Genetic Polymorphisms of Transforming Growth Factor Receptors (TGF-\u03b2RI, TGF-\u03b2RII) and Risk Factors Associated with Keloid Scars in Burkina Faso: A Cross-Sectional Study

Background The mechanism and prediction of stricturing Crohn’s disease (CD) to guide medical therapy remains elusive. Formation of hypertrophic and keloid skin scars have not been previously studied in IBD. This study aimed to evaluate the nature of skin scars in patients with CD, and their association with different phenotypes. Methods Adult patients attending a tertiary IBD service were invited to participate in this cross-sectional observational study. Patients were divided into three groups (non-IBD controls, non-structuring/non-penetrating CD and stricturing/penetrating CD). A patient survey tool, consisting of 8 questions was administered to assess for the presence of scars and their type. Disease characteristics were ascertained using electronic medical records. The primary outcome of the study was the prevalence of either hypertrophic or keloid scarring across the 3 groups. Secondary outcomes included the rates of scarring overall as well as keloid and hypertrophic scars individually between the three groups. The chi-square test along with pair-wise comparison differences was used to assess for differences between groups. Results 189 patients were recruited for this study with baseline characteristics presented in table 1 below. Control patients were older than the other groups (p = 0.013). Patients with stricturing/penetrating CD were more likely to be male. Patients with stricturing/penetrating CD (36.5%) were more likely to have either hypertrophic or keloid scars than those with non-stricturing/non-penetrating CD (11.1%) and non-IBD controls (6.3%), p = <0.0001. Pairwise comparisons revealed differences individually between the stricturing/penetrating and non-stricturing/non-penetrating (p = 0.001) and healthy controls (p < 0.001), but not between the latter two groups (p = 0.52). 8 patients with structuring/penetrating CD had a history of keloid scars, but none in the other 2 groups (p < 0.001). Across the 3 groups, there was no difference in the rates of normal scars (p = 0.112). Conclusion In this pilot cross-sectional study, a higher rate of hypertrophic or keloid scarring was demonstrated in patients with stricturing or penetrating disease in comparison to healthy controls and those with non-stricturing/non-penetrating disease, suggesting potential underlying common pathophysiology. Future studies confirming this novel association may provide an avenue for further study of predictive and therapeutic targets for IBD.

Keloid disease: clinical relevance of single versus multiple site scars

The potential connection between blood types and cancer risk has become a puzzling yet intriguing area of research in recent years; while its association is widely assessed with cancer occurrence, it is not well assessed in case of hereditary cancers. This study aimed to assess the relation between ABO blood groups and the occurrence of familial cancers among the patients with cancer in Kirkuk city, Iraq. This cross-sectional study was carried out in Kirkuk city from the period 15th October 2023 to 15th April 2024 on 398 patients diagnosed with cancer in Kirkuk oncology center. The study included three groups: patients with a family history of cancer in first-degree relatives (n=90), no family history (n=288), and second-degree relatives (n=20). Data was collected from both medical records and patient interviews, focusing on demographic details, lifestyle habits, medical history, and exposure to potential risk factors. In-depth interviews provided further details on lifestyle patterns, including smoking habits, alcohol use, eating habits, medical conditions, medication use, viral infections, and exposure to chemicals or radiation. Data was analyzed using SPSS V.26 comparing based on the familial cancer history. Significant differences were noted in age distribution, with the 38-47 and 48-57 age groups being more common in the first-degree and no family history groups, respectively. Sex distribution showed a higher proportion of females in the first-degree and second-degree groups (P=0.017). There were higher proportions of high socioeconomic status in the second-degree group and married individuals in the first-degree and second-degree groups compared to non-familial cancers. Breast cancer was more prevalent in the first-degree and second-degree groups. Adjusting for demographic, socioeconomic, Body-Mass-Index (BMI), cancer type, and health related habits, O- blood group was significantly associated with a higher likelihood of familial cancer history in first-degree relatives (OR: 14.083, P=0.024) compared to non-familial ones. Other blood groups did not show significant associations. There might be an association between familial cancer O- blood group that needs to be re-evaluated in further studies.

