Genetic architecture of retinoic-acid signaling-associated ocular developmental defects.

Ocular developmental anomalies are among the most common causes of severe visual impairment in newborns (combined incidence 1-2:10,000). They comprise a wide range of inborn errors of eye development with a spectrum of overlapping phenotypes and they are frequently associated with extraocular malformations, neuropsychomotor developmental delay and/or intellectual disabilities. Many studies from model organisms have demonstrated the role of retinoic acid (RA) during organogenesis, including eye development, and have revealed the wide spectrum of malformations that can arise from defective RA signaling. However, genes coding for homeobox proteins and morphogenetic factors were implicated in anomalies of ocular development long before genes coding for RA-signaling proteins. The purpose of this review is to discuss current knowledge about the highly complex genetic architecture of RA-signaling-associated ocular developmental anomalies in humans. Despite less than a dozen genes identified thus far, all steps of RA-signaling, from vitamin A transport to target cells to transcriptional activation of RA targets, have been implicated. Furthermore, the majority of these genetic disorders are associated with both dominant and recessive inheritance patterns and a wide spectrum of ocular malformations, which can dominate the phenotype or represent one of many features. Although some genotype-phenotype correlations are described, in many cases, the variability of clinical expression cannot be accounted for by the genotype alone. This observation and the large number of unsolved cases suggest that the relationship between RA signaling and eye development deserves further investigation.

Autologous bilayered self-assembled skin substitutes (SASSs) as permanent grafts: a case series of 14 severely burned patients indicating clinical effectiveness.

Split-thickness skin autografts (AGs) are the standard surgical treatment for severe burn injuries. However, the treatment of patients with substantial skin loss is limited by the availability of donor sites for skin harvesting. As an alternative to skin autografts, our research group developed autologous self-assembled skin substitutes (SASSs), allowing the replacement of both dermis and epidermis in a single surgical procedure. The aim of the study was to assess the clinical outcome of the SASSs as a permanent coverage for full-thickness burn wounds. Patients were recruited through the Health Canada's Special Access Program. SASSs were grafted on debrided full-thickness wounds according to similar protocols used for AGs. The graft-take and the persistence of the SASS epithelium over time were evaluated. 14 patients received surgical care with SASSs. The mean percentage of the SASS graft-take was 98 % (standard deviation = 5) at 5 to 7 d after surgery. SASS integrity persisted over time (average follow-up time: 3.2 years), without noticeable deficiency in epidermal regeneration. Assessment of scar quality (skin elasticity, erythema, thickness) was performed on a subset of patients. Non-homogeneous pigmentation was noticed in several patients. These results indicated that the SASS allowed the successful coverage of full-thickness burns given its high graft-take, aesthetic outcome equivalent to autografting and the promotion of long-term tissue regeneration. When skin donor sites are in short supply, SASSs could be a valuable alternative to treat patients with full-thickness burns covering more than 50 % of their total body surface area.

Skin characteristics: normative data for elasticity, erythema, melanin, and thickness at 16 different anatomical locations.

BACKGROUND: The clinical use of non-invasive instrumentation to evaluate skin characteristics for diagnostic purposes and to evaluate treatment outcomes has become more prevalent. The purpose of this study was to generate normative data for skin elasticity, erythema (vascularity), melanin (pigmentation), and thickness across a broad age range at a wide variety of anatomical locations using the Cutometer(®) (6 mm probe), Mexameter(®) , and high-frequency ultrasound in a healthy adult sample. METHODS: We measured skin characteristics of 241 healthy participants who were stratified according to age and gender. Sixteen different anatomical locations were measured using the Cutometer(®) for maximum skin deformation, gross elasticity, and biological elasticity, the Mexameter(®) for erythema and melanin, and high-frequency ultrasound for skin thickness. Standardized measurement procedures were applied for all participants. RESULTS: The means and standard deviations for each measured skin characteristic for females and males across five different age groups (20-29, 30-39, 40-49, 50-59, 60-69, and 70-85 years old) are presented. As previously described, there were variations in skin characteristics across age groups, anatomical locations, and between females and males highlighting the need to use site specific, age and gender matched data when comparing skin characteristics. CONCLUSION: The reported data provides normative data stratified by anatomical location, age, and gender that can be used by clinicians and researchers to objectively determine whether patients' skin characteristics vary significantly from healthy subjects.

