ΟΡΓΑΝΩΤΙΚΗ ΕΠΙΤΡΟΠΗ2) «“ΞΕΡΩ ΤΙ ΖΗΤΑΩ” - Πρόγραμμα πρόληψης της χρήσης εξαρτησιογόνων ουσιών στα
Σύνθεση, χαρακτηρισμός και αποδέσμευση βιοδραστικών...
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Transcript of Σύνθεση, χαρακτηρισμός και αποδέσμευση βιοδραστικών...
-
,
.. 861
:
, 2013
-
3
,
.
.
.
-
(-TCP) . ,
( - ibuprofen)
(
- FITC) .
, ,
. ,
. ,
,
.
XRD, SEM, ATR, UV/Vis Raman.
,
.
.
: SEM
-TCP.
-
Synthesis, characterization and release of bioactive compounds from calcium
phosphate bone cements
Abstract
Use of bone cements is beneficial for the treatment of many bone diseases caused by
injuries or other factors. There is great interest in using bone cements in clinical
practice due to their composition and bioactivity. The presence of bioactive
compounds can help the clinical management of the diseased area. The present
Diploma Thesis is on the synthesis and characterization of bone cements composed
of a-tricalcium phosphate (-TCP) and gypsum. Furthermore the release of a
sparingly soluble drug (ibuprofen) and a water soluble pigment (FITC) from cement
matrices were studied. In the first chapter, the different types, the classification and
some applications of biomaterials are presented. Next chapter is on the topical and
controlled drug delivery systems. In the third and fourth chapters the specifications
of calcium phosphate bone cements and gypsum and the bioactive substances used
in the present study are presented. In the fifth chapter the fundamental principles of
instrumental analytical methods used in the present work such as XRD, SEM, UV/Vis
spectroscopy and Raman spectroscopy are described. Finally, in the last chapter all
the experimental results are shown and discussed. The results are on the synthesis
and structural, mechanical and morphological characterization of different cements
formulations. Finally this chapter reports on the ability of the prepared formulations
as a controlled release system of two bioactive compounds.
-
5
, .
. ,
,
.
. . (
/) Raman
SEM XRD . . (
... /).
.
.
,
,
. , .
-
7
1
1. ............. ................................................................................................................ 15
1.1 ....................................................................... 15
1.2 ....................................................................................................... 16
1.3 ................................................................................... 18
1.4 ..................................................................................... 19
2
2. 21
2.1 CDDS ............................................... 22
2.2 ....................................... 23
2.3 CDDS ................................................................................... 25
2.4 CDDS ................. 26
3
3. .................................................................................. 29
3.1 (CPCs) ........................................................... 30
3.1.1 CPCs ............................................................................................. 31
3.1.2 CPCs ..................................... 33
3.1.2.1 ...................................................................................... 34
3.1.2.2 .................................................................................................. 36
3.1.3 CPCs .............................................................. 37
3.1.4 CPCs ................................................................. 37
3.1.5 CPCs .................................................. 39
3.1.6 CPCs ..................................................... 40
3.1.7 CPCs ..................................................................... 41
3.1.7.1 CPCs ....................................................... 41
3.1.7.2 CPC ................................. 43
-
3.1.7.3 CPCs ............................. 43
3.2 ........................................................................................................................... 45
3.2.1 ......................................................... 45
3.2.2 ................................................................................................ 46
3.2.2.1 ............................................................................................... 46
3.2.2.2 .................................................................................... 46
3.2.2.3 ..................................... 46
3.2.2.4 ............................................................................................................ 47
3.2.2.5 ................................................................................................ 47
4
4. ........................................................................................................ 49
4.1 ............................................................................................................ 49
4.1.1 ..................................................................................................... 50
4.1.2 ............................................................................................... 51
4.2 ............................................................................ 51
4.2.1 ................................................................................................ 52
4.2.1.1 ......................................................................... 52
4.2.1.2 ............................................................... 53
5
5. .................................................................................... 55
5.1 - (XRD) ................................................................................ 55
5.2 (SEM) ...................................................... 58
5.3 (ATR) ......... 64
5.4 / (UV/Vis) .. 67
5.5 RAMAN....................................................................................... 70
6
6. - ................................................................ 75
6.1 ......................................................................................... 75
-
9
6.1.1 -TCP ....................................................................................... 75
6.1.2 ............................ 77
6.1.3 / .. 79
6.2 -TCP - ...................................... 80
6.3 -TCP Raman 82
6.4 ..................................................................................... 83
6.4.1 - (XRD) 85
6.4.2 SEM 88
6.5 ................................... 90
6.5.1 .. 90
6.5.1.1 .................................................................... 90
6.5.1.2 FTIR/ATR ............................... 93
6.5.1.3 - (XRD) ............................ 96
6.5.1.4 SEM ............................................................................................................ 98
6.5.2 .....................................................................100
6.5.2.1 - (XRD) .......................... 102
6.6 ....................................................................................................... 104
6.7 .......................................................................105
..................................................................................................................... 107
-
1: . ................................................ 20
2: )
) CDDS .................................................................................. 26
3: CDDS )
, ) , )
, ) ............................................................... 28
4: CPCs ............................................................................ 33
5: Ca(OH)2 -
H3PO4 - H2O 37oC.