Objective This study aims to explore the influence of the serine and arginine rich splicing factor 2 (SRSF2) on the biological characteristics of keloid fibroblast, by comparing the expression levels of SRSF2 in normal skin and keloid, with the purpose to provide a new method to study the pathogenesis of keloid. Methods Samples of normal skin were derived from excess tissue of skin grafts, collected from 8 patients, aged 8-53 years old, while the specimens of keloid were from 12 keloid patients, aged 18-24 years old. All the patients were admitted in the Plastic Surgery Hospital, Chinese Academy of Medical Sciences. The expression of SRSF2 was assessed by immunohistochemistry and immunofluorescent staining in both normal and keloid tissue. Keloid fibroblasts were cultured in vitro to detect the relationship between TGF-β1 stimulation and SRSF2 expression. After constructing the lentiviral sh-RNA-expression vector, targeting SRSF2 and infecting keloid fibroblast, the apoptosis and proliferation of cells were analyzed by the MuseTM Cell Analyzer. The expression of CyclinE1 was analyzed by real-time PCR and western blot, and the secretion and expression of extracellular matrix and TGF-β1 were detected by real-time PCR and ELISA.The software SPSS 21.0 was used to do the statistical analysis. Results The expression of SRSF2 was significantly higher in the keloid samples than in the normal skin samples. The percentage of SRSF2 positive cells in keloid was (77.04±4.37)% , while the percentage of SRSF2 positive cells in normal skin was (25.10±1.24)%. TGF-β1 promotes the expression of SRSF2 to (159.73±17.03)% times in keloid fibroblasts. After the SRSF2 knock-out, the apoptosis rate of the keloid fibroblasts increased, its cell cycle arrest was observed at the G0/G1 phase, and the expression of Cyclin E1 decreased. Apoptotic cells in control group was about (18.83±1.24)% , while (25.81±7.09)% and (26.71±6.14)% were in the two knock-out groups respectively. Cells in G0/G1 phase was about (58.97±1.73)% in control group, while (63.95±2.07)% and (64.65±3.23)% were in the two knock-out groups respectively. Compared to the control group, the expression of Cyclin E1 mRNA in the two knock-out groups were (31.60±6.81)% and (33.01±11.39)% respectively. Additionally, the expression of COL3A1 FN1 mRNA decreased, and the expression and secretion of TGF-β1 was declined. Compared to the control group, the expression of COL3A1 mRNA in the two knock-out groups were (64.90±23.71)% and (67.97±13.50)%, the expression of FN1 mRNA in the two knock-out groups were (59.10±8.11)% and (70.70±18.26)%, the expression of TGFB1 mRNA in the two knock-out groups were (53.37±15.51)% and (67.53±19.33)% respectively. The concentration of TGF-β1 in the medium of control group was (115.60±18.17)pg/ml, while (75.35±12.25) pg/ml and (72.06±14.66) pg/ml were in the two knock-out groups respectively. Conclusions The expression of SRSF2 increased in keloid. The inhibition of SRSF2 in keloid fibroblast resulted in the growth restriction and apoptosis of cells, and decreased the secretion of extracellular matrix and TGF-β1, which providing a new insight into keloid pathogenesis, and suggesting that SRSF2 could be a new therapeutic target for keloid conditions. Key words: SRSF2; Keloid; Fibroblast; Biological characteristics