What work means to people with work disability: a scoping review.

PURPOSE: As paid work is the occupation that people spend the most amount of their time doing, it is an important provider of personal meaning in their lives. This meaning has been shown to vary from person to person and to be important to health and well being. When a person is unable to work due to a disabling condition, it is unclear whether this meaning remains or is replaced by other meanings. The purpose of this scoping review was to explore what was known in the existing literature on what work means to those with work disability. METHODS: The review involved identifying and selecting relevant studies, charting the data and collating and summarizing the results. RESULTS: Fifty-two studies explored the meaning of work for those with cancer, mental illness, musculoskeletal disorders, brain injuries, paraplegia, and AIDS. The studies revealed that, for most, work continued to be meaningful and important. Common themes across all types of disability included work being a source of identity, feelings of normality, financial support, and socialization. These meanings were found to be both motivating for return to work and health promoting. Conversely, a small number of studies found that the meanings and values ascribed to work changed following disability. New meanings, found either at home or in modified work, replaced the old and contributed to new identities. CONCLUSIONS: The exploration of the meaning of work has been shown to provide important understanding of the experience of work and disability. This understanding can guide rehabilitation professionals in their interventions with the work disabled.

Resistance of human hair cuticle after a shaking process in wet conditions: comparison between Chinese and Caucasian hair.

Hair cuticle is the first protection shield of hair against external aggressions such as daily combing or brushing that induce friction, mechanical stress leading to ageing process. The identification of alterations of the hair surface induced by shaking process of various hair fibres, such as virgin and chemically treated Chinese and Caucasian hair, sheds some light on some specific features, e.g. Chinese cuticle is torn out in smaller particles than Caucasian, which seems to be related to geometrical rather than ethnic reasons. The present work also shows that two geometrical parameters of human hair, the inter-scale distance and the scale angle are directly linked to fibre dimensions, regardless of ethnic origin. Representative of hair with an average larger cross-sectional area, Chinese hair shows a slightly smaller inter-scale distance as well as a higher scale angle. On the other hand, Chinese hair cuticle seems to be more sensitive to chemical treatment than Caucasian hair, at least for straightening treatment taken here as an example. Such an approach can be of interest in designing cosmetic products more suited to specific needs of human hair.

Naltrexone for the management of post-burn pruritus: A preliminary report.

Severe pruritus after burn continues to be a clinical challenge that is inadequately addressed by traditional therapies. The success of naltrexone, an opioid antagonist, in treating pruritus in other patient populations, supported the concept that it may also be effective in burn survivors. Thus, burn patients who were dissatisfied with itch relief obtained after a course of traditional therapy were offered naltrexone. Of the 15 subjects included in this case series, 2 discontinued treatment because of side-effects. Of the remaining 13, 72% reported that they were satisfied with the itch relief provided by naltrexone, 69% were able to decrease or stop taking other medications to reduce itch, 85% would recommend it to other burn patients, and 62% divulged that it improved their quality of life. A sub-group (n=8) completed the French-Canadian version of the Questionnaire for Pruritus Assessment before and after a 2-week course of naltrexone treatment. Comparisons of individual responses before and after naltrexone treatment revealed that the affective impact of itch appeared to be less burdensome. The majority reported either a reduction or no change in the frequency of itch experienced. All reported either a decrease or no change in the duration of itch experienced after naltrexone treatment. All items that previously increased itch were less likely to affect itch or there was no change, except with fatigue, physical effort, sweating and eating. There was a statistically significant reduction in their visual analogue scale intensity ratings of itch "at its worst". Thus, naltrexone positively impacted the itch experienced by burn patients in association with a high percentage of patient-satisfaction. Further controlled trials are recommended.

Cross-cultural adaptation and validation of the Questionnaire for Pruritus Assessment for use in the French Canadian burn survivor population.