pH ............................................................. 34
6: ........................ 35
7: DCPD .......................... 36
8: CPCs. 38
9: CPCs .......................................... 42
10: .. 44
11: . ..................................................................... 50
12: ................................................................ 51
13: . ............................................................................. 52
14: . ............................................................................... 53
15: .................. 53
16: - ............................. 56
17: - ................................................ 57
18: Bruker D8. ............................................................................ 58
19: LEO SUPRA 35VP. ......................... 59
20: ..................................... 60
21: SEM. ................................................ 62
-
11
22: sputtering .
sputtering. .............................................................................................. 63
23: R ...................................................................... 65
24:
. .......................................................................................................... 65
25: FTIR ATR ....................................................... 67
26: / ........................ 69
27: UV/VIS . 69
28: . ............................................................................ 71
29: Raman. ....................................... 72
30: Raman. .................................................... 72
31: Micro-Raman (Labram HR-800) ............................... 74
32: , 1cm ................ 76
33: . ................................ 76
34:
() ........ 77
35: . ................................ 78
36:
. .............................................................................................................. 78
37: . ........................... 79
38: . .......... 80
39: M - Hounsfield H20K-W. .................................. 84
40: . ....................................................... 84
41: SEM -TCP
(A) Ringer
7 () 340 () . ................................................................................. 88
42: SEM 90% -TCP 10%
Ringer 7 (,)
340 () .................................................................................................. 89
43: SEM
(A) Ringer 340 ()
...................................................................................................................... 90
-
44: SEM -TCP 2%
PBS. .............. 99
45: SEM 88% -TCP 10%
2%
PBS ........................................................................................................................ 99
46: SEM 2%
PBS ................. 99
1: :
25C ........................ 32
2: .................. 78
3: Ringer ................................................................... 78
4: .......................... 79
5: PBS ........................................................................ 80
-
1: -
-TCP, -TCP (HAP). ... 81
2: Raman -TCP -TCP
-TCP (200-825 cm-1). ..................................................................... 82
3: Raman -TCP -TCP
-TCP (825-1200 cm-1) .................................................................... 83
4:
............................................................... 85
5: - 100% -
TCP Ringer 0, 7 340
-TCP, -TCP HAP ........................................ 86
6: - 90% -
TCP 10% Ringer 0, 7
340 -TCP, -TCP, HAP
. .................................................................................................................. 87
7: - 100%
Ringer 0, 7 340
(CSH) ................................................................................................ 87
8: . . 90
9:
3 120 ppm. ......................................................................... 91
10: (c) (1%)
(: 99% -TCP, : 89% -TCP 10% , : 99% ).92
11: (c) (2%)
(: 98% -TCP, : 88% -TCP 10% , : 98% ).93
12: FTIR/ATR -
TCP, 0% 2% , ............ 94
13: FTIR/ATR -TCP 10%
, 0% 2% , . ....... 95
14: FTIR/ATR
, 0% 2% , . ....... 95
-
15: - 98% -
TCP 2%
-TCP, -TCP, CDHA ........................................... 96
16: - 88% -
TCP, 10% 2%
-TCP, , CDHA ........................................... 97
17: - 98%
2%
,
(CSH) . ...................... 98
18:
............................................................................................... 100
19:
1 - 12ppm ...................... 101
20: (c)
(0,2%) (: 99,8% -TCP, : 89,8% -TCP 10%
, : 99,8% ) .................................................................................... 102
21: - 99,8% -
TCP 0,2% (FITC)
-TCP, -TCP, HAP FITC .............................................. 103
22: - 89,8% -
TCP, 10% 0,2% (FITC)
-TCP, , HAP FITC. ........................... 103
23: - 99,8%
0,2% (FITC)
, (CSH) FITC.104
-
15 1:
,
. ,
: (
) , ,
,
, . 1986,
,
:
[1][2].
1.1
,
[2]:
1. :
.
Williams 1987:
.
,
. , ,
, .
, ,
1
-
1990
. ,
, ,
,
.
2.
3.
4. :
:
5.
.
6. .
,
in vivo in vitro .
.
1.2
,
[2][3]:
:
, ,
( , ,
, ..)
. ,
-
17 1:
, , ...
,
. ,
,
, ,
.
: ,
,
.
,
. , , ,
.
