Development of an allogeneic cultured dermal substitute using a standard human fibroblast bank

Fibroblasts are mesenchymal cells that can be readily cultured in the laboratory and play a significant role in epithelialmesenchymal interactions, secreting various growth factors and cytokines that have a direct effect on epidermal proliferation, differentiation and formation of extracellular matrix. They have been incorporated into various tissue-engineered and used for a variety of clinical applications, including the treatment of burns, chronic venous ulcers and several other clinical applications in dermatology and plastic surgery. Isolated fibroblasts by the enzymatic process from foreskin were cultivated successively in a culture medium to establish cell banking. Foreskin and the last subcultured cells were checked for HBV, HCV, HIV, HSV I, HSV II, HTLV I, HTLV II, EBV, CMV, Treponema Pallidum, Mycoplasma sp. and Clamydia. The 1[st], 5[th] and 10[th] subcultured cells were processed for immunocytochemistry studies using a panel of monoclonal antibodies including antibodies to MHC class I and II antigens for ensuring the elimination of superficial cell antigens during cultivation. Subcultured cells were karyotyped to find any chromosomal abnormalities. The best passages were chosen for culturing on silicone sheets provided by the Iran Polymer and Petrochemical Institute. Evaluation for bacteria and viruses by molecular methods was negative. Karyotyping of cultured fibroblasts after the 10[th] passage showed some abnormalities. HLA expression was imperceptible in the cells obtained from the 10[th] sub-culture. The best passages were from 5th to 10[th] for banking and culturing on silicone sheets. Expression of HLA on fibroblast surfaces was diminished during subculturing. To prevent chromosomal abnormalities in fibroblast passaging, we should select the best colony that is expected to be chromosomally stable with the least antigenicity. In our study, the 5[th] to 10[th] sub-cultures were the best cells for the purpose of grafting and acceleration of the wound healing

Staphylococcus aureus carriage in patients with psoriasis

The aggravating role of Staphylococcus aureus is well known in atopic dermatitis but has not yet been proven in psoriasis. The role of Staphylococcus aureus superantigens is emphasized in the initiation, maintenance and complications of psoriasis. We investigated the frequency of nasal, axillary, and perineal carriage of Staphylococcus aureus [SA] in patients with psoriasis and its possible influence on the severity of the disease. One hundred patients with the clinical diagnosis of psoriasis participated in the study. Cultures of the bacterial flora were obtained from the right and left axilla and nasal nares and perineum, inoculated on standard bacterial medium [blood agar], and incubated at 37°C degrees for 48 h. One hundred patients with the clinical diagnosis of psoriasis [42% female and 58% male] comprised the study group. Mean age of the patients was 41.1 +/- 17.1 years. About 42% of the patients carried S. aureus; of these, 32% were from the nose, 13% from axilla, and 11% from the perineum. Three patients were carriers in all 3 sites. There was no significant difference in the severity of the disease between the carriers and non-carriers measured by the psoriasis area and severity index [PASI] score. According to our findings, S.aureus carriage in psoriasis had no significant influence on disease severity. It might be relevant for a subgroup of patients only when superantigen productions are found

Establishment of a primary cell culture of human fibroblast in Iran

Human fibroblasts are the part of the dermis that secrete extracellular matrix for the purpose of tissue repair. Culturing fibroblasts, which leads to formation of a monolayer of these cells, is used for treating various conditions including thermal burns and other skin defects such as diabetic and varicose vein leg ulcers. Therefore, we aimed at developing a fibroblast bank to accomplish multiple goals including skin repair in defects such as burns and ulcers and also performing various research projects on these cells in order to further study of the mechanisms involved in wound healing, rejuvenation and medication effects. We initially developed primary cultures of skin fibroblasts in a DMEM medium. These primary cultures were formed by washing and trypsinizing foreskin specimens followed by separation of epidermis from dermis and cutting the dermis into small pieces. In about 10 days, a monolayer of fibroblasts was formed. We were able to develop the fibroblast bank successfully and to initiate other projects utilizing this bank. With these cultured cells, we would be able to perform different research projects including studying the mechanisms of wound healing, rejuvenation, drug affects, inflammatory mediators, growth factors, etc. Moreover, further progress in this field will result in our independence from requesting these cells from external sources

Evaluation of the efficacy of intermittent isotretinoin doses in acne patients

Oral isotretinoin is the only treatment that has an effect on all the major etiological factors involved in acne [increased sebum production, alterations in microbial flora, hyperkeratinization of pilosebaceous duct, and inflammation]. Considering complications and relative expense, several treatment regimens have been suggested. The objective of this study was to evaluate the efficacy of intermittent doses of isotretinoin in acne patients. We made a quasi-experimental clinical trial in the acne patients without nodular and cystic lesions that were recalcitrant to conventional therapy. Therapeutic regimen included 0.5 mg/kg isotretinoin per day for a week every month; so we use 21 mg/kg as total dosage. Acne severity decreased in entire patients at the end of treatment course. After six months follow up recurrence rate was 19.3% and partially recurrence rate was 44%. It seems that intermittent isotretinoin treatment with a total dose of 21 mg/kg has led to good therpeutic results in patients without nodulocystic lesions