ABSTRACT
Objective: Human amniotic membrane [HAM] is used as a supporter for limbal stem cell [LSC] expansion and corneal surgery. The aim of study is to use HAM extracts from healthy donors to enhance proliferation of LSCs in vitro and in vivo
Materials and Methods: In this interventional experimental study, the effective and cytotoxic doses of the amniotic membrane extract eye drops [AMEED] was assessed by adding different concentrations of AMEED [0-2.0 mg/ml] to LSC cultures for 14 days. Subsequently, the expression levels of ATP-binding cassette sub-family G member 2 [ABCG2, a putative stem cell marker], cytokeratin 3 [K3, corneal maker], K12 and K19 [corneal-conjunctival cell makers] were assessed by real-time polymerase chain reaction [PCR]. In the second step, the corneal epithelium of 10 rabbits was mechanically removed, and the right eye of each rabbit was treated with 1 mg/ml AMEED [every 2 hours [group 1] or every 6 hours [group 2]]. The left eyes only received an antibiotic. The corneal healing process, conjunctival infection, degree of eyelid oedema, degree of photophobia, and discharge scores were evaluated during daily assessments. Finally, corneal tissues were biopsied for pathologic evidences
Results: In comparison to the positive control [10% foetal bovine serum [FBS]], 0.1-1 mg/ml AMEED induced LSC proliferation, upregulated ABCG2, and downregulated K3. There were no remarkable differences in the expression levels of K12 and K19 [P>0.05]. Interestingly, in the rabbits treated with AMEED, the epithelium healing duration decreased from 4 days in the control group to 3 days in the two AMEED groups, with lower mean degrees of eyelid oedema, chemosis, and infection compared to the control group. No pathologic abnormalities were observed in either of the AMEED groups
Conclusion: AMEED increases LSCs proliferation ex vivo and accelerates corneal epithelium healing in vivo without any adverse effects. It could be used as a supplement for LSC expansion in cell therapy
ABSTRACT
Objective: Dermal papilla and hair epithelial stem cells regulate hair formation and the growth cycle. Damage to or loss of these cells can cause hair loss. Although several studies claim to reconstitute hairs using rodent cells in an animal model, additional research is needed to develop a stable human hair follicle reconstitution protocol. In this study, we have evaluated hair induction by injecting adult cultured human dermal papilla cells and a mixture of hair epithelial and dermal papilla cells in a mouse model
Materials and Methods: In this experimental study, discarded human scalp skins were used to obtain dermal papilla and hair epithelial cells. After separation, cells were cultured and assessed for their characteristics. We randomly allocated 15 C57BL/6 nude mice into three groups that received injections in their dorsal skin. The first group received cultured dermal papilla cells, the second group received a mixture of cultured epithelial and dermal papilla cells, and the third group [control] received a placebo [phosphate-buffered saline [PBS-]]
Results: Histopathologic examination of the injection sites showed evidence of hair growth in samples that received cells compared with the control group. However, the group that received epithelial and dermal papilla cells had visible evidence of hair growth. PKH tracing confirmed the presence of transplanted cells in the new hair
Conclusion: Our data showed that injection of a combination of adult human cultured dermal papilla and epithelial cells could induce hair growth in nude mice. This study emphasized that the combination of human adult cultured dermal papilla and epithelial cells could induce new hair in nude mice
ABSTRACT
The aim of the study was to assess the effectiveness of vitamin D[3] [1, 25[OH][2]D[3]] treatment in IBD with regard to tumor necrosis factor-alpha [TNF-alpha] serum level and clinical disease activity index [CDAI]. Vitamin D has immune-regulatory functions in experimental inflammatory bowel disease [IBD] and vitamin D deficiency is common in IBD patients. This was a randomized clinical trial on 108 IBD patients with serum 25-OHD levels less than 30ng/ml, which divided into vitamin D and control groups. Vitamin D group received 50000 IU vitamin D[3] for 12 weeks. Before and after the study, TNF-alpha and 25-OHD serum levels were measured by ELISA method. Data were analyzed using paired t-test, chi-square test and Spearman correlation coefficient. P-values less than 0.05 were considered statistically significant. Before the intervention no significant difference was found between baseline characteristics and TNF-alpha serum level of two groups. After intervention TNF-alpha serum level reduced but this reduction was not statistically significant [p=0.07, 95% CI: -0.45 to 8.14]. The mean serum 25-OHD level of vitamin D increased from 15.54 to 67.89, which was statistically significant [p= 0.00, 95% CI: -61.40 to -43.30]. TNF-alpha level was also associated significantly with CDAI before [Spearman's rho: 0.3, p<0.0001] and after [Spearman's rho: 0.27, P=0.01] intervention. Oral supplementation vitamin D[3] significantly increased serum vitamin D levels and insignificantly reduced serum TNF-alpha level. More studies with larger samples would be beneficial to assess vitamin D[3] supplementation efficient effect in IBD