:
,
.
,
, , .
CDDS.
:
. ,
,
(). , ,
, ,
.
.
:
.
,
,
-
. ,
. ,
.
, , .
1.3
,
.
. ,
[4]:
:
. ,
, ,
.
:
.
.
, , ,
(.. HAp),
Pt- HAp
. , ,
.
:
( )
. -
(Ca3(PO)4)
-
19 1:
- .
, o
.
:
,
.
, ,
.
1.4
,
,
.
.
[4]:
:
.
(, ,
..) , ,
(..
),
. ,
,
. ,
.
:
, .
:
.
,
,
-
.
, ,
.
:
(.. ,
, ..).
: .
.
.
.
: , ,
,
.
.
1: [2].
-
21 2:
, .
(, , ,
)
.
, (.. , , ,
, ..)
. ,
,
,
.
(control drug
delivery system, CDDS). CDDS
, ,
.
.
,
, .
,
[5].
2
-
2.1 CDDS
,
.
CDDS
[6]:
.
.
.
.
,
.
.
.
.
.
,
,
, ,
.
.
,
.
.
.
-
23 2:
, .
.
,
.
.
.
.
.
.
.
:
.
.
.
.
.
2.2
[5]:
:
.
.
.
.
-
.
in vitro in vivo [7].
:
,
,
. ,
. ,
.
CDDS :
.
,
.
,
[8].
: ,
, CDDS
.
.
5 CDDS
: ,
, , ,
,
. ,
CDDS
, ,
. ,
CDDS , ,
,
-
25 2:
, ,
.
:
.
.
: (prodrugs)
, .
.
2.3 CDDS
CDDS
.
CDDS [6]:
,
,
, .
, , CDDS :
,
,
,
,
,
,
,
.
-
2.4 CDDS
CDDS
. 2:
.
(, ..),
,
.
. , CDDS,
,
CDDS
[10].
2: )
) CDDS [9]
CDDS ,
, ,
,
.
:
-
27 2:
.
. ,
,
.
,
. ,
.
.
[10]:
.
( 3:).
( 3:).
.
.
,
.
pH, ( 3:).
,
, ,
( 3:).
-
3: CDDS )
, ) , ) , )
[10].
-
29 3:
,
, .
( )
.
, , ,
.
.
, , .
:
1. (Osteointegration):
.
2. (Osteoconduction):
.
3. O (Osteoinduction):
.
4. O (Osteogenesis):
.
,
3
-
.
.
,
.
. ,
/ .
,
. , ,
[7].
3.1 (CPCs)
R.Z. LeGeros
1982 W.E. Brown
L.C. Chow 1986 [1].
1990
.
, , ,
,
.
in vivo .
CPC
,
,
.
. CPCs
,
, , ,
.
, CPCs
:
-
31 3:
,
(
).
, CPCs ,
,
.
CPCs,
.
CPCs
.
,
CPCs
.
3.1.1 CPCs
CPCs
() .
.
(Na2HPO4)
(NaH2PO4) .
, ,
CPCs
( 4:).
:
, ,
,
pH
. p
-
[11].
,
. :
(p), (CA), - (-TCP), -
(-TCP), (TTCP),
(DCPA), (MCPM),
(DCPD)
(OCP). HAp, -TCP, CA
. a-TCP, TCP, DCPD, OCP
CPCs [12].
1: :
25C [13].
/ Ca/P Log(Kip)
Ca(H2PO4)2H2O MCPM 0,50 1,14
CaHPO42H2O DCPD 1,00 6,59
CaHPO4 DCPA 1,00 6,9
-
-Ca3(PO4)2 -TCP 1,50 25,5
-
-Ca3(PO4)2 -TCP 1,50 28,9
Ca8H2(PO4)65H2O OCP 1,3 96,6
Ca10(PO4)6(OH)2 p 1,6 118,8
Ca4P2O9 TTCP 2 38-40
Ca10(PO4)6-x(CO3)x(OH)2 CA - -
, CPCs
:
(p) (DCPD).
-
33 3:
p > 4.2
pH < 4.2.
4: CPCs [11].
3.1.2 CPCs
(HAp)
,
DCPD OCP
HAp. pH,
. , pH 4 6
DCPD, pH 6 7
OCP.
pH
-
. pH (7.08.5)
HAp ( 5:).
5: Ca(OH)2 -H3PO4 -
H2O 37oC.
pH
.
DCPD OCP ,
HAp. ,
. ,
. ,
10-3 mol/L
OCP.
.
DCPD OCP.
.
(, , )
DCPD, OCP,
HAp [14].
3.1.2.1
(p)
,
-
35 3:
Ca10(PO4)6(OH)2.
.
6: [16][17].
O .
X ,
.
, . ,
. , ,
[14].
