Page 1
2555
สำนกัหอ
สมุดกลาง
Page 2
2555
สำนกัหอ
สมุดกลาง
Page 3
PREPARATION AND PROPERTIES OF ELECTROSPUN CHITOSAN/ POLY (LACTIC
ACID) NANOFIBER FOR POTENTIAL APPLICATION AS WOUND DRESSING
By
Miss Amornrat Tantithumrongwut
A Thesis Submitted in Partial Fulfillment of the Requirements for the Degree
Master of Engineering Program in Polymer Science and Engineering
Department of Materials Science and Engineering
Graduate School, Silpakorn University
Academic Year 2012
Copyright of Graduate School, Silpakorn University
สำนกัหอ
สมุดกลาง
Page 4
“
”
…….…...........................................................
(
(
( (
( (
สำนกัหอ
สมุดกลาง
Page 5
52402240 :
100
(Unhydrolyzed chitosan Hydrolyzed chitosan)
H
20%
20 19
10, 20 40
2555
สำนกัหอ
สมุดกลาง
Page 6
52402240 : MAJOR : POLYMER SCIENCE AND ENGINEERINGKEY WORD : POLY(LACTIC ACID) CHITOSAN ELECTROSPINNING NANOFIBER
AMORNRAT TANTITHUMRONGWUT : PREPARATION AND PROPERTIES OF ELECTROSPUN CHITOSAN/ POLY (LACTIC ACID) NANOFIBER FOR POTENTIAL APPLICATION AS WOUND DRESSING. THESIS ADVISORS : NUTTAWUT CHAIYUT, Ph.D., ASSOC. PROF. KALYANEE JIRASRIPONGPUN, Ph.D. 100 pp.
Chitosan/poly(lactic acid) (PLA) nanofibers were successfully prepared by electrospinning process. Electrospinnability and morphology of the electrospun fibers were investigated by adjusting electrospinning parameters, such as spinning electric field, spinning distance, types of chitosan (unhydrolyzed and hydrolyzed), concentration of PLA solution and solution blend ratios of chitosan and PLA. Morphological study of the obtained fibers was carried out by scanning electron microscope (SEM). The suitable condition for producing chitosan/PLA fibers as follows: applied voltage is 19 kV and collection distance is 20 cm. Hydrolyzed chitosan and chitosan/PLA at ratio of 40/60 were suitable to electrospinning uniform chitosan/PLA nanofibers.
Hydrolyzed chitosan/PLA and hydrolyzed chitosan/PLA by adding gentamicin sulfate at 10, 20 and 40 mg/ml nanofibers was prepared by electrospinning process which tested and observed for physical and mechanical properties changes. This study found that the more gentamicin sulfate added to hydrolyzed chitosan/PLA solution, the less water absorbed, resulting in low vapor transmission rate of the electrospuns. Hydrolyzed chitosan/PLA and hydrolyzed chitosan/PLA by adding gentamicin sulfate showed no cytotoxicity toward growth of Vero cell.
Department of Materials Science and Engineering Graduate School, Silpakorn University Student's signature ........................................ Academic Year 2012Thesis Advisor's signature 1. ........................... 2. ...........................
สำนกัหอ
สมุดกลาง
Page 7
Scanning Electron Microscope (SEM)
สำนกัหอ
สมุดกลาง
Page 8
1 1
1
2
3
4
2 5
5
5
7
Low-adherent Dressing 8
8
Hydrogel 8
Hydrocolloid 8
Alginate dressing 9
Non-adhesive-semi-permeable Polyurethand Foam Dressing 9
9
9
10
10
สำนกัหอ
สมุดกลาง
Page 9
10
10
11
11
12
12
12
13
13
13
14
14
14
14
15
16
16
16
16
17
Degree of deacetylation 17
17
Coagulation ability 17
Molecular conformation 17
18
19
19
19
สำนกัหอ
สมุดกลาง
Page 10
20
20
21
21
24
24
24
25
25
26
26
26
2
29
29
30
33
34
35
3 36
36
37
38
39
hydrolyzed chitosan 39
39
40
สำนกัหอ
สมุดกลาง
Page 11
41
41
43
44
44
44
45
45
46
46
47
49
4 51
51
51
53
55
Hydrolyzed chitosan PLA
56
58
59
60
สำนกัหอ
สมุดกลาง
Page 12
61
62
63
64
5 66
66
67
69
73
74
76
82
85
93
95
(Drug release)
97
100
สำนกัหอ
สมุดกลาง
Page 13
1 LPL
–
21
2 23
3 25
4 30
5 (Permeability)
(PET) (OPS)
(OPLA)
33
6 Unhydrolyzed chitosan Hydrolyzed chitosan 55
7
56
8
58
9 60
10 61
11 S. aureus /
64
12 /
24
65
13 (Intrinsic viscosity)
75
14
77
15
78
16
Unhydrolyzed
สำนกัหอ
สมุดกลาง
Page 14
chitosan hydrolyzed chitosan hydrolyzed
79
17
80
18
81
19 83
20
10
83
21
20
84
22
30
84
23 (Water vapor
transmission rate: WVTR)
86
24 (Water vapor
transmission rate: WVTR)
10
87
25 (Water vapor
transmission rate: WVTR)
20
88
26 (Water vapor
transmission rate: WVTR)
40
89
27 (Water vapor rate: WVTR)
92
สำนกัหอ
สมุดกลาง
Page 15
28 (Water adsorption)
94
29 (Cytotoxicity)
96
30 max = 400
24
98
31 max = 400
98
สำนกัหอ
สมุดกลาง
Page 16
1 (a) neutrophils
(b) (Epithelial
cells)(c) (d)
6
2 15
3 16
4 27 5 28 6 (T
g) (Tm)
31 7 (Tg) -
31
8 (Tm) -
33
9 (LDPE)
(PS) (PLA) L:D
96:4 (PET)
34
10 43
11 44
12 46
13 UV-VIS spectrophotometer max = 400
50
14 Hydrolyzed chitosan/PLA
20
0.01
a) 13 b) 15 c) 17 d) 19
52
15 Hydrolyzed chitosan/PLA
สำนกัหอ
สมุดกลาง
Page 17
19
0.01
a) 8 b) 12 c) 16 d) 20
54
16
a) Unhydrolyzed chitosan b) Hydrolyzed
chitosan
55
17
Hydrolyzed chitosan/PLAa) PLA
15%wtb) PLA 18%wt c) PLA 20%wt
57
18
Hydrolyzed chitosan/PLA a) 40/60 b)
50/50
59
19 /
/
62
20
24
63
21 Clear zone
Staphylococcus aureus (S. aureus)
64
22
65
23
Hydrolyzed chitosan
75
24
90
25
10%
90
สำนกัหอ
สมุดกลาง
Page 18
26
20%
91
27
30%
91
สำนกัหอ
สมุดกลาง
Page 19
1
1
1.1
(Wound dressing)
30
[1]
สำนกัหอ
สมุดกลาง
Page 20
2
[2,3]
(Poly(lactic acid) ;PLA) (Polylactide)
Aliphatic polyester (Lactic acid)
PLA
PLA
[4]
(Gentamicin sulphate)
(Aminoglycosides)
(Gram
negative aerobic bacteria) Staphylococcus aureus [5,6]
Electrospinning
process)
1.2
1.2.1
1.2.2
สำนกัหอ
สมุดกลาง
Page 21
3
1.2.3
1.2.4
1.3
(Unhydrolyzed chitosan Hydrolyzed chitosan)
1.4
1.4.1 1.4.2 1.4.3
1.4.3.1
(Unhydrolyzed chitosan Hydrolyzed chitosan)
1.4.3.2
(Water vapor permeability)
สำนกัหอ
สมุดกลาง
Page 22
4
(Water absorption) (Cytotoxicity)
(Antibacterial test) (Drug releasability)
1.4.4
1.4.5
1.4.6
1.5
1.5.1
1.5.2
1.5.3
สำนกัหอ
สมุดกลาง
Page 23
5
2
2.1 [7-9]
2.1.1 rocess)
(Platelets) (Fibrin)
“Haemostatic phase”
“Inflammatory phase”
(Epidermal cell)
(Epithelialization) (Fibrobrast)
“Proliferative phase”
“Maturation phase” “Remodeling phase”
สำนกัหอ
สมุดกลาง
Page 24
6
1 (a) Neutrophils
(b) (Epithelial cells) (c)
(d)
[8]
Inflammatory
phase
(Infection)
Senescent cell
1.
