Adipose Stem Cells with Conditioned Media for Treatment of Acne Vulgaris Scar

This study was to investigate the effect of subcutaneous injection of the adipose stem cells (ASCs) with conditioned media (CM) in the treatment of acne vulgaris scar. We used Adult male New Zealand white rabbit ears as an animal model and induced acne formation by Kignman method. Adipose tissue was isolated and harvested from the scapula of rabbits, and ASCs were cultured and expanded until passage 1. There have four groups in our experiment, include phosphate buffered saline (PBS), ASCs with PBS (ASC + PBS), CM, and ASCs with CM (ASC + CM) group. This solution of 0.6 ml injected to subcutaneous in each group. ASC + PBS and ASC + CM groups were containing ASCs of 5.0 × 106 cells/ml. We analyzed the treatment of 4 groups to scar tissue after 2 and 4 weeks by hematoxylin and eosin stain, immunohistochemistry, and RNA expression level of tumor necrosis factor-α (TNF-α), interleukin-1α (IL-1α), and matrix metalloproteinase-2 (MMP-2). Also, the expression of keratin 16 (K16) was detected by western blot analysis. H&E stain showed that infiltration of inflammation cells was significantly reduced at 2 and 4 weeks, as well as re-epithelialization was improved in the ASC + CM group. The ASC + CM gourp was reduced both expression levels of TNF-α, IL-1α, and MMP-2 and K16 protein level. In conclusion, the ASCs with CM has a significant curative effect on acne vulgaris scar, more to the point, the CM has a key role on treatment. It could be applied to a therapeutic approach to regenerate to treat acne vulgaris scar.

Platycodin D May Improve Acne and Prevent Scarring by Downregulating SREBP-1 Expression Via Inhibition of IGF-1R/PI3K/Akt Pathway and Modulating Inflammation with an Increase in Collagen

BACKGROUND: Although many therapeutic agents have been developed, only a few drugs are known to target multiple pathogenic factors in the treatment of acne. OBJECTIVE: The purpose of this study was to identify a new drug candidate, platycodin D, which is a substance extracted from the root of Platycodon grandiflorum. METHODS: Using western blotting and Cell Counting Kit-8 assay, we studied the effects of platycodin D on SEB-1 sebocytes, fibroblasts, and keratinocytes. We investigated its effects in view of lipogenesis, collagen production, anti-inflammatory activity, and dyskeratinization. RESULTS: In SEB-1 sebocytes, platycodin D showed a sebosuppressive effect by downregulating ERK and insulin- like growth factor-1R/PI3K/Akt/sterol-regulatory element binding protein-1 signaling pathways. In addition, adiponectin, one of the adipokines responsible for sebum production, was decreased in platycodin D-treated SEB-1 sebocytes. In fibroblasts, platycodin D increased collagen production and reduced inflammation by inhibiting nuclear factor kappa B and matrix metalloproteinases. Platycodin D also showed anti-inflammatory effects on keratinocytes. It also suppressed keratin 16 expression induced by lipopolysaccharide. Furthermore, platycodin D showed no cytotoxicity on both SEB-1 sebocytes and fibroblasts. CONCLUSION: Our data demonstrate the clinical feasibility of platycodin D for acne treatment and the prevention of acne scarring by sebosuppressive and anti-inflammatory effects, as well as through an increase in collagen levels.

Is Ki-67, keratin 16, involucrin, and filaggrin immunostaining sufficient to diagnose inflammatory linear verrucous epidermal nevus? A report of eight cases and a comparison with psoriasis vulgaris