Keloid disease is a benign, quasineoplastic disease with a high recurrence rate. Mesenchymal-like stem cells (MLSC) have previously been demonstrated in keloid scars and may be involved in keloid pathobiology. However, as these cells have only been examined by single colour fluorescence activated cell sorting (FACS) alone, they need to be more comprehensively characterized so that the key cellular contributors to keloid scars can be better understood. To identify and characterize MLSC in intralesional and extralesional keloid, and to distinguish haematopoietic stem cells (HSC) from mesenchymal stem cells (MSC). Punch biopsies from intralesional (top, middle and margin) and extralesional keloid scar sites were obtained from 17 patients with a keloid. Multicolour FACS analysis using antibodies specific for HSC markers CD34 and CD117 and MSC markers CD13, CD29, CD44 and CD90 was performed on freshly isolated keloid scar cells and on passage 0 and 1 cells. This was complemented by real-time quantitative polymerase chain reaction (PCR) and immunohistological in situ analyses. Keloid scars contain distinct subpopulations of MLSCs. Cells positive for CD13, CD29, CD44 and CD90 were found to be significantly (P<0·05) higher in the top and middle compartments of keloid scars compared with extralesional skin, where cells positive for CD34, CD90 and CD117 (representing HSCs) predominated. A unique population of CD34+ cells (cells positive for CD13, CD29, CD34, CD44 and CD90) were found in keloid scars and in extralesional skin. FACS and quantitative PCR analysis showed that many of the MSC markers were progressively downregulated and all HSC markers were lost during extended keloid fibroblast culture up to passage 1. We have found distinct subpopulations of haematopoietic and nonhaematopoietic MSC in keloid scars, whereby HSC accumulate extralesionally, while keloids seem to provide a niche environment for nonhaematopoietic MSC. Future therapy of keloids may have to target differentially both stem cell populations in order to deprive these tumours of their regenerative cell pools.

A novel RHCE*cE allele identified in a Chinese individual, encodes weakened expression of c and E antigens

To investigate the genome structure variation (SV) related with keloid using the whole-gene resequencing technology. We studied a keloid pedigree containing 4 generation of 27 people. 5 people (4 cases of keloid patients, and 1 case of normal) were selected to extract the genomic DNA. Then the whole-gene resequencing technique was used to check the variations. Through database comparison and variation annotation analysis, we obtained 2 SVs associated with keloid formation. We used DAVID software to do the gene ontology and pathway analysis. We found a 168 bp inversion in gene tetraspanin 8 (TSPAN8) in all keloid patients, which contained the forth exon of TSPAN8. There was no report about SVs related to keloid. In this study, we found 2 SVs associated with keloid, especially TSPAN8. The tumor cells express the TSPAN8 can up-regulate the vascular endothelial growth factor and its receptors, promote the adjacent fibroblasts secrete matrix metalloproteinases and uridylyl phosphate adenosine. So we hypothesis that the inversion of the forth exon in TSPAN8 may lead to the signal transduction disorder in the keloid patients. This study was a preliminary research. It needs a further study containing large sample to confirm.

Aim: to investigate the association between vitamin D receptor polymorphisms and circulating 1,25-Dihydroxyvitamin D (1,25(OH)2D) levels with keloid scar (KS) risk. Methods: A total of 261 patients with KS and 261 normal healthy individuals were enrolled. VDR gene polymorphisms were determined. Circulating 1,25(OH)2D levels were detected. Results: In this study, we investigated the role of four loci of VDR gene polymorphisms in determining the risk of KS in a Chinese cohort. We found that the TaqI C>T polymorphism was closely associated with the KS incidence. Carriers with CC genotype of TaqI had a higher chance of developing KS. Stratification analyses by sex showed that this trend exists only in female subjects but not in male subjects. Furthermore, the TaqI C>T polymorphism affects the circulating 25-Hydroxyvitamin D levels. The CC carriers had a significantly lower mean circulating 1,25(OH)2D level than TT carriers and CT carriers. KS subjects had significantly lower mean serum circulating 1,25(OH)2D level than controls. The receiver operating characteristic curve analysis revealed that the serum circulating 1,25(OH)2D level at the cut-off point of 16.1 ng/ml could discriminate KS subjects from controls. Conclusion: Collectively, these data provide evidence that the vitamin D/VDR pathway plays an important role in the development of KS. The TaqI gene polymorphisms of VDR and circulating 1,25(OH)2D levels may thus be used as potential markers for prediction of KS development.