,
, , .
1970
. p
. ,
-
400-700C p.
- .
- , CPCs,
p in vivo.
.
3.1.2.2
,
(DCPD) , ,
CPCs
, in vivo
.
.
, -TCP
, (MCPM)
( 4:). -TCP
-TCP [11].
DCPD ,
.
DCPD
( ).
b a c,
7: [14].
7: DCPD.
-
37 3:
,
800 . DCPD
. ,
(3 5 )
.
.
3.1.3 CPCs
3 :
1) , 2) (
) 3) . ,
-. ,
.
,
.
, .
,
,
.
3.1.4 CPCs
CPCs
, .
,
. ,
.
-
.
/ (L/P). L/P,
( 8:2). ,
.
,
( 8:1 82 ). ,
CPCs
[11].
) ) /
8: CPCs.
-
39 3:
3.1.5 CPCs
CPCs .
,
.
,
. CPCs 2
: )
, ,
p - )
.
p 5.5,
.
,
.
,
pH. , (
pH)
.
CPCs
. ,
,
.
CPCs :
, , , ,
.
, CPC ,
in vivo.
-
,
,
() [11].
3.1.6 CPCs
( ). CPCs
,
,
, .
( CPCs
)
,
.
filter-pressing phenomenon (
)
-
, . .
.
.
.
/ (L/P).
L/P = 1,3-
2,5 g/ml , ( 2-3 mm)
.
. L/P
.
CPCs
. (TTCP)
(DCPA)
L/P = 3,3 g/ml
60%. 500 mM
-
41 3:
()
(injectability > 95%)
800 m.
.
HA. pH (500 mM, pH=1.32)
( )
.
,
p, 10,93 11,73 [18].
3.1.7 CPCs
3.1.7.1 CPCs
CPC
. , ,
,
. CPCs
o
.
,
.
, :
(.. , , )
CPCs,
( 9:).
-
9: CPCs [11].
,
.
,
.
CPCs.
,
,
. ,
-
.
.
,
. ,
( 4:). ,
,
.
.
o CPC.
-
43 3:
: )
(.. ) )
[11].
3.1.7.2 CPC
CPC
,
. ,
, ,
CPCs CPCs.
CPC.
-
pH
,
. ,
CPCs
[11].
3.1.7.3 CPCs
CPCs -
. ,
- - ,
, CPCs ,
.
.
CPC.
,
.
CPC. ,
-
(p ).
,
, 10:
1. ,
2.
3. ,
.
.
10: CPCs
[11].
,
, pH
,
CPC.
,
-
45 3:
( 10:).
CPCs Higuchi,
,
. [20]:
1.
,
2.
,
3.
4. (
).
3.2
,
. ,
CSO42H2O. 3/2
1/2 [23].
3.2.1
,
, ,
[23]:
500C: .
1000 - 1200C: .
1800C: 20,9% 6,2%
, CSO4. H2O,
.
1900 - 2200C:
.
-
3000C: .
5000 - 6000C: , CSO4,
.
10000C:
, CO, CSO4 , :
CSO4 CO + SO3
3.2.2
,
, .
3.2.2.1
.
,
. silos,
.
3.2.2.2
,
,
.
.
, ,
[23]:
1. ,
2. ,
3.
4. .
3.2.2.3
:
-
47 3:
CaSO4
H2O +
H2O CaSO4 2H2O + Q
H
. ,
,
,
.
CO2
. , ,
.
5 30 min :
1. ,
2. ,
3.
4. .
.
,
, , [23].
3.2.2.4
:
1. .
2. .
3. .
4. .
5. .
3.2.2.5
, ,
, .
-
, ,
. ,
[23].
,
1892 [24]. ,
( ) .
in vivo.
[25].
[26].
.
,
.
.
[27].
[26].
[28].
-
49 4:
,
(Ibuprofen)
, (Fluorescein
Isothiocyanate - FITC).
.
4.1
1969 .
- -
() .
) , ,
, )
,
, , )
) (ductus
arteriosus) , .
,
.
.
,
, , .
74-77 C.
[29].
4
-
1960
Boots . Boots, ,
6
.
BHC 1991
3 .
[30].
4.1.1
.
1: Friedel-Crafts ,
.
2: A Darzens -
.
3: A .
4: A .
5: M .
6: Y 2,4--
, .
60% .
11: .
-
51 4:
4.1.2
.
1: Friedel-Crafts
.
2: Raney,
-, .
3: ,
,
.
1% .
12: .
4.2
(FITC)
.
.
.
:
1. (FITC), .
2. (TMRITC),
.
3. (R), .
-
4.2.1
.
[31].
4.2.1.1
(IFA)
:
1:
.
2:
.
3: .
4:
.
5: .
13: .
(DFA)
:
1:
.
2:
.