Page 25
7
2.
(Epithelialization) 2
3.
4.
Exudates
5.
6.
7.
8.
9.
10.
11.
12.
2.1.2
2.1.2.1 s)
Page 26
8
Debride Pack
Polyester Rayon
Hypertonic Saline Sponge
2.1.2.2 -
Exudates
2.1.2.3 (Semi-
(Adhesives)
Autolytic
debridement
2.1.2.4
2.1.2.5
(Granule)
Page 27
9
2.1.2.6
(Ion
exchange) (Ca+)
2.1.2.7 Non- -semi-
2.2 ) [10-13]
2
(Reinforcing fibers)
2.2.1
“Taylor
cone”
(Electrostatic repulsion)
Page 28
10
Bending instability)
(Unbalance force)
2.2.2
3
2.2.2.1
2.2.2.1.1 (Concentration)
(Beads)
(Bending instability)
Power law
2.2.2.1.2
Dielectric strength
Page 29
11
(Chain entanglement)
(Beads)
2.2.2.1.3 ( iscosity)
1-215
poise [11]
Stress relaxation
2.2.2.1.4
Page 30
12
2.2.2.1.5 (
(Bending instability)
2.2.2.2
2.2.2.2.1
(Electrostatic force)
Page 31
13
2.2.2.2.2
2.2.2.2.3
(Dimple)
2.2.2.2.4distance) (Flight time)
(Electric field strength)
(Merge) (Intra
and inter layer bonding) Scaffold
Page 32
14
2.2.2.3
2.2.2.3.1 (Humidity)
Atomic force
2.2.2.3.2
Freon ®-12 2
2.2.2.3.3 (Pressure)
Page 33
15
2.2.3
(
2
2 [13]
Page 34
16
2.3 [14-19]
2.3.1
(Deacetylation)
(-NHCOCH3) (-NH2) 2
D-glucosamine(2-amino-2-deoxy-D-glucose)
Poly( (1 4)-2-amino-2-deoxy-D-glucose) 3
3 [14]
2.3.2
2.3.2.1
(Organic solvent)
pH 6
-
(Non-Newtonian)
(pKa) pKa
6.2-6.8
Page 35
17
2.3.2.2 1×105 - 2×106
2.3.2.3 n -
- N-acetyl-D-glucosamine
D-glucosamine Degree of
deacetylation
Degree of deacetylation
2.3.2.4
Degree of deacetylation Ionic strength
- (pH)
pH
pH
pH
2.3.2.5
(Flocculating and coagulating agent)
Cheese whey
(Coordinate)
Degree of deacetylation
2.3.2.6
(Cationic Polyelectrolyte)
–NH+3 Conformation Mark-
Houwink Exponent ( a) a 0, 0.5-0.8 1.8
Page 36
18
(Sphere) Random coil (Rod) Conformation
Ionic Strength pH
Degree of deacetylation
2.3.2.7
(Oligomer) (Oligosaccharide)
(Monomer) (Monosaccharide)
/ N-Acetyl-
Chitooligosaccharide Chitooligosaccharides /
N-Acetyl-D-Glucosamine D-Glucosamine
2.3.2.7.1
Random
2.3.2.7.2
Peeling Reaction
2.3.2.7.3sonication)
2.3.2.7.4
- Chitinase (EC 3.2.1.14)
(Random) (-1,4-Linkage) N-acetyl-chitooligosaccharide
Chitosanase (EC 3.2.1.132) (Random)
(-1, 4-Linkage) Chitooligosaccharide Lysozyme (EC 3.2.1.17)
Chitinase N-acetylglucosaminidase (EC3.2.1.30) N-
acetylglucosaminidase (EC3.2.1.50) N-acetylchitooligo saccharides N-
acetyl-glucosamine (Non-reducing end)
Page 37
19
2.3.2.7.5
(Dry heat) 80
Glass transition temperature (Tg)
120
160 2
(0.2 M)/ (0.1M)
(Saturated steam)
115 2
120 1
120
2.3.2.8 3
(-NH2) 2 (C-2) Primary
alcohol (-CH2OH) 6 (C-6) Secondary alcohol (-CHOH)
3 (C-3) (Chemical modification)
2.2.1 [15-16]
(Biomaterials) (Biocompatibility)
(Biodegradable)
(-NH2) (-OH)
(Derivatives)
-
2.2.1.1 -
-
Page 38
20
-
-
2.2.1.1.1 -
(Non–woven fabrics)
(Hydrogel) -
-
Chitosan–collagen [17] (Contact lenses) Partially
depolymerisation Spin casting
(Blood dialysis membrane)
Antibacterial, Antivirus Antitumor
-
Periodontal tissue
(Foreign body reaction)
2.2.1.1.2 , Anti- Heparin
(N-sulphate
group) Heparin - -
Lipoprotein lipase (LPL)
1 Chitin-3, 6-sulphate
Heparin 2 LPL 0.1
O-Sulphated Chitin (Toxicity, LD50 = 1.25–3.25 g/kg)
Haparin (LD50 = 1.59–2.00 g/kg) [17] -
N-Hexanoyl N-Octanoychitosan
[16]
Page 39
21
1 LPL
– [16]
MW
(x103) -
Chitin 26 2.0 355 (2.0) 100 (0.1)
Chitosan 12-22 1.7 239 (3.4) 3200 (3.4)
N-Desulphated chitosan 22 0.7 n.d.c Inactive
N-Hexanoy chitosan 27 1.8 n.d.c Inactive
CM-chitosan 245-540 0.6 26 (0.10) 700 (0.7)
Heparin 21 - 174 (1.0) 950 (1.0) a The dosage was 0.1 mgkg-1 of body weight, and maximum LPL activity is shown in mole
equivalents of free acids per litre of plasma after incubation at 37 C for 30 min, and the activity
relative to heparin is shown in parentheses b With respect to the activated thromboplastin time (APTT); the activity relative to heparin is
shown in parentreses. c Not determined.