BACKGROUND: Severe pruritus after burn continues to be a clinical challenge. To address this challenge, thorough documentation of burn survivors' itch experience is required. A "Questionnaire for Pruritus Assessment" has been developed to capture the sensory and affective experiences associated with itch as well as its functional and quality of life implications, but to evaluate the Quebec burn survivor population, a Canadian French version is required. Thus, the purpose of this study was to generate a Canadian French cross-cultural adaptation of the questionnaire suitable for the burn survivor population. METHODS: Cross-cultural adaptation followed six steps: forward translation into Canadian French, back translations into English, resolution between the two versions, synthesis of the Canadian French versions, revision by a committee of experts, and testing of the pre-final version. RESULTS: The adaptation process resolved issues surrounding translation, content validity, burn survivor-specific modifications, and format, resulting in a Canadian French pre-final version that was pilot-tested among 32 burn survivors. Content validity of the Canadian French version was assessed by a committee of experts and the participants. The results showed good item completion and adequate distribution of scores without a ceiling or floor effect. CONCLUSIONS: This study resulted in a Canadian French version of the "Questionnaire for Pruritus Assessment" that can confidently be compared to other investigations utilizing the same tool. The adapted questionnaire also provides a valuable data collection means, enabling more thorough documentation of the burn survivor itch experience.

Understanding and building upon effort to return to work for people with long-term disability and job loss.

BACKGROUND: Effort is a concept that underlies programs assisting people with work disability to re-enter the labour force. During re-entry, attention is paid to the effort invested by the worker with an injury. However, for those with chronic work disability, the motivation to return to work (RTW) may be questioned by benefit service providers and healthcare professionals. OBJECTIVE: The objective of this paper is to describe the efforts made by people with long term work-disability to regain a foothold on the labour market. METHODS: This phenomenological study explored the meaning of work for people with long-term work disability and job loss. Twenty-seven interviews were conducted with nine participants. A thematic analysis was completed of the collected data. RESULTS: A key finding of this study is the variety and degree of effort exerted by participants to regain employment, despite time away from the workplace and system barriers. Effort was exerted to retain pre-accident employment; to obtain new work following job loss; and, to remain in a new job. CONCLUSIONS: This study suggests that if the RTW effort of people with long-term work disability is not fully acknowledged or supported, this population will remain unemployed where their strengths as competent, experienced workers will continue to be wasted.

Collagenase production is lower in post-burn hypertrophic scar fibroblasts than in normal fibroblasts and is reduced by insulin-like growth factor-1.

We recently demonstrated that the accumulation of extracellular matrix in post-burn hypertrophic scarring (HSc) tissues is, in part, caused by an overexpression of mRNA for fibronectin, type I, and type III procollagen. Here, we report that five different fibroblast cell strains derived from HSc tissues are deficient in collagenase activity relative to paired fibroblasts from normal skin of the same patients. Quantitative analysis demonstrated significantly lower (52.5 +/- 16.8% vs 100 +/- 8.3%; n = 9; p < 0.05) collagenase activity in conditioned medium from HSc fibroblasts relative to that obtained from the control. The expression of collagenase mRNA was also significantly depressed (51 +/- 7% vs 100 +/- 11%; n = 5; p < 0.05) in four of five strains of HSc fibroblasts examined. The level of mRNA for collagenase in both HSc and normal fibroblasts increased with serial passage, but at any given passage number, the expression of this transcript was lower in HSc fibroblasts. Insulin-like growth factor 1 (IGF-1), which is present at the site of HSc in high quantity, reduced collagenase mRNA but increased type I collagen mRNA expression in a time-dependent manner. The collagenase activity in conditioned medium derived from IGF-1-treated normal dermal fibroblasts was reduced (23.1 +/- 7.81% vs 100 +/- 6.6%; n = 7; p < 0.05). A significant reduction in collagenase mRNA and activity was also found in HSc fibroblasts following IGF-1 treatment. These findings suggest that IGF-1-induced suppression of collagenase mRNA and activity may be a mechanism by which IGF-1 promotes the development of post-burn HSc.

Myofibroblasts and apoptosis in human hypertrophic scars: the effect of interferon-alpha2b.

