Scientific Research and Essays Vol. 6(14), pp.

3049-3057, 18 July, 2011

Available online at http://www.academicjournals.org/SRE
ISSN 1992-2248 2011 Academic Journals

Full Length Research Paper

Clinical/computational investigation on post traumatic

heterotopic ossification in head injury patients; the
positive and negative role of temperature in
physical treatment
Babak Khalili Hadad1*, Masoud Ramin Mirza Zadeh Javaheri2 and Sara Nouroozi3
1

Department of Biological Sciences, Roudehen Branch, Islamic Azad University, Roudehen, Iran.
2
Department of Orthopedics, Kashan University of Medical Sciences, Kashan, Iran.
3
Department of Biological Sciences, Pishva Branch, Islamic Azad University, Varamin, Iran.
Accepted 10 June 2011

Bone morphogenetic proteins (BMPs) induce bone formation and healing. Bone can form in
extraskeletal tissue in response to trauma and elevating BMPs. Several BMPs stimulated to increase
due to neuroprotective effect. To get the injured zone of fraction warm is one of the suggested methods
to prevent heterotopic ossification (HO). But about half of patients get the worst clinical situation.
Montecarlo simulation was done with Hyperchem 8 and Ramachandran plot were designed with VMD
1.8.2. pdb files of both 1 tfg and 2 tgi proteins were selected from Protein Data Bank. and of all 110
amino acid were calculated in both proteins after Montecarlo simulation at 290, 310 and 315 K for 200
ps, surrounded by water. 1 tfg is affected by increasing the temperature. The favored and allowed
regions were decreased in population of dihedral angles. Increasing the temperature more than room
temperature can cause more allowed dihedrals for 2 tgi. It is suggested that utilization of
thermogenerator lamp progress the HO in such patients.
Key words: Transforming growth factor, bone morphogenic proteins, post traumatic heterotopic ossification.

INTRODUCTION
Bone formation and healing study is considered as a
multidisciplinary field that applies principles of
Biochemistry, Cell biology, Histology and Orthopedics
surgery sciences, towards the development of biological
substitutes for the restoration, maintenance or
improvement of tissue form and function (Sanders et al.,
2010; Ruimerman, 2005). Bone is a living tissue and its
contains proteins, calcium phosphate and is capable of
regeneration and remodeling. Bone formation occurs in a

*Corresponding author. E-mail: khalili@riau.ac.ir.

+989123177387. Fax: +9821772956278.

Tel:

Abbreviations: BMPs, Bone morphogenetic proteins; HO,

heterotopic ossification; TGF-, transforming growth factor ;
MC, monte carlo.

dynamic integration of biochemical, cellular and hormonal

processes that are facilitated by states of bone
deposition, bone resorption and bone remodeling
(Ruimerman, 2005). Specific bone-inductive proteins can,
induce bone formation and healing in vivo. Bone can form
in extraskeletal tissue in response to trauma (Sanders et
al., 2010; Atkinson et al., 2010). This initiated research
into bone regeneration by putative soluble signals, led to
the discovery and identification of the effect of
temperature on the 1tfg (Figure 1a) and 2 tgi (Figure 1b)
growth factors entirely novel family of protein initiators,
collectively called the BMPs, which belong to the
transforming growth factor (TGF-) super family (Betts
and Sternberg, 1999).
BMPs affect no biological matrices; make them to have
the capacity to induce bone, cartilage, ligament and
tendon at both heterotopic and orthotropic sites. Release
of such factors cause heterotopic ossification (Figure 2). In

3050

Sci. Res. Essays

Figure 1. Structure of 1tfg and 2tgi, inducible proteins, the effective proteins in heterotopic ossification.

www.pdb.org
Figure 1. Structure of 1tfg and 2tgi, inducible proteins, the effective proteins in hetrotopic ossification.

cerebral ischemia and Parkinson's disease and may

therefore have direct clinical applications for the
treatment of central nervous system disorders. Our
clinical trial shows that in some cases after the head
trauma, the movement limitations in some skeleton
regions are caused by heterotopic ossification. BMPs
have a grand potential in bone formation at fracture sites
and unpredicted zones. This will cause the bone hinges
to be locked in an ectopic regions. Limited human studies
have been performed to discuss the reasons. The action
of BMP is mediated through receptor kinases and
transcription factors called Smads that regulate the
expression of target genes. Heterotopic ossification is a
type of bone formation in the soft tissues. Soft tissue
bone deposition may vary from the minimal and
inconsequential to massive and clinically significant
(Ruimerman, 2005).
EXPERIMENTAL
Molecular modeling methods

Figure. 2 Heterotopic ossification in the injury zone, cause

limitation in movement.

addition, several BMPs have been shown to have a

neuroprotective effect in animal models of head injury,

The modeling of biological materials requires conformational

sampling. Low-energy geometries are obtained from multiple
searching methods and dynamics minimizations. Mathematical
methods that use random numbers for solving quantitative
problems are commonly called Monte Carlo methods. Monte Carlo
method solves problem by randomly selecting a large number of
points within the square and determining how many of these points
fall within the circle. Refereeing to steric hindrances, the main chain
of a polypeptide usually has energetically favorable conformations
(Ruimerman et al., 2001; Prosinecki et al., 2007). These
conformations can be characterized by the value of two torsion

Hadad et al.