Previous attempts to explain the presence of focal hyperintensities in the cerebral white matter on MR images have focused on the patient's age and cardiovascular risk factors. The purpose of this study was to survey many variables in subjects' social, medical, and surgical histories to identify those factors associated with focal hyperintensities in asymptomatic persons. Asymptomatic volunteers were examined with MR imaging of the head and questioned concerning smoking history; chemical dependence; alcohol consumption; medical history (e.g., high blood pressure, kidney disease); surgical history (e.g., for appendix, gallbladder); medications (e.g., antihypertensives, analgesics); and other historical family and social variables. Vital signs were recorded. Focal hyperintensities were counted by observers who were not provided any information about the subjects other than that they were either volunteers or patients. Each of the variables in the history was tested for an association with the number of focal hyperintensities seen on MR images. A statistically significant positive association was found between both age and the use of antihypertensive medications and the number of focal hyperintensities. Associations were also found between the number of focal hyperintensities and diastolic and systolic blood pressures at the time of MR imaging, but these associations did not reach statistical significance. No other variables in the medical, surgical, or social histories were found to be significantly related to the number of focal hyperintensities. Age and the use of antihypertensive medications were the only significant risk factors identified in respect to focal hyperintensities of the cerebral white matter.

Epidemiological and clinical features of hypertrophic scar and keloid in Chinese college students: A university-based cross-sectional survey

Objective To study the effect of miR-194-3p on the migration of keloid fibroblasts. Methods Differentially expressed miRNA were screened by gene chip in 8 human keloid and normal tissues. The down regulated miR-194-3p was selected for study and its binding to RUNX2 was predicted by MiRDB, and verified by fluorescent reporter gene in human keloid fibroblasts (HKFs) and passage 3 keloid cells, respectively. The effect of miR-194-3p on the migration of fibroblasts was detected by transwell assay. Western blot and real-time PCR were used to analyze the effect of miR-194-3p on RUNX2 and MMP2 expression in HKFs. The results were analyzed by SPSS 19.0 software and compared by non-paired t test. P<0.05 was considered statistically significant. Results There were abnormal expressions of miR-721, miR-21-3p, miR-382-3p, miR-194-3p, miR-3107-5p and miR-144-5p in keloid. miRDB software analysis showed that miR-194-3p and RUNX2 had targeted binding sites. Reporter gene experiments showed that miR-194-3p inhibited the activity of RUNX2 by about 50% compared with the control group (t=2.7764, P=0.0005). MiR-194-3p could significantly inhibit the expression of RUNX2 and MMP2, and inhibited the migration of keloid fibroblasts (t=2.7764, P<0.05). Conclusion MiR-194-3p may inhibit the migration of fibroblasts by inhibiting the activity of RUNX2 in keloid. Key words: Keloid; Fibroblast; Migration; miR-194-3p; RUNX2

