Σύνθεση, χαρακτηρισμός και αποδέσμευση βιοδραστικών...

110
ΠΑΝΕΠΙΣΤΗΜΙΟ ΠΑΤΡΩΝ ΤΜΗΜΑ ΕΠΙΣΤΗΜΗΣ ΤΩΝ ΥΛΙΚΩΝ ΔΙΠΛΩΜΑΤΙΚΗ ΕΡΓΑΣΙΑ «Σύνθεση, χαρακτηρισμός και αποδέσμευση βιοδραστικών ουσιών από οστικά τσιμέντα με βάση το φωσφορικό ασβέστιο» ΜΥΣΤΗΡΙΔΟΥ ΕΜΜΑΝΟΥΕΛΑ Α.Μ. 861 Επιβλέπων Καθηγητής: Νικόλαος Μπουρόπουλος Επίκουρος Καθηγητής Πάτρα, Οκτώβριος 2013

description

Σύνθεση, χαρακτηρισμός και αποδέσμευσηβιοδραστικών ουσιών από οστικά τσιμένταμε βάση το φωσφορικό ασβέστιο

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

    [1] . , IV -

    , , ,

    2008.

    [2] N. Patel, P. Gohil, A Review on Biomaterials: Scope, Applications & Human

    Anatomy Significance, International Journal of Emerging Technology and

    Advanced Engineering, 2 (2012) 91-101.

    [3] B. Ratner, A. Hoffman, F. Schoen, J. Lemons, Biomaterials Science An

    introduction to materials in medicine, Academic Press, 2nd Edition, 2004.

    [4] A.M.M. Abdulrazzaq, An in vitro study of genotoxicity of locally produced

    bovine pericardium, School of Dental Sciences, Health Campus, University of

    Sains Malaysia, 2007.

    [5] Drug Delivery Systems: http://www.biobasics.gc.ca/english/View.asp?x=785

    [6] http://www.mddionline.com/article/polymers-controlled-drug-delivery

    [7] . , , ,

    , 2011.

    [8] C. Vogelson, Advances in drug delivery systems, Modern Drug Discovery, 4

    (2001) 49-50.

    [9] Drug Delivery Solutions:http://landec.com/applications/drug-delivery-solutions

    [10] . ,

    , , ,

    , 2012.

    [11] E. Verron, I. Khairoun, J. Guicheux, J. Bouler, Calcium phosphate biomaterials

    as bone drug delivery systems: a review, Drug Discovery Today, 15 (2010) 547-

    552.

    [12] . ,

    , , ,

    , 2008.

    [13] . , ,

    , , ,

    2011.

    [14] V. Budzynski, M. Chen, D. Hakimi-Mehr, M. Landy, M. Tsui, A. Tsvetkov, Q.

    Yang, Lipid coatings for implantable medical devices, Pattent # WO

    2009048645 A2.

    [15] M. Bohner, Design of ceramic-based cements and putties for bone graft

    substitution, European Cells and Materials, 20 (2010) 1-12.

  • [16] M. Sikiric, H. Milhofer, The influence of surface active molecules on the

    crystallization of biominerals in solution, Advances in Colloid and Interface

    Science, 128 (2006) 135-158.

    [17] . , A

    , ,

    , , 2006.

    [18] . ,

    , ,

    , , 2006.

    [19] M. Ginebra, C. Canal, M. Espanol, D. Pastorino, E. Montufar, Calcium

    phosphate cements as drug delivery materials, Advanced Drug Delivery Reviews,

    64 (2012) 10901110.

    [20] M. Ginebra, T. Traykova, J. Planell, Calcium phosphate cements as bone drug

    delivery systems: A review, Journal of Controlled Release, 113 (2006) 102110.

    [21] . , in situ

    , ,

    , , 2006.

    [22] H. Catherine, W. Skinner, Studies in the Basic Mineralizing System, CaO-P2Os-

    H2O, Calcified Tissue Research, 14 (1974) 3-14.

    [23] ., . , - 1,

    , 2006.

    [24] S. Parikh, Bone graft substitutes: past, present, future, Journal of Postgraduate

    Medicine, 48 (2002) 142-148.

    [25] . ,

    , ,

    , , 2010.

    [26] C. Kelly, R. Wilkins, S. Gitelis, C. Hartjen, J. Watson, P. Kim, The use of a

    surgical grade calcium sulfate as a bone graft substitute: results of a multicenter

    trial, Clinical Orthopaedics, 382 (2001) 42-50.

    [27] W. Walsh, P. Morgberg, Y. Yu, J. Yang, W. Haggard, P. Sheath, M. Svehla,

    Response of a calcium sulfate bone graft substitute in a confined cancellous

    defect, Clinical Orthopaedics, 406 (2003) 228-36.