3051

Figure.3 The computational method algorithm.

angles, and , . The third angle is restricted to values of 180

for trans-peptide, and 0 for cis-peptides). The angle is defined by
the torsion Ci1-Ni-Ci-Ci, and y by the torsion Ni-Ci-Ci-Ni+1. The
Ramachandran plot is defined as the distribution of and . More
than ten years ago, such plots were used to remove two structures
from a database of high-resolution structures that had been created
for model building in experimental maps. The Ramachandran plot is
one of the simplest and most sensitive methods to assess the
protein model in the absence of experimental data. It will clearly
show how well the find , angles cluster (Podtelezhnikov, 2005).
In recent study, 1 tfg and 2 tgi protein crystallography concluded
pdb file selected from protein data bank as the main targets. and
of all 110 amino acid were calculated in both proteins after
Montecarlo simulation with Hyperchem7.4 (2005), and the
Ramachandran plot was designed, using VMD software, to
compare the amino acid positions before and after Monte Carlo
(MC) simulation at 290, 310 and 315 K for 200 ps Both two proteins
were included sufficient water in a box around to make the
simulation more real (Ruimerman, 2005; Betts and Sternberg,
1999). The conformational changes and dihedrals of and were
analyzed step by step for both proteins, as it is shown in Figure 3.

RESULTS
MC simulation were done at 290, 310 and 315 K for 200
ps. Due to the crystallographic data the 1tfg has 104/110
(94.50%) amino acid in favored region and
109/110(99.10%) of them are in allowed zone. Where, 2
tgi the 106/110 (96.40%) amino acid in favored region
and 110/110 (100%) (Table 3). Increasing in temperature
of Montecarlo simulation reveals the decreasing in both

favored and allowed and dihedrals for 1tfg, although

some data were not obtained because of protein
complicity during simulation. In 315 K, the structure of
protein was not calculable by software (Tables 1 and 3).
2 tgi, shows another behavior, increasing the temperature
cause the decreasing in and dihedral angels, where
increasing up to 290 K decrease the allowed dihedral and
up to this temperature the allowed dihedrals decreased
(Tables 2 and 3).
DISCUSSION
Nowadays, to get the injured zone of fraction warm, by
different methods in Physiotherapy centers, one of the
suggested methods to prevent heterotopic ossification
(Ruimerman, 2005). The present study will prove the
accuracy of this method, biochemically. There are two
effective growth hormone, in the super family of
Transforming growth factor , 1t fg and 2 tgi. Primary
studies revealed some differences between these two
peptides (Figure 4). The behavior of these two factors is
different in various temperatures due to the result.

CONCLUSIONS
Heterotopic ossification develops when soft tissue
mesenchymal cells undergo metaplasia. This is caused

3052

Sci. Res. Essays

Table 1. and dihedral analysis of 110 amino acid, 1tfg. Results were obtained by ramachandran plot, after
montecarlo simulation.

No
1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
17
18
19
20
21
22
23
24
25
26
27
28
29
30
31
32
33
34
35
36
37
38
39
40
41
42
43
44
45
46
47
48

Amino acid
ALA
LEU
ASP
ALA
ALA
TYR
CYS
PHE
ARG
ASN
VAL
GLN
ASP
ASN
CYS
CYS
LEU
ARG
PRO
LEU
TYR
ILE
ASP
PHE
LYS
ARG
ASP
LEU
GLY
TRP
LYS
TRP
ILE
HIS
GLU
PRO
LYS
GLY
TYR
ASN
ALA
ASN
PHE
CYS
ALA
GLY
ALA
CYS

Phi
0.00
-74.01
-43.06
-72.75
53.93
0.00
89.61
-161.34
-179.96
0.00
-131.16
-127.89
-26.79
-167.52
-21.84
-56.14
-6.09
137.58
30.48
153.80
-142.61
-108.52
-20.03
-40.05
-54.70
-43.81
-170.14
-63.28
-58.64
140.40
-158.54
-43.86
29.18
-1.66
-170.14
-

273 K
Psi
152.38
177.83
-38.97
-22.78
0.00
92.34
-34.19
0.00
165.94
61.08
160.06
137.93
-117.11
74.91
-55.72
72.84
159.62
108.53
0.00
58.02
-36.66
145.73
-59.66
0.00
76.11
-35.32
126.09
-174.76
0.00
177.47
166.01
105.91
-37.53
-170.94
161.52
-

