Development of an osteochondral cartilage defect model: = Entwicklung eines osteochondralen Knorpeldefektmodells
Gespeichert in:
| 1. Verfasser: | |
|---|---|
| Format: | Abschlussarbeit Buch |
| Sprache: | English |
| Veröffentlicht: |
Würzburg
2017
|
| Schlagworte: | |
| Online-Zugang: | Inhaltsverzeichnis Volltext Inhaltsverzeichnis |
| Beschreibung: | XX, 129, 11 Blätter Illustrationen, Diagramme 30 cm |
Internformat
MARC
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| 003 | DE-604 | ||
| 005 | 20180413 | ||
| 007 | t | ||
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| 084 | |a 610 |2 sdnb | ||
| 100 | 1 | |a Schwab, Andrea |d ca. 21. Jh. |e Verfasser |0 (DE-588)1155201396 |4 aut | |
| 245 | 1 | 0 | |a Development of an osteochondral cartilage defect model |b = Entwicklung eines osteochondralen Knorpeldefektmodells |c submitted by Andrea Schwab born in Würzburg |
| 246 | 1 | 1 | |a Entwicklung eines osteochondralen Knorpeldefektmodells |
| 264 | 1 | |a Würzburg |c 2017 | |
| 300 | |a XX, 129, 11 Blätter |b Illustrationen, Diagramme |c 30 cm | ||
| 336 | |b txt |2 rdacontent | ||
| 337 | |b n |2 rdamedia | ||
| 338 | |b nc |2 rdacarrier | ||
| 502 | |b Dissertation |c Julius-Maximilians-Universität Würzburg |d 2017 | ||
| 546 | |a Zusammenfassung in deutscher und englischer Sprache | ||
| 650 | 0 | 7 | |a Hyaliner Knorpel |0 (DE-588)4266348-9 |2 gnd |9 rswk-swf |
| 650 | 0 | 7 | |a Modell |0 (DE-588)4039798-1 |2 gnd |9 rswk-swf |
| 650 | 0 | 7 | |a Regeneration |0 (DE-588)4048983-8 |2 gnd |9 rswk-swf |
| 653 | |a hyaline cartilage | ||
| 653 | |a Test system | ||
| 653 | |a cartilage regeneration | ||
| 655 | 7 | |0 (DE-588)4113937-9 |a Hochschulschrift |2 gnd-content | |
| 689 | 0 | 0 | |a Hyaliner Knorpel |0 (DE-588)4266348-9 |D s |
| 689 | 0 | 1 | |a Regeneration |0 (DE-588)4048983-8 |D s |
| 689 | 0 | 2 | |a Modell |0 (DE-588)4039798-1 |D s |
| 689 | 0 | |5 DE-604 | |
| 776 | 0 | 8 | |i Erscheint auch als |n Online-Ausgabe |o urn:nbn:de:bvb:20-opus-155617 |a Schwab, Andrea |t Development of an osteochondral cartilage defect model |d Würzburg : Universität Würzburg, 2017 |h Online-Ressource |
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| 856 | 4 | 2 | |m DNB Datenaustausch |q application/pdf |u http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=030277966&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA |3 Inhaltsverzeichnis |
| 912 | |a ebook | ||
| 999 | |a oai:aleph.bib-bvb.de:BVB01-030277966 | ||
Datensatz im Suchindex
| _version_ | 1804178424124669952 |
|---|---|
| adam_text | C ONTENTS
1 INTRODUCTION 1
1.1 ARTICULAR CARTILAGE
............................................................................................
2
1.1.1 BIOLOGY OF ARTICULAR CARTILAGE T I S S U E
....................................................
2
1.1.2 CARTILAGE D E F E C T S
...................................................................................
4
1.2 CARTILAGE TRE A TM E N TS
.........................................................................................
5
1.2.1 MARROW STIMULATING TR E A TM E N TS
..........................................................
6
1.2.2 MATERIAL ASSISTED TRE A TM E N
TS................................................................. 6
1.2.3 CELL BASED TREATM
ENTS.............................................................................
7
1.2.4 OSTEOCHONDRAL G R A F T S
.............................................................................
8
1.2.5 LIMITATIONS OF CARTILAGE TR E A TM E N TS
....................................................
8
1.3 CARTILAGE IMPLANT DEVELOPMENT: FROM THE MATERIAL TO AN ADVANCED
THERAPY
MEDICINAL P RO D U
CT...............................................................................................