3: .
-
53 4:
14: .
IFA, DFA
.
.
15: .
4.2.1.2
1.
, , , , ,
.
2.
.
-
3.
: , , ,
, , , ,
.
.
4. .
.
.
5.
.
.
-
55 5:
5.1 - (XRD)
, ,
, -
, . , XRD
.
-
,
.
,
.
.
.
.
,
( ),
( )
( ).
,
,
.
, n, .
5
-
16: - .
Bragg
, Bragg:
n
, , d
.
-,
Cu Mo, ,
.
.
Bragg .
,
,
.
- ,
.
-
57 5:
,
.
2.
2
[13].
17: -.
,
, ,
. ,
.
, ,
.
-
(Bruker D8)
(/) Bragg-Brentano
0 160 ( 18:).
, .
Cu ( = 1.5421 ) Ni.
0.35 sec/step.
- 40 kV
40 mA.
-
18: Bruker D8.
5.2 (SEM)
(Scanning Electron Microscopy, SEM)
.
,
.
(.. ).
.
.
Auger.
. SEM
.
.
-
59 5:
,
.
SEM
(/) LEO SUPRA 35VP ( 19:)
.
Al
(PELCO
Image Tabs).
, .
sputtering (~510-2 mbar) BAL-TEC SCD-004.
19: LEO SUPRA 35VP.
.
,
[7].
-
)
, ) , )
) .
20: [32].
20: :
1. : .
.
2. .
3. : )
)
180,
.
4. : )
)
.
.
-
61 5:
5. - (X-Rays): ) )
)
, ( )
- Bremsstahlung.
-
.
.
)
-
.
,
-.
.
6. Auger : Auger
-
.
SEM
.
, ,
.
:
1.
.
2. ,
,
.
3.
.
.
-
21: SEM.
(
), .
(filament current). ,
1-30
kV (accelerating voltage). , ,
.
,
. ,
.
(filament saturation).
, ,
. ,
.
.
.
SEM
.
-
63 5:
, .
.
(--).
.
Sputtering ( 22:).
, ,
.
Sputtering.
22: sputtering ().
(-) (-) sputtering.
sputtering:
1.
.
.
2. .
3. .
4. .
5. ( ) .
-
6. (
)
.
7. ,
.
(coating)
10-30 nm [1].
5.3
(ATR)
IR
.
. ,
(KBr)
().
KBr
.
,
.
, , ,
.
(Attenuated Total
Reflectance, ATR).
,
, ,
.
.
75 psi ( 23:).
-
65 5:
23: R.
, ATR
1 .
,
[35].
( 24:),
c (critical angle), :
(5.1)
n2 n1
. (penetration depth), dp,
Io/e (
/ e
2,718).
n1 n2,
, :
(
) (5.2)
24:
.
-
ATR : ,
o n2 n1,
, dp .
(5.3)
ev , z
, o z=0.
.
ATR IR ,
. ATR
,
.
ATR
, >1.
.
, (effective penetration depth)
dp .
.
FT-ATR FT-IR
.
,
ATR, ,
,
, ,
[35].
,
,
-
67 5:
.
CPCs ,
FTIR (Excalibur Digilab) ATR (Pike
Magic) 450 cm-1 4000 cm-1,
( 25:).
25: FTIR (Digelab Excalibur) ATR (Pike Magic).
5.4 /
(UV/Vis)
/ (UV/Vis)
, ,
,
.
b cm.
,
.
r, b Beer-
Lambert Beer:
(5.4)
-
, Po , P
, Tr , b
, ,
c .
Beer :
,
,
,
.
.
,
( )
( ). Beer
.
,
.
[13].
: (i)
, (ii) , ,
, (iii)
, (iv)
, (v)
(vi)
.
(160-340 nm).
-
69 5:
(340-780 nm)
. , ,
,
.
.
.
/
,
Perkin Elmer, (Model Lambda 35 UV/VIS
Spectrophotometer).
1 cm.
26: / (Perkin Elmer, Model
Lambda 35 UV/VIS Spectrophotometer).
27:.
27: UV/VIS .
-
,
( ).
() .
.
.
, .
,
, .
.
.
,
.
5.5 RAMAN
Raman
, .
Raman
Raman
.
h.
.
.
cm /c, c
.
,
, .
(, ),
,
,
.
,
.
-
71 5:
h. ,
, ,
Rayleigh ( 28:).
Rayleigh . ,
,
. Rayleigh
,
.
28: .
Rayleigh
( Raman).
.
,
( ).
,
,
( ).
( -
Stokes)
Rayleigh ( - anti-Stokes),
, ( 29:).
Raman
,
.
-
29: Raman.
Raman .
30: Raman.
-
73 5:
:
(i) .
(ii) ()
( ).
.
