Calcific Aortic Stenosis: A Disease of the Valve and the Myocardium
,”
J. Am. Coll. Cardiol.
,
60
(
19
), pp.
1854
1863
.10.1016/j.jacc.2012.02.093
5.
Chen
,
J. H.
, and
Simmons
,
C. A.
,
2011
, “
Cell-Matrix Interactions in the Pathobiology of Calcific Aortic Valve Disease: Critical Roles for Matricellular, Matricrine, and Matrix Mechanics Cues
,”
Circ. Res.
,
108
(
12
), pp.
1510
1524
.10.1161/CIRCRESAHA.110.234237
6.
Mohler
III,
E. R.
,
Gannon
,
F.
,
Reynolds
,
C.
,
Zimmerman
,
R.
,
Keane
,
M. G.
, and
Kaplan
,
F. S.
,
2001
, “
Bone Formation and Inflammation in Cardiac Valves
,”
Circulation
,
103
(
11
), pp.
1522
1528
.10.1161/01.CIR.103.11.1522
7.
Langanay
,
T.
,
Flecher
,
E.
,
Fouquet
,
O.
,
Ruggieri
,
V. G.
,
De La Tour
,
B.
,
Felix
,
C.
,
Lelong
,
B.
,
Verhoye
,
J. P.
,
Corbineau
,
H.
, and
Leguerrier
,
A.
,
2012
, “
Aortic Valve Replacement in the Elderly: The Real Life
,”
Ann. Thorac. Surg.
,
93
(
1
), pp.
70
77
; Discussion 77–78.10.1016/j.athoracsur.2011.07.033
8.
Thomas
,
M.
,
Schymik
,
G.
,
Walther
,
T.
,
Himbert
,
D.
,
Lefevre
,
T.
,
Treede
,
H.
,
Eggebrecht
,
H.
,
Rubino
,
P.
,
Michev
,
I.
,
Lange
,
R.
,
Anderson
,
W. N.
, and
Wendler
,
O.
,
2010
, “
Thirty-Day Results of the Sapien Aortic Bioprosthesis European Outcome (Source) Registry: A European Registry of Transcatheter Aortic Valve Implantation Using the Edwards Sapien Valve
,”
Circulation
,
122
(
1
), pp.
62
69
.10.1161/CIRCULATIONAHA.109.907402
9.
Hutcheson
,
J. D.
,
Aikawa
,
E.
, and
Merryman
,
W. D.
,
2014
, “
Potential Drug Targets for Calcific Aortic Valve Disease
,”
Nat. Rev. Cardiol.
,
11
(
4
), pp.
218
231
.10.1038/nrcardio.2014.1
10.
Hutcheson
,
J. D.
,
Ryzhova
,
L. M.
,
Setola
,
V.
, and
Merryman
,
W. D.
,
2012
, “
5-Ht(2b) Antagonism Arrests Non-Canonical Tgf-Beta1-Induced Valvular Myofibroblast Differentiation
,”
J. Mol. Cell. Cardiol.
,
53
(
5
), pp.
707
714
.10.1016/j.yjmcc.2012.08.012
11.
Yip
,
C. Y.
,
Blaser
,
M. C.
,
Mirzaei
,
Z.
,
Zhong
,
X.
, and
Simmons
,
C. A.
,
2011
, “
Inhibition of Pathological Differentiation of Valvular Interstitial Cells by C-Type Natriuretic Peptide
,”
Arterioscler. Thromb. Vasc. Biol.
,
31
(
8
), pp.
1881
1889
.10.1161/ATVBAHA.111.223974
12.
Walker
,
G. A.
,
Masters
,
K. S.
,
Shah
,
D. N.
,
Anseth
,
K. S.
, and
Leinwand
,
L. A.
,
2004
, “
Valvular Myofibroblast Activation by Transforming Growth Factor-Beta: Implications for Pathological Extracellular Matrix Remodeling in Heart Valve Disease
,”
Circ. Res.
,
95
(
3
), pp.
253
260
.10.1161/01.RES.0000136520.07995.aa
13.
Yip
,
C. Y.
,
Chen
,
J. H.
,
Zhao
,
R.
, and
Simmons
,
C. A.
,
2009
, “
Calcification by Valve Interstitial Cells is Regulated by the Stiffness of the Extracellular Matrix
,”
Arterioscler. Thromb. Vasc. Biol.
,
29
(
6
), pp.
936
942
.10.1161/ATVBAHA.108.182394
14.
Jian
,
B.
,
Narula
,
N.
