Fong Siu Lai. / Thesis (M.Phil.)--Chinese University of Hong Kong, 1997. / Includes bibliographical references (leaves 187-196). / Acknowledgement --- p.v / Abstract --- p.vii / Chapter Chapter One --- INTRODUCTION --- p.1 / Chapter Chapter Two --- LITERATURE REVIEW --- p.4 / Chapter 1 --- STRUCTURE OF SKIN --- p.4 / Chapter 1.1 --- epidermis / Chapter 1.1.1 --- stratum corneum / Chapter 1.1.2 --- stratum lucidum / Chapter 1.1.3 --- stratum granulosum / Chapter 1.1.4 --- stratum spinosum / Chapter 1.1.5 --- stratum germinativum / Chapter 1.2 --- dermis / Chapter 1.2.1 --- collagen / Chapter 1.2.2 --- elastin / Chapter 1.2.3 --- reticulin / Chapter 1.2.4 --- fibroblasts / Chapter 1.2.5 --- ground subsatnce / Chapter 1.3 --- dermo-epidermal junction / Chapter 1.4 --- skin appendages / Chapter 1.4.1 --- hair / Chapter 1.4.2 --- nails / Chapter 1.4.3 --- glands / Chapter 1.5 --- cutaneous vascular system / Chapter 1.5.1 --- cutaneous blood flow and its significance / Chapter 1.5.2 --- cutaneous lymphatic flow / Chapter 2 --- FUNCTIONS OF SKIN --- p.24 / Chapter 2.1 --- protection / Chapter 2.2 --- sensation / Chapter 2.3 --- thermal regulation / Chapter 2.4 --- absorption / Chapter 2.5 --- protection against ultraviolet radiation / Chapter 2.6 --- storage / Chapter 3 --- BIOMECHANICS OF SKIN --- p.28 / Chapter 3.1 --- skin elasticity and the physical variation / Chapter 3.2 --- mechanical properties / Chapter 3.2.1 --- tensile strength / Chapter 3.2.2 --- distensibility / Chapter 3.2.3 --- Young's modulus / Chapter 3.2.4 --- visco-elastic character / Chapter 3.2.5 --- hysteresis / Chapter 3.3 --- fibre orientation / Chapter 3.4 --- mechanical considerations / Chapter 3.5 --- physiological factors / Chapter 3.6 --- clinical application / Chapter 4 --- PHYSIOLOGICAL RESPONSE OF HUMAN SKIN --- p.47 / Chapter 4.1 --- response to mechanical loading / Chapter 4.1.1 --- triple response / Chapter 4.1.2 --- reactive hyperaemia / Chapter 4.2 --- thermal response / Chapter 4.2.1 --- skin temperature / Chapter 4.2.2 --- response to heat / Chapter 4.2.3 --- response to cold / Chapter 4.3 --- local tissue response to burn / Chapter Chapter Three --- BACKGROUND OF THE PRESENT STUDY --- p.55 / Chapter 1 --- BURN INJURIES --- p.55 / Chapter 1.1 --- nature / Chapter 1.2 --- depth / Chapter 1.3 --- extent / Chapter 1.4 --- location of burn / Chapter 1.5 --- age / Chapter 1.6 --- "associated major trauma, inhalation injury" / Chapter 1.7 --- general health status / Chapter 2 --- WOUND HEALING PROCESS --- p.65 / Chapter 2.1 --- role of collagen in wound healing / Chapter 2.2 --- role of oxygen in wound healing / Chapter 2.3 --- role of fibroblasts and myofibroblasts in wound healing / Chapter 2.4 --- role of mast cells in wound healing / Chapter 3 --- HYPERTROPHIC SCAR --- p.71 / Chapter 3.1 --- aetiological factors / Chapter 3.1.1 --- age / Chapter 3.1.2 --- time for wound healing / Chapter 3.1.3 --- racial factor / Chapter 3.1.4 --- depth of injury / Chapter 3.1.5 --- location / Chapter 3.1.6 --- tension / Chapter 3.2 --- characteristics / Chapter 3.3 --- pathogenesis of hypertrophic scar / Chapter 3.3.1 --- blood flow / Chapter 3.3.2 --- tissue gas / Chapter 3.3.3 --- filamentous material / Chapter 3.3.4 --- mast cells / Chapter 3.3.5 --- chondroitin sulfate / Chapter 3.3.6 --- enzyme proline hydroxylase / Chapter 3.4 --- histopathology / Chapter 3.5 --- response towards pressure / Chapter 4 --- TREATMENT OF POST-BURN HYPERTROPHIC SCAR AND THEIR RESPONSE --- p.92 / Chapter 4.1 --- surgery / Chapter 4.2 --- radiotherapy / Chapter 4.3 --- ultrasonics / Chapter 4.4 --- chemotherapy/ intralesional injection