Abstract: Inflammatory linear verrucous epidermal nevus and linear psoriasis are sometimes hard to differentiate clinically and pathologically. Although immunohistochemical expression of keratin 10 (K10), K16, Ki-67, and involucrin may be useful for differentiating both entities, these results have been reported in only a few cases. We collected data from 8 patients with inflammatory linear verrucous epidermal nevus, 11 with psoriasis vulgaris, and 8 healthy controls and evaluated immunohistochemical expression of Ki-67, K16, involucrin, and filaggrin among them. Ki-67 and K16 overexpression was similar in inflammatory linear verrucous epidermal nevus and psoriasis vulgaris compared with normal skin. Although staining for involucrin showed discontinuous expression in parakeratotic regions in 4 inflammatory linear verrucous epidermal nevus cases, it was continuous in the other 4 cases and in all psoriasis vulgaris cases. Filaggrin expression was present in hyperkeratotic regions but scarce in parakeratotic areas in both inflammatory linear verrucous epidermal nevus and psoriasis vulgaris. The immunostaining pattern of Ki-67, K16, involucrin, and filaggrin may be insufficient to discriminate inflammatory linear verrucous epidermal nevus from psoriasis vulgaris.

Up-regulation of Hsp72 and keratin16 mediates wound healing in streptozotocin diabetic rats

BACKGROUND: Impaired wound healing is a complication of diabetes and a serious problem in clinical practice. We previously found that whey protein (WP) was able to regulate wound healing normally in streptozotocin (STZ)-dia-betic models. This subsequent study was designed to assess the effect of WP on heat shock protein-72 (Hsp72) and keratin16 (Krt16) expression during wound healing in diabetic rats. METHODS: WP at a dosage of 100 mg/kg of body weight was orally administered daily to wounded normal and STZ-diabetic rats for 8 days. RESULTS: At day 4, the WP-treated diabetic wound was significantly reduced compared to that in the corresponding control. Diabetic wounded rats developed severe inflammatory infiltration and moderate capillary dilatation and regeneration. Treated rats had mild necrotic formation, moderate infiltration, moderate to severe capillary dilatation and regeneration, in addition to moderate epidermal formation. Hsp72 and Krt16 densities showed low and dense activity in diabetic wounded and diabetic wounded treated groups, respectively. At day 8, WP-treatment of diabetic wounded animals revealed great amelioration with complete recovery and closure of the wound. Reactivity of Hsp72 and Krt16 was reversed, showing dense and low, or medium and low, activity in the diabetic wounded and diabetic wounded treated groups, respectively. Hsp72 expression in the pancreas was found to show dense reactivity with WP-treated diabetic wound rats. CONCLUSION: This data provides evidence for the potential impact of WP in the up-regulation of Hsp72 and Krt16 in T1D, resulting in an improved wound healing process in diabetic models.

Comparative Analysis of the Expression of Involucrin, Filaggrin and Cytokeratin 4, 10, 16 in Cholesteatoma

BACKGROUND AND OBJECTIVES: The aim of this study is to determine whether the hyperproliferative and hyperkeratotic characters of cholesteatoma are associated with differentiation of keratinocytes in cholesteatoma by examining the localization of marker proteins, such as involucrin, filaggrin, and cytokeratins. MATERIALS AND METHODS: Immunohistochemical study was carried out in 30 cholesteatoma tissues and 10 retroauricular skins to examine the expression of involucrin, filaggrin, cytokeratin 4, 10 and 16. The staining results were graded as negative, weakly positive (70%). RESULTS: Involucrin was strongly expressed in upper spinous, granular, and corneal layer of cholesteatoma. Filaggrin was strongly expressed in granular and corneal layer of cholesteatoma. Cytokeratin 4 was expressed in basal layer of retroauricular skin, but occasionally expressed in suprabasal layer of cholesteatoma. Cytokeratin 10 was homogenously expressed in all suprabasal layer of retroauricular skin, whereas pattern of shift to surface layer was showed in cholesteatoma. Cytokeratin 16 was moderately expressed at suprabasal layer in cholesteatoma. CONCLUSIONS: It can be suggested that early differentiation of suprabasal layer may lead to hyperdifferentiation and hyperkeratosis. Different expression of cytokeratins possibly indicates the altered differentiation of cholesteatoma.