2.2.1.2
2.2.1.2.1 [18]2.2.1.2.1.1
2
(Cholesterol)
Page 40
22
20
1-2 25–50 %
–
2.2.1.2.1.2 -
2
2 Glucosamine ( )
pH 6 -
Interaction
Chelating agent
(Toxin)
Page 41
23
2 [18]
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
2.2.1.2.1.3
2.2.1.2.1.4
Gelatin, Bentonite, Silica sol,
Tannins, Potassium caseinate Polyvinyl pyrrolidone
Grapefruit Fining
agent
Page 42
24
2.2.1.2.2 (Drug delivery and controlled release systems)
Glutaraldehyde cross–linked chitosan
2.2.1.2.3
Hyaluronic acid
(Water binding capacity)
(Wella Inc.)
2.2.1.3 [15,17] -
1981 -
- 3
-
( )
Page 43
25
3 [16]
Chitosana
N-Acetylchitosanb
N-Propionlychitosanb
3.74-9.02
3.18
4.71
0.78–1.24
0.79
0.52
13.0–28.1
27.1
25.9 aA 5% chitosan solution in aqueous 2% CH2COOH was span into an aqueous 10% NaOH
containing 30%CH3COONa. b Each solution (5–8%) of sodium N-acetylchitosan salt and sodium N-propionylchitosan salt in
aqueous 14% NaOH as span into aqueous 10% H2SO4 containing 25-33% Na2SO4 and 1.3%
ZnCl2
(Anionic inks) – Hydroxymethyl chitin
[17]
2.2.1.4 [15,19
2.2.1.5 [15]
-
(Tree-protecting agents) (Animal feed)
Page 44
26
(Fertilizers/Soil strabilizers)
2.2.1.6 [15]
Support
Crystalline cellulose (Polyamide) Cross–
linked
Gas chromatography ( Stationary phase) High performance liquid chromatography
Bio-reactor, Bio-sensor Ligand carrier Affinity chromatography
Carrier ( Glucose isomerase)
2.3 [20-23]
(Poly(lactic acid), PLA)
(Aliphatic polyester) (Lactic acid)
2.3.1
( -Hydroxy acids)
2-
(2-Hydroxy propionic acid)
( 4)
(Enantiomer) (Optical active)
- - -
(Racemic mixture, = 1:1 DL)
(meso-compound) (Optically inactive)
(Optical
purity)
Page 45
27
4 [21]
(Azeotropic dehydrative condensation) (Direct
condensation polymerization) / (Lactide for-mation)
( 5) ( 100,000 )
(Ring-opening polymerization)
- ( -
(PLLA)) ( -
(PDLLA)) -
- (PDLA)
( ) ( )
- ( ) ,
185
175
Page 46
28
5 [21]
2.3.2
(Optical purity)
(Barrier pro-
perties) - 90
(Semicrystalline polymer) -
(Amorphous)
-
(PET) (PS)
Page 47
29
2.3.2.1
(Degree of crystallinity)
(Hexane) (Heptane)
(Poly(L-lactic acid), PLLA)
(Chlorinatedor fluorinated organic solvents) (Dioxane) (Dioxolane)
(Furane)
(Poly(rac-lactic acid)
(Acetone) (Pyridine) (Ethyl lactate) (Ethyl
acetate) (Tetrahydrofuran) (Xylene)
(Dimethylsul-foxide) , - (N,N- dimethylformamide)
(Methyl ethyl ketone)
2.3.2.2
1.25 (Unoriented PLA)
(Stiffness) (Strength) (Oriented)
(Polyethylene terepthalate, PET)
(Oriented PS) (Tensile and flexural modulus)
(HDPE) (PP) (PS)
(Izod impact strength) (Elongation at break)
4
Page 48
30
[20]
(Ultimate tensile strength, psi)6,900-7,700 6,900-24,000
(Tensile yield strength, psi)6,600-8,900 N/A
(Tensile modulus, psi) 500,000-580,000 564,000-600,000
(Notched Izod impact, ftlb in-1)0.3-0.4 N/A
Elongation at break (%) 3.1-5.8 15-160
(Rockwell hardness) 82-88 82-88
* (Degree of orientation)
2.3.2.3 (Tg)
(Tm) ( 6)
(L:D ratio, 7)
-
50:50
Page 49
31
6 (Tg) (Tm)
[20]
7 (Tg)
- [20]
Page 50
32
Flory-Fox 1 =
= Tg
PLLA PDLA 57-58
K = (Excess free volume)
(End groups) PLLA PDLA
(5.5-7.3) x 104
=
(Thermal history)
(Tm)
( PLLA PDLA) 180 40-50
-
50 8
(meso-lactide)
130-
160
Page 51
33
8 (Tm)
[20]
2.3.2.4
PLA
(PS) (PET) ( 9)
( )
-
(D-linonene)
(Permeability) (PET)
(OPS) (OPLA) [20]
( .)