9
1.3.1 ADVANCED THERAPY MEDICINAL PRODUCTS (A T M P
S)............................... 9
1.3.2 MARKETING AUTHORIZATION OF A T M P S
....................................................
10
1.3.3 ATMPS FOR CARTILAGE REGENERATION
.......................................................
11
1.4 TE OF C A R TILA G E
..................................................................................................
11
1.4.1 CELL
SOURCE...............................................................................................
11
1.4.1.1 CHONDROCYTES ( C Z S )
............................................................... 11
1.4.1.2 MULTIPOTENT STROMAL CELLS (M SC S)
.........................................
12
1.4.1.3 CELL
CO-CULTURES........................................................................
12
1.4.2 MATERIALS
...............................................................................................
13
1.4.2.1 SYNTHETIC M
ATERIALS..................................................................
13
1.4.2.2 NATURAL M
ATERIALS.....................................................................
14
1.4.3 B IO RE A C TO
RS............................................................................................
15
1.5 TEST SYSTEMS: AN ALTERNATIVE TO ANIMAL E X P ERIM EN TS
.....................................
16
1.5.1 IN VITRO M O D E LS
......................................................................................
17
1.5.2 EX VIVO M O D E LS
......................................................................................
18
2 AIM 20
3 M ATERIALS 22
3.1 BIOLOGIC M A TE R IA
L...............................................................................................
22
3.2 LIST OF CHEMICALS
...............................................................................................
22
3.3 LIST OF EN ZY M
ES...................................................................................................
25
3.4 CELL CULTURE TECHNIQUES: CULTURE MEDIA AND B U FFE R
.........................................
26
3.5 HYDROGEL MATERIALS AND
PEPTIDES.......................................................................
28
3.6 BIOCHEMICAL AND PROTEIN ANALYSIS: BUFFERS AND SOLUTIONS
................................
30
3.7 HISTOLOGY: ANTIBODIES AND
SOLUTIONS.................................................................
31
3.8 GENE EXPRESSION ANALYSIS: PRIMERS AND SO LU TIO N
S............................................ 33
3.9 K I T S
.....................................................................................................................
34
3.10 LABORATORY DEVICES AND SOFTW
ARE.......................................................................
35
3.11 LABORATORY E Q U IP M E N
T......................................................................................
36
4 M ETHODS 37
4.1 ESTABLISHMENT OF EX VIVO OSTEOCHONDRAL DEFECT MODEL WITH CRITICAL
SIZE DEFECTS 37
4.1.1 ISOLATION OF OSTEOCHONDRAL TISSUE E X P LAN TS
...........................................
37
4.1.2 STATIC C U LTU R E
.........................................................................................
38
4.1.3 DYNAMIC CULTURE UNDER CONSTANT F LO W
.................................................
39
4.2 CARTILAGE DEFECT C RE A TIO N
...................................................................................
40
4.2.1 CLINICAL STANDARD: BIOPSY P U N CH
..........................................................
40
4.2.2 AUTOMATED, STANDARDIZED METHOD: ARTIFICIAL TISSUE C U TT E R
...............
41
4.3 CELL CULTURE METHODS: CELL ISOLATION, EXPANSION AND CHARACTERIZATION
.... 42
4.3.1 CELL IS O LA TIO N
.........................................................................................
42
4.3.1.1 CZ ISOLATION................................................. 42
4.3.1.2 MESENCHYMAL STROMAL CELL (MSC) ISO LATIO N
......................... 42
4.3.2 CELL C O U N TIN G
.........................................................................................
43
4.3.3 FREEZING AND THAWING OF
CELLS.................................................................
43
4.3.4 EMBEDDING OF CELLS INTO HYDROGEL MATERIAL
........................................
44
4.3.5 CZ
CHARACTERIZATION................................................................................
44
4.3.5.1 CZ POPULATION D O U B LIN G
.......................................................
44
4.3.5.2 CZ PELLET C U LTU RE
.................................................................... 44
4.4 PREPARATION OF HYDROGEL B IO M A TE RIA LS
.............................................................
45
4.4.1 COLLAGEN I
HYDROGEL................................................................................
45
4.4.1.1 ISOLATION OF COLLAGEN I FROM RAT TA ILS
.....................................
45
4.4.1.2 PROTEIN CHARACTERIZATION OF ISOLATED COLLAGEN I FROM RAT TAIL
BY WESTERN B L O T
.......................................................................
