UNASSIGNED: Keloids form as a pathological response to skin wound healing, and their etiopathology is poorly understood. Myofibroblasts, which are cells transformed from normal fibroblasts, are believed to contribute to pathological scar formation in wounds.
UNASSIGNED: We carried out a double-blinded randomized controlled trial (RCT) comparing the efficacy of intralesional 5-fluorouracil (5-FU) and triamcinolone (TAC) injections in treating keloids. A total of 43 patients with 50 keloids were treated with either intralesional TAC or 5-FU injections, and their clinical response was evaluated. Biopsies were collected before, during, and after injection therapy from the active border of a keloid. To understand the role of myofibroblasts in keloids, we conducted an immunohistochemical analysis to identify myofibroblasts [α-smooth muscle actin (αSMA)] from the biopsies. We first defined the three histologically distinct regions-superficial, middle, and deep dermis-in each keloid.
UNASSIGNED: We then demonstrated that myofibroblasts almost exclusively exist in the middle dermis of the keloids as 80% of the cells in the middle dermis were αSMA positive. However, both the percentage of myofibroblasts as well as the area covered by them was substantially lower in the superficial and deep dermis than in the middle dermis of the keloids. Myofibroblasts do not predict the clinical response to intralesional injection therapies. There is no difference in the myofibroblast numbers in keloids or in the induced change in myofibroblasts between the responders and non-responders after treatment.
UNASSIGNED: This study demonstrates that myofibroblasts reside almost exclusively in the middle dermis layer of the keloids, but their numbers do not predict the clinical response to intralesional injection therapies in the RCT.

Background Plaque myofibroblasts are critical players in the initiation and advancement of atherosclerotic disease. They are involved in the production of extracellular matrix, the formation of the fibrous cap, and the underlying lipidic core via modulation processes in response to different environmental cues. Despite clear phenotypic differences between myofibroblast cells and healthy vascular smooth muscle cells, smooth muscle cells are still widely used as a cellular model in atherosclerotic research. Methods and Results Here, we present a conditioned outgrowth method to isolate and culture myofibroblast cells from plaques. We obtained these cells from 27 donors (24 carotid and 3 femoral endarterectomies). We show that they keep their proliferative capacity for 8 passages, are transcriptionally stable, retain donor-specific gene expression programs, and express extracellular matrix proteins (FN1, COL1A1, and DCN) and smooth muscle cell markers (ACTA2, MYH11, and CNN1). Single-cell transcriptomics reveals that the cells in culture closely resemble the plaque myofibroblasts. Chromatin immunoprecipitation sequencing shows the presence of histone H3 lysine 4 dimethylation at the MYH11 promoter, pointing to their smooth muscle cell origin. Finally, we demonstrated that plaque myofibroblasts can be efficiently transduced (>97%) and are capable of taking up oxidized low-density lipoprotein and undergoing calcification. Conclusions In conclusion, we present a method to isolate and culture cells that retain plaque myofibroblast phenotypical and functional capabilities, making them a suitable in vitro model for studying selected mechanisms of atherosclerosis.

UNASSIGNED: Oral squamous cell carcinoma (OSCC) is the most common malignancy of the oral cavity, with multifactorial etiopathogenesis. Data from the past literature suggest that myofibroblasts (MFs) can also contribute significantly to the pathogenesis of the disease. Hence, the present study was undertaken for assessing the expression of MF in well-differentiated OSCC (WDOSCC), moderately differentiated OSCC (MDOSCC), poorly differentiated OSCC (PDOSCC) and healthy controls by immunohistochemistry using alpha-smooth muscle actin (α-SMA) antibody.
UNASSIGNED: Forty cases each of WDOSCC, MDOSCC, PDOSCC and healthy controls were included. 4-μm thick sections from each tissue sample were stained with routine hematoxylin and eosin as well as immunohistochemically using α-SMA. Among different grades of OSCC, expression of MFs was compared. All the results were subjected to statistical analysis.
UNASSIGNED: While comparing the expression of MFs in between different grades of OSCC, nonsignificant results were obtained. While comparing the expression of MF in between OSCC cases and normal controls, significant results were obtained.
UNASSIGNED: MFs are one of the vital pathogenetic components in OSCC cases in predicting their invasive behaviors. We advocate the use of MFs as a stromal marker for visualizing invasion and progression in OSCC patients.