Region
N.A.
A.
A.
A.
N.A.
N.A.
N.A.
N.A.
A.
F.
F.
F.
N.A.
A.
N.A.
N.A.
N.A.
N.A.
N.A.
N.A.
F.
N.A.
N.A.
A.
A.
A.
A.
N.A.
N.A.
A.
A.
N.A.
N.A.
A.
-

Temperature (k)
290
Phi
Psi
Region
0.00
170.92
N.A.
-85.90
45.74
A.
-96.83
-171.98
A.
-71.95
-19.39
A.
-103.41
-28.96
A.
-109.17
-23.49
A.
-81.29
-12.91
N.A.
-92.28
-9.39
N.A.
-176.87
114.47
A.
-70.45
122.65
F.
-102.08
-174.74
A.
-97.11
-1.63
N.A.
-98.05
-176.06
A.
-64.37
97.94
F.
-127.98
154.33
F.
-73.75
137.29
F.
-119.97
149.46
F.
-81.23
121.44
F.
-140.48
89.94
F.
-80.17
119.90
F.
-107.84
106.98
F.
-75.45
131.01
F.
-68.22
-43.71
F.
-87.60
-27.70
A.
-85.96
-20.77
A.
-115.35
-22.60
N.A.
-115.72
-10.85
N.A.
76.68
0.29
N.A.
-74.48
117.40
F.
-108.86
2.18
N.A.
-85.03
-15.41
N.A.
-89.47
109.16
F.
-78.12
-14.50
N.A.
-168.68
133.06
A.
-81.30
-171.98
A.
-88.19
-1.56
N.A.
170.17
176.10
N.A.
-157.03
174.03
A.
-102.11
65.50
A.
-64.33
-38.18
F.
74.18
-173.37
N.A.
-167.54
165.96
A.
-113.31
135.43
F.
-145.36
-176.56
A.
91.31
174.32
N.A.
-88.42
161.57
F.
-116.47
78.86
A.

Phi
0.00
-62.98
-89.05
-35.37
-19.92
-75.17
-92.88
-54.96
-72.49
171.24
-32.53
-87.19
-80.70
-61.47
-20.06
-134.45
-50.49
-134.88
-68.15
-138.69
-48.71
-99.34
-61.21
-6.29
-80.66
-84.74
-120.94
-111.83
53.08
-82.47
-104.81
-90.40
-76.67
-36.53
177.58
-57.89
-45.37
173.05
-163.50
-95.65
-54.01
63.20
-161.33
-123.62
-153.13
106.48
-65.14
-124.30

310 K
Psi
157.28
35.79
173.43
-63.16
-52.72
-57.02
-37.08
-42.64
-7.71
80.67
103.57
177.52
-30.61
167.93
88.51
137.33
139.19
131.43
123.40
78.75
98.69
99.74
100.11
-76.92
-32.34
-17.85
-29.22
-4.86
15.40
121.70
5.44
-29.42
75.55
-17.33
125.68
171.59
-27.93
-179.60
176.31
46.18
-30.39
-173.02
165.22
129.96
149.60
173.58
174.66
53.09

Region
N.A.
N.A.
F.
N.A.
N.A.
F.
F.
F.
N.A.
N.A.
N.A.
A.
A.
F.
N.A.
F.
A.
F.
F.
A.
A.
F.
F.
N.A.
A.
N.A.
N.A.
N.A.
N.A.
F.
N.A.
A.
A.
N.A.
N.A.
F.
A.
N.A.
A.
A.
A.
N.A.
A.
F.
A.
N.A.
F.
A.

Hadad et al.

3053

Table 1. Contnd

49
50
51
52
53
54
55
56
57
58
59
60
61
62
63
64
65
66
67
68
69
70
71
72
73
74
75
76
77
78
79
80
81
82
83
84
85
86
87
88
89
90
91
92
93
94
95
96
97
98
99
100

PRO
TYR
LEU
TRP
SER
SER
ASP
THR
GLN
HIS
SER
ARG
VAL
LEU
SER
LEU
TYR
ASN
THR
ILE
ASN
PRO
GLU
ALA
SER
ALA
SER
PRO
CYS
CYS
VAL
SER
GLN
ASP
LEU
GLU
PRO
LEU
THR
ILE
LEU
TYR
TYR
ILE
GLY
LYS
THR
PRO
LYS
ILE
GLU
GLN

76.36
-87.11
62.62
44.62
37.11
-21.53
0.00
0.00
0.00
-85.89
-145.89
-132.60
-48.67
-144.43
-105.10
-95.52
-147.12
-47.82
-116.97
-64.43
84.98
-63.32
-90.65
-68.97
-71.27
-45.90
-106.36
-143.57

-15.51
133.68
-6.93
169.25
0.00
-58.09
174.27
-80.76
-104.73
150.93
152.31
137.89
163.32
131.66
107.21
167.73
38.33
159.59
119.20
32.95
-98.14
-24.91
105.25
74.25
84.12
109.10
168.72
69.30

N.A.
F.
N.A.
N.A.
N.A.
N.A.
N.A.
N.A.
N.A.
F.
A.
F.
A.
F.
F.
F.
A.
A.
F.
N.A.
N.A.
A.
F.
A.
A.
A.
F.
A.