BACKGROUND: Hypertrophic scars (HSc) are a dermal fibroproliferative disorder that leads to considerable morbidity. Preliminary evidence suggests that interferon (IFN) may improve HSc clinically. The aims of this study were (1) to compare the cell density in HSc and in wounds that heal without the development of HSc (normotrophic scars), (2) to examine the presence of myofibroblasts and apoptosis in normotrophic and HSc scars over time, and (3) to determine if the systemic administration of IFN-alpha2b can induce apoptosis. METHODS: Two groups of patients underwent serial tissue biopsies. Six burn patients were studied prospectively by obtaining biopsy specimens from wound granulation tissue, normal skin, post-burn HSc, and normotrophic scars (healed donor sites). A second patient group with HSc was treated with systemic IFN-alpha2b and had biopsy material taken before, during, and after IFN therapy. The tissue was analyzed by immunohistochemical staining for alpha-smooth muscle actin (alpha-SMA) and in situ DNA fragmentation terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling (TUNEL) assay for apoptosis. RESULTS: The total numbers of fibroblasts in HSc were found to be similar to granulation tissue and twice that of normal skin and normotrophic scar. Over time the numbers of cells in HSc tissue decreased toward normal skin levels. There was a significantly higher percentage of fibroblasts staining for alpha-SMA in HSc as compared with normotrophic scar or normal skin obtained from the same patient (P >.05). Serial biopsy specimens of resolving HSc tissue obtained from the patients who received systemic IFN-alpha2b showed a general reduction in total number of fibroblasts and myofibroblasts associated with a significant increase in the percentage of apoptotic cells compared with normal dermis from the same patient. CONCLUSIONS: HSc tissues have greater numbers of fibroblasts and myofibroblasts than normal skin and normotrophic scars. As HSc remodels, the numbers of fibroblasts and myofibroblasts reduces, possibly by the induction of apoptosis. Systemic IFN-alpha2b may contribute to the resolution of HSc in part by the enhanced induction of apoptosis.

Hypertrophic scars, keloids, and contractures. The cellular and molecular basis for therapy.

Keloids, hypertrophic scars, and contractures are a result of aberrations of the normal wound healing process. An understanding of the cellular and molecular events that are implicated in the development of these fibroproliferative disorders will allow for optimization of wound healing. In turn, treatment choices can be based on the most current scientific information available.

Transforming growth factor-beta in thermally injured patients with hypertrophic scars: effects of interferon alpha-2b.

Hypertrophic scarring is a common dermal fibroproliferative disorder that leads to poor quality wound healing, prolongs rehabilitation, and increases morbidity following major thermal and other injuries to the deep dermis. Local and systemic transforming growth factor (TGF)-beta has been implicated as a fibrogenic cytokine in the pathogenesis of many fibrotic disorders, whereas interferon (IFN) alpha-2b may improve the pathologic features of dermal fibrosis directly or by antagonizing the effects of TGF-beta and histamine. Nine patients with severe hypertrophic scarring were evaluated for 8 weeks before treatment with subcutaneous recombinant IFN alpha-2b, 2 x 10(6) IU three times per week for 24 weeks. Clinical assessment was performed using standardized photography, a burn scar assessment tool, and serial scar volume measurements. Monthly measurements of serum TGF-beta and plasma Ntau-methylhistamine were made prior to, during, and after IFN alpha-2b therapy and compared with 27 age-matched controls. Serial biopsies of the hypertrophic scars and normal skin were performed for evaluation of mast cell numbers. Significant improvement in scar assessment occurred in 7 of 9 patients, and 3 of 9 demonstrated significant reductions in scar volume with interferon therapy beyond that occurring during the 8-week control period. For the entire group, mean rates of improvement were significantly better during interferon therapy with no recurrence following treatment. Before interferon therapy, serum TGF-beta was significantly higher in the burn patients with hypertrophic scarring than in a control population (123.04 +/- 36.48 vs. 56.85 +/- 8.38 ng/ml, p < 0.05). Within 3 months of IFN alpha-2b therapy, serum TGF-beta levels fell significantly and remained within the normal range during therapy and after interferon therapy was stopped. Plasma Ntau-methylhistamine levels were also significantly elevated in the hypertrophic scar patients as compared with age and sex-matched controls (153.6 +/- 92.07 vs. 48.3 +/- 28.9 pg/ml, p < 0.05), and significant reductions were achieved with interferon therapy and maintained after interferon was discontinued. Paired biopsies of hypertrophic scarring and normal tissue demonstrated increased numbers of mast cells in hypertrophic scars compared with normal uninjured skin from the same patients (2.65 +/- 1.63 vs. 1.04 +/- 0.62 cells/high power field, p < 0.001); however, no significant change in mast cell content of the hypertrophic scars accompanied interferon therapy. Patients with severe hypertrophic scarring demonstrate increased levels of serum TGF-beta and plasma Ntau-methylhistamine following thermal injury. A significant clinical improvement in scar quality and volume occurred during IFN alpha-2b therapy, which was associated with normalization of serum TGF-beta and plasma Ntau-methylhistamine levels. A double-blind, placebo-controlled trial will be required to further assess the usefulness of subcutaneous treatment with IFN alpha-2b for the treatment of hypertrophic scarring.