For the optimal management of patients with both allograft kidneys and native kidney diseases, the recognition of the histological features associated with older age is important. This is because most pathological findings are non-specific. Central fibrous areas (CFAs) have recently been proposed to be age-related. However, the components of CFAs and whether CFAs are observed in various kidney diseases remain undetermined. This cross-sectional study was undertaken to clarify the histological features, epidemiology, and clinicopathological features of CFAs.
One hundred and one consecutive kidney needle biopsy specimens were retrospectively collected from seven facilities in the Hokuriku region and diagnosed at the Kanazawa University Hospital in 2015. First, the components of CFAs were analyzed using normal histostaining, immunostaining, and electron microscopy. Second, the patients were divided into two groups (CFA [+] or CFA [-]) according to the presence of CFA in the obtained samples. Clinical and histological features were compared between the two groups, and factors associated with CFA formation were determined using univariate and multivariate analyses. The number of CFAs per specimen was counted in the CFA (+) group. Third, the presence of myofibroblasts in CFA was examined by immunostaining.
CFAs were observed in 56 of 101 patients (55.4%) with various kidney diseases. CFAs consist of fibrillar collagens (collagen I and III) in addition to non-fibrillar collagens (collagen IV and VI), as confirmed by electron microscopy. Clinically, the CFA (+) group was older and had a significantly higher prevalence of hypertension and hyperlipidemia than the CFA (-) group. Histologically, elastofibrosis of the interlobular artery, arteriolar hyalinosis, and membranous nephropathy were significantly more evident in the CFA (+) group than in the CFA (-) group. Multivariate analysis revealed that older age was the sole factor associated with CFA formation. Finally, 27 of 58 (46.6%) CFA-containing glomeruli in 26 cases included alpha-smooth muscle actin-positive cells in or adjacent to the CFA.
CFAs consist of fibrous collagens in addition to matrix collagens. CFA formation is associated with older age and was observed in various kidney diseases.

To clarify the characteristics and origin of the cellular components in atherosclerosis, carotid atherosclerotic plaques (CAPs) of four patients were studied by light microscopy using hematoxylin-eosin, Congo red and alpha-smooth-muscle actin stains, and by transmission electron microscopy of different regions of CAPs. By light microscopy, CAPs were composed of 1) a fibrous cap; 2) an atherosclerotic core presenting focal fibrosis, neovascularization, hemorrhage, necrosis, chondrification and ossification; and 3) a basal band composed of a hyperplasic pseudo-media and affected tunica media. Ultrastructurally, the CAPs contained a diversity of cells including fibroblasts, myofibroblasts, osteochondrocytes, vascular smooth-muscle cells, foam cells and other myoid cells characterized by varied features of the above mentioned cells. The results indicated that CAPs were derived from a proliferation of multipotential mesenchymal stem cells, leading to the presence of degenerated foam cells and lipid-laden cells.

BACKGROUND: Dermatomyofibroma (DMF) is a rare, benign tumour that is little-known among clinicians. However, it has typical clinical, histological and immunohistochemical features that distinguish it from other fibrous tumours.
METHODS: We report herein on the clinical, histological and immunohistochemical aspects of eight cases of DMF identified between 2008 and 2019 at the dermatopathology laboratory of Strasbourg.
RESULTS: Five men and three women of average age at diagnosis of 21 years and 9 months (range: 9 to 54 years) were included. Lesions ranged in size from 1 to 11cm. Most cases involved the upper body (6 cases), with one case on the abdomen and one on the side. The lesions presented as a solitary asymptomatic red or reddish brown nodule or plaque that gradually developed. The plaques were hard and caused functional discomfort on movement of the neck. Well-circumscribed spindle cell proliferation was noted in the reticular dermis parallel to the epidermis, without mitotic figures or cytological atypia. The subcutis was infiltrated in 5 cases. Expression of calponin was positive in all cases but one, while that of caldesmon, PS100 and desmin was negative. Expression of smooth muscle actin was positive in 2 cases, and both cases were also positive for stromylesin-3. CD34 was positive in 2 cases.
CONCLUSIONS: DMF is an extensive tumour capable of attaining large diameters and must be completely excised. The main differential diagnoses of DMF are dermatofibrosarcoma protuberans, dermatofibroma, fibrous hamartoma, myofibromatosis and cheloid. It can be identified based on various factors, whether clinical (young age, extensive lesion), histological (horizontal proliferation in the reticular dermis) or immunohistochemical (positive expression of calponin).

OBJECTIVE: To investigate, in vivo and in vitro, the fibroblast-to-myofibroblast transition in patients with hypermobile Ehlers-Danlos Syndrome (EDS). To analyze the dermis of patients with classical form of EDS (cEDS) and with hEDS, to identify qualitative and/or quantitative differences in ECM component and ultrastructural changes in collagen.
METHODS: Seven subjects, aged over 18, two with cEDS and five with hEDS underwent two skin biopsy. One sample was prepared for transmission electron microscopy (TEM), the other for immunofluorescence. The diameter of collagen fibers was measured with TEM. Fibrils were analyzed in four patients: the two with cEDS and two with hEDS. For each patient, the diameter of n=250 collagen fibrils was measured. αSMA was used as specific marker for myofibroblast to highlight their presence in vivo in the skin of patients with hEDS.
RESULTS: IF observation could not assess an increased expression of αSMA in hEDS patients, which showed no statistical difference compared to classic form patients. The major result from the analysis of TEM images is the clear difference in ECM composition between the two forms of EDS: ECM in hEDS is optically more dense and more prominently composed of elastic fibers.
CONCLUSIONS: Our study provides the following important evidence: 1) the absence in vivo of dermal fibroblasts in patients with hEDS, demonstrated by αSMA negativity; 2) the presence of statistically significant changes in the diameter of collagen fibrils between the classic and the hypermobile forms.