-73.18
-64.64
73.56
-86.52
67.79
-65.09
-68.40
-133.01
-71.98
-65.09
-71.13
-83.28
-73.43
-86.85
-78.07
-101.18
-70.18
-75.69
-78.63
-92.99
-123.71
-64.61
-83.27
-89.86
67.62
-92.60
-75.37
-71.51
-98.92
-86.61
-89.27
-78.37
-109.41
-113.92
-136.06
-133.94
-81.31
-136.80
-92.43
-111.39
-141.05
-77.88
-115.31
-88.73
82.14
-104.22
-100.64
-75.11
-90.88
-84.95
-124.57
-131.32

176.43
117.70
12.98
83.77
35.40
110.39
-34.87
179.68
-23.45
110.39
-26.10
-37.46
-29.65
-18.49
-10.69
-31.30
-30.71
-17.99
-36.16
-31.16
57.52
-13.02
-4.56
-13.52
18.86
29.70
150.21
150.78
154.69
81.50
177.40
143.65
-53.16
141.29
162.76
155.04
157.20
126.82
126.34
169.92
68.28
163.21
124.71
57.90
-96.34
6.76
114.78
90.15
101.87
131.29
166.87
63.76

A.
F.
N.A.
A.
N.A.
F.
A.
A.
A.
F.
A.
A.
N.A.
A.
A.
N.A.
A.
A.
A.
N.A.
N.A.
N.A.
N.A.
A.
F.
F.
F.
A.
A.
F.
F.
F.
F.
F.
F.
F.
F.
F.
A.
F.
F.
A.
N.A.
N.A.
F.
F.
F.
F.
F.
A.

-55.44
-34.33
82.39
-65.95
61.53
-13.40
-44.90
-125.74
-58.63
-51.38
-30.70
-42.29
-56.24
-76.56
-37.40
-111.06
-56.05
-39.25
-33.50
-73.67
-119.39
-48.57
-55.12
-74.37
74.67
-57.91
-53.50
-50.58
-91.91
-59.36
-17.32
-51.64
-101.22
-77.72
-130.23
-127.26
-77.63
-128.12
-97.66
-92.40
-145.91
-50.76
-95.23
-73.85
82.54
-107.62
-92.27
-69.48
-75.35
-27.47
-123.96
-131.97

171.26
100.09
-10.63
90.36
9.60
80.75
-51.48
189.86
-29.11
-66.65
-56.92
-62.92
-37.65
-51.05
-16.63
-31.59
-54.53
-58.26
-67.14
-31.88
43.16
-34.01
-28.31
-21.88
4.58
-2.61
144.88
151.92
139.27
44.04
141.19
130.79
-71.56
140.82
161.52
143.88
164.70
132.85
120.89
167.85
41.39
148.68
123.97
37.66
-87.30
-0.01
113.94
73.12
68.76
124.44
157.56
54.28

F.
N.A.
N.A.
A.
N.A.
N.A.
A.
F.
A.
A.
N.A.
A.
A.
F.
N.A.
A.
B-G
N.A.
N.A.
A.
A.
A.
A.
A.
N.A.
N.A.
F.
A.
F.
N.A.
N.A.
A.
A.
F.
F.
F.
F.
F.
F.
F.
A.
A.
F.
N.A.
N.A.
N.A.
F.
A.
N.A.
F.
A.

3054

Sci. Res. Essays

Table 1. Contnd

101
102
103
104
105
106
107
108
109
110

LEU
SER
ASN
MET
ILE
VAL
LYS
SER
CYS
LYS

-67.59
-57.38
90.63
-43.33
-126.02
-69.36
-91.20
-168.77
0.00

112.59
136.34
5.35
-37.81
95.16
111.39
-2.05
-125.55
0.00

F.
F.
N.A.
A.
F.
F.
A.
N.A.
N.A.

-59.32
-80.29
82.11
-79.29
-134.36
-80.92
-97.01
-163.06
-104.92
-170.52

115.32
145.24
18.58
-5.75
108.95
116.11
-9.42
155.11
-177.57
151.30

F.
F.
N.A.
N.A.
F.
F.
N.A.
A.
A.
A.

-46.34
-35.43
85.98
-70.06
-136.90
-63.75
-84.65
-174.69
-74.53
175.17

84.93
134.28
8.25
-8.45
84.32
109.47
-6.50
126.12
-167.73
134.18

N.A.
N.A.
N.A.
N.A.
A.
F.
N.A.
A.
A.
N.A.