Rating the resolving hypertrophic scar: comparison of the Vancouver Scar Scale and scar volume.

The increased focus of research interests and clinical documentation on outcomes demands that evaluation tools provide reliable and valid data. The Vancouver Scar Scale (VSS) was developed to provide a more objective measurement of burn scars; however, the validity (a test's ability to measure the phenomenon for which it was designed) of the VSS has not been tested. To examine the construct validity of the VSS, we compared it with scar volume, which has established face validity. Burn scars were evaluated monthly for a minimum of 7 months. Three scar volume measurements were performed on each scar. In addition, 3 independent examiners completed the VSS for the same scar. The data generated by these 2 measurements were used to establish the following: (1) the interrater agreement estimated by interclass correlation coefficient, (2) convergence validity, (3) the sensitivity of the assessments to discriminate changes in the scar over time, and (4) the prevalence of related parameters that are not currently being captured by the VSS. In an attempt to address some of the deficiencies of the VSS, we propose several modifications. We anticipate that these changes will increase the reliability and validity of the VSS through an increase in the awareness that training in the use of this scale is required, through improvement in the quality of the subscales, and through the documentation of additional pertinent information.

Control of wound contraction. Basic and clinical features.

Although a substantial amount of molecular and cellular data have been generated in an effort to understand the process of wound contraction and scar contracture formation, questions remain. What seems apparent is that the myofibroblast is not the only cell that generates contractile forces within wounds, but it does appear to be intrinsically linked to the development of hypertrophic scars. The supposition that the formation of scar contractures is solely the result of a continuation of wound contraction is an oversimplification. Figure 4 provides a model of the possible evolution of contractile forces during the wound healing process and their role in the development of scar contractures. Migration of fibroblasts into and through the extracellular matrix during the initial phase of wound healing, prior to the expression of alpha-SMA, appears to be a fundamental component of wound contraction. During this migration, the pulling of collagen fibrils into a streamlined pattern in their wake, and the associated production of collagenase, may facilitate a more normal arrangement of collagen. Once the wound has been repopulated and the chemotactic gradient that was established by inflammatory cells is decreased, fibroblast migration will cease. It is at this point that myofibroblasts appear and play a key role in the production of hypertrophic scars, given that their prolonged presence and over-representation are hallmarks of this pathology. One of the pivotal differences between wounds that proceed to normal scar compared with those that develop hypertrophic scars and scar contractures may be a lack (or late induction) of myofibroblast apoptotic cell death. The combined contribution of fibroblasts and myofibroblasts to abnormal extracellular matrix protein production results in an excessive and rigid scar. The isometric application of contractile forces by myofibroblasts probably contributes to the formation of the whorls, nodules, and scar contractures characteristic of hypertrophic scars. Because the prolonged presence of myofibroblasts, producing an imbalance in extracellular matrix proteins and proteases, probably exacerbates hypertrophic scars and wound contraction, accelerating the rate of apoptotic cell death to reduce the cell number to that seen in normal scar may be a useful strategy for providing effective and efficient treatment of scar contracture.

Decreased serum insulin-like growth factor-I in burn patients: relationship with serum insulin-like growth factor binding protein-3 proteolysis and the influence of lipid composition in nutritional support.