( 2 )) PET 18 2.82 x 10-15 ± 1.50x10-17
OPS 18 4.18 x 10-15 ± 2.07x10-17
OPLA 20 1.34 x 10-14 ± 3.61x10-17
Page 52
34
9 (LDPE)
(PS) (PLA) L:D 96:4
(PET) [20]
2.3.3 [22]
(Oligomer)
60
60 2
-
Page 53
35
(Autocatalyzed) (Carboxylic acid)
2.3.4 [23]
-
(Biocompatible) (Bioresorbable)
2 (Sutures) (Stables)
(Wound dressing) (Surgical implants)
(Orthopedic fixation devices)
-
-
- Non-woven
- (Bumpers) (Floor mats)
-
-
Page 54
36
3
3.1
1. Poly(lactic acid) (PLA) 2002D NatureWorks®
2. Chitosan (CS) Seafresh Chitosan (LAB) CO.,Ltd
150,000 Degree of deacetylation (%) 90
3. Gentamicin sulfate Vesco Pharmaceutical CO.,Ltd Gentamicin
40 mg/ml
4. Acetic acid Glacial 100% suitable for biopharmaceutical production
EMPROVE® bio Merck
5. Acetone Analysis EMSURE® ACS, ISO, Reag. PhEur Merck
6. Chloroform Commercial Labscan
7. Deionized water
8. Dimethylsulfoxide (DMSO) Analysis EMSURE® ACS Merck
9. 3-(4,5-Dimethylthiazol-2-yl)-2,5-Diphenyltetrazolium Bromide (MTT)
Invitrogen
10. Distilled water
11. Ethanol
12. Ethylen diamine tetra acetic acid (EDTA: C10H16N2O8) Ameresco
13. Fetal Bovine Serum (FBS) HyClone
14. Modified Eagle’s Medium (MEM) Gibco
15. Ninhydrin Ajax
16. Nutrient agar (NA) Lab008 Lab M
17. Nutrient broth (NB) Lab014 Lab M
18. Potassium chloride (KCl) Plant cell culture test Sigma
19. Potassium phosphate (KH2PO4) Plant cell culture test Sigma
Page 55
37
20. Sodium chloride (NaCl) Cell culture Reagent Sigma
21. Sodium hydroxide (NaOH) American Chemical Society certified (ACS)
Carlo Erba
22. Sodium phosphate (Na2HPO4) Plant cell culture test Sigma
23. Trypan blue Sigma
24. Trypsin Fluka
3.2
1. (Aluminium foil)
2. (Autoclave) Tommy SS-325
3. Autopipette 20-200 ml
4. Autopipette 100-1000 ml
5. Breaker)
6. Buchner funnel)
7. Centrifuge) Hettrich
8. (Centrifuge tubes) 15 ml
9. (Clamp/Clamp holder)
10. (Condenser)
11. Conical flask with joint)
12. Cotton)
13. –80 (Deep freezer) NuAire
14. Dropper)
15. (Drum collector)
16. Electrospinning machine)
17. Filter paper)
18. Forceps)
19. Graduated cylinder)
20. Hemocytometer
21. Hot plate with stirrer)
22. CO2 Incubator) NuAire
Page 56
38
23. (Incubator) Memmert
24. Laminar air flow class II) NuAire
25. (Light microscope) Olympus
26. (Magnetic bar)
27. 0.2 μm (Membrane filter)
28. Metal plate collector)
29. 3 3-Necked round bottom flask)
30. (Oil bath)
31. (Petri dish)
32. pH indicator paper)
33. pH-Meter) Schott
34. (Pipette) 5 10 ml
35. Shaker) GFL 3017
36. Spatula)
37. 26 (Spinneret)
38. (Stop watch)
39. (Suction flask)
40. (Syringe)
41. (Temperature controller)
42. T-flask 25 cm2
43. Thermometer)
44. Vacuum oven with pump)
45. (Vial tube)
46. (Viscometer) Brookfield DV-I+ prime
47. Volumetric flask)
48. 96-well plate
3.3
1. (Microplate reader TECAN
ELISA
Page 57
39
2. Scanning electron microscope : SEM)
CamScan MX-2000
3. Ubbelohde viscometer
4. Universal testing machine Lloyd
5. UV-VIS spectroscopy (UV-VIS) PG Instruments
T80+UV/VIS Spectrometer
3.4
3.4.1 H -CS)
Hydrolyzed chitosan (h-CS) 1
NaOH 50% 25 95 24
Deionized water
100% Deionized water Hydrolyzed chitosan
60 16
3.4.2 [35]
Hydrolyzed chitosan
(Intrinsic viscosity) 90%
Hydrolyzed chitosan 5
0.5-5.0 g/l
Mark-Houwink equation 2
[ ] = (2) K a Mark-Houwink parameter
1.8×10-3 0.93 [ ]
(Intrinsic viscosity) Huggins equation 3)
Kramer Equation 4)
Huggins equation : = [ ] + [ ] (3)
Kramer equation : = [ ] [ ] (4)
Page 58
40
c = (g/l)
= relative viscosity
= ( ) ( )
= 1 = Specific viscosity
Hydrolyzed chitosan
plot
(C) (g/l) y
Huggins Kramer equation [ ] [ ]
2
3.4.3
Hydrolyzed chitosan (h-CS) Hydrolyzed chitosan
3.4.1 90% 5%
(PLA solution)
(CHCl3) ((CH3)2CO)
70:30 15, 18 20%
48
(chitosan : poly(lactic acid) blend ratio) 50/50 40/60
Hydrolyzed chitosan
Unhydrolyzed chitosan (CS)
5% 90%
(PLA solution) 20%
(chitosan : poly(lactic acid)) 50/50
Page 59
41
3.4.4
3.4.3
Brookfield viscometer (Model DV-
I+prime) Spindle 2 Switch power
Spindle 2 Select spindle
Spindle 3.4.3
0.05-200
%Torque
Option Time to stop 1 Enter Set speed
Enter 2
3.4.5
2
1. hydrolyzed
chitosan/PLA Hydrolyzed chitosan/PLA
2.