of steroid / Chapter 4.5 --- pressure therapy / Chapter 4.6 --- topical silicone gel / Chapter 4.6.1 --- mechanics / Chapter 4.6.2 --- bacteriology / Chapter 4.6.3 --- water-vapour transmission rate / Chapter 4.6.4 --- appearance in the Scanning Electronic Microscope / Chapter 4.7 --- prevention of hypertrophic scar and scar contracture / Chapter 5 --- ASSESSMENT TOOLS FOR HYPERTROPHIC SCAR AND THE CLINICAL APPLICATION --- p.105 / Chapter 5.1 --- clinical observation of the appearance / Chapter 5.2 --- ultrasonography and thickness / Chapter 5.2.1 --- ultrasound / Chapter 5.2.2 --- pulse-echo distance measurement / Chapter 5.2.3 --- echo generation / Chapter 5.2.4 --- transducer beam pattern / Chapter 5.2.5 --- ultrasound instrumentation / Chapter 5.2.6 --- application of ultrasound in the study of hypertrophic scar thickness / Chapter 5.3 --- elastometry (Cutometer) and elasticity / Chapter 5.4 --- application of elastometry / Chapter Chapter Four --- OBJECTIVES & METHODOLOGY OF THE STUDY --- p.123 / Chapter 1 --- Objectives of the study --- p.123 / Chapter 2 --- Study subjects --- p.123 / Chapter 3 --- Methodology --- p.125 / Chapter 4 --- Assessment of thickness of hypertrophic scar --- p.128 / Chapter 5 --- Assessment of visco-elasticity of hypertrophic scar --- p.130 / Chapter 6 --- Clinical rating scale --- p.133 / Chapter 7 --- Study of normal skin as control --- p.133 / Chapter 8 --- Reliability of the ultrasound and cutometer measurement --- p.134 / Chapter Chapter Five --- RESULTS --- p.136 / Chapter 1 --- Inter- and intra- examiner variations of the ultrasound and cutometer measurement --- p.136 / Chapter 2 --- Comparison with normal skin control --- p.138 / Chapter 3 --- Results of ultrasonographic measurements of thickness of hypertrophic scar and its correlation with the clinical grading --- p.139 / Chapter 4 --- Results of Cutometer reading (visco-elastic properties) and the correlation with clinical grading --- p.142 / Chapter 5 --- Observation from raw data --- p.152 / Chapter Chapter Six --- DISCUSSION --- p.154 / Chapter 1 --- Measuring thickness with ultrasonography and clinical grading --- p.157 / Chapter 2 --- Elastic properties of hypertrophic scar and clinical grading --- p.158 / Chapter 3 --- The predictive value of the ultrasonography and elastometry through monthly longitudinal measurement --- p.161 / Chapter 4 --- Inter- and intra- examinar reliability of the ultrasonography and elastometry in the assessmetn of post-burn hypertrophic scar --- p.164 / Chapter 5 --- "The use of a composite ""Visco-elasticity-Thickness Chart"" and case studies" --- p.165 / Chapter 6 --- Limitations of the study --- p.182 / Chapter Chapter Seven --- CONCLUSION AND RECOMMENDATION FOR FURTHER STUDY --- p.184 / REFERENCES --- p.187 / APPENDICES --- p.197 / Appendix 1 Patients' record --- p.197 / Appendix 2 Record of the scars --- p.200 / Appendix 3 Clinical Grading of the hypertrophic scar --- p.202 / Appendix 4 Measurement of the visco-elastic properties --- p.204 / Appendix 5 Ultrasonic measurements of the hypertrophic scars --- p.235 / Appendix 6 List of graphs --- p.237 / Appendix 7 List of figures --- p.238 / Appendix 8 List of tables --- p.240