(edge filter)
Raman ,
Rayleigh
Raman (Stokes), > LASER.
Raman
,
Rayleigh
Raman.
(iii)
.
(iv) .
(v) CCD .
(vi)
.
, Raman
Micro-Raman (Labram HR-800)
o (/) (
31:). laser HeCd (Kimmon
Electric Co., Model: IK5651R-G)
441,6 nm ( 80 mW) 325 nm ( 20 mW).
HeCd. ,
,
.
Rayleigh .
-
Raman
1800 nm 2D CCD
.
[36].
31: Micro-Raman (Labram HR-800) o
(/).
-
75 6: -
6.1
:
100% -TCP
90% -TCP 10%
100%
(CSH) :
CaSO3
H2O +
H2O CaSO4 2H2O + Q (6.1)
Q
[37].
1%, 2% 0,2%
.
.
,
3.2.2.5.
6.1.1 -TCP
-TCP, ,
:
6 -
-
(6.2)
, (CaCO3)
(Ca2P2O7)
(ball milling) ( 32:) 450
20 3 .
32: (Pulverisette 5, Fritsch), 1cm
.
, (Alumina
Crucible, Sigma Aldrich) 100 ml,
.
( 33:)
1350C 12
164C/h .
33: .
,
, ( 34:).
4 20
a-TCP
-
77 6: -
.
[38] 100 m.
34:
() ().
-TCP
.
-TCP
(CDHA:
Calcium Deficient Hydroxy Apatite)
-TCP, :
(6.3)
1000 mL 20C 75%
12 mg . -TCP
600 mg CDHA [39].
6.1.2
,
2:. ,
100% -TCP 100%
90% -TCP 10% , 4%
w/v Na2HPO4. ,
,
12 mm 6 mm ( 35:).
-
2: .
a-TCP 4% w/v Na2HPO4
100% - 0,32 ml/gr
90% 10% 0,35 ml/gr
- 100% 0,50 ml/gr
35: .
24h 100%
37C (setting). ,
( 36:)
15 ml Ringer (Fresenius Kabi -
Lactated Ringers) 37C
(hardening). Ringer ,
3:.
36: .
3: Ringer.
/ 100 ml
CH3CH(OH)COONa 0,31 gr
NaCl 0,60 gr
KCl 0,03 gr
CaCl22H2O 0,02 gr
( Ringer),
1, 2, 8, 10 15
-
79 6: -
. XRD
SEM .
6.1.3
,
4:.
( 37:).
4: .
a-TCP 4% w/v Na2HPO4
IBUPROFEN
99% - 1% 0,32 ml/gr
89% 10% 1% 0,35 ml/gr
- 99% 1% 0,50 ml/gr
98% - 2% 0,32 ml/gr
88% 10% 2% 0,35 ml/gr
- 98% 2% 0,50 ml/gr
FITC
99,8% - 0,2% 0,32 ml/gr
89,8% 10% 0,2% 0,35 ml/gr
- 99,8% 0,2% 0,50 ml/gr
37: .
24h 100%
. ,
,
25 ml PBS (Phosphate Buffered Saline)
-
pH 7,4. PBS
5:.
37C.
5: PBS.
1 lt :
NaCl KCl Na2HPO4 KH2PO4
8 gr 0,2 gr 1,44 gr 0,27 gr
15 ml
.
PBS ( 38:). ,
UV/Vis
PBS
. , Microsoft Excel
.
38: .
6.2 -TCP -
-TCP
-. ,
,
.
Cu ( = 1.5421 ).
0,35 sec/step. (2) 10 40.
-
81 6: -
,
ICDD (International Centre for Diffraction Data). ,
29-359 -TCP, 9-169 -TCP
9-432 HAP,
.
10 15 20 25 30 35 40
HAP # 9-432
,
,
-TCP
(
a.u
.)
2
-TCP # 29-359
-TCP # 9-169
1: -
-TCP, -TCP (HAP).
-
-CP
-TCP.
-TCP -TCP 2 30,75 31,09
.
(1 7 0) -TCP (0 2 10) -TCP [18].
.
-TCP
1300C
, 1180C
-. , Ginebra et. al.
-TCP 15% [40]. ,
-
,
-TCP,
[41].
6.3 -TCP
Raman
Raman ( 2: 3:)
-TCP
430, 580,
592, 609 960 cm-1 , [42][43][44][45],
.
2: -TCP, -TCP
O-P-O [44][46].
-TCP 406, 440, 481,512, 548, 586, 610, 623 737
cm-1 -TCP 420, 448, 564, 591 623 cm-1.
.
200 300 400 500 600 700 800
43
0
60
9
58
05
92
62
3
73
761
0
58
6
48
1
51
2
54
8
40
6
44
04
48
59
1
56
4
62
3
(a
.u.)