    [28] J. Lane, S. Khan, Bone grafts of the 20th century: multiple purposes, materials

    and goals, Special to Orthopaedics Today, Jan. (2000) 1-12.

    [29] http://www.nsaids-list.com/nsaids-list/ibuprofen/

    [30] http://synthesisofibuprofen.wikispaces.com/

    [31] http://users.teiath.gr/petef/Web_Lessons/Lessons/IEK_Notes/IEK_Diagnoseis/IE

    K_Diagnoseis_IFA_Karkalousos.pdf

    [32] http://www.vcbio.science.ru.nl/en/fesem/eds/

  • 109

    [33] . ,

    , , -

    , , 2005.

    [34] R. Engelberg, Silk encapsulated films for sustained release of Buprenorphine,

    Bachelor Thesis in Biomedical Engineering, Tufts University, 2010.

    [35] . ,

    (Bicalutamide) ,

    , , ,

    , 2013.

    [36] . ,

    ,

    , , , 2009.

    [37] N. Singh, B. Middendorf, Calcium sulphate hemihydrate hydration leading

    togypsum crystallization, Progress in Crystal Growth and Characterization of

    Materials, 53 (2007) 57-77.

    [38] A. Ebrahimpour, M. Johnsson, F. Richardson, H. Nancollas, The

    Characterization of Hydroxyapatite Preparations, Journal of Colloid and Interface

    Science, 159 (1993) 158-163.

    [39] S. Dorozhkin, M. Epple, Biological and Medical Significance of Calcium

    Phosphates, Angew. Chem Int. Ed, 41 (2002) 3130-3146.

    [40] M. Ginebra, E. Fernadez, E. De Maeyer, R. Verbeeck, M. Boltong, J. Ginebra, F.

    Driessens, J. Plannel, Setting reaction and hardening of an apatitic calcium

    phosphate cement, Journal of Dental Research, 76 (1997) 905-12.

    [41] M. Ginebra, F. Driessens, J. Plannel, Effect of the particle size on the micro and

    nanostructural features of a calcium phosphate cement: a kinetic analysis,

    Biomaterials, 25 (2003) 3453-3462.

    [42] A. Antonakos, E. Liarokapis, T. Leventouri, Micro-Raman and FTIR studies of

    synthetic and natural apatites, Biomaterials, 28 (2007) 3043-3054.

    [43] G. Sauer, W. Zunic, J. Durig, R. Wuthier, Fourier Transform Raman

    Spectroscopy of Synthetic and Biological Calcium Phosphates, Calcified Tissue

    International, 54 (1994) 414-420.

    [44] H. Li, B. Ng, K. Khor, P. Cheang, T. Clyne, Raman spectroscopy determination

    of phases within thermal sprayed hydroxyapatite splats and subsequent in vitro

    dissolution examination, Acta Materialia, 52 (2004) 445-453.

    [45] C. Silva, A. Sombra, Raman spectroscopy measurements of hydroxyapatite

    obtained by mechanical alloying, Journal of Physics and Chemistry of Solids, 65

    (2004) 10311033.

  • [46] A. Jillavenkatesa, R. Condrate, The Infrared and Raman Spectra of - and -

    Tricalcium Phosphate (Ca3(P4)2), Spectroscopy Letters, 31 (1998), 1619-1634.

    [47] S. Koutsopoulos, Synthesis and characterization of hydroxyapatite crystals: A

    review study on the analytical methods, Journal of Biomedical Materials Research,

    62 (2002) 600-612.

    [48] S. Ramukutty, E. Ramachandran, Growth, spectral and thermal studies of

    ibuprofen crystal, Crystal Research and Technology, 47 (2012) 31-38.

    [49] B. Fowler, Infrared studies of apatites. I. Vibrational assignments for calcium,

    strontium and barium hydroxyapatites utilizing isotopic substitution, Inorganic

    Chemistry, 13 (1974) 194-207.

    [50] P. Regnier, A. Lasaga, R. Berner, O. Han, K. Zilm, Mechanism of carbonate

    substitution in francolite: Evidence from FTIR, 13C NMR and quantum

    mechanical calculations, American Mineralogist, 79 (1994) 809-818.

    [51] M. Lanzn, P. Garca-Ruiz, Effect of citric acid on setting inhibition and

    mechanical properties of gypsum building plasters, Construction and Building

    Materials, 28 (2012) 506511.

    [52] T. Gong, Z. Wang, Y. Zhang, C. Sun, Q. Yang, T. Troczynski, U. Hafeli,

    Preparation, characterization, release kinetics and in vitro cytotoxicity of

    calcium silicate cement as a risedronate delivery system, Journal of Biomedical

    Materials Research, Part A (2013), doi: 10.1002/jbm.a.34908.