A. =Allowed, F. = Favored, N.A. = Not allowed.

Table 2. and dihedral analysis of 110 amino acid, 2 tgi. Results were obtained by ramachandran plot, after montecarlo simulation.

No
1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
17
18
19
20
21
22
23
24
25
26
27
28
29
30
31
32
33

Amino acid
ALA
LEU
ASP
ALA
ALA
TYR
CYS
PHE
ARG
ASN
VAL
GLN
ASP
ASN
CYS
CYS
LEU
ARG
PRO
LEU
TYR
ILE
ASP
PHE
LYS
ARG
ASP
LEU
GLY
TRP
LYS
TRP
ILE

Phi
0.00
-45.23
-119.47
-46.42
-41.73
-46.90
-80.12
-29.29
43.66
-136.57
-95.74
-107.03
-75.45
-75.49
-12.22
-146.74
-57.14
-121.42
-69.66
-158.56
-92.39
-91.58
-73.19
-42.04
-18.85
-35.77
-83.33
-91.30
92.09
-55.74
-55.10
-65.05
-56.50

273
Psi
142.92
63.25
-179.82
-53.01
-69.08
-65.39
-60.07
157.73
26.92
130.12
95.55
147.69
-30.90
154.34
101.05
145.41
130.77
137.56
145.97
97.22
105.93
106.98
119.18
-87.14
-80.05
-51.32
-61.16
-23.94
1.43
72.07
-42.70
-28.28
109.28

Region
N.A.
A.
A.
A.
N.A.
A.
A.
N.A.
A.
F.
F.
F.
A.
F.
N.A.
F.
F.
F.
F.
A.
F.
F.
F.
N.A.
N.A.
N.A.
A.
A.
N.A.
N.A.
F.
A.
F.

Phi
0.00
-65.04
-118.53
-37.11
-48.48
-43.72
-65.42
-57.84
70.84
-116.94
-87.03
-112.57
-39.46
-30.07
-165.57
-24.34
-119.60
-61.73
-172.05
-62.55
-102.77
-41.33
-28.64
-55.84
-16.69
-89.28
-70.82
81.26
-65.56
-60.23
-72.56
-36.39

290
Psi
153.84
55.48
-176.72
-56.81
-58.34
-78.45
-53.73
147.42
-0.52
141.50
98.32
149.07
151.07
120.39
138.18
151.07
132.93
152.84
80.12
115.73
95.40
110.01
-73.07
-81.43
-63.68
-57.89
-35.48
12.00
86.08
-33.72
-37.38
100.39

Temperature (K)
310
Region
Phi
Psi
N.A.
0.00
120.04
N.A.
-55.96
50.40
A.
-117.15 166.27
N.A.
-46.81
-49.01
A.
-34.10
-51.32
N.A.
-50.03
-79.65
F.
-72.38
-48.82
F.
-56.56
163.51
N.A.
56.41
18.29
F.
137.37 125.59
F.
-85.72
73.67
F.
-110.71 136.16
-59.81
-35.13
N.A.
-71.74
152.92
N.A.
-31.41
114.64
A.
-149.28 160.59
N.A.
-49.03
109.58
F.
-105.71 128.84
F.
-56.22
142.21
A.
-162.95 94.89
F.
-82.24
115.82
F.
-90.62
108.31
N.A.
-70.69
107.02
N.A.
-34.62
-67.85
N.A.
-61.12
-58.83
N.A.
-50.35
-47.84
F.
-88.45
-48.03
A.
-93.83
-32.46
N.A.
101.73 -12.88
A.
-68.99
86.61
A.
-43.24
-36.60
A.
-90.01
-18.24
N.A.
-57.90
117.53

Region
N.A.
N.A.
F.
A.
N.A.
N.A.
F.
F.
A.
F.
A.
F.
A.
F.
N.A.
A.
A.
F.
F.
A.
F.
F.
F.
N.A.
F.
A.
F.
A.
N.A.
A.
A.
N.A.
F.

Phi
0.00
-69.14
-118.25
-59.35
-53.43
-88.27
-66.86
-128.56
-78.63
-76.18
-35.53
-38.18
-172.82
-46.51
-112.86
-71.46
-161.12
-74.63
-73.99
-85.74
-58.64
-40.30
-87.89
-103.59
79.22
-78.61
-38.66
-81.73
-74.42

315
Psi
141.21
76.42
-174.35
-44.68
-51.01
-46.98
148.45
117.57
111.46
-31.91
151.88
124.96
135.62
124.95
142.73
145.14
102.31
105.92
117.87
107.23
-65.31
-57.62
-36.21
-39.65
24.59
84.94
35.54
33.03
109.53

Region
N.A.
A.
A.
F.
A.
F.
F.
F.
F.
A.
N.A.
N.A.
A.
A.
F.
F.
A.
F.
F.
F.
A.
N.A.
A.
F.
N.A.
A.
N.A.
A.
F.