Unhydrolyzed chitosan Hydrolyzed chitosan
hydrolyzed chitosan/PLA
5%wt/v
Hydrolyzed chitosan/20%wt/v
50/50 10 26
10
(Feed rate) 0.01
Page 60
42
20 20 13, 15, 17
19
5%wt/v Hydrolyzed chitosan/20%wt/v f
50/50 10
26 10
(Feed rate) 0.01 19
20
8, 12, 16 20
5%wt/v Hydrolyzed chitosan/20%wt/v
5%wt/v Unhydrolyzed chitosan/20%wt/v
50/50 10
26
10 (Feed rate) 0.01
19 20
20
5%wt/v Hydrolyzed chitosan/ 15, 18
20%wt/v 50/50
10 26
10 (Feed rate)
0.01 19
20 20
5%wt/v Hydrolyzed chitosan/20%wt/v
50/50 40/60 10
26
10 (Feed rate) 0.01
Page 61
43
19 20
20
10
3.4.6
5%wt/v h-CS/20%wt/v PLA
3.4.4
10 26
11 0.01
(Drum collector)
20
19
15
10 6
60 C 12
Page 62
44
11
3.4.7
3.4.6
400
3.4.8
3.4.8.1
3.4.4 60
12
(Scanning electron microscope: MX-2000, CamScan)
Stub
Sputtering
Page 63
45
2,000
ImageTool
3.4.8.2
3.4.5 3.4.6
5×65 Tensile test)
ASTM D882-83 Universal testing machine (Lloyd)
Load cell 1 Gage length 20 10
3.4.8.3
) -37]
3.4.6 3.4.7
3×3
(Parafilm)
(%RH)
6
12 WVTR ( ) = ( ) × 24
w t = 1 (g/hr)
A = (1.51×10-4 m2)
Page 64
46
12
3.4.8.4 [29]
24 3.4.6 3.4.7
3×3 (W0)
24 We)
%
(%) = × 100
We
W0
3.4.8.5 )
3.4.7.5.1
Vero cell
African green monkey kidney cell
w t
Page 65
47
3.4.7.5.2
Vero cell
Minimum essential medium (MEM) Fetal bovine serum (FBS) 10%
37 5%
log phase (Conference) 80-90%
3.4.7.5.3
3.4.5 3.4.6 1
100 20 deionized
water
200 rpm 3 0.2
3.4.7.5.4
MTT assay ISO
10993-5 2×105
96 well plate 100 μl 24 37
5% 3.4.7.5.3
0.25, 0.5, 1, 2 4 mg/ml 100 μl
24
Phosphate-buffered saline (PBS) MTT 1.0 mg/ml 50 μl
4 MTT
DMSO 50 μl 570
3.4.8.6 (
Agar diffusion
Page 66
48
3.4.7.6.1
Staphylococcus aureus (S. aureus)
3.4.7.6.2 Nutrient agar (NA) Nutrient broth (NB)
Growth media
(Sterile)
3.4.7.6.2.1 Nutrient broth (NB)
Nutrient Broth 1.25 50 ml
pH NaOH 1 M pH 6.8 - 7.2
10 5 ml
(Sterilize) Autoclave 121
15
3.4.7.6.2.2 Nutrient agar (NA)
Nutrient Agar 4.2 150 ml
pH NaOH 1 M pH 6.8 - 7.2
Autoclave 121 15
Nutrient agar (Plate) Nutrient agar
3.4.7.6.3
3.4.5 3.4.6 Aseptic technique Laminar air
flow class II
3.4.7.6.4
(NB) 5 ml 1 loop 37 24
Nutrient agar plate (NA plate)
3.4.7.6.3 NA plate 37
24 Clear zone
Caliper
Page 67
49
3.4.8.7
(Drug release) [38-39]
3.4.6 1
100 Phosphate-buffered saline (PBS) pH 7.4
10 100 37
1, 2, 3, 6, 12, 24, 48 72
5 Ninhydrin
1.25% 1.5 95 15
UV-VIS
400 Phosphate-buffered saline (PBS) pH 7.4
Phosphate-buffered saline (PBS)
pH 7.4 Ninhydrin 5:1
(Standard curve)
13
Page 68
50
13
UV-VIS = 400
y = 0.0163x + 0.219 R² = 0.9844
0
0.2
0.4
0.6
0.8
1
0 10 20 30 40 50
Page 69
51
4
4.1
Hydrolyzed chitosan PLA
Hydrolyzed chitosan PLA
Unhydrolyzed chitosan Hydrolyzed chitosan
4.1.1 H PLA
Hydrolyzed
chitosan PLA
Hydrolyzed chitosan (h-CS) 5%
(PLA solution) 20%
(chitosan : poly(lactic acid) blend ratio)
20
0.01 13,
15, 17 19 14
Page 70
52
14 Hydrolyzed chitosan PLA
20
0.01
a) 13 b) 15 c) 17 d) 19
14
Hydrolyzed chitosan PLA
237.39±59.45, 210.92±45.27, 181.74±49.02
137.07±25.53 13, 15, 17 19
Hydrolyzed chitosan PLA
Hydrolyzed chitosan PLA
Hydrolyzed chitosan PLA
b
d
SEI 15 kV 10 μm
SEI 15 kV 10 μm SEI 15 kV 10 μm
SEI 15 kV 10 μm
a
c
Page 71
53
[10-12]
Hydrolyzed chitosan PLA 19
4.1.2
Hydrolyzed chitosan
(h-CS) 5% (PLA solution)
20%
(chitosan : poly(lactic acid) blend ratio)
19 0.01
8, 12, 16 20
15
Page 72
54
15 Hydrolyzed chitosan PLA
19 0.01
a) 8 b) 12 c) 16 d) 20
15
256.58±38.88, 233.14±21.92, 205.48±24.04
137.07±25.53 8, 12, 16 20
(Beads)
a b
d
SEI 15kV 10 μm SEI 15kV 10 μm
SEI 15kV 10 μm SEI 15kV 10 μm
c
Page 73
55
[10-12]
Hydrolyzed chitosan PLA 20
4.1.3
Unhydrolyzed
chitosan Hydrolyzed chitosan ( 6)
(chitosan : poly(lactic acid) blend ratio)
19 20
0.01
16
6 Unhydrolyzed chitosan Hydrolyzed chitosan
( )
Unhydrolyzed chitosan 150,000
Hydrolyzed chitosan 50,318
16
a) Unhydrolyzed chitosan b) Hydrolyzed chitosan
a b
SEI 15kV 10 μm SEI 15kV 10 μm
Page 74
56
16
Unhydrolyzed chitosan
Hydrolyzed chitosan
Unhydrolyzed chitosan
[10-12] Hydrolyzed chitosan
4.1.4
PLA
H
(chitosan : poly(lactic acid) blend ratio)
19
20 0.01
17
7
( (
15 220.33
18 573.20
20 985.76
Page 75
57
17
Hydrolyzed chitosan PLA
a) PLA 15%wt b) PLA 18%wt c) PLA 20%wt
17
(Beads)
133.37±15.21, 134.87±12.11 137.07±25.53
15, 18 20 %
( 7)
(Entanglement)
b c
SEI 15kV 10 μm SEI 15kV 10 μm
a
SEI 15kV 10 μm
Page 76
58
[10-12]
20%
4.1.5
Hydrolyzed chitosan (h-CS)
5% (PLA solution)
20%
(chitosan : poly(lactic acid) blend ratio) 0 5 50
19 20
0.01
18
8
(
1,243.48
985.76
Page 77
59
18
Hydrolyzed chitosan PLA a) 4 60 b) 0
18
137.07±25.53
369.10±45.39 50 60 %
60
50
4.2
(Tensile test)
ASTM D882 Load cell 1 Gage
length 20 10
a b
SEI 15kV 10 μm SEI 15kV 10 μm
Page 78
60
9
)
33.18 ± 0.64 8.92 ± 0.36 3.59 ± 0.09
10 30.36 ± 0.90 9.70 ± 0.48 3.64 ± 0.21
29.68 ± 1.16 9.73 ± 0.67 3.82 ± 0.17
40 27.70 ± 0.55 9.87 ± 0.21 4.33 ± 0.14
9
(Tensile strength)
(%Elongation)
4.3
WVTR)
10
Page 79
61
10
2
1,796.03
10 1,764.24
20 1,732.45
40 1,494.04
10
4.4
19
Page 80
62
19
4.5
90-100 %
10-
10-
297.89 291.33 262.87
239.42
0.00
50.00
100.00
150.00
200.00
250.00
300.00
350.00
Page 81
63
20
24
4.6 (
Agar diffusion test
(Clear zone)
Staphylococcus aureus (S. aureus
21 S. aureus
Clear zone
S. aureus 10-40
Clear zone 11)
0.00
20.00
40.00
60.00
80.00
100.00
120.00
140.00
No GS Add GS Add GS Add GS
()
Page 82
64
21 Clear zone
Staphylococcus aureus (S. aureus)
11 S. aureus
9.50
18.25
20 22.50
40 25.00
4.7
release)
C+
C-
C+
C-
4
4
4
2
2
+2
Page 83
65
12
10 7.12
20 18.04
