Connective Tissue Formation in Wound Healing - E-thesis

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Supervised by Professor Juhani Ahonen Department of Surgery Helsinki University Central Hospital Helsinki, Finland Reviewed by Professor Jorma Keski-Oja Departments of Pathology and Virology Biomedicum and Haartman Institute University of Helsinki Helsinki, Finland and Docent Erkki Koivunen Department of Biosciences Division of Biochemistry University of Helsinki, Finland Opponent Docent Jouko Lohi Department of Pathology University of Helsinki Helsinki, Finland ISBN 952-91-6240-5 (paperback) ISBN 952-10-1313-3 (PDF) Yliopistopaino Helsinki 2003
CONTENTS ORIGINAL PUBLICATIONS 5 ABBREVIATION 6 ABSTRACT 7 INTRODUCTION 9 REVIEW OF THE LITERATURE 11 WOUND HEALING 11 Blood clot formation 12 Inflammation 12 Epithelialization 13 Neovascularization 13 Formation of Granulation Tissue 15 Matrix Remodeling 16 Viscose cellulose sponges as a wound healing model 17 EXTRACELLULAR MATRIX 18 Collagens 18 Structure and biosynthesis of collagens 21 Fibrillar collagens 22 The gene structure of the fibril-forming collagens 23 Regulation of collagen genes 25 Type I collagen 28 Type III collagen 28 Type V collagen 29 Cell and ECM interaction 31 DEGRADATION OF EXTRACELLULAR MATRIX 33 Matrix metalloproteinases 33 Gelatinases 35 Other MMPs 35 Regulation of MMP activity 38 The role of MMPs and TIMPs in wound healing 40 GROWTH FACTORS IN WOUND HEALING 41 TGF-β 41 PDGF 46 CTGF 48 OUTLINE OF THE PRESENT STUDIES 53 MATERIALS AND METHODS 54 Animals 54 Experimental wound healing model 54 Histological staining for collagens 54 Hemoglobin 54 Determination of total tissue collagen and DNA 55 Collagen extraction, identification, and quantitation 55 Extraction of metalloproteinases and gelatinolytic activity assay 56 Primary antibodies 56
Page 1: Connective Tissue Formation in Woun
Page 5 and 6: ORIGINAL PUBLICATIONS This thesis i
Page 7 and 8: ABSTRACT A tight balance between co
Page 9 and 10: INTRODUCTION The regulation of extr
Page 11 and 12: REVIEW OF THE LITERATURE WOUND HEAL
Page 13 and 14: days, and neutrophils are themselve
Page 15 and 16: Formation of Granulation Tissue New
Page 17 and 18: emodeled by contraction of the woun
Page 19 and 20: Table I Collagen types, their genes
Page 21 and 22: Structure and biosynthesis of colla
Page 23 and 24: A B COL1A1 COL3A1 COL5A1 C COL1A1 C
Page 25 and 26: TATA box GC box (Sp1/Sp3 binding el
Page 27 and 28: y sequences in their 3’ untransla
Page 29 and 30: espectively (160). Liver has as muc
Page 31 and 32: in the matrix as compared to type I
Page 33 and 34: is the need to generate mechanical
Page 35 and 36: and the catalytic domain, which con
Page 37 and 38: specific binding and cleavage of th
Page 39 and 40: constitutively expressed and contro
Page 41 and 42: inflammatory cells (283). MMP-9 may
Page 43 and 44: TG F-β ligand Receptor II Receptor
Page 45 and 46: and the core proteins of the ECM pr
Page 47 and 48: hydrophobic stretch of the C-termin
Page 49 and 50: similarities to insulin-like growth
Page 51 and 52: CTGF and TGF-β distinguishes them
Page 53 and 54:
OUTLINE OF THE PRESENT STUDIES The
Page 55 and 56:
were determined: reduced hemoglobin
Page 57 and 58:
(Minneapolis, MN, USA). Anti-PDGF A
Page 59 and 60:
solution, 10% dextran sulphate and
Page 61 and 62:
sequencer, Model 373A (Applied Bios
Page 63 and 64:
histology. The hemoglobin content o
Page 65 and 66:
especially at the later stages. The
Page 67 and 68:
owing to the high standard deviatio
Page 69 and 70:
A Staining index A 50 45 40 35 30 2
Page 71 and 72:
the proliferative phase of wound re
Page 73 and 74:
type V collagen expression is assoc
Page 75 and 76:
healing model presented here lacks
Page 77 and 78:
PDGF-A and -B protein was observed
Page 79 and 80:
CTGF functions as a downstream medi
Page 81 and 82:
I am also grateful for all my frien
Page 83 and 84:
19. Brown LF, Yeo KT, Berse B, et a
Page 85 and 86:
56. Phillips C, Wenstrup RJ. Biosyn
Page 87 and 88:
89. Lohi J, Lehti K, Valtanen H, Pa
Page 89 and 90:
124. Wiestner M, Krieg T, Horlein D
Page 91 and 92:
159. Gay S, Vijanto J, Raekallio J,
Page 93 and 94:
196. Saelman EU, Nieuwenhuis HK, He
Page 95 and 96:
237. Knäuper V, Cowell S, Smith B,
Page 97 and 98:
273. Rajavashisth TB, Liao JK, Gali
Page 99 and 100:
307. Geng Y, Weinberg RA. Transform
Page 101 and 102:
346. Heldin CH, Östman A, Rönnstr
Page 103 and 104:
382. Frazier KS, Grotendorst GR. Ex
Page 105 and 106:
416. Glumoff V, Mäkelä JK, Vuorio
Page 107:
454. Pierce GF, Tarpley JE, Tseng J
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