Cultured allogeneic fibroblast injection vs. fibroblasts cultured on amniotic membrane scaffold for dystrophic epidermolysis bullosa treatment.

BACKGROUND: Different methods of fibroblast application have been examined to treat recessive dystrophic epidermolysis bullosa (RDEB). OBJECTIVES: To compare the effects of intradermal injection of cultured allogeneic fibroblasts in healing RDEB wounds with those of fibroblasts seeded on amniotic membrane scaffolds (FAMS) or standard wound care (SWC) with Vaseline® gauze as controls. METHODS: Seven patients were recruited, and seven wounds were assessed in each patient: three wounds were treated with injection of intradermal fibroblasts, three were treated with FAMS and one was dressed with SWC. Changes in wound size were assessed after 2 and 12 weeks of treatment. Qualitative wound scores (QWS) were used to assess wound severity. Additionally, biopsies and antigen mapping were performed to detect type VII collagen in the dermoepidermal junction. RESULTS: In both treated areas, the QWS and wound size were significantly decreased (P < 0·001), whereas there were no changes in the control group (P = 0·29). After 2 and 12 weeks of treatment, the wound size was significantly decreased in wounds that were treated with fibroblast injection compared with those treated with FAMS (P < 0·001); but no significant changes were found in the control group. CONCLUSIONS: Fibroblast injection has been shown to promote healing of RDEB wounds and is superior to FAMS or the control treatment.

Serum 25-hydroxy vitamin D level in patients with pemphigus and its association with disease severity.

BACKGROUND: There is a growing number of studies suggesting a key role for vitamin D in the pathogenesis and progression of autoimmune disorders. AIM: To determine the serum levels of 25-hydroxy vitamin D [25(OH)D] in patients with pemphigus and association of 25(OH)D with clinical features. METHODS: We enrolled 52 patients with pemphigus and 56 age- and sex-matched healthy controls (HCs). Only patients newly diagnosed with pemphigus or patients who had relapsed after discontinuing their treatment for > 6 months were included. Serum 25(OH)D and parathyroid hormone (PTH) levels were measured. Data on demographics, body mass index (BMI), disease severity (based on Pemphigus Area and Activity Score; PAAS) and involved surface body area (BSA) were obtained. RESULTS: Suboptimal levels of vitamin D (< 30 ng/mL) were observed in 78.8% and 91.0% of patients and HCs, respectively (P = 0.13). Univariate and multivariate ordinal logistic regression models showed that low vitamin D status was more likely to occur in patients with higher BSA (OR = 1.07, 95% CI 1.01-1.13) and those with higher cutaneous (OR = 1.36, 95% CI 1.11-1.66) and total (OR = 1.24, 95% CI 1.08-1.41) PAAS. However, there was no significant association between vitamin D levels and presence of pemphigus, season of sampling, age, BMI or smoking habit. CONCLUSION: Vitamin D deficiency is common both in patients with pemphigus and in HCs. Patients with more severe disease (higher PAAS) are likely to have lower vitamin D levels.

Acellular amniotic membrane: an appropriate scaffold for fibroblast proliferation.

BACKGROUND: Various materials have been investigated as possible skin substitutes to repair skin defects such as burns. Because of its unique characteristics, acellular amniotic membrane seems to provide a good scaffold for cell cultures. AIM: To investigate the proliferation of fibroblasts on an amniotic membrane scaffold, and the preparation of a temporary skin substitute using this method. METHODS: Neonatal foreskin tissue was harvested after circumcision and used for isolation of skin fibroblasts. The skin sample was refrigerated in cell-culture solution, and later treated with trypsin, minced, and incubated in the same solution at 37 °C with in an atmosphere of 95% O2 /5% CO2 . The confluent cultures were treated with trypsin, and fibroblasts were subcultured up to the 10th passage. Cells were tested for microbial contamination, presentation of major histocompatibility complex, and karyotype changes. Amniotic membrane was harvested after elective caesarean section from donors who had been screened for infection. The membrane was washed and then subjected to three freeze-thaw cycles, before having the cells removed. The fibroblasts were seeded onto the scaffold, and after 24 h, the prepared skin substitute was ready. This was examined under electron microscopy. RESULTS: The skin substitute showed excellent growth of fibroblasts on the amniotic membrane scaffold. CONCLUSIONS: Fibroblasts had excellent adherence to and viability on the acellular amniotic membrane, which seems to provide an acceptable temporary skin substitute that can be used for wounds.