40 37.42
3 (
33
[40]
0.00
2.00
4.00
6.00
8.00
10.00
0 10 20 30 40 50 60 70 80
( )
Page 84
66
5
5.1
/
2
Unhydrolyzed chitosan Hydrolyzed chitosan
Hydrolyzed chitosan/PLA Hydrolyzed
chitosan/PLA PLA 20%
40/60
20 19
0.01
369.10±45.39
10 20 40
24 Universal testing machine
Page 85
67
/
(Tensile strength)
(%Elongation)
(0-40 mg/ml)
Staphylococcus aureus (S. aureus)
S. aureus 10-40 mg/ml
3
5.2
1. /
Page 87
69
[1]
cited 1 2553]; Available from:
[2] Novel chitin and chitosan
nanofibers in biomedical applications -
[3]
cited 1 2553]; Available from:
C K Preparation and characterization of biodegradable
poly(l- -
[5] cited 1 2553]; Available
-topical-
[6] Gentamicin Cream
- A review
on wound dressings with an emphasis on electrospunnanofibrous polymeric bandages
-95
[9]
-
C An Introduction to
Electrospinning and Nanofibers
[11] Electrospinning: A fascinating fiber fabrication technique
-
[12] Z A review on polymer nanofibers by
electrospinning and their applications in nanocomposites
-
Page 88
[13] Bending instability of electrically
charged liquid jets of polymer solutions in electrospinning
-
-
- -
[15] -
- -
[16] Chitin and chitosan as novel biotechnological materials
-
Chitin Biotechnology Applications -
-
F Kamil Food applications of chitin and chitosans
-
[19] - cited 1 2553]; Available from:
:
-
[21] Poly(lactic acid) fiber: An overview
-
[22] Large-scale production, properties and commercial applications of polylactic
acid polymers – -
[23] Polylactides “NatureWorks® PLA -
C C C Stretching-induced crystallinity and orientation of
polylactic acid nanofibers with improved mechanical properties using an electrically
charged rotating viscoelastic jet -
[25] Chitosan bicomponentnanofibers and nanoporous fibers
-
[26] Influence of acetic acid concentration on the
solubilization of chitosan -
Page 89
X Electrospinning of chitosan dissolved in concentrated
acetic acid solution -
Che K
as wound dressing
– -
[29] Electrospinning of gelatin an -
lactide) blend and its characteristics for wound dressing
-
Preparation and
characterization of electrospun
system -
[31]
-
[32] K Preparation of fish gelatin and fish
-lactide) nanofibers by electrospinning
-
[33] A Electrospinning of chitosan
nanofibers: Processing optimization -
- In vitro gentamicin
sustained and controlled release from chitosan cross-linked films
-
[35] Calculation of Mark–Houwink–Sakurada (MHS) equation viscometric
constants for chitosan in any solvent–temperature system using experimental reported
viscometric constants data -
[36] In vitro assesment of water vapour
transmission of synthesic wound dressing -
Application of benzyl
hyaluronate membranes as potential wound dressing: evaluation of water vapour and
gas permeabilities -
Page 90
ez- A validated quantitative colorimetric
assay for gentamicin -
[39]
In vitro elution and dissolution of tobramycin and gentamicin from
calcium phosphate -
Amoxycillin release from a floating dosage form
based on alginates - -
Page 93
75
13 (Intrinsic viscosity)
Concentration (g/l) 1 2 3 4 5
Hydrolyzed
chitosan
(ln rel)/c : Kraemer 0.0600 0.0592 0.0588 0.0587 0.0582
sp/c : Huggins 0.0618 0.0628 0.0643 0.0661 0.0676
23
Hydrolyzed chitosan
y 2 Kraemer Huggins [ ] [ ] Mark-Houwink parameter
(1) (2)
K = 1.8×10-3 a = 0.93 (2)
[ ] = (1.8 × 10 / ) . (1)
= [ ] /. × / × . (2)
y = 0.0015x + 0.0601
R² = 0.9914
y = -0.0004x + 0.0602
R² = 0.9281 0.0550
0.0570
0.0590
0.0610
0.0630
0.0650
0.0670
0.0690
0 2 4 6
intr
insic
visc
osity
concentration (g/l)
Huggin eq
Kraemer eq
(Huggin eq)
(Kraemer eq)
Page 95
77
14
(nm)
13 kV 20 cm 15 kV 20 cm 17 kV 20 cm 19 kV 20 cm
1 209.52 171.43 206.03 78.54
2 188.56 209.52 202.48 133.33
3 209.52 171.43 296.31 133.33
4 205.15 231.72 202.48 120.47
5 285.71 310.07 170.37 157.07
6 286.35 247.62 134.69 133.33
7 217.18 190.48 161.62 152.38
8 198.86 210.39 171.43 137.35
9 198.86 153.57 138.67 152.38
10 374.23 212.96 133.33 172.48
Mean 237.39 210.92 181.74 137.07
Std. Dev. 59.45 45.27 49.02 25.53
Page 96
78
15
(nm)
8 cm 19 kV 12 cm 19 kV 16 cm 19 kV 20 cm 19 kV
1 342.86 231.72 175.61 78.54
2 232.51 206.03 170.37 133.33
3 247.62 259.08 232.51 133.33
4 285.71 229.36 215.50 120.47
5 217.18 250.53 217.18 157.07
6 217.18 266.67 243.93 133.33
7 283.16 229.36 191.43 152.38
8 264.62 205.15 188.56 137.35
9 242.44 209.52 202.48 152.38
10 232.51 243.93 217.18 172.48
Mean 256.58 233.14 205.48 137.07
Std. Dev. 38.88 21.92 24.04 25.53
Page 97
79
16
Unhydrolyzed chitosan Hydrolyzed chitosan
(nm)
PLA/h-CS PLA/CS
1 78.54 170.37
2 133.33 247.62
3 133.33 250.53
4 120.47 230.15
5 157.07 230.15
6 133.33 222.13
7 152.38 235.61
8 137.35 222.13
9 152.38 222.13
10 172.48 247.62
Mean 137.07 227.84
Std. Dev. 25.53 23.02
Page 98
80
17
Hydrolyzed chitosan/PLA
(nm)
PLA 15%wt/h-CS PLA 18%wt/h-CS PLA 20%wt/h-CS
1 111.07 148.77 78.54
2 121.96 127.78 133.33
3 145.06 127.78 133.33
4 153.57 145.06 120.47
5 133.33 127.78 157.07
6 111.07 121.96 133.33
7 153.57 134.69 152.38
8 134.69 120.47 137.35
9 134.69 137.35 152.38
10 134.69 157.07 172.48
Mean 133.37 134.87 137.07
Std. Dev. 15.21 12.11 25.53
Page 99
81
18
Hydrolyzed chitosan/PLA
(nm)
PLA:h-CS (50:50) PLA:h-CS (60:40)
1 78.54 383.33
2 133.33 396.35
3 133.33 325.49
4 120.47 419.48
5 157.07 307.13
6 133.33 355.84
7 152.38 361.40
8 137.35 434.35
9 152.38 400.45
10 172.48 307.13
Mean 137.07 369.10
Std. Dev. 25.53 45.39
Page 101
83
19
Sample
No.