by Cheng Chi Wa. / Thesis (M.Phil.)--Chinese University of Hong Kong, 1998. / Includes bibliographical references (leaves 110-120). / Abstract also in Chinese. / Title --- p.i / Abstract --- p.ii / Acknowledgement --- p.iv / Abbreviations --- p.v / Abbreviation Table for Amino Acids --- p.vi / Table of Contents --- p.vii / List of Figures --- p.xii / List of Tables --- p.xv / Chapter Chapter 1 --- Introduction --- p.1 / Chapter Chapter 2 --- Literature review --- p.2 / Chapter Part I --- Hypertrophic Scar / Chapter 2.1 --- Definition of hypertrophic scar --- p.2 / Chapter 2.2 --- Pathology --- p.2 / Chapter 2.3 --- Epidemiology findings --- p.3 / Chapter 2.3.1 --- Ethnicity --- p.3 / Chapter 2.3.2 --- Age --- p.3 / Chapter 2.3.3 --- Body location --- p.3 / Chapter 2.4 --- Mechanism of cutaneous wound healing --- p.4 / Chapter 2.4.1 --- Phase I - Haemostasis and inflammation --- p.4 / Chapter 2.4.1.1 --- Haemostasis --- p.6 / Chapter 2.4.1.2 --- Early phase of inflammation --- p.6 / Chapter 2.4.1.3 --- Late phase of inflammation --- p.7 / Chapter 2.4.2 --- Phase II - Re-epithelialization --- p.7 / Chapter 2.4.2.1 --- Migration of epidermal keratinocytes --- p.8 / Chapter 2.4.2.2 --- Migration of fibroblasts --- p.8 / Chapter 2.4.2.3 --- Angiogenesis --- p.9 / Chapter 2.4.3 --- Phase III - Tissue remodeling --- p.10 / Chapter 2.4.3.1 --- Cell maturation and apoptosis --- p.10 / Chapter 2.4.3.2 --- Exrtracellular matrix remodeling --- p.10 / Chapter 2.5 --- Alteration of wound healing - Possible pathogenic factors of hypertrophic scar --- p.11 / Chapter 2.5.1 --- Changes in Phase I-Inflammation --- p.13 / Chapter 2.5.2 --- Changes in Phase II - Re-epithelialization/ tissue formation --- p.14 / Chapter 2.5.3 --- Changes in Phase III - Tissue remodeling --- p.15 / Chapter 2.6 --- The Role of fibroblasts in the formation of hypertrophic scar --- p.16 / Chapter 2.6.1 --- Functions of fibroblasts in wound healing --- p.16 / Chapter 2.6.2 --- Suggested aetiological role in the formation of hypertrophic scar fibroblasts --- p.16 / Chapter 2.6.2.1 --- Fibroproliferation disorder --- p.18 / Chapter 2.6.2.2 --- Extracellular Matrix remodeling disorder --- p.18 / Chapter a) --- CoUaqen --- p.18 / Chapter b) --- Proteoglycan --- p.19 / Chapter 2.6.2.3 --- Other differentially expressed factors --- p.20 / Chapter 2.7 --- Treatment of hypertrophic scar --- p.21 / Chapter Part II --- Differential Display / Chapter 2.8 --- Current approaches for the studies of differential gene expression --- p.23 / Chapter 2.9 --- Comparison amongst different approaches --- p.23 / Chapter 2.10 --- The strategy of Differential Display RT-PCR (DDRT-PCR) --- p.24 / Chapter 2.11 --- The application of DDRT-PCR to identify differentially expressed genes --- p.26 / Chapter Chapter 3 --- Aims and Strategies --- p.27 / Chapter Chapter 4 --- Methods and Materials --- p.29 / Chapter 4.1 --- Materials --- p.29 / Chapter 4.2 --- Clinical specimen collection --- p.31 / Chapter 4.3 --- Primary explant culture --- p.31 / Chapter 4.4 --- Immunohistochemical staining --- p.32 / Chapter 4.5 --- Total RNA extraction --- p.32 / Chapter 4.6 --- DNase I digestion --- p.33 / Chapter 4.7 --- Differential display-RTPCR (DD-RTPCR) --- p.33 / Chapter 4.8 --- Polyacrylamide gel electrophoresis --- p.34 / Chapter 4.9 --- Reamplification of the differentially expressed fragments --- p.35 / Chapter 4.10 --- Molecular cloning of the DNA fragments --- p.35 / Chapter 4.11 --- Screening and miniprep of the plasmid DNA --- p.36 / Chapter 4.12 --- Cycle sequencing --- p.38 / Chapter 4.13 --- Data analysis --- p.38 / Chapter 4.14 --- RT-PCR --- p.39 / Chapter 4.15 --- Probe labeling by PCR with DIG-dUTP --- p.40 / Chapter 4.16 --- Southern blotting --- p.41 / Chapter Chapter5 --- p.42 / Chapter 5.1 --- Clinical Specimen --- p.42 / Chapter 5.2 --- Primary explant culture --- p.42 / Chapter 5.3 --- The total RNA extraction from the cultured fibroblast --- p.45 / Chapter 5.4 --- Differential display RT-PCR --- p.47 / Chapter 5.5 --- Reamplification of the DNA fragments --- p.49 / Chapter 5.6 --- Molecular cloning of the DNA fragment --- p.53 / Chapter 5.7 --- DNA sequencing of the inserts --- p.58 / Chapter 5.8 --- Analysis and identification of the DNA sequences --- p.62 / Chapter 5.9 --- Semi-quantitative RT-PCR analysis of the differentially expressed genes --- p.76 / Chapter Chapter6 --- p.87 / Chapter Part I --- Validity of the Findings / Chapter 6.1 --- The Limitation of Tissue Sampling --- p.87 / Chapter 6.2 --- Tissue Culture model --- p.88 / Chapter 6.3 --- Differential Display RT-PCR --- p.89 / Chapter 6.3.1 --- Identification of the differentially expressed genes --- p.89 / Chapter 6.3.2 --- Confirmation of the differentially expressed genes --- p.91 / Chapter 6.4 --- Technical difficulties and Limitations --- p.92 / Chapter 6.4.1 --- Sampling --- p.92 / Chapter 6.4.2 --- Primary tissue culture --- p.93 / Chapter Part II --- Significance and Future Studies / Chapter 6.5 --- Down-regulation of thrombospondin 1 (TSP 1) in the hypertrophic scar fibroblasts --- p.94 / Chapter 6.6 --- Biochemical and biological functions of TSP1 --- p.96 / Chapter 6.6.1 --- The biochemical functions of TSP1 --- p.96 / Chapter 6.6.2 --- The biochemical functions of TSP1 --- p.97 / Chapter 6.7 --- The role of TSP 1 in the pathogenesis of hypertrophic scar --- p.98 / Chapter 6.7.1 --- Down-regulation of TSP 1 may be responsible for the excessive microvessels in hypertrophic scar --- p.98 / Chapter 6.7.2 --- Down-regulation of TSP 1 may be responsible for the failure of the apoptosis of the fibroblasts in the hypertrophic scar --- p.101 / Chapter 6.8 --- Expression of TSP 1 during wound healing --- p.103 / Chapter 6.9 --- Expression of TSP 1 in hypertrophic scarring --- p.107 / Chapter 6.10 --- Cytochrome b561 and its biological function --- p.109 / Chapter 6.11 --- Future studies --- p.108 / Chapter 6.11.1 --- The expression of TSP 1 in hypertrophic scarring and normal wound healing --- p.108 / Chapter 6.11.2 --- The expression of cytochrome b561 --- p.109 / Chapter 6.11.3 --- A full scale study of differential display RT-PCR --- p.109 / References --- p.110 / Appendices --- p.121 / Chapter I --- The complete mRNA sequence of thrombospondin1 precursor --- p.121 / Chapter II --- The mRNA sequence of cytochrome b561 --- p.123

O processo de reparação tecidual evolui tardiamente com alteração das propriedades físicas da pele, cuja gravidade varia, entre outros fatores, em função da profundidade original da lesão. Clinicamente a pele é seca, descamativa, discrômica e com diminuição da elasticidade, especialmente em cicatrizes hipertróficas. As características do tegumento pós-queimadura podem ser melhoradas com agentes de uso tópico normalmente utilizados em patologias como acne e fotoenvelhecimento. Foram avaliados 77 pacientes portadores de seqüela de queimadura peri-oral, entre seis e 46 anos de idade. O tipo de tegumento foi classificado como restaurado, quando epitelizado espontaneamente, ou enxertado. O tratamento tópico consistiu de tretinoína (0,01-0,05%) e ácido glicólico (5-7%) durante três meses. Duas distâncias foram medidas com paquímetro digital em abertura bucal máxima, a inter-dentária (D) e a inter-labial (L). Houve aumento significativo (p£0,01) de D e L nos grupos tratados quando comparados aos respectivos controles, tanto nos pacientes com tegumento restaurado como nos enxertados. O aumento de L foi significativamente maior que D. Sugerimos a utilização da medida inter-labial como parâmetro complementar à medida inter-incisal como método de avaliação indireta da elasticidade cutânea no tegumento pós-queimadura. / Tissue repair processes evolve with