,
Li
,
Q. Y.
,
Mohler
III,
E. R.
, and
Levy
,
R. J.
,
2003
, “
Progression of Aortic Valve Stenosis: Tgf-Beta1 is Present in Calcified Aortic Valve Cusps and Promotes Aortic Valve Interstitial Cell Calcification Via Apoptosis
,”
Ann. Thorac. Surg.
,
75
(
2
), pp.
457
465
; Discussion 465–466.10.1016/S0003-4975(02)04312-6
15.
Benton
,
J. A.
,
Kern
,
H. B.
,
Leinwand
,
L. A.
,
Mariner
,
P. D.
, and
Anseth
,
K. S.
,
2009
, “
Statins Block Calcific Nodule Formation of Valvular Interstitial Cells by Inhibiting Alpha-Smooth Muscle Actin Expression
,”
Arterioscler. Thromb. Vasc. Biol.
,
29
(
11
), pp.
1950
1957
.10.1161/ATVBAHA.109.195271
16.
Mohler
III,
E. R.
,
Chawla
,
M. K.
,
Chang
,
A. W.
,
Vyavahare
,
N.
,
Levy
,
R. J.
,
Graham
,
L.
, and
Gannon
,
F. H.
,
1999
, “
Identification and Characterization of Calcifying Valve Cells from Human and Canine Aortic Valves
,”
J. Heart Valve Dis.
,
8
(
3
), pp.
254
260
.
17.
Gu
,
X.
, and
Masters
,
K. S.
,
2011
, “
Role of the Rho Pathway in Regulating Valvular Interstitial Cell Phenotype and Nodule Formation
,”
Am. J. Physiol. Heart Circ. Physiol.
,
300
(
2
), pp.
H448
H458
.10.1152/ajpheart.01178.2009
18.
Gu
,
X.
, and
Masters
,
K. S.
,
2009
, “
Role of the Mapk/Erk Pathway in Valvular Interstitial Cell Calcification
,”
Am. J. Physiol. Heart Circ. Physiol.
,
296
(
6
), pp.
H1748
H1757
.10.1152/ajpheart.00099.2009
19.
Fisher
,
C. I.
,
Chen
,
J.
, and
Merryman
,
W. D.
,
2013
, “
Calcific Nodule Morphogenesis by Heart Valve Interstitial Cells Is Strain Dependent
,”
Biomech. Model. Mechanobiol.
,
12
(
1
), pp.
5
17
.10.1007/s10237-012-0377-8
20.
Hutcheson
,
J. D.
,
Chen
,
J.
,
Sewell-Loftin
,
M. K.
,
Ryzhova
,
L. M.
,
Fisher
,
C. I.
,
Su
,
Y. R.
, and
Merryman
,
W. D.
,
2013
, “
Cadherin-11 Regulates Cell-Cell Tension Necessary for Calcific Nodule Formation by Valvular Myofibroblasts
,”
Arterioscler. Thromb. Vasc. Biol.
,
33
(
1
), pp.
114
120
.10.1161/ATVBAHA.112.300278
21.
Bowler
,
M. A.
, and
Merryman
,
W. D.
,
2015
, “
In Vitro Models of Aortic Valve Calcification: Solidifying a System
,”
Cardiovasc. Pathol.
25
(
1
), pp.
1
10
.10.1016/j.carpath.2014.08.003
22.
Benton
,
J. A.
,
Kern
,
H. B.
, and
Anseth
,
K. S.
,
2008
, “
Substrate Properties Influence Calcification in Valvular Interstitial Cell Culture
,”
J. Heart Valve Dis.
,
17
(
6
), pp.
689
699
.
23.
Valente
,
M.
,
Bortolotti
,
U.
, and
Thiene
,
G.
,
1985
, “
Ultrastructural Substrates of Dystrophic Calcification in Porcine Bioprosthetic Valve Failure
,”
Am. J. Pathol.
,
119
(
1
), pp.
12
21
.
24.
Zhao
,
Y.
,
Urganus
,
A. L.
,
Spevak
,
L.
,
Shrestha
,
S.
,
Doty
,
S. B.
,
Boskey
,
A. L.
, and
Pachman
,
L. M.
,
2009
, “
Characterization of Dystrophic Calcification Induced in Mice by Cardiotoxin
,”
Calcif. Tissue Int.
,
85
(
3
), pp.
267
275
.10.1007/s00223-009-9271-5
25.
Rajamannan
,
N. M.
,
Subramaniam
,
M.
,
Rickard
,
D.
,
Stock
,
S. R.
,
Donovan
,
J.
,
Springett
,
M.
,
Orszulak
,
T.
,
Fullerton
,
D. A.
,
Tajik
,
A. J.
,
Bonow
,
R. O.
, and
Spelsberg
,
T.