Comparative Analysis of the Expression of Involucrin, Filaggrin and Cytokeratin 4, 10, 16 in Cholesteatoma

BACKGROUND AND OBJECTIVES: The aim of this study is to determine whether the hyperproliferative and hyperkeratotic characters of cholesteatoma are associated with differentiation of keratinocytes in cholesteatoma by examining the localization of marker proteins, such as involucrin, filaggrin, and cytokeratins. MATERIALS AND METHODS: Immunohistochemical study was carried out in 30 cholesteatoma tissues and 10 retroauricular skins to examine the expression of involucrin, filaggrin, cytokeratin 4, 10 and 16. The staining results were graded as negative, weakly positive (70%). RESULTS: Involucrin was strongly expressed in upper spinous, granular, and corneal layer of cholesteatoma. Filaggrin was strongly expressed in granular and corneal layer of cholesteatoma. Cytokeratin 4 was expressed in basal layer of retroauricular skin, but occasionally expressed in suprabasal layer of cholesteatoma. Cytokeratin 10 was homogenously expressed in all suprabasal layer of retroauricular skin, whereas pattern of shift to surface layer was showed in cholesteatoma. Cytokeratin 16 was moderately expressed at suprabasal layer in cholesteatoma. CONCLUSIONS: It can be suggested that early differentiation of suprabasal layer may lead to hyperdifferentiation and hyperkeratosis. Different expression of cytokeratins possibly indicates the altered differentiation of cholesteatoma.

The Expressions of Cytokeratin 16, Involucrin and PCNA in Clear Cell Acanthoma on Areola / 대한피부과학회지

BACKGROUND: Clear cell acanthoma usually appears as an asymptomatic nodule on the leg. It has an unusual clinical feature in that it is presented as chronic eczema on the areola. The origin of clear cell acanthoma is not yet clear, although many hypotheses have been proposed, including a benign neoplasm or an inflammatory dermatosis. OBJECTIVE: In this study, clear cell acanthoma on the areola showing clinically eczematous features were analysed by immunohistochemical techniques, using antibodies against cytokeratin 16, involucrin and PCNA and compared with psoriasis and squamous cell carcinoma. METHODS: Using the immunohistochemical method with formalin-fixed, paraffin-embedded sections, we analysed the expression of cytokeratin 16, involucrin and PCNA in biopsy specimens of 6 cases of clear cell acanthoma on the areola, 5 cases of psoriasis, and 5 cases of squamous cell carcinoma. RESULTS: The expression of cytokeratin 16 was detected in spinous and granular layers in all cases of clear cell acanthoma and psoriasis and three cases of squamous cell carcinoma. Psoriasis showed slightly higher immunoreactivity than clear cell acanthoma and squamous cell carcinoma, but this difference was not statistically significant (p=0.142). The expression of involucrin was detected in spinous and granular layers in all cases of clear cell acanthoma, psoriasis, and squamous cell carcinoma. The immunoreactivities were similar. The expression of PCNA was detected in basal and spinous layers in two cases of clear cell acanthoma, four cases of psoriasis, and five cases of squamous cell carcinoma. The expression of PCNA was higher in psoriasis and squamous cell carcinoma than in clear cell acanthoma, and this difference was statistically significant (p=0.034, p=0.004). CONCLUSION: Clear cell acanthoma on the areola may result from increased psoriasis-like inflammatory proliferation and accelerated differentiation of keratinocytes.

Hyperproliferative Characteristics in Human Deep Meatal Epidermis / 대한이비인후과학회지

In benign hyperproliferative epidermal diseases(eq. warts, psoriasis) and squamous carcinoma, some molecular markers of hyperproliferative keratinocyte such as cytokeratin 16 and PCNA were expressed predominantly. However, all healthy epidermis including the meatal epidermis are nonreactive to those molecular markers except some of thick skin. Recently, there are several reports which show unusal proliferative capacity around the annular region of the ear drum. Our study has concentrated on the characteristics of the differentiation in healthy deep meatal epidermis using immunohistochemistry with cytokeratins and PCNA. Our investigation has demonstrated that the deep meatal epidermis around the annular region in contrast to the other region of the meatus exhibited unusal proliferative capacity. This result suggests a pathology link such as invasion mechanism and hyperkeratinization between the cholesteatoma and deep meatal skin.