Tensile Strength
(MPa)
% Elongation
at Break (%)
Tensile Strength
at Yield (MPa)
% Elongation
at Yield (%)
Young’s
modulus (MPa)
1 33.886 3.5323 33.886 3.8364 8.833
2 33.162 3.6537 32.162 3.7350 8.611
3 33.259 3.6989 32.259 3.3876 9.523
4 32.159 3.598 32.159 3.5982 8.938
5 33.447 3.4822 33.447 3.8582 8.669
Mean 33.183 3.59302 32.783 3.6831 8.915
S.D. 0.636 0.087871 0.823 0.1945 0.364
20
10
Sample
No.
Tensile Strength
(MPa)
% Elongation
at Break (%)
Tensile Strength
at Yield (MPa)
% Elongation
at Yield (%)
Young’s
modulus (MPa)
1 30.352 3.8615 30.831 3.1998 9.635
2 31.858 3.4113 31.776 3.4450 9.224
3 29.885 3.6503 29.445 2.8580 10.303
4 30.193 3.4447 30.193 3.2526 9.283
5 29.492 3.8490 30.492 3.0286 10.068
Mean 30.356 3.6434 30.547 3.1568 9.702
S.D. 0.902 0.2140 0.856 0.2235 0.475
Page 102
84
21
20
Sample
No.
Tensile Strength
(MPa)
% Elongation
at Break (%)
Tensile Strength
at Yield (MPa)
% Elongation
at Yield (%)
Young’s
modulus (MPa)
1 30.975 3.6317 30.752 3.3452 9.193
2 30.905 3.6553 29.395 3.3223 8.848
3 28.845 3.8869 29.346 2.8342 10.354
4 28.997 4.0078 28.997 2.8864 10.046
5 28.674 3.9090 29.174 2.8612 10.196
Mean 29.679 3.8181 29.533 3.0499 9.727
S.D. 1.157 0.1660 0.699 0.2599 0.666
22
30
Sample
No.
Tensile Strength
(MPa)
% Elongation
at Break (%)
Tensile Strength
at Yield (MPa)
% Elongation
at Yield (%)
Young’s
modulus (MPa)
1 26.750 4.5520 26.030 2.6454 9.840
2 27.808 4.2270 30.808 3.0495 10.103
3 27.866 4.2983 28.866 3.0032 9.612
4 27.867 4.3793 28.200 2.8010 10.068
5 28.201 4.2115 29.139 2.9954 9.728
Mean 27.698 4.3336 28.609 2.8989 9.870
S.D. 0.552 0.1390 1.732 0.1708 0.213
Page 103
85
(Water vapor transmission rate: WVTR)
Page 104
86
23 (Water vapor transmission rate:
WVTR)
time
(h) g) g) g)
0
1 23.5262 23.5262 0.0000
0.0000 2 23.5261 23.5261 0.0000
3 23.5262 23.5262 0.0000
1
1 23.5262 23.5150 0.0112
0.0113 2 23.5261 23.5147 0.0114
3 23.5262 23.5150 0.0112
2
1 23.1829 23.1601 0.0228
0.0224 2 23.1824 23.1601 0.0223
3 23.1826 23.1604 0.0222
3
1 23.4129 23.3813 0.0316
0.0323 2 23.4129 23.3805 0.0324
3 23.4128 23.3800 0.0328
4
1 23.5112 23.4666 0.0446
0.0447 2 23.5105 23.4662 0.0443
3 23.5112 23.4660 0.0452
5
1 23.5161 23.4600 0.0561
0.0562 2 23.5163 23.4600 0.0563
3 23.5162 23.4599 0.0563
6
1 22.8973 22.8289 0.0684
0.0685 2 22.8970 22.8286 0.0684
3 22.8972 22.8285 0.0687
Page 105
87
24 (Water vapor transmission rate:
WVTR) 10
time
(h) g) g) g)
0
1 23.3740 23.3740 0.0000
0.0000 2 23.3738 23.3738 0.0000
3 23.3738 23.3738 0.0000
1
1 23.3740 23.3623 0.0117
0.0116 2 23.3738 23.3621 0.0117
3 23.3738 23.3623 0.0115
2
1 23.4635 23.4410 0.0225
0.0225 2 23.4635 23.4410 0.0225
3 23.4633 23.4408 0.0225
3
1 23.6099 23.5766 0.0333
0.0336 2 23.6104 23.5766 0.0338
3 23.6103 23.5765 0.0338
4
1 23.4530 23.4098 0.0432
0.0435 2 23.4535 23.4097 0.0438
3 23.4533 23.4098 0.0435
5
1 23.4788 23.4245 0.0543
0.0541 2 23.4789 23.4247 0.0542
3 23.4785 23.4248 0.0537
6
1 22.8966 22.8292 0.0674
0.0678 2 22.8966 22.8290 0.0676
3 22.8975 22.8290 0.0685
Page 106
88
25 (Water vapor transmission rate:
WVTR) 20
time
(h) g) g) g)
0
1 23.6374 23.6374 0.0000
0.0000 2 23.6372 23.6372 0.0000
3 23.6378 23.6378 0.0000
1
1 23.6374 23.6253 0.0121
0.0123 2 23.6372 23.6250 0.0122
3 23.6378 23.6253 0.0125
2
1 23.5549 23.5315 0.0234
0.0234 2 23.5547 23.5313 0.0234
3 23.5547 23.5313 0.0234
3
1 23.4577 23.4228 0.0349
0.0348 2 23.4575 23.4227 0.0348
3 23.4573 23.4225 0.0348
4
1 23.3639 23.3197 0.0442
0.0440 2 23.3638 23.3198 0.0440
3 23.3638 23.3199 0.0439
5
1 23.4010 23.3472 0.0538
0.0536 2 23.4012 23.3477 0.0535
3 23.4008 23.3473 0.0535
6
1 23.5773 23.5102 0.0671
0.0670 2 23.5772 23.5102 0.0670
3 23.5772 23.5103 0.0669
Page 107
89
26 (Water vapor transmission rate:
WVTR) 40
time
(h) g) g) g)
0
1 23.4589 23.4589 0.0000
0.0000 2 23.4593 23.4593 0.0000
3 23.4588 23.4588 0.0000
1
1 23.4589 23.4464 0.0125
0.0125 2 23.4593 23.4465 0.0128