OBJECTIVES: To test the effects of the amount and type of fat in the nutritional support on serum insulin-like growth factor (IGF)-I concentrations in burn patients and to test the hypothesis that the serum proteolytic activity for insulin-like growth factor binding protein (IGFBP)-3 is a major mechanism for the decreased serum IGF-I observed in these patients. DESIGN: Randomized, double-blind trial of three different nutritional supports and analysis of serum IGF-I, IGFBP-3, and serum IGFBP-3 proteolysis. SETTING: Burn center in a university hospital. PATIENTS: A total of 23 severely burned (>25% total body surface area burned) adult patients. INTERVENTIONS: Patients were randomly assigned to three types of nutritional support differing in the amount of energy derived from fat and the presence or absence of fish oil: Group I (control), 35% fat; Group II, 15% fat; Group III, 15% fat with 50% as fish oil. Nutritional support was both parenteral and enteral and was started within 24 hrs of admission. MEASUREMENTS AND MAIN RESULTS: Serum IGF-I and IGFBP-3 were measured by radioimmunoassay every 3 days for 28 days in 23 severely burned adults. In six patients, IGFBP-3 was measured by ligand binding assay and the serum proteolytic activity for rhIGFBP-3 was measured as well. Serum IGF-I concentration was low in all subjects throughout the study period, but did increase with time (p < .01); significantly higher values were found in Group III (p < .05). Multivariate analysis showed that fish oil and low fat solutions were significantly correlated to serum IGF-I concentrations. Serum IGFBP-3 (radioimmunoassay) was higher than normal throughout the study with no difference between the groups. Between days 4 and 16, IGFBP-3 was cleaved into two fragments in all patients studied, and the molecular weights of the fragments were equal to those observed in the serum of a woman late in pregnancy. During this period of time, serum proteolytic activity for rhIGFBP-3 was >30% in 24 of the 30 samples measured, whereas 20 of the 28 samples measured thereafter were normal (<25%). Serum IGFBP-3 concentration from ligand binding assay was correlated with serum proteolytic capacity in all subjects (mean r2 = 0.77; p < .01) and with serum IGF-I concentrations in five of six subjects (mean r2 = 0.81; p < .01). CONCLUSIONS: In burn injury, serum IGF-I concentrations are sensitive to the amount and type of fat in their nutritional support. The presence of fish oil allowed for a more rapid recovery of serum IGF-I levels. The proteolysis of IGFBP-3 may be an important cause of the decreased serum IGF-I values and the protease(s) responsible for this seem to be similar to those observed in late pregnancy.

Upregulation of rat P23 (a member of the YjgF protein family) by fasting, glucose diet and fatty acid feeding.

In a previous study, we identified and purified a 99-amino-acid rat liver-kidney perchloric-acid-soluble 23-kDa protein (P23) which displays 30% identity with a highly conserved domain of heat shock proteins (HSPs), as well as an AT-rich 3' untranslated region, which has also been described to play a role in H70 mRNA life span and protein expression. An identical perchloric-acid-soluble protein inhibiting protein synthesis in a rabbit reticulocyte lysate system was also found 2 years later by another group. More recently, the novel, the YjgF, protein family has been described, comprising, 24 full-length homologues, including P23, highly conserved through evolution, and consisting of approximately 130 residues each and sharing a common ternary structure. Independent studies from different laboratories have provided various hypothetical functions for each of these proteins. The high degree of evolutionary conservation may suggest that these proteins play an important role in cellular regulation. Although the function of none of these proteins is known precisely, we present experimental evidence which, combined with the relationship to glucose-regulating protein revealed here, and the relationship to fatty-acid-binding protein revealed by others, allow us to propose a role for P23. In rat liver, P23 expression is developmentally regulated and modulated by dietary glucose, and its mRNA is induced by starvation, in the presence of fatty-acids and in 3-MeDAB-induced hepatomas. The mRNA encoding mouse liver P23 is also hormonally modulated in a mouse line AT1F8. These data indicate that P23 protein might be a key controller of intermediary metabolism during fasting.

The effect of interferon alpha 2b on the expression of cytoskeletal proteins in an in vitro model of wound contraction.

Wound contraction is an essential component of wound healing. However, the development of scar contractures in tissues and organs disrupts normal organ integrity and produces functional deformities. Although interferons alpha and gamma inhibit extracellular matrix protein production by fibroblasts, their effects on cytoskeletal protein mediated-wound contraction are as yet unclear. The fibroblast-populated collagen lattice is an in vitro assay that simulates wound contraction. When matched pairs of human hypertrophic scar and normal dermal fibroblast cultures established from patients recovering from a thermal injury were used, interferon-alpha 2b exposure before lattice formation was found to significantly inhibit contraction in a treatment time-dependent manner (p < 0.05). Fibroblasts generated contractile forces that were triphasic and serum sensitive (p < 0.01). Comparison of hypertrophic scar and normal dermal fibroblasts revealed no significant differences in ability to induce lattice contraction. Northern blot analysis of mRNAs for the intracellular contractile proteins revealed that interferon-alpha 2b significantly down-regulated mRNA levels of the actin isoforms beta and gamma (50% to 60%) but had no significant effect on alpha-tubulin, vimentin, and alpha-actinin. Fibroblast-populated collagen lattices were stained with rhodamine-labeled phalloidin to reveal filamentous actin proteins. Marked morphologic alterations of the stress fibers were associated with reductions in lattice contraction after interferon-alpha 2b treatment. Thus interferon-alpha 2b's inhibition of wound contraction in vitro is associated with reductions in mRNA for beta and gamma actin and distinct morphologic alterations in fibroblast stress fiber morphology.

