ABSTRACT
BACKGROUND@#Hair loss is a prevalent medical problem in both men and women. Maintaining the potential hair inductivityof dermal papilla cells (DPCs) during cell culture is the main factor in hair follicle morphogenesis and regeneration. Thepresent study was conducted to compare the effects of different concentrations of human hair outer root sheath cell(HHORSC)and platelet lysis (PL) exosomes to maintain hair inductivity of the human dermal papilla cells (hDPCs). @*METHODS@#In this study, hDPCs and HHORSCswere isolated from healthy hair samples. Specific markers of hDPCs (versican,a-SMA) and HHORSCs (K15) were evaluated using flow cytometric and immunocytochemical techniques. The exosomes wereisolated fromHHORSCsand PL with ultracentrifugation technique.Western blot was used to detect specific markers of HHORSCsand PL exosomes. Particle size and distribution of the exosomes were analyzed by NanoSight dynamic light NanoSight DynamicLight Scattering. Different methods such as proliferation test (MTS assay), migration test (Transwell assay) were used to evaluatethe effects of different concentrations of exosomes (2,550,100 lg/ml) derived from HHORSC and PL on hDPCs. Expression ofspecific genes in the hair follicle inductivity, including ALP, versican and a-SMA were also evaluated using real time-PCR. @*RESULTS@#The flow cytometry of the specific cytoplasmic markers of the hDPCs and HHORSCs showed expression ofversican (77%), a-SMA (55.2%) and K15 (73.2%). The result of particle size and distribution of the exosomes wereanalyzed by NanoSight dynamic light NanoSight Dynamic Light Scattering, which revealed the majority of HHORSC andPL exosomes were 30–150 nm. For 100 lg/ml of HHORSC exosomes, the expressions of ALP, versican and a-SMAproteins respectively increased by a factor of 2.1, 1.7and 1.3 compared to those in the control group. @*CONCLUSION@#In summary, we applied HHORSC exosomes as a new method to support hair inductivity of dermalpapilla cells and improve the outcome for the treatment of hair loss.
ABSTRACT
BACKGROUND@#Hair loss is a prevalent medical problem in both men and women. Maintaining the potential hair inductivityof dermal papilla cells (DPCs) during cell culture is the main factor in hair follicle morphogenesis and regeneration. Thepresent study was conducted to compare the effects of different concentrations of human hair outer root sheath cell(HHORSC)and platelet lysis (PL) exosomes to maintain hair inductivity of the human dermal papilla cells (hDPCs). @*METHODS@#In this study, hDPCs and HHORSCswere isolated from healthy hair samples. Specific markers of hDPCs (versican,a-SMA) and HHORSCs (K15) were evaluated using flow cytometric and immunocytochemical techniques. The exosomes wereisolated fromHHORSCsand PL with ultracentrifugation technique.Western blot was used to detect specific markers of HHORSCsand PL exosomes. Particle size and distribution of the exosomes were analyzed by NanoSight dynamic light NanoSight DynamicLight Scattering. Different methods such as proliferation test (MTS assay), migration test (Transwell assay) were used to evaluatethe effects of different concentrations of exosomes (2,550,100 lg/ml) derived from HHORSC and PL on hDPCs. Expression ofspecific genes in the hair follicle inductivity, including ALP, versican and a-SMA were also evaluated using real time-PCR. @*RESULTS@#The flow cytometry of the specific cytoplasmic markers of the hDPCs and HHORSCs showed expression ofversican (77%), a-SMA (55.2%) and K15 (73.2%). The result of particle size and distribution of the exosomes wereanalyzed by NanoSight dynamic light NanoSight Dynamic Light Scattering, which revealed the majority of HHORSC andPL exosomes were 30–150 nm. For 100 lg/ml of HHORSC exosomes, the expressions of ALP, versican and a-SMAproteins respectively increased by a factor of 2.1, 1.7and 1.3 compared to those in the control group. @*CONCLUSION@#In summary, we applied HHORSC exosomes as a new method to support hair inductivity of dermalpapilla cells and improve the outcome for the treatment of hair loss.
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
As a biological tissue material, amniotic membrane [AM] has low immunogenicity and to date has been widely adopted in clinical practice. However, some features such as low biomechanical consistency and rapid biodegradation is limited the application of AM. Therefore, in this study, we fabricated a novel three-dimensional [3D] spongy scaffold made of the extracellular matrix [ECM] of denuded AM. Due to their unique characteristics which are similar to the skin, these scaffolds can be considered as an alternative option in skin tissue engineering. In this experimental study, cellular components of human amniotic membrane [HAM] were removed with 0.03% [w/v] sodium dodecyl sulphate [SDS]. Quantitative analysis was performed to determine levels of Glycosaminoglycans [GAGs], collagen, and deoxyribonucleic acid [DNA]. To increase the low efficiency and purity of the ECM component, especially collagen and GAG, we applied an acid solubilization procedure hydrochloridric acid [HCl 0.1 M] with pepsin [1 mg/ml]. In the present experiment 1-ethyl-3-[3-dimethyl aminopropyl] carbodiimide hydrochloride [EDC]/N-hydroxysuccinimide [NHS] cross linker agent was used to improve the mechanical properties of 3D lyophilized AM scaffold. The spongy 3D AM scaffolds were specified, by scanning electron microscopy, hematoxylin and eosin [H and E] staining, a swelling test, and mechanical strength and in vitro biodegradation tests. Human fetal fibroblast culture systems were used to establish that the scafolds were cytocompatible. Histological analysis of treated human AM showed impressive removal of cellular components. DNA content was diminished after treatment [39 +/- 4.06 micro g/ml vs. 341 +/- 29.60 micro g/ml]. Differences were observed between cellular and denude AM in matrix collagen [478 +/- 18.06 micro g/mg vs. 361 +/- 27.47 micro g/mg].With the optimum concentration of 1 mM NHS/EDC ratio1:4, chemical cross-linker agent could significantly increase the mechanical property, and resistance to collagenase digestion. The results of 2, 4, 6-Trinitrobenzenesulfonic acid [TNBS] test showed that cross-linking efficiency of AM derived ECM scaffolds was about 65% +/- 10.53. Scaffolds treated with NHS/EDC cross-linker agent by 100 micro g/ml collagenase, lost 75% of their dry weight after 14 days. The average pore size of 3D spongy scaffold was 160 micro m measured from scanning electron microscope [SEM] images that it is suitable for cell penetration, nutrients and gas change. In addition, the NHS/ EDC cross-linked AM scaffolds were able to support human fetal fibroblast cell proliferation in vitro. Extracts and contact prepared from the 3D spongy scaffold of AM showed a significant increase in the attachment and proliferation of the human fetal fibroblasts cells. The extra-cellular matrix of denuded AM-based scaffold displays the main properties required for substitute skin including natural in vitro biodegradation, similar physical and mechanical characterization, nontoxic biomaterial and no toxic effect on cell attachment and cell proliferation