3 23.4588 23.4465 0.0123
2
1 23.5168 23.4991 0.0177
0.0178 2 23.5172 23.4992 0.0180
3 23.5169 23.4991 0.0178
3
1 23.5638 23.5348 0.0290
0.0289 2 23.5637 23.5349 0.0288
3 23.5636 23.5348 0.0288
4
1 23.3977 23.3599 0.0378
0.0376 2 23.3975 23.3599 0.0376
3 23.3973 23.3598 0.0375
5
1 23.3839 23.3342 0.0497
0.0496 2 23.3838 23.3343 0.0495
3 23.3837 23.3342 0.0495
6
1 23.4952 23.4390 0.0562
0.0562 2 23.4953 23.4391 0.0562
3 23.4953 23.4391 0.0562
Page 108
90
24
25
10
y = 0.0113x - 0.0004
R² = 0.9992
-0.0200
0.0000
0.0200
0.0400
0.0600
0.0800
0 2 4 6 8
()
( )
y = 0.011x + 0.0002
R² = 0.9986
0.0000
0.0200
0.0400
0.0600
0.0800
0 2 4 6 8
()
( )
Page 109
91
26
20
y = 0.0109x + 0.001
R² = 0.9979
0.0000
0.0200
0.0400
0.0600
0.0800
0 2 4 6 8
()
( )
y = 0.0094x + 0.0008
R² = 0.9944
0.0000
0.0100
0.0200
0.0300
0.0400
0.0500
0.0600
0 1 2 3 4 5 6 7
()
( )
Page 110
92
27
40
27 (Water vapor transmission rate: WVTR)
Sample w/ t
(g/h)
time
(h/day) (m2)
WVTR
(g/m2 day)
PLA/h-CS 0.0113 24 0.000151 1796.03
PLA/h-CS
Add gentamicin 10 mg/ml 0.0111 24 0.000151 1764.24
PLA/h-CS
Add gentamicin 20 mg/ml 0.0109 24 0.000151 1732.45
PLA/h-CS
Add gentamicin 40 mg/ml 0.0094 24 0.000151 1494.04
Page 111
93
(Water adsorption)
Page 112
94
28 (Water adsorption)
Sample info Sample
No. g) g)
%water
absorption SD
PLA/h-CS
1 0.0031 0.0122 293.55
297.89 4.99 2 0.0031 0.0123 296.77
3 0.003 0.0121 303.33
PLA/h-CS
Add GS 10 mg/ml
1 0.003 0.0119 296.67
291.33 9.25 2 0.003 0.0119 296.67
3 0.0031 0.0118 280.65
PLA/h-CS
Add GS 20 mg/ml
1 0.0031 0.0114 267.74
262.87 8.44 2 0.0032 0.0113 253.13
3 0.0031 0.0114 267.74
PLA/h-CS
Add GS 40 mg/ml
1 0.0032 0.0107 234.38
239.42 4.37 2 0.0031 0.0106 241.94
3 0.0031 0.0106 241.94
Page 113
95
(Cytotoxicity)
Page 114
96
29 (Cytotoxicity)
(mg/ml)
OD %
1 2 3 4
GS 0
0 0.287 0.336 0.271 0.323 0.304 100.00
0.25 0.312 0.331 0.296 0.296 0.309 101.48
0.5 0.376 0.348 0.300 0.316 0.335 110.11
1 0.345 0.339 0.335 0.308 0.332 109.04
2 0.281 0.303 0.254 0.259 0.274 90.14
4 0.310 0.337 0.299 0.300 0.312 102.38
GS 10 mg/ml
0 0.277 0.345 0.318 0.318 0.315 100.00
0.25 0.357 0.377 0.334 0.334 0.351 111.45
0.5 0.313 0.330 0.271 0.319 0.308 98.01
1 0.311 0.345 0.307 0.325 0.322 102.38
2 0.310 0.317 0.301 0.317 0.311 98.97
4 0.300 0.315 0.306 0.341 0.316 100.32
GS 20 mg/ml
0 0.302 0.312 0.337 0.329 0.320 100.00
0.25 0.340 0.362 0.387 0.381 0.368 114.84
0.5 0.325 0.311 0.284 0.271 0.298 93.05
1 0.313 0.322 0.368 0.346 0.337 105.39
2 0.282 0.315 0.339 0.322 0.315 98.28
4 0.292 0.319 0.337 0.362 0.328 102.34
GS 40 mg/ml
0 0.318 0.317 0.322 0.368 0.331 100.00
0.25 0.366 0.318 0.354 0.339 0.344 103.92
0.5 0.296 0.345 0.394 0.334 0.342 103.32
1 0.300 0.307 0.374 0.352 0.333 100.60
2 0.313 0.324 0.359 0.389 0.346 104.53
4 0.270 0.316 0.320 0.317 0.306 92.30
Page 115
97
(Drug release)
Page 116
98
30 max = 400
24
GS load (mg/ml) Absorbance Actual GS (mg/m)
10 0.335 7.12
20 0.513 18.04
40 0.829 37.42
31 max = 400
GS-load (mg/ml) Time (hr) Absorbance Drug release (mg/ml)
10
1 0.262 2.64
2 0.251 1.96
3 0.237 1.10
6 0.231 0.74
12 0.222 0.18
24 0.221 0.12
72 0.22 0.06
20
1 0.284 3.99
2 0.277 3.56
3 0.267 2.94
6 0.251 1.96
12 0.242 1.41
24 0.235 0.98
72 0.223 0.25
40
1 0.357 8.47
2 0.323 6.38
3 0.286 4.11
Page 117
99
31 ( ) max = 400
GS-load (mg/ml) Time (hr) Absorbance Drug release (mg/ml)
40
6 0.277 3.56
12 0.269 3.07
24 0.256 2.27
72 0.242 1.41
Absorbance
UV-VIS spectrophotometer max 400 y =
0.0618x + 0.219
y Absorbance x
Page 118
100
-
20/3
92000
(080) 430-1458
E-mail address [email protected]
2548
2552
2552
2552-2554
1. Amornrat Tantithumrongwut and Nattawut Chaiyut, “Preparation of electrospun
chitosan/poly(lactic acid) nanofibers”, The 3rd National Research Symposium on Petroleum,
Petrochemicals, and Advanced Materials and The 18th PPC Symposium on Petroleum,
Petrochemicals, and Polymer, April 24, 2012, Bangkok, Thailand.