Hadad et al.

3055

Table 2. Contd.

34
35
36
37
38
39
40
41
42
43
44
45
46
47
48
49
50
51
52
53
54
55
56
57
58
59
60
61
62
63
64
65
66
67
68
69
70
71
72
73
74
75
76
77
78
79
80
81
82
83
84
85

HIS
GLU
PRO
LYS
GLY
TYR
ASN
ALA
ASN
PHE
CYS
ALA
GLY
ALA
CYS
PRO
TYR
LEU
TRP
SER
SER
ASP
THR
GLN
HIS
SER
ARG
VAL
LEU
SER
LEU
TYR
ASN
THR
ILE
ASN
PRO
GLU
ALA
SER
ALA
ALA
PRO
CYS
CYS
VAL
SER
GLN
ASP
LEU
GLU
PRO

-68.88
-161.14
-60.01
-79.79
-163.08
-163.80
-94.52
-32.32
177.68
-112.90
-164.75
154.23
-42.28
-106.78
-52.57
-4.86
87.60
-54.34
72.54
-34.32
-66.90
-124.62
-42.14
-34.10
-2.37
-43.66
-17.50
-68.70
-51.34
-24.17
-20.94
-41.87
-53.27
-73.82
-118.65
-43.23
-61.14
-24.32
77.88
-79.91
-35.26
-66.49
-95.68
-53.33
-131.01
-32.28
-127.46
-78.25
-129.91
-107.54
-69.33

-51.26
120.79
174.15
-25.65
-163.50
163.36
60.01
-49.70
142.10
141.87
118.38
165.84
142.30
73.83
148.87
81.26
-7.38
86.50
17.11
116.38
-61.62
165.98
-55.07
-81.38
-74.63
-82.76
-64.06
-28.21
-64.23
-83.41
-67.96
-41.66
-57.11
-41.46
59.69
-43.04
-27.11
-50.64
0.45
3.12
146.23
132.17
133.54
107.15
147.48
144.94
-68.39
131.04
153.43
159.93
174.16

F.
A.
F.
A.
A.
A.
A.
N.A.
N.A.
F.
A.
N.A.
N.A.
A.
A.
N.A.
N.A.
N.A.
N.A.
N.A.
A.
F.
N.A.
N.A.
N.A.
N.A.
N.A.
A.
A.
N.A.
N.A.
N.A.
A.
F.
A.
A.
A.
N.A.
N.A.
N.A.
N.A.
F.
F.
A.
F.
N.A.
A.
F.
F.
F.
F.

-69.26
-153.98
-79.53
-57.97
165.20
-163.76
-52.94
-51.77
60.25
-155.70
-97.65
-139.28
123.33
-29.32
-109.31
-73.18
-39.74
77.41
-38.53
58.57
-17.99
-40.84
-99.56
-26.74
-30.91
-46.66
-44.56
-39.59
-48.65
-56.60
-36.40
-5.18
-28.26
-22.72
-67.62
-165.57
-64.18
-58.05
-8.77
53.02
-78.92
-31.22
-62.64
-110.98
-49.74
-137.01
-35.54
-92.09
-113.17
-124.84
-73.73

-52.00
107.42
-179.28
-25.31
149.01
142.39
68.23
40.08
-177.57
152.17
125.50
158.33
146.86
136.09
75.14
-179.63
105.62
-13.43
81.36
16.48
92.14
-84.12
169.13
-65.47
-62.81
-52.86
-74.03
-63.88
-35.91
-55.59
85.25
-83.73
-55.71
-72.68
-44.74
138.18
-30.72
-43.92
-49.09
22.26
-1.78
152.25
195.05
132.27
154.02
152.60
113.71
-80.81
162.15
149.94
177.31

F.
F.
A.
A.
N.A.
A.
N.A.
A.
N.A.
A.
F.
F.
N.A.
N.A.
A.
A.
N.A.
N.A.
N.A.
N.A.
N.A.
N.A.
F.
N.A.
N.A.
A.
N.A.
N.A.
A.
F.
N.A.
N.A.
N.A.
N.A.
F.
A.
A.
F.
N.A.
N.A.
N.A.
N.A.
F.
F.
A.
F.
N.A.
N.A.
F.
F.
A.

