Alopecia areata is a usually reversible hair loss condition that often occurs in clearly defined areas. The use of growth factors to treat hair loss has shown quite interesting activity in promoting hair growth. In this concept, VEGF (vascular endothelial growth factor), as a biomarker of angiogenesis, can stimulate hair growth by promoting nutrient supply to hair follicles, resulting in an increase in hair follicle diameter. The purpose of this study is to evaluate the stimulation of VEGF liposome rich local gel on hair growth and explore its toxicological performance.
method
Randomly divide the experimental subjects into three groups. The control group used Aristoflex ® The gel was used for treatment. The 1% group used the same gel with 1% VEGF, and the 3% group used the gel with 3% VEGF. Subsequently, biochemical, hematological, and histological analyses were conducted.
result
At the end of the experiment (i.e. the 15th day of VEGF treatment), the therapeutic effect can be visually observed through hair density skin microscopy analysis, and the effect can also be seen under a microscope through hair diameter analysis. The results all indicate that the hair in the VEGF group grows faster and denser compared to the control group. On the other hand, biochemical and hematological results indicate that VEGF is not completely inactive (not 100% inert).
conclusion
VEGF increases hair follicle area, but further research is needed to confirm its toxicity.
Peer review report
background
Hair loss is a general term used to describe hair loss. There are different types of hair loss, which can be caused by immunity, metabolism, or unknown reasons. The pathological problem of hair loss is also a social issue. In history, there have been various methods for treating hair loss, which can even be traced back to Egyptian papyrus records dating back to 4000 AD. Currently, numerous treatments and cosmetics for skin diseases are being developed with the goal of increasing hair density and preventing or at least slowing down hair loss. Accurately identifying the type of hair loss to be treated is crucial for achieving good clinical outcomes. At present, treatment plans are formulated based on the clinical type of hair loss that patients suffer from [4].
Recently, two new types of active cosmetics have been launched. The first category is defined as “growth factors”. These proteins are produced through genetic engineering technology, where the encoding genes are introduced into the DNA of Escherichia coli, allowing for large-scale production of these peptides. One major advantage of this method is that the recombinant protein obtained during this process is 100% homologous to human protein, reducing the risk of allergic reactions caused by the product [5]. The product undergoes a purification process to obtain pure peptides. Then, this peptide is nano encapsulated and encapsulated in liposomes to form a protective barrier, which not only increases the stability of the product, preventing it from being damaged by endogenous proteases, but also enhances its penetration ability into the skin [5].
The use of growth factors to treat hair loss has shown promising activity in promoting hair growth. Under this concept, VEGF (vascular endothelial growth factor) serves as a biomarker of angiogenesis, stimulating hair growth and increasing hair follicle diameter by promoting nutrient supply to hair follicles. Research has shown that compared to normal hair follicles, the expression of VEGF in human hair loss follicles is significantly reduced [6]. Studies have observed that minoxidil, as one of the approved drugs for treating hair loss, can promote hair growth by upregulating the expression of VEGF in dermal papilla cells [7].
Based on the above description, the purpose of this study is to quantitatively evaluate the stimulation of VEGF liposome rich local gel on hair growth, as well as its toxicological performance in changes of biochemical and hematological parameters, and to conduct histopathological analysis.
method
This study has been approved by the Ethics and Animal Experimentation Committee of FMABC (protocol number 001/2010). The feeding and handling of animals follow the guidelines of the National Health Research Institute. Select 18 hamsters (Mesocrecetus auratus) and randomly divide them into three groups, with 6 hamsters in each group. The control group used Aristoflex ® For gel treatment, 1% group used the same gel with 1% VEGF, and 3% group used the gel with 3% VEGF. All animals are shaved off their back hair with a regular razor before treatment. Apply 650 μ L of the formulation for local treatment on an area of 2 cm ², twice daily for 15 days.
On the 13th day, shave the animals again and use FotoFinder dermoscope ® Medicam ® Take back photos for 500. Repeat the operation on the 14th and 15th days. All collected images are from Dermoscope ® 3.8 Conduct analysis using software.
At the end of the experiment (day 15), animals were anesthetized with a mixture of xylazine (10mg/kg/ip) and ketamine (100mg/kg/ip). Collect blood from the aorta for hematological and biochemical analysis. After animal sacrifice, skin was collected and fixed in 10% phosphate buffered formalin.
Hematology and Biochemical Analysis
Hematological analysis in ABX Pentra 60 Horiba ® Conducted in a cell counter. Microscope slides are also used for qualitative analysis of blood cells. Using Biot é cnica ® The reagent kit evaluates aspartate aminotransferase (AST), alanine aminotransferase (ALT), gamma glutamyl transferase (GGT), and alkaline phosphatase. All analysis procedures were conducted in accordance with the standards of clinical laboratory analysis.
Organizational analysis
Perform graded dehydration and paraffin embedding treatment on fixed specimens. Cut 5 μ m thick paraffin sections and stain with hematoxylin eosin (HE) for image analysis. These images were captured using a NIKON ECLIPSE E800 microscope. Measure epidermal, dermal, and subcutaneous thickness in digital images of HE stained sections using Micrometrics Plus software. In HE stained sections, the thickness of hair follicles was measured at the level of the maximum diameter of the hair follicles clearly visible in the papilla (50 hair follicles were measured at each time point). Using ImageJ ® Software quantifies hair diameter.
statistical analysis
Statistical analysis was conducted using SPSS 17.0 software. All variables were subjected to descriptive analysis. Quantitative values are represented by median, minimum, and maximum values. For the comparison of the three groups, non parametric Kruskal Wallis test was used, and multiple comparisons were conducted using Dunn test. A level of p<0.05 is considered statistically significant.
result
Skin microscopy images and capillary density: FotoFinder dermoscope ® Medicam ® 500 and dermscope ™ The 3.8 software is used to analyze the hair density of three groups (control group, 1% VEGF group after 15 days of treatment, and 3% VEGF group). Figure 1 shows the FotoFinder dermscope ® Medicam ® Hair density evaluated at 500. Table 1 shows the analysis area: 0.65 cm ²; Density unit: 1/cm ².