Raman Shift / cm-1
-TCP
-TCP
42
0
2: Raman -TCP -TCP
-TCP (200-825 cm-1).
-
83 6: -
3: -TCP 948
970 cm-1
-P-O. -TCP 966 cm-1
960 cm-1. 1045 cm-1
O-P-O.
.
850 900 950 1000 1050 1100 1150 1200
10
45
97
0
94
8
(a
.u.)
Raman Shift / cm-1
-TCP
-TCP
96
09
66
3: Raman -TCP -TCP
-TCP (825-1200 cm-1).
6.4
( )
-
Hounsfield H20K-W ( 39:),
1,54 mm/min.
.
-
-.
( ).
-
39: M - Hounsfield H20K-W.
40: .
, ,
MPa
:
(6.4)
, F
mm2.
,
.
.
.
. 24
37oC,
(setting).
4:
-TCP (40,68 4,14) MPa.
[40].
10%
(34,61 2,50) MPa. (14,52 2,64) MPa.
-
85 6: -
,
.
, 24
.
0 1 2 8 1 0 1 5
0
5
10
15
20
25
30
35
40
45
50
(
MP
a)
/
100% -TCP
90% -TCP & 10%
100%
4:
.
6.4.1 - (XRD)
,
-. T
, Ringer
,
XRD.
ICDD: 29-359 (-TCP), 9-169 (-TCP), 9-432
( - HAP) 33-311 ().
5:
100% -TCP Ringer, -TCP 7
.
-
-TCP [40].
-TCP
HAP [41].
-TCP .
20 25 30 35 40
-TCP # 9-169
HAP # 9-432
-TCP # 29-359
(
a.u
.)
2
340h
7h
0h
5: - 100% -TCP
Ringer 0, 7 340
-TCP, -TCP HAP.
.
6: -TCP
90% -TCP 10%
Ringer. -TCP
.
(0h, 7h 340h)
.
(HAP)
(2 1 1), (1 1 2), (3 0 0) 31,60, 32,02 32,72
[47].
.
-
87 6: -
20 25 30 35 40
(
a.u
.)
2
# 33-311
340h
7h
0h
-TCP # 9-169
HAP # 9-432
-TCP # 29-359
6: - 90% -TCP
10% Ringer 0, 7 340
-TCP, -TCP, HAP .
.
10 15 20 25 30 35 40 45 50
# 33-311
(
a.u
.)
2Theta
340h
7h
0h
CSH
7: - 100%
Ringer 0, 7 340
(CSH).
-
( 7:) ,
(CSH) 14,7
0 , 25,5 31,9.
.
( 4:).
6.4.2 SEM
(SEM)
LEO SUPRA 35VP /.
41: -TCP
1m. 41:
-TCP. 41:
TCP.
41: SEM -TCP
(A) Ringer 7 ()
340 () .
-
89 6: -
42:
() . 340
42: SEM 90% -TCP 10%
Ringer 7 (,) 340 () .
62 m
( 43:). 340
.
6.4
( 4:).
-
43: SEM
(A) Ringer 340 () .
6.5
6.5.1
6.5.1.1
UV/Vis
=264 nm
-
( 8:).
8: .
-
91 6: -
(stock) 120 ppm
PBS. , stock
(3ppm, 6ppm, 12ppm,
16ppm, 20ppm, 30ppm, 40ppm 60ppm). PBS
:
Cstock Vstock = C V (6.5)
, Cstock =
120ppm, C
V ,
stock
.
0 20 40 60 80 100 120
0,00
0,05
0,10
0,15
0,20
A
C (ppm) 9:
3 120 ppm.
:
(6.6)
-
9:
. ,
.
1%
2%. 10: -TCP () 194
(99,48% 6,22%)
() ()
.
44,33% 4,20% 38,04% 1,13% .
0 50 100 150 200 250 300 350 400
0
20
40
60
80
100
C (
%)
/
A
10: (c) (1%)
(: 99% -TCP, : 89% -TCP 10% , : 99% ).
2%
( 11:) 414
-
TCP () 90,23% 5,06%. , ()
() 194
, 30,69% 0,39% 40,13% 3,25%
.
-
93 6: -
0 50 100 150 200 250 300 350 400
0
20
40
60
80
100
C (
%)
/
Z
E
11: (c) (2%)
(: 98% -TCP, : 88% -TCP 10% , : 98% ).
, ,
10% 89% () 88% () -TCP
-TCP ( )
( ).
6.5.1.2 FTIR/ATR
FTIR/ATR.
-TCP
668 cm-1
.
1019 cm-1 -TCP. 12:
779, 1231 1721 cm-1.
(2%)
,
.
-
,
FTIR. , 1721 cm-1
C=O, 1231 cm-1 C-C 779 668
cm-1 -C-H
[48].