Enhanced expression of mRNA for insulin-like growth factor-1 in post-burn hypertrophic scar tissue and its fibrogenic role by dermal fibroblasts.

Hypertrophic scarring (HSc) which frequently develops in patients following severe thermal injury is characterized by accumulation of extracellular matrix (ECM) proteins including type I and type III collagen. In this study, we examined the presence and quantity of IGF-1 mRNA transcripts in post-burn HSc. The results of dot blot experiments showed a 77.5% (100 +/- 8.15 vs 177.5 +/- 19, p < 0.01) increase in expression of IGF-1 IIIRNA in HSc tissue relative to normal dermis obtained from the same patients. A Northern blot analysis confirmed the specificity of the IGF-1 cDNA. This cDNA visualized four different transcripts with apparent sizes of 7.0, 3.9, 1.8 and 1.0 kb, similar to those previously reported. The possible fibrogenic role of IGF-1 was examined by analyzing the effect of this growth factor on the expression of mRNA for the pro alpha 1(I) chain of type I procollagen and the pro alpha 1(III) chain of type III procollagen in dermal fibroblasts. IGF-1 increased the expression of these transcripts as early as 6 h and the effect was maximal at 24 h. Quantitative analysis by densitometry showed a 149 and 166% increase in pro alpha 1(I) and pro alpha 1(III) mRNA after 24 h of IGF-1 treatment, respectively. This effect seems to be specific as the abundance of mRNA for the pro alpha 2(I) chain of type I procollagen or TIMP-II was unchanged.(ABSTRACT TRUNCATED AT 250 WORDS)

Effect of interferon-alpha2b on guinea pig wound closure and the expression of cytoskeletal proteins in vivo.

Scar contraction following the healing of deep partial-thickness or full-thickness dermal injury is a leading cause of functional and cosmetic morbidity. The therapeutic use of interferon for the treatment of fibroproliferative disorders associated with scar contraction, including hypertrophic scar, has been suggested because of its antifibrotic properties. Treatment of fibroblasts with interferon has been shown to reduce the rate and extent of contraction using the in vitro fibroblast-populated collagen lattice model. In order to establish the effect of interferon-alpha2b on full-thickness wound contraction in vivo, osmotic pumps loaded with interferon or sterile saline were implanted intraperitoneally in guinea pigs. Seven days following implantation, six full-thickness punch biopsy wounds were created and were monitored by daily assessment of the wound. There was a significant reduction in the rate of wound contraction in the interferon-treated animals after day 3 (p < 0.01). Western blot analysis was used to quantitate selected cytoskeletal proteins in the normal skin and tissue biopsied from the wound at days 7, 14, and 21 postinjury. The amount of vimentin in the contracted wound increased following injury as compared with the amount present in normal skin (p < 0.0001); however, the relative amounts of the myofibroblast-associated cytoskeletal proteins alpha-smooth muscle actin and smooth muscle myosin were less than those found in normal, uninjured skin. By using vimentin to adjust the levels of cytoskeletal proteins for the increase in cellularity in the wounds, both alpha-smooth muscle actin and smooth muscle myosin significantly increased after closure of the wounds on day 14, as compared with the open-wound stage (day 7), before further reductions occurred with remodeling on day 21. Measurement of apoptotic cells using the terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling assay revealed an increase in apoptosis in the interferon-alpha2b-treated animals at 21 days following wounding (p < 0.001), which did not colocalize with alpha-smooth muscle actin staining. Taken together, these findings suggest that interferon-alpha2b inhibits wound contraction in vivo, not through an appreciable alteration in myofibroblast number or cytoskeletal protein expression, but possibly through a reduction in fibroblast cellularity by the induction of apoptosis.