Contracting Mycobacterium tuberculosis exogenously through intralesional steroid injection for keloid is an unique phenomenon not so far mentioned in literature.Primary cutaneous tuberculosis developing at keloid following intralesional steroid(ILS) which is negative for routine bacterial cultures & insensitive to routine antimicrobials warrants high index of suspicion for Mycobacterial infection. known for its varied clinical presentation, with many of them diagnosed years after the onset of symptoms. The occurrence of cutaneous nontuberculous mycobacterial (NTM) infections is being increasing reported. Although most pathogenic species of NTM may cause skin and soft tissue infections, Mycobacterium marinum, Mycobacterium ulcerans and the rapidly growing mycobacteria such as Mycobacterium fortuitum, Mycobacterium chelonae and Mycobacterium abscessus are the most frequent causes.(1) However there is rarely any report of infection with Mycobacterium tuberculosis in patients of keloid following intralesional steroid injection. Cutaneous infections by NTM usually occur following contact of surgical or traumatic wounds with water or other contaminated source although cutaneous infections with Mycobacterium tuberculosis following surgical and traumatic wounds is very rare. Among the immunosuppressed it can occur as a consequence of a disseminated mycobacterial disease. A high index of suspicion about Mycobacterium tuberculosis and NTM infections is necessary, in those with chronic infections of the skin and soft tissue, to ensure prompt and appropriate treatment. (3) Abscess formation at the site of the puncture wound is most often due to rapidly growing mycobacteria. Wallace and coworkers noted that M. fortuitum and M. chelonae were the predominant isolates when they reviewed 125 cases of human infections caused by rapidly growing mycobacteria. Among these 59% had cutaneous infections following surgery, accidental trauma or needle injections. (4) We report a case of subcutaneous abscesses due to M. tuberculosis following intralesional steroid in a patient with keloid. Literature from our subcontinent suggests association of mycobacterial infection following yellow jacket sting and recurrent soft tissue infections. (5,6) However, we were unable to retrieve literature regarding association of keloids with M.tuberculosis following a PubMed search, which emphasizes the uniqueness of our report. II. Case Report A 43-year-old man who is a non diabetic and non hypertensive, presented with keloid lesions of 10- years duration. He noticed that the size increased gradually over a period of time which prompted him to consult a dermatologist, who administered intralesional steroid injection given once a month for 7 months. The patient noticed progressive increase in size of the lesion followed by drainage of pus when he was referred to our facility at Dermatology OPD of School of Tropical Medicine,Calcutta. A diagnosis of pyogenic infection in a keloid was made and he was advised oral and topical antibiotics, but there was no improvement. On examination he was afebrile and there were no palpable lymph nodes. Single keloid was observed over the manubrium, (3 × 1.5 × 1.5 cm) The lesion was associated with sinuses which was discharging pus. He had no family history of keloids. No abnormal findings were seen in his blood picture and chest X-ray. Sputum for acid-fast bacilli (AFB) and Mantoux test were negative. Serology for HIV and VDRL were non- reactive.Histopathology was not performed apprehending flaring up of keloid following punch biopsy induced trauma. Pus was aspirated from the lesions for microbiological studies. Gram stain showed plenty of pus cells but no bacteria, while Ziehl-Neelsen stain revealed plenty of acid-fast bacilli in a background of polymorphs raising the possibility of mycobacteria. Routine bacterial culture of the pus showed growth of Klebsiella pneumonia and MRSA for which he was given antibiotics according to sensitivity, but still there was no improvement. BACTEC was done which showed growth of Mycobacterium tuberculosis and line probe assay did not show any resistance to INH or Rifampicin. Following this Interferon Gamma Release Assay was done

Objective To study the expression of gravin in the keloid and to investigate its probable role in the pathogenesis of keloid. Methods Skin biopsy was performed and skin samples were obtained from 18 normal human controls and 25 patients with keloid. The mRNA expression of gravin in specimens was detected by real-time fluorescent RT-PCR, and double immunofluorescence analysis was used to investigate the distribution of gravin protein in skin tissues. Results As detected by real time PCR, the relative expression of gravin mRNA was remarkably reduced in comparison with normal skin (0.4565±0.1728 vs 0.0953±0.0664, P<0.01). Results of double staining indicated that, in normal skin, gravin was primarily distributed in the cytoplasm of fibroblasts; while in keloid, it was observed mainly in macrophages and sporadically in fibroblasts. Conclusions Compared with the normal skin, keloid shows a different expression intensity and distribution of gravin, which might contribute to the development of keloid by regulating the proliferation of fibroblasts and activation of macrophages. Key words: Keloid; Genes, gravin; Immunohistochemistry