-74.42
-153.98
-75.99
-64.79
-142.23
-174.99
-79.10
-45.30
73.26
-157.53
-100.04
-154.78
143.64
-52.92
-110.86
-63.53
-15.89
80.40
-51.24
60.57
-36.14
-64.99
-64.68
-41.86
-32.12
-56.26
-4.49
-14.22
-50.82
-41.85
-31.38
-53.51
-47.60
-18.51
-124.22
-67.01
-22.85
-50.72
64.69
-101.46
-53.75
-37.05
-139.42
-53.14
125.47
-43.40
-97.51
-93.59
-122.11
-111.66
-

-52.14
116.77
172.29
-50.18
-145.80
154.44
72.61
-54.99
168.38
151.74
124.44
126.12
170.14
138.74
58.29
156.87
104.64
-14.84
95.73
19.42
119.78
-56.18
-41.41
-55.51
57.33
-71.95
-92.01
-60.56
-51.35
-78.45
-58.12
-35.61
-76.18
-61.69
69.75
-40.56
47.07
-39.05
37.23
7.21
128.01
167.91
131.33
91.50
159.61
119.64
70.67
131.47
152.95
160.96
-

F.
F.
F.
F.
N.A.
A.
A.
A.
N.A.
A.
F.
F.
N.A.
A.
A.
F.
N.A.
N.A.
A.
A.
N.A.
F.
F.
N.A.
N.A.
A.
N.A.
N.A.
A.
N.A.
N.A.
A.
N.A.
N.A.
A.
F.
N.A.
A.
N.A.
N.A.
A.
N.A.
F.
A.
F.
A.
A.
F.
F.
F.
-

-55.65
-131.14
-48.59
-69.11
-149.12
-163.82
-85.92
-51.63
79.52
-161.61
-112.79
-159.23
117.49
-11.08
-131.59
-37.31
-42.55
87.42
-63.80
63.91
-36.17
-82.79
-106.66
-44.68
-29.19
-57.63
-43.09
-36.97
-47.84
-48.87
-34.20
-59.97
-14.59
-41.67
-109.16
-49.81
-42.64
-40.66
73.00
-96.96
-43.85
-35.54
-90.12
-61.68
-116.48
-49.08
-120.59
-84.23
-141.55
-134.01
-64.41

-69.15
143.49
171.37
-48.57
161.97
166.82
79.19
-62.38
176.99
165.81
136.18
143.22
149.74
144.73
63.78
155.45
118.54
-86.14
93.73
16.59
-161.97
-56.82
169.16
-60.37
52.55
-58.99
-71.03
-47.52
-54.33
-63.91
-52.61
-49.57
85.31
-59.40
67.92
-41.47
-41.52
-41.26
16.52
7.87
129.45
146.05
133.87
92.21
177.29
148.25
-73.61
138.84
156.98
152.59
177.99

A.
F.
A.
F.
N.A.
A.
A.
A.
N.A.
A.
F.
A.
N.A.
N.A.
A.
N.A.
N.A.
N.A.
A.
N.A.
N.A.
F.
F.
A.
N.A.
F.
N.A.
N.A.
A.
A.
N.A.
F.
N.A.
N.A.
A.
A.
N.A.
N.A.
N.A.
N.A.
A.
N.A.
F.
A.
A.
A.
N.A.
F.
A.
F.
F.

3056

Sci. Res. Essays

Table 2. Contd.

86
87
88
89
90
91
92
93
94
95
96
97
98
99
100
101
102
103
104
105
106
107
108
109
110

LEU
THR
ILE
LEU
TYR
TYR
ILE
GLY
LYS
THR
PRO
LYS
ILE
GLU
GLN
LEU
SER
ASN
MET
ILE
VAL
LYS
SER
CYS
LYS

-135.07
-80.44
-109.81
-129.44
-120.07
-92.22
-95.07
71.33
-129.01
-39.42
-42.46
-114.30
-59.18
-133.62
-87.37
-96.80
58.11
68.94
-71.01
-104.09
-69.19
-102.81
-164.80
-119.08
-149.01

134.26
126.80
151.76
115.26
159.99
131.52
84.87
-92.72
-0.03
79.15
126.35
95.58
130.38
134.63
109.16
110.51
151.25
14.24
-41.25
117.22
121.15
-25.04
136.03
166.89
150.07

F.
F.
F.
F.
F.
F.
A.
N.A.
N.A.
N.A.
N.A.
F.
F.
F.
F.
F.
F.
N.A.
F.
F.
F.
A.
A.
F.
A.

-153.79
-66.13
-114.74
-121.54
-134.98
-58.95
-114.30
70.12
-123.51
-59.62
-50.51
-103.02
-86.72
-146.43
-70.56
-87.17
-47.77
62.91
-79.55
-108.13
-54.95
-103.53
-127.19
-132.09
-142.19

113.08
125.95
156.91
122.58
162.06
140.01
92.84
-103.27
-10.49
94.95
108.15
104.10
138.16
102.17
98.44
109.91
154.02
28.66
-52.58
117.02
112.71
36.50
166.96
162.37
151.24