46
4.4.1.3 COLLAGEN I HYDROGEL
PREPARATION........................................... 47
4.4.1.4 FREE SWELLING CULTURE OF PCZ LOADED COLLAGEN I HYDROGEL . . 47
4.4.2 FIBRIN
GLUE...............................................................................................
47
4.4.3 FOUR ARMED POLYETHYLENE GLYCOL) POLYMERIZED WITH HEPARIN
(STARPEGH) 48
4.4.3.1 PEPTIDE AUGMENTED STA RP E G H
..............................................
48
4.4.4 PEG AND PARTIALLY METACRYLATED POLY[N-(2-HYDROXYPROLYL) METHACRY
LAMIDE MONO/DILACTATE (M P /H A )]
.......................................................
49
4.4.5 THIOL FUNCTIONALIZED HYALURONIC ACID/ALLYL FUNCTIONALIZED POLY
(GLYCI-
DOL) (P(A G E /G )-H A -SH
).................................................................... 50
4.5 VIABILITY S TA IN IN G
...............................................................................................
51
4.5.1 LIVE-DEAD S TA IN IN G
................................................................................
51
4.5.2 3-(4,5-DIMETHYL-2-THIAZOL)-2,5-DIPHENYL-2H-TETRAZOLIUM BROMIDE
(MTT) S TA IN IN G
......................................................................................
51
4.6 BIOCHEMICAL CHARACTERIZATION: PROTEOGLYCAN AND DNA C O N T E N T
..................
51
4.6.1 PAPAIN DIGESTION OF CARTILAGE TISSUE AND H Y D RO G ELS
............................
52
4.6.1.1 BLYSCAN* SULFATED GLYCOSAMINOGLYCAN ASSAY
.
.....................
52
4.6.1.2 DIMETHYLMETHYLENE BLUE ASSAY
..............................................
52
4.6.2 DNA
QUANTIFICATION................................................................................
53
4.7 HISTOLOGICAL, IMMUNO-HISTOLOGICAL AND IMMUNO-FLUORESCENT STAININGS
.... 53
4.7.1 SAMPLE PREPARATION AND EM BEDDING
....................................................
53
4.7.1.1 PLASTIC E M B E D D IN G
................................................................. 54
4.7.1.2 PARAFFIN EMBEDDING AND DECALCIFICATION
...............................
54
4.7.2 SAFRANIN-0
STAINING................................................................................
54
4.7.3 IMMUNO-HISTOLOGICAL STA IN IN G S
.............................................................
55
4.7.4 IMMUNO-FLUORESCENT S TA IN IN G S
.............................................................
55
4.8 GENE EXPRESSION A N A LY
SIS...................................................................................
55
4.8.1 RNA
ISOLATION.........................................................................................
56
4.8.2 CDNA
SYNTHESIS......................................................................................
56
4.8.3 QUALITATIVE REALTIME POLYMERASE CHAIN REACTION (QRT P G R )
............
56
4.9 MIGRATION OF CELLS IN HYDROGELS: TIME LAPSE EX P ERIM E N
TS............................... 58
4.9.0.1 STRUCTURAL CHARACTERIZATION BY SCANNING ELECTRON MICROSCOPY 58
4.10 STATISTICS AND DATA ANALYSIS
.............................................................................
58
5 RESULTS 59
5.1 ESTABLISHMENT OF EX VIVO OSTEOCHONDRAL DEFECT MODEL WITH CRITICAL
SIZE DEFECTS 59
5.1.1 INFLUENCE OF ISOLATION, DEFECT CREATION AND EX VIVO CULTURE ON
EXPLANT
V IA B ILITY
..................................................................................................
59
5.1.2 EX VIVO CRITICAL SIZE DEFECT: VIABILITY, PROTEOGLYCAN CONTENT AND
CELL
RE C RU ITM E N
T............................................................................................
61
5.1.3 PROTOTYPE BIOREACTOR: INFLUENCE OF DYNAMIC CULTURE ON VIABILITY .
. . 65
5.2 CHARACTERIZATION OF COLLAGEN I ISOLATED FROM RAT T A I L
..................................... 67
5.3 CHARACTERIZATION OF PORCINE ARTICULAR CARTILAGE AND PORCINE
CHONDROCYTES . . 68
5.3.1 PHENOTYPICAL CHARACTERIZATION OF PCHONDROCYTES AND ARTICULAR
CARTILAGE 68
5.3.2 CHONDROCYTE PELLET CULTURE AND FREE SWELLING CULTURE OF COLLAGEN I
LOADED
HYDROGEL......................................................................................
69
5.4 EX VIVO EVALUATION OF NOVEL BIOMATERIALS
..........................................................