To observe the effects of conditioned medium from keloid fibroblasts under hypoxia on angiogenesis, and to investigate the role of hypoxic microenvironment in invasive growth of keloid. Primary keloid fibroblasts and human umbilical endothelial cells (HUVEC) were cultured as conventional method. Keloid fibroblasts were cultured either in a hypoxic incubator (2% O2) for 48 h or in a normoxic incubator (20% O2) as control. Then those cell culture mediums were collected and mixed with endothelial cell medium by the proportion of 1:1 as conditioned medium. The mRNA and secreted protein of pro-angiogenic factors such as vascular endothelial growth factor (VEGF), angiopoietin-1 (Ang-1) and periostin of keloid fibroblasts under hypoxia were detected by real time PCR and ELISA. The proliferation, migration and invasion, tube formation of HUVEC cultured with conditioned medium were evaluated by CCK-8 assay, Transwell assay and matrigel tube formation assay, respectively. Hypoxia increased the expression of VEGF, Ang-1 and periostin in both mRNA (increased by 75%, 43% and 118% respectively, P < 0.05) and secreted protein (increased by 30.2%, 14.2% and 19.5% respectively, P < 0.05) levels; the proliferations of HUVEC in hypoxic conditioned medium in 1, 2 and 3 d were 0.67 +/- 0.07, 0.84 +/- 0.09 and 1.08 +/- 0.10 respectively, which were higher compared to those in control group (0.52 +/- 0.08, 0.72 +/- 0.10 and 0.91 + 0.14, P < 0.05); the numbers of migration, invasion and tube formation of HUVEC were (73.2 +/- 8.9), (56.3 +/- 12.5), (9.66 +/- 1.96) cells/HP, which were higher compared to those in control group [(59.0 +/- 8.0), 35.5 +/- 8.5), (6.5 +/- 1.87) cells/HP, P < 0.05]. Hypoxia increases the expression of pro-angiogenic factors of keloid fibroblasts, and its conditioned medium under hypoxia could promote angiogenesis. The results suggest hypoxic microenvironment may play a significant role in the invasive growth of keloid by inducing angiogenesis.

To construct and characterize the TGF-β1, induced epithelial-mesenchymal transition (EMT) model of keloid epithelial cells in vitro, and to investigate the expression of epithelial stem cells related surface markers in keloid epithelial cells during EMT induction. The epithelial cells from 3 keloid samples of ears were cultured in vitro and induced by transforming growth factor betal (TGF-β1, 1 ng/ml) for 5 days, which was the experimental group, the same cells untreated were considered as the negative control group. The expressions of EMT-associated markers and regulative genes were detected using immunofluorescence staining, real-time PCR and western blot analysis. Then the surface markers of epithelial stem cells were detected using real-time PCR. Statistical significance was determined using Independent-Samples t Test, a p value less than 0. 05 was considered statistically significant. The mRNA expression of transcription factor snail2 and mesenchymal-specific marker vimentin increased significantly in TGF-β1, induced keloid epithelial cells (P < 0. 05), in which snail2 increasing from 0. 91 ± 0. 23 to 1. 69 ± 0. 10, and vimentin from 5. 86 ± 2. 07 to 24. 29 ± 5. 39. Whereas the mRNA expression of epithelial-specific marker E-cadherin decreased from 1. 06 ± 0. 19 to 0. 65 ± 0. 09. The mRNA expression of CD29 and Lgr6, two surface markers of epithelial stem cells, significantly increased after induction of the TGF-β1, (P < 0. 05), from 0. 55 ± 0. 14 and 1. 61 ± 0. 31 to 1. 19 ± 0. 12 and 3. 84 t 0. 62 respectively. In induced cells, the immunofluorescence results showed staining of E- cadherin became faint, but the number of positive staining cells of vimentin increased. Western blot confirmed the protein expression of E-cadherin weakened, and the vimentin and p-Smad3 enhanced (P < 0. 05). TGF-β1, initiated EMT in keloid epithelial cells by inducing the up-regulation of snail2, and TGF-β1,/Smad3 signaling pathway was involved in EMT. EMT could change the phenotype of epithelial stem cells in keloid.