alterations of the cutaneous physical properties, which are proportional, among other factors, to the original depth of the lesion. At examination, the skin is dry, dyschromic and less elastic, specially in hypertrophic scars. Post-burn skin characteristics can be improved by topical agents regularly used in pathologies like acne or photodamage. We evaluated 77 patients who had peri-oral burn sequela. The ages ranged from six to 46 years old. The tegument was classified as restored, when spontaneously healed, or skin grafted. The topical treatment consisted of tretinoin (0,01-0,05%) and glycolic acid (5-7%) for three months. Inter-incisal (D) and inter-labial (L) distances were measured at maximal mouth opening with a digital pachymeter. Both inter-incisal and inter-labial distances increased significantly in the groups treated when compared to respective controls (p£0,01). This fact was similar in both types of tegument. Inter-labial distance (L) increased significantly more than D. We suggest its use as a complementary measure to inter-incisal distance for indirect evaluation of skin elasticity in the post-burn tegument.

O processo de reparação tecidual evolui tardiamente com alteração das propriedades físicas da pele, cuja gravidade varia, entre outros fatores, em função da profundidade original da lesão. Clinicamente a pele é seca, descamativa, discrômica e com diminuição da elasticidade, especialmente em cicatrizes hipertróficas. As características do tegumento pós-queimadura podem ser melhoradas com agentes de uso tópico normalmente utilizados em patologias como acne e fotoenvelhecimento. Foram avaliados 77 pacientes portadores de seqüela de queimadura peri-oral, entre seis e 46 anos de idade. O tipo de tegumento foi classificado como restaurado, quando epitelizado espontaneamente, ou enxertado. O tratamento tópico consistiu de tretinoína (0,01-0,05%) e ácido glicólico (5-7%) durante três meses. Duas distâncias foram medidas com paquímetro digital em abertura bucal máxima, a inter-dentária (D) e a inter-labial (L). Houve aumento significativo (p£0,01) de D e L nos grupos tratados quando comparados aos respectivos controles, tanto nos pacientes com tegumento restaurado como nos enxertados. O aumento de L foi significativamente maior que D. Sugerimos a utilização da medida inter-labial como parâmetro complementar à medida inter-incisal como método de avaliação indireta da elasticidade cutânea no tegumento pós-queimadura. / Tissue repair processes evolve with alterations of the cutaneous physical properties, which are proportional, among other factors, to the original depth of the lesion. At examination, the skin is dry, dyschromic and less elastic, specially in hypertrophic scars. Post-burn skin characteristics can be improved by topical agents regularly used in pathologies like acne or photodamage. We evaluated 77 patients who had peri-oral burn sequela. The ages ranged from six to 46 years old. The tegument was classified as restored, when spontaneously healed, or skin grafted. The topical treatment consisted of tretinoin (0,01-0,05%) and glycolic acid (5-7%) for three months. Inter-incisal (D) and inter-labial (L) distances were measured at maximal mouth opening with a digital pachymeter. Both inter-incisal and inter-labial distances increased significantly in the groups treated when compared to respective controls (p£0,01). This fact was similar in both types of tegument. Inter-labial distance (L) increased significantly more than D. We suggest its use as a complementary measure to inter-incisal distance for indirect evaluation of skin elasticity in the post-burn tegument.