Prolonged Scar-in-a-Jar: an in vitro screening tool for anti-fibrotic therapies using biomarkers of extracellular matrix synthesis.

BACKGROUND: Idiopathic pulmonary fibrosis (IPF) is a rapidly progressing disease with challenging management. To find novel effective therapies, better preclinical models are needed for the screening of anti-fibrotic compounds. Activated fibroblasts drive fibrogenesis and are the main cells responsible for the accumulation of extracellular matrix (ECM). Here, a prolonged Scar-in-a-Jar assay was combined with clinically validated biochemical markers of ECM synthesis to evaluate ECM synthesis over time. To validate the model as a drug screening tool for novel anti-fibrotic compounds, two approved compounds for IPF, nintedanib and pirfenidone, and a compound in development, omipalisib, were tested. METHODS: Primary human lung fibroblasts from healthy donors were cultured for 12 days in the presence of ficoll and were stimulated with TGF-ß1 with or without treatment with an ALK5/TGF-ß1 receptor kinase inhibitor (ALK5i), nintedanib, pirfenidone or the mTOR/PI3K inhibitor omipalisib (GSK2126458). Biomarkers of ECM synthesis were evaluated over time in cell supernatants using ELISAs to assess type I, III, IV, V and VI collagen formation (PRO-C1, PRO-C3, PRO-C4, PRO-C5, PRO-C6), fibronectin (FBN-C) deposition and α-smooth muscle actin (α-SMA) expression. RESULTS: TGF-ß1 induced synthesis of PRO-C1, PRO-C6 and FBN-C as compared with unstimulated fibroblasts at all timepoints, while PRO-C3 and α-SMA levels were not elevated until day 8. Elevated biomarkers were reduced by suppressing TGF-ß1 signalling with ALK5i. Nintedanib and omipalisib were able to reduce all biomarkers induced by TGF-ß1 in a concentration dependent manner, while pirfenidone had no effect on α-SMA. CONCLUSIONS: TGF-ß1 stimulated synthesis of type I, III and VI collagen, fibronectin and α-SMA but not type IV or V collagen. Synthesis was increased over time, although temporal profiles differed, and was modulated pharmacologically by ALK5i, nintedanib, pirfenidone and omipalisib. This prolonged 12-day Scar-in-a-Jar assay utilising biochemical markers of ECM synthesis provides a useful screening tool for novel anti-fibrotic compounds.

Depressed scar after filler injection successfully treated with pneumatic needleless injector and radiofrequency device.

Fillers are known to be associated with a number of side effects, one of the most severe being skin necrosis. The most vulnerable areas are those that are supplied by a single arterial branch; for example, the glabellar and nasolabial folds are susceptible. In this study, we report good cosmetic outcomes were produced by utilizing the pneumatic needleless injector and radiofrequency device to treat depressed scars that occurred after necrosis following filler injection. Initially, applying light-emitting diode treatment and following through with the two devices appears to have synergistic effects for scar remodeling when dealing with treatment of depressed scars with irregular borders.

Pulsed dye laser in treatment of steroid-induced atrophy.

BACKGROUND: One of the important and distressing cutaneous side effects of steroid therapy is skin atrophy, which has no definite and effective treatment. To the best of our knowledge, laser therapy for steroid-induced atrophic scars has not been investigated to date. OBJECTIVE: The aim of this study was to evaluate the efficacy and safety of pulsed dye laser in the treatment of steroid-induced atrophic scars. METHODS: In this pilot study, 15 patients with at least one atrophic patch were treated with the 585-nm pulsed dye laser at 4-week interval sessions until achieving complete improvement or until patient were lost to follow-up. Clinical outcome was assessed via standard photographic method before each treatment session and after the final visit. An independent dermatologist evaluated the photographs. RESULT: All of the patients (13 females and two males) with 25-59 years of age experienced some degree of improvement, except one patient who withdrew from the treatment after three sessions. The treatment was well tolerated. CONCLUSION: The results of our study indicated that pulsed dye laser therapy could be employed as a new method in the treatment of steroid-induced atrophic scars. Pulsed dye laser might affect the lesions through inducing collagen deposition and production of more superficial dermal elastin as well as less unidirectional collagen in clusters.

Alopecia secondary to mesotherapy.

Mesotherapy has recently become an advertised method for the treatment of different types of alopecia despite the lack of any data regarding its efficacy and possible side effects. The substances injected into the scalp include "cocktails" of natural plant extracts, homoeopathic agents, vitamins, vasodilators, and drugs that may stimulate hair growth, such as finasteride and minoxidil. We report two cases of patchy alopecia that developed after mesotherapy for the treatment of androgenetic alopecia. In the first patient, alopecia developed after injections of the heparinoid vasodilator mesoglycan; the 3-month follow-up examination revealed a small residual area of cicatricial alopecia. The second patient developed reversible alopecia after multiple scalp injections of homeopathic agents. These cases underline the possible risks of mesotherapy as a therapeutic technique for hair loss.

Anti-inflammatory and cicatrizing activities of a carbohydrate fraction isolated from sugary kefir.

Kefir is an association of microrganisms generally grown in milk, with known probiotic activities identified from its soured suspensions. Aqueous media are also able to grow kefir, but little is known about the probiotic properties of its fermented products. This work aimed to evaluate some probiotic properties of a carbohydrate fraction isolated from sugary kefir (sugary kefir carbohydrate [SKC]). Anti-inflammatory activity of the isolated fraction of carbohydrate was tested both in vitro (cellular respirometry and macrophage culture) and in vivo (50% effective dose, rat paw edema, vascular permeability, and cicatrizing test). The results indicated no significant difference for oxygen uptake or macrophage culture between control and test groups. Rat paw edema, however, showed a significant inhibitory activity by 30 +/- 4% and 54 +/- 8% (P < .001) for carrageenan and dextran, respectively. In the cicatrizing test, animals treated with SKC cream also presented less trauma after treatments as compared to the negative control group (P < .05). The overall data suggested the SKC as a natural product that could be used as a constituent of an anti-inflammatory compound.

Multifocal scalp abscess with subcutaneous fat necrosis and scarring alopecia as a complication of scalp mesotherapy.

Over the past several years, there has been a growing interest in the treatment method termed mesotherapy. Marketed for nonsurgical fat melting, skin rejuvenation, and hair regrowth, this technique has become increasingly popular and, in the public's view, it is considered to be a relatively benign intervention method. Mesotherapy was introduced over 50 years ago by M. Pistor, a French physician who utilized this technique initially as a novel analgesic therapeutic method for a variety of rheumatologic disorders. Since its introduction, the basic principal of locally injecting subcutaneous doses of varying chemicals has been expanded and is now utilized for the aforementioned cosmetic concerns. With its increased popularity, there has been an increase in the number of reported side effects resulting from mesotherapeutic intervention. We report multifocal scalp abscesses with subcutaneous fat necrosis as a direct result of mesotherapy; therefore, requiring extensive surgical repair.