71
5.4.1 CELL FREE HYDROGEL MATERIAL TE STIN G
.......................................................
71
5.4.2 CHONDROCYTE LOADED HYDROGEL MATERIAL T E S T I N G
.................................. 74
5.4.2.1 CHARACTERIZATION OF HYDROGEL MATERIALS: HISTOLOGY AND PRO
TEOGLYCAN CO
NTENT....................................................................
74
5.4.2.2 GENE EXPRESSION ANALYSIS
.......................................................
80
5.4.2.3 INFLUENCE OF PEPTIDE AUGMENTED STARPEGH MATERIAL ON
ECM SY N TH E
SIS.......................................................................
83
5.4.3 MIGRATION OF CELLS IN HYDROGELS: TIME LAPSE E X P ERIM EN TS
..................
83
5.5 INFLUENCE OF DEFECT DEPTH ON CARTILAGE REGENERATION
........................................
85
5.5.1 VIABILITY AND CELL MIGRATION IN CHONDRAL DEFECTS
............................... 85
6 DISCUSSION 87
6.1 OSTEOCHONDRAL EXPLANT MODEL: RELEVANCE OF SUBCHONDRAL BONE AND
TISSUE
SPECIFIC MEDIA S U P P LY
.........................................................................................
87
6.1.1 ROLE OF SUBCHONDRAL BONE IN CARTILAGE
REGENERATION............................ 88
6.1.2 TISSUE SPECIFIC MEDIA IS CRITICAL TO MAINTAIN CARTILAGE ECM DURING
EX VIVO
CULTURE.........................................................................................
89
6.1.3 BIOREACTOR FOR DYNAMIC CULTURE OF E X P LA N TS
........................................ 91
6.2 EX VIVO EVALUATION OF CARTILAGE REGENERATION IN THE OSTEOCHONDRAL
DEFECT MODEL 92
6.2.1 DEFINITION OF EX VIVO CRITICAL SIZE DIAMETER IN OSTEOCHONDRAL
DEFECT
M O D E L
.....................................................................................................
92
6.2.2 STIMULATIVE EFFECT OF OSTEOCHONDRAL EXPLANT ON CARTILAGE MATRIX
PRO
DUCTION
..................................................................................................
94
6.2.3 HYDROGEL MATERIALS REVEALED DIFFERENCES IN REPAIR OF FULL
THICKNESS
CARTILAGE
DEFECTS......................................................................................
96
6.2.4 PARTIAL THICKNESS DEFECTS: REPAIR AND CELL REC RU ITM EN T
........................
102
6.2.5
CONCLUSION.................................................................................................
103
7 OUTLOOK 105
7.1 CLINICAL RELEVANCE: CARTILAGE THERAPIES AND SIMULATED O A
...............................
105
7.1.1 DEFECT GEOMETRY AND TREATMENT APPROACHES
..........................................
105
7.1.2 INFLAMMATORY CO
NDITIONS.........................................................................106
7.2 INFLUENCE OF CULTURE CONDITIONS: LOW OXYGEN TENSION AND CARTILAGE
BIOMECHANICS 107
7.2.1 OXYGEN TE N S IO N
.........................................................................................107
7.2.2 CARTILAGE
BIOMECHANICS............................................................................