Background. Oral squamous cell carcinoma (OSCC) is among the ten most frequent malignant tumors, with SCC accounting for 94% of oral malignancies. Myofibroblasts and macrophages are multifunctional cells that have a crucial role in the biological behavior of these tumors. This study aimed to comparatively evaluate the frequency of myofibroblasts and macrophages between oral and cutaneous squamous cell carcinomas. Methods. Sixty paraffin blocks, consisting of 20 cases of OSCC, 20 cases of CSCC, 10 cases of normal skin, and 10 cases of normal oral mucosa, were selected for this descriptive-analytical cross-sectional study. To evaluate the prevalence of myofibroblasts, α-SMA staining and CD163 markers for macrophages were used. In this study, the data were analyzed with Wilk-Shapiro test and t-test using SPSS 19. Statistical significance was set at P<0.05. Results. The mean myofibroblast scores in CSCC and OSCC were 20.05 and 20.95, respectively, with no significant difference between the means (P>0.05). The mean macrophage scores in the skin and oral cavity were 28.125 and 49.67, respectively, with a statistically significant difference (P<0.05), indicating that the mean oral macrophage score was significantly higher than that in the skin. There was no significant difference between the presence and accumulation of macrophages and myofibroblasts between the oral and cutaneous SCCs; however, the intensity of accumulation and color pattern in OSCC and CSCC were higher than those in the normal skin and mucosa (P<0.05). Conclusion. According to the results of this study, it appears the biological behavior of OSCC and CSCC does not depend on myofibroblasts, and other factors might be involved.

BACKGROUND: Myofibroblasts (MFs) are fibroblasts with smooth muscle-like features characterized by the presence of a contractile apparatus. Alpha-smooth muscle actin (α-SMA) is the actin isoform that predominates within vascular smooth muscle cells and plays an important role in fibrogenesis. MFs are metabolically and morphologically distinctive fibroblasts expressing α-SMA, and their activation plays a key role in development of the fibrotic response.
OBJECTIVE: The aim of this study is to demonstrate the frequency, distribution and expression of α-SMA-positive MFs in odontogenic keratocyst (OKC), dentigerous cyst (DC) and ameloblastoma and correlate it to their aggressive biological behavior.
METHODS: A retrospective study of 45 diagnosed cases, which includes 15 cases of OKC, 15 cases of DC and 15 cases of ameloblastoma, was undertaken to demonstrate expression of α-SMA retrieved from archives of our department.
METHODS: α-SMA mouse anti-human antibody and horseradish peroxidase detection system were used in this study.
METHODS: Descriptive statistical analysis and ANOVA test were used for statistical analysis.
RESULTS: The difference in mean α-SMA count was found to be statistically significant between ameloblastoma and DC group (P < 0.001) as well as OKC and DC group (P < 0.001). No significant difference is observed between ameloblastoma and OKC group (P > 0.05). Results showed that mean number of stromal MFs in OKC and ameloblastoma were significantly higher than DC.
CONCLUSIONS: The present study has shown that the mean number of MFs was higher in OKC and ameloblastoma, while the mean number of MFs in DC was quite low and significantly different from that of ameloblastoma and OKC.

BACKGROUND: Invasion of epithelial cells into the connective tissue brings about massive morphological and architectural changes in the underlying stroma. Myofibroblasts reorganize the stroma to facilitate the movement of tumor cells leading to metastasis. The aim of this study was to determine the number and pattern of distribution of myofibroblasts and the qualitative and quantitative change that they cause in the collagen present in the stroma in various grades of oral squamous cell carcinoma (OSCC).
METHODS: The study was divided into two groups with group I (test group, 65 cases) consisting of 29 cases of well-differentiated squamous cell carcinoma, 25 moderately differentiated SCC, and 11 poorly differentiated SCC, and group II (control group) consisting of 11 cases of normal mucosa. Sections from each sample were stained with anti-α-smooth muscle actin (α-SMA) antibodies, hematoxylin and eosin, and Picrosirius red. Several additional sections from each grade of OSCC were stained with Masson\'s trichrome to observe the changes in collagen. For the statistical analysis, Fisher\'s exact test, Tukey\'s post hoc honest significant difference test, ANOVA, and the chi-square test were used, and p < .05 was considered statistically significant.
RESULTS: As the tumor stage progressed, an increase in the intensity α-SMA expression was seen, and the network pattern dominated in more dedifferentiated carcinomas. The collagen fibers became thin, loosely packed, and haphazardly aligned with progressing cancer. Additionally, the mean area fraction decreased, and the fibers attained a greenish yellow hue and a weak birefringence when observed using polarizing light microscopy.
CONCLUSIONS: Myofibroblasts bring about numerous changes in collagen. As cancer progresses, there isincrease in pathological collagen,which enhances the movement of cells within the stroma.