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.

Non-cultured melanocyte-keratinocyte transplantation for the treatment of vitiligo: a clinical trial in an Iranian population.

BACKGROUND: Non-cultured cellular grafting as a surgical procedure is indicated to treat stable vitiligo, refractory to medical treatment, and is gaining wider acceptance among dermatologists. OBJECTIVE: The aim of this study was to assess the efficacy of non-cultured melanocyte-keratinocyte transplantation (MKT) for the treatment of generalized vitiligo in Iranian patients. METHODS: In this clinical trial, a total of 14 vitiligo patches in eight patients were treated; eight patches with non-cultured MKT and six patches dermabraded alone without application of keratinocyte-melanocyte suspension. Repigmentation was compared at about 4 months post-transplantation. RESULTS: Of the eight lesions treated with non-cultured MKT, four lesions showed 96-100% repigmentation, one lesion 65-95% and three lesions 0-25%. Of the patients who showed excellent results, only one showed a post-inflammatory hyperpigmentation in recipient and control areas. Of the six control lesions, five showed failed repigmentation and one showed post-inflammatory hyperpigmentation. CONCLUSION: Non-cultured MKT is an effective method to treat stable vitiligo. Studies on larger series of vitiligo patients are required to confirm its efficacy.

The epidemiology of Candida species associated with vulvovaginal candidiasis in an Iranian patient population.

OBJECTIVES: Vulvovaginal candidiasis is a common infection among women worldwide. According to previous epidemiological studies, Candida albicans is the most common species of Candida. The prevalence of non-Candida species, however, is increasing. Identification of Candida species among the population will not only help health professionals to choose suitable antifungal treatments, but also prevent development of drug resistance. The aim of this study was to identify, using chromogenic agar medium, the Candida species associated with vulvovaginal candidiasis among a sample of the Iranian population. STUDY DESIGN: In a prospective cohort study during a two year period from March 2006 to March 2008, swab samples of vaginal discharge/secretion were taken from 200 patients admitted to the gynecology clinic of Mahdieh Hospital (Tehran, Iran) with a clinical presentation suggestive of vulvovaginal candidiasis. The isolates obtained were cultured on Sabouraud dextrose agar and chromogenic agar medium. Candida species were also identified by germ tube formation in serum, chlamydospore production on Corn Meal Agar and carbohydrate absorption using the API 20C-AUX kit. Participants were asked to complete a questionnaire investigating the risk factors associated with candidiasis. An assessment of the different species of recurrent and non-recurrent candidiasis was also made. Descriptive statistics, chi-square test, and t-test were used to analyze the data. RESULTS: A total of 191 isolates were obtained from 175 vaginal specimens. Candida albicans accounted for 67% of the strains including single and mixed infections. The other identified species were Candida glabrata (18.3%), Candida tropicalis (6.8%), Candida krusei (5.8%), Candida parapsilosis (1.6%), and Candida guilliermondii (0.5%) respectively. Mixed infection with two or more species of Candida was seen in 10.3% of patients. The most common mixed cause was the combination of Candida albicans and Candida glabrata. Participants who were sexually active and those who had orogenital sex were more likely to suffer recurrent vulvovaginal candidiasis. CONCLUSIONS: Candida albicans was the most common cause of recurrent and non-recurrent vulvovaginitis. The second most common species was Candida glabrata. This study suggests CHROMagar method as a convenient and cost effective yet reliable method to isolate the species of Candida especially in cases where more than one species is present.

Identification of leishmania species isolated from human cutaneous leishmaniasis using PCR method.

BACKGROUND: To determine the epidemiological status of cutaneous leishmaniasis outbreak, isolation and identification of the agent parasite, Leishmania, using PCR method in Gonbad-e Qabus County, north Iran, during 2006-2007. METHODS: Data were collected on the prevalence of scars and ulcers over a period of 3 months among 6990 inhabitants of five villages around Gonbad-e Qabus County, north Iran, during 2006-2007. Cultured promastigotes were identified using PCR technique. Its1 and its2 of Non Coding Transcribed region at ribosomal DNA of 46 Leishmania isolates were amplified and the PCR products were separated by electrophoresis in 1.5% agarose gel (200 mA, 140 V), visualized by staining with ethidium bromide, and photographed. RESULTS: Among 6990 inhabitants of 5 villages, 62.9% were identified as scars and 1.5% as active lesions. Individuals 11 to 20 years were the most highly infected age group. All the parasite isolates were Leishmania major. CONCLUSION: Cutaneous leishmaniasis due to L. major is endemic in Gonbad-e Qabus County, north Iran.