,
2003
, “
Human Aortic Valve Calcification is Associated With an Osteoblast Phenotype
,”
Circulation
,
107
(
17
), pp.
2181
2184
.10.1161/01.CIR.0000070591.21548.69
26.
Cloyd
,
K. L.
,
El-Hamamsy
,
I.
,
Boonrungsiman
,
S.
,
Hedegaard
,
M.
,
Gentleman
,
E.
,
Sarathchandra
,
P.
,
Colazzo
,
F.
,
Gentleman
,
M. M.
,
Yacoub
,
M. H.
,
Chester
,
A. H.
, and
Stevens
,
M. M.
,
2012
, “
Characterization of Porcine Aortic Valvular Interstitial Cell 'Calcified' Nodules
,”
PLoS One
,
7
(
10
), p.
e48154
.10.1371/journal.pone.0048154
27.
Chen
,
J. H.
,
Yip
,
C. Y.
,
Sone
,
E. D.
, and
Simmons
,
C. A.
,
2009
, “
Identification and Characterization of Aortic Valve Mesenchymal Progenitor Cells With Robust Osteogenic Calcification Potential
,”
Am. J. Pathol.
,
174
(
3
), pp.
1109
1119
.10.2353/ajpath.2009.080750
28.
Bertazzo
,
S.
,
Gentleman
,
E.
,
Cloyd
,
K. L.
,
Chester
,
A. H.
,
Yacoub
,
M. H.
, and
Stevens
,
M. M.
,
2013
, “
Nano-Analytical Electron Microscopy Reveals Fundamental Insights into Human Cardiovascular Tissue Calcification
,”
Nat. Mater.
,
12
(
6
), pp.
576
583
.10.1038/nmat3627
29.
Merryman
,
W. D.
,
Liao
,
J.
,
Parekh
,
A.
,
Candiello
,
J. E.
,
Lin
,
H.
, and
Sacks
,
M. S.
,
2007
, “
Differences in Tissue-Remodeling Potential of Aortic and Pulmonary Heart Valve Interstitial Cells
,”
Tissue Eng.
,
13
(
9
), pp.
2281
2289
.10.1089/ten.2006.0324
30.
Wang
,
H.
,
Haeger
,
S. M.
,
Kloxin
,
A. M.
,
Leinwand
,
L. A.
, and
Anseth
,
K. S.
,
2012
, “
Redirecting Valvular Myofibroblasts into Dormant Fibroblasts through Light-Mediated Reduction in Substrate Modulus
,”
PLoS One
,
7
(
7
), p.
e39969
.10.1371/journal.pone.0039969
31.
Engler
,
A. J.
,
Sen
,
S.
,
Sweeney
,
H. L.
, and
Discher
,
D. E.
,
2006
, “
Matrix Elasticity Directs Stem Cell Lineage Specification
,”
Cell
,
126
(
4
), pp.
677
689
.10.1016/j.cell.2006.06.044
32.
Tse
,
J. R.
, and
Engler
,
A. J.
,
2010
, “
Preparation of Hydrogel Substrates With Tunable Mechanical Properties
,”
Curr. Protoc. Cell. Biol.
Chapter 10.10.1002/0471143030.cb1016s47
33.
Gregory
,
C. A.
,
Gunn
,
W. G.
,
Peister
,
A.
, and
Prockop
,
D. J.
,
2004
, “
An Alizarin Red-Based Assay of Mineralization by Adherent Cells in Culture: Comparison With Cetylpyridinium Chloride Extraction
,”
Anal. Biochem.
,
329
(
1
), pp.
77
84
.10.1016/j.ab.2004.02.002
34.
Sewell-Loftin
,
M. K.
,
Brown
,
C. B.
,
Baldwin
,
H. S.
, and
Merryman
,
W. D.
,
2012
, “
A Novel Technique for Quantifying Mouse Heart Valve Leaflet Stiffness With Atomic Force Microscopy
,”
J. Heart Valve Dis.
,
21
(
4
), pp.
513
520
.
35.
Goldstein
,
J.
,
2003
,
Scanning Electron Microscopy and X-Ray Microanalysis
,
Kluwer Academic/Plenum Publishers
,
New York
.
36.
Bonucci
,
E.
,
2007
,
Biological Calcification: Normal and Pathological Processes in the Early Stages
,
Springer
,
Berlin New York
.
37.
Benjamin Kramer
,
M. J. S.
,
1928
, “
Composition of Bone: Iv. Primary Calcification
,”
J. Biol. Chem.
,
79
, pp.
147
160
.
38.
Gilbert
,
S. F.
,
2000
,
Developmental Biology
,
Sinauer Associates
,
Sunderland, MA
.
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