107
8 APPENDIX A -L
|
| any_adam_object | 1 |
| author | Schwab, Andrea ca. 21. Jh |
| author_GND | (DE-588)1155201396 |
| author_facet | Schwab, Andrea ca. 21. Jh |
| author_role | aut |
| author_sort | Schwab, Andrea ca. 21. Jh |
| author_variant | a s as |
| building | Verbundindex |
| bvnumber | BV044883825 |
| collection | ebook |
| ctrlnum | (OCoLC)1136279239 (DE-599)DNB1148797130 |
| discipline | Medizin |
| format | Thesis Book |
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| oclc_num | 1136279239 |
| open_access_boolean | 1 |
| owner | DE-384 DE-473 DE-BY-UBG DE-703 DE-1051 DE-824 DE-29 DE-12 DE-91 DE-BY-TUM DE-19 DE-BY-UBM DE-1049 DE-92 DE-739 DE-898 DE-BY-UBR DE-355 DE-BY-UBR DE-706 DE-20 DE-1102 DE-860 DE-2174 |
| owner_facet | DE-384 DE-473 DE-BY-UBG DE-703 DE-1051 DE-824 DE-29 DE-12 DE-91 DE-BY-TUM DE-19 DE-BY-UBM DE-1049 DE-92 DE-739 DE-898 DE-BY-UBR DE-355 DE-BY-UBR DE-706 DE-20 DE-1102 DE-860 DE-2174 |
| physical | XX, 129, 11 Blätter Illustrationen, Diagramme 30 cm |
| psigel | ebook |
| publishDate | 2017 |
| publishDateSearch | 2017 |
| publishDateSort | 2017 |
| record_format | marc |
| spelling | Schwab, Andrea ca. 21. Jh. Verfasser (DE-588)1155201396 aut Development of an osteochondral cartilage defect model = Entwicklung eines osteochondralen Knorpeldefektmodells submitted by Andrea Schwab born in Würzburg Entwicklung eines osteochondralen Knorpeldefektmodells Würzburg 2017 XX, 129, 11 Blätter Illustrationen, Diagramme 30 cm txt rdacontent n rdamedia nc rdacarrier Dissertation Julius-Maximilians-Universität Würzburg 2017 Zusammenfassung in deutscher und englischer Sprache Hyaliner Knorpel (DE-588)4266348-9 gnd rswk-swf Modell (DE-588)4039798-1 gnd rswk-swf Regeneration (DE-588)4048983-8 gnd rswk-swf hyaline cartilage Test system cartilage regeneration (DE-588)4113937-9 Hochschulschrift gnd-content Hyaliner Knorpel (DE-588)4266348-9 s Regeneration (DE-588)4048983-8 s Modell (DE-588)4039798-1 s DE-604 Erscheint auch als Online-Ausgabe urn:nbn:de:bvb:20-opus-155617 Schwab, Andrea Development of an osteochondral cartilage defect model Würzburg : Universität Würzburg, 2017 Online-Ressource B:DE-101 application/pdf http://d-nb.info/1148797130/04 Inhaltsverzeichnis https://nbn-resolving.org/urn:nbn:de:bvb:20-opus-155617 Resolving-System kostenfrei Volltext DNB Datenaustausch application/pdf http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=030277966&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA Inhaltsverzeichnis |
| spellingShingle | Schwab, Andrea ca. 21. Jh Development of an osteochondral cartilage defect model = Entwicklung eines osteochondralen Knorpeldefektmodells Hyaliner Knorpel (DE-588)4266348-9 gnd Modell (DE-588)4039798-1 gnd Regeneration (DE-588)4048983-8 gnd |
| subject_GND | (DE-588)4266348-9 (DE-588)4039798-1 (DE-588)4048983-8 (DE-588)4113937-9 |
| title | Development of an osteochondral cartilage defect model = Entwicklung eines osteochondralen Knorpeldefektmodells |
| title_alt | Entwicklung eines osteochondralen Knorpeldefektmodells |
| title_auth | Development of an osteochondral cartilage defect model = Entwicklung eines osteochondralen Knorpeldefektmodells |
| title_exact_search | Development of an osteochondral cartilage defect model = Entwicklung eines osteochondralen Knorpeldefektmodells |
| title_full | Development of an osteochondral cartilage defect model = Entwicklung eines osteochondralen Knorpeldefektmodells submitted by Andrea Schwab born in Würzburg |
| title_fullStr | Development of an osteochondral cartilage defect model = Entwicklung eines osteochondralen Knorpeldefektmodells submitted by Andrea Schwab born in Würzburg |
| title_full_unstemmed | Development of an osteochondral cartilage defect model = Entwicklung eines osteochondralen Knorpeldefektmodells submitted by Andrea Schwab born in Würzburg |
| title_short | Development of an osteochondral cartilage defect model |
| title_sort | development of an osteochondral cartilage defect model entwicklung eines osteochondralen knorpeldefektmodells |
| title_sub | = Entwicklung eines osteochondralen Knorpeldefektmodells |
| topic | Hyaliner Knorpel (DE-588)4266348-9 gnd Modell (DE-588)4039798-1 gnd Regeneration (DE-588)4048983-8 gnd |
| topic_facet | Hyaliner Knorpel Modell Regeneration Hochschulschrift |
| url | http://d-nb.info/1148797130/04 https://nbn-resolving.org/urn:nbn:de:bvb:20-opus-155617 http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=030277966&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA |
| work_keys_str_mv | AT schwabandrea developmentofanosteochondralcartilagedefectmodelentwicklungeinesosteochondralenknorpeldefektmodells AT schwabandrea entwicklungeinesosteochondralenknorpeldefektmodells |
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