ABSTRACT
Hypertrophic scar involves excessive amounts of collagen in dermal layer and may be painful. Nowadays, we can't be sure about effectiveness of procedure for hypertrophic scar management. The application of stem cells with natural scaffold has been the best option for treatment of burn wounds and skin defect, in recent decades. Fibrin glue [FG] was among the first of the natural biomaterials applied to enhance skin deformity in burn patients. This study aimed to identify an efficient, minimally invasive and economical transplantation procedure using novel FG from human cord blood for treatment of hypertrophic scar and regulation collagen synthesis. In this case series study, eight patients were selected with hypertrophic scar due to full-thickness burns. Human keratinocytes and fibroblasts derived from adult skin donors were isolated and cultured. They were tested for the expression of cytokeratin 14 and vimentin using immunocytochemistry. FG was prepared from pooled cord blood. Hypertrophic scars were extensively excised then grafted by simply placing the sheet of FG containing autologous fibroblast and keratinocytes. Histological analyses were performed using Hematoxylin and eosin [H and E] and Masson's Trichrome [MT] staining of the biopsies after 8 weeks. Cultured keratinocytes showed a high level of cytokeratin 14 expression and also fibroblasts showed a high level of vimentin. Histological analyses of skin biopsies after 8 weeks of transplantation revealed re-epithelialization with reduction of hypertrophic scars in 2 patients. These results suggest may be the use of FG from cord blood, which is not more efficient than previous biological transporters and increasing hypertrophic scar relapse, but could lead to decrease pain rate
ABSTRACT
The aim of this study was to determine the prevalence of HDV infection between HBV chronic patients referred to gastroenterology ward of Taleghani hospital Tehran, Iran and also investigating the risk factors in acquiring the HDV infection. Hepatitis B virus [HBV] and Hepatitis D virus [HDV] are major public health issues. Worldwide there are approximately 350 million individuals chronically infected with the HBV. A significant part of them, including 15 to 20 million coinfected with HDV. Hepatitis Delta virus is transferred mostly through blood and body fluids. HBV and HDV infections were evaluated by Enzyme-linked immunosorbent assay [ELISA]. Liver functional tests were assessed through auto analyzer. Patients were interviewed and data along the test results were entered into SPSS program. We used chi-square, independent t-test and logistic regression for statistical analysis. 278 [54.6%] patients of the study group were male and 231 [45.4%] were female and the mean age of patients was 40.03 +/- 14.93. From 509 patients, 39[7.7%] had anti-HDV antibody. In a uni-variable analysis, age [p=0.001], periodontal procedures [p=0.015], endoscopy [p=0.024] and colonoscopy [p=0.012] were significantly related to HDV seropositivity. After adjustment by logistic regression, age remained the only significant factor in acquiring HDV infection. We highly recommend the health care workers to strictly follow the disinfection protocols of medical instruments. Since HDV seroprevalence changes over time, regular epidemiological studies are necessary to monitor the epidemiological trend of infection
ABSTRACT
The aim of this study is to create an ex vivo model to examine the expression of two heat shock protein 70 [HSP70] family members, heat shock protein 72 [HSP72] and heat shock constitute protein 70 [HSC70], at the mRNA and protein levels in differentiating corneal cells from air exposed limbal stem cells. Limbal biopsies were cultured as explants on a cellular amniotic membrane for 14 days. The cells were then exposed to air for 16 extra additional days. The proposed expression of limbal stem cell markers [p63, ABCG2], corneal markers [K3/12, connexin 43], as well as HSP72 and HSC70 were analyzed by reverse transcription-polymerase chain reaction [RT-PCR] at the mRNA level, and by immunocytochemistry and flow cytometry at the protein level both pre and post air exposure. Fresh limbal and corneal tissues were used as control group. Air exposure decreased expression of p63 and increased expression of K3/ K12 indicating an increase in the number of corneal cells. Our data showed that HSP72 and HSC70 were expressed at the mRNA level before and after air exposure while their expression significantly increased post air exposure at the protein level. We assume HSC70 expression may be related to early and terminal stages of differentiation in cultured limbal stem cells. In addition, limbal stem cells were protected during normal development against oxidative stress thru increased HSP72 expression. These findings may have broader implications in development of therapeutic strategies for treating wound healing disorders by induction of HSPs