F.
F.
F.
F.
F.
F.
F.
N.A.
N.A.
A.
A.
F.
F.
F.
F.
F.
A.
N.A.
F.
F.
F.
A.
F.
F.
F.

-151.93
-61.25
-111.32
-100.82
-151.14
-72.64
-112.46
57.61
-107.75
-53.24
-50.51
-138.71
-108.76
-127.50
-43.34
-100.41
-58.25
59.21
-96.57
-120.68
-85.62
-115.32
-139.97
-111.92
-142.68

118.11
117.46
134.68
132.83
144.99
139.12
92.43
-114.84
-11.43
100.95
143.52
116.81
110.78
114.77
105.59
105.76
149.18
47.92
-30.54
118.93
142.93
-25.09
167.84
157.22
162.58

F.
F.
F.
F.
A.
F.
F.
N.A.
N.A.
A.
A.
F.
F.
F.
A.
F.
F.
A.
A.
F.
F.
N.A.
A.
F.
A.

-114.17
-48.23
-102.63
-144.72
-148.68
-54.51
-113.41
75.87
-133.23
-85.14
-46.85
-103.43
-85.29
-119.25
-95.92
-53.02
-68.37
64.36
-79.01
-90.66
-66.09
101.33
-162.90
-125.21
-176.51

108.47
114.13
167.55
122.16
131.31
129.48
71.00
-86.14
12.65
95.84
120.46
117.95
119.31
123.34
86.03
121.06
163.48
18.99
-61.31
127.13
120.84
-20.57
149.01
-176.54
142.73

F.
A.
F.
F.
F.
F.
A.
N.A.
N.A.
F.
A.
F.
F.
F.
A.
A.
F.
N.A.
A.
F.
F.
A.
A.
A.
A.

A. =allowed, F. = favored, N.A. = not allowed.

Table 3. 1tfg and 2tgi, and dihedral analysis after Montecarlo simulation at a glance.

2 tgi
Crystallographic
data
Simulted at 273 K
Simulted at 290 K
Simulted at 310 K
Simulted at 315 K

Ratio
Percent
Ratio
Percent
Ratio
Percent
Ratio
Percent
Ratio
Percent

Favored
106/110
96.40%
41/110
37.27%
40/110
36.36%
44/110
40%
34/110
30.91%

1 tfg
Allowed
110/110
100%
61/110
55.45%
60/110
60%
70/110
60.64%
74/110
67.27%

Favored
104/110
94.50%
m.d.
m.d.
38/110
34.54%
31/110
28.18%
m.d.
m.d.

Allowed
109/110
99.10%
m.d.
m.d.
78/110
70.90%
62/110
58.18%
m.d.
m.d.

m.d. = missed data.

to form bone. The phenomenon is not well known.

Neurogenic, traumatic and myositis ossification
progressiva are the main roots of HO formation. In closed
head injuries Neurogenic HO, spinal cord trauma,
strokes, tumors and central nervous system infections

HO has been described. It always happens after the

neurologic lesion, also it may be mediated by a humoral
factor. Neurogenic HO can be severe and may lead to
ankylosis (Sawyer et al., 1991). This phenomenon makes
some movement limitation for patients although it

Hadad et al.

3057

Figure. 4 Sequence comparisons of 1 tfg, and 2 tgi, data were collected from protein data bank. www.pdb.org

depends the class of HO. Both 2 tgi and 1tfg are doing
inducible activities on connective tissue around the hurt
zone of bone fracture, or a different zone, especially in
head trauma patients which these factors elevated base
on a neuroprotective role. The affected humoral factors
are members of TGF- super family. These factors are 1
tfg and 2 tgi, where the crystallographic data are
available in protein data bank. Results reveal that, 1tfg is
affected by increasing the temperature. The favored and
allowed regions were decreased in population of dihedral
angles. Therefore it is though that to keep warm the
region by some equipment, with short frequencies radii,
may affect this factor to inhibit its function, therefore the
heterotropic ossification will decrease and it will be
suggested as a role in treatment. The result is more
complicated for 2tgi, increasing the temperature up to
room temperature cause the protein to find more
unfavored dihedral, but the more temperature cause
more allowed ones. It is clear that utilization of IR lamp
does not affect on such patients. Therefore it offered that
in patients with head trauma, before any invasive
physical therapy, the type of transforming growth factors
has to be clarified biochemically.

REFERENCES
Atkinson GJ, Lee MY, Mehta MK. (2010) Heterotopic Ossification in the
Residual Lower Limb in an Adult Nontraumatic Amputee Patient. Am
J. Phy. Med. Rehabil., 89 (3): 245-248.
Betts MJ, Sternberg JE. (1999). An analysis of conformational changes
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