564 603 cm-1
-P-O
[49]. 965 cm-1
O-P-O.
1380-1480 cm-1
.
, 2%,
. ,
1114 cm-1 [49].
1800 1700 1600 1500 1400 1300 1200 1100 1000 900 800 700 600
2% ibuprofen
& 0%
0%
(a
.u.)
/ cm-1
10
19
ibuprofen
66
8
77
9
12
30
17
21
1114 965603
564
1380 - 1480
12: FTIR/ATR -TCP,
0% 2% , .
-
95 6: -
1800 1700 1600 1500 1400 1300 1200 1100 1000 900 800 700 600
2% ibuprofen
& 10%
(a
.u.)
/ cm-1
ibuprofen
10% 66
8
10
19
66877912311721
13: FTIR/ATR -TCP 10% ,
0% 2% , .
1800 1700 1600 1500 1400 1300 1200 1100 1000 900 800 700 600
66
7
(a
.u.)
/ cm-1
2% ibuprofen
& 100%
ibuprofen
100%
59
7
10
98
16
82
16
19
14: FTIR/ATR ,
0% 2% , .
-
( 14:)
. 597
667 cm-1 1098
cm-1 S-O.
- 1619 1682 cm-1.
,
(CSH) [51].
6.5.1.3 - (XRD)
-
-TCP
2%.
1%
.
15: -TCP
-TCP 2% PBS
. 414
,
,
TCP .
10 15 20 25 30 35 40
(a.u
.)
2
ibuprofen
-TCP
-TCP
HAP
HAP
15: - 98% -TCP
2%
-TCP, -TCP, CDHA
.
.
-
97 6: -
( 16:)
-TCP,
PBS,
, 11,63 20,72,
(10%) . ,
-TCP .
10 15 20 25 30 35 40
HA
P
(
a.u
.)
2
ibuprofen
-TCP
HAP
HA
P
16: - 88% -TCP,
10% 2%
-TCP, , CDHA
.
.
, 2%
( 17:) ,
(CSH) 25,5 32,13.
-
10 15 20 25 30 35 40C
SH
CS
H
(
a.u
.)
2
ibuprofen
CSH
17: - 98%
2%
,
(CSH) .
6.5.1.4 SEM
2%
,
(SEM),
.
.
44: ()
() -TCP. 3.1.4,
-TCP.
-TCP,
.
-
99 6: -
44: SEM -TCP 2%
PBS.
45:
.
.
45: SEM 88% -TCP 10%
2% PBS.
46:
.
46: SEM 2%
PBS.
B
-
6.5.2
UV/Vis
=493 nm.
18:
.
(stock) 12 ppm
PBS. ,
stock
(1ppm, 1,6ppm, 4ppm, 5ppm, 6,4ppm, 8ppm 10ppm).
:
(6.7)
-
101 6: -
0 2 4 6 8 10 12
0,0
0,2
0,4
0,6
0,8
1,0
A
C (ppm) 19:
1 - 12ppm.
( 20:)
-TCP () 55
(95,31% 7,49%)
()
69,70% 7,46%. ()
(5,84% 0,33%)
.
. ,
28
10% [52].
. ,
,
-TCP,
.
-
0 20 40 60 80 100 120 140
0
20
40
60
80
100
c (
%)
/
20: (c)
(0,2%) (: 99,8% -TCP, : 89,8% -TCP 10% , :
99,8% ).
6.5.2.1 - (XRD)
-TCP
(FITC) 0,2%
-.
21: -TCP
-TCP 0,2%
PBS
. 145
,
-TCP -TCP .
22:, -TCP,
PBS, ,
11,63, (10%) .
, -TCP .
FITC
XRD .
-
103 6: -
10 15 20 25 30 35 40
-TCP
FITC
(
a.u
.)
2
-TCP
HAP
21: - 99,8% -TCP
0,2% (FITC)
-TCP, -TCP,
HAP FITC.
10 15 20 25 30 35 40
(a.u
.)
2
FITC
-TCP
HAP
22: - 89,8% -TCP,
10% 0,2% (FITC)
-TCP,
, HAP FITC.
-
, 0,2%
,
(CSH) 25,5 32,13.
10 15 20 25 30 35 40
CSH
(
a.u
.)
2
FITC
23: - 99,8%
0,2% (FITC)
,
(CSH) FITC.
6.6
.
1. -TCP,
-
.
Ringer PBS.
Raman FTIR/ATR, XRD SEM.
2. 10% w/w
-TCP.
3. , Ringer
PBS,
-
105 6: -
. 24
37C.
4. SEM
-TCP .
5. -TCP
.
-TCP,
.
6.
-TCP
.
6.7
1.
.
2.
.
3. (setting time) (injectability)
.
.
-
107
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