Park, Yu Gil
(Department of Plastic and Reconstructive Surgery, Bucheon Hospital, Soonchunhyang University College of Medicine)
,
Lee, In Ho
(Onjin-gun Public Health Center)
,
Park, Eun Soo
(Department of Plastic and Reconstructive Surgery, Bucheon Hospital, Soonchunhyang University College of Medicine)
,
Kim, Jin Young
(Department of Plastic and Reconstructive Surgery, Bucheon Hospital, Soonchunhyang University College of Medicine)
Background Platelet-rich plasma (PRP) contains high concentrations of growth factors involved in wound healing. Hydrogel is a 3-dimensional, hydrophilic, high-molecular, reticular substance generally used as a dressing formulation to accelerate wound healing, and also used as a bio-applicable scaffo...
Background Platelet-rich plasma (PRP) contains high concentrations of growth factors involved in wound healing. Hydrogel is a 3-dimensional, hydrophilic, high-molecular, reticular substance generally used as a dressing formulation to accelerate wound healing, and also used as a bio-applicable scaffold or vehicle. This study aimed to investigate the effects of PRP and hydrogel on wound healing, in combination and separately, in an animal wound model. Methods A total of 64 wounds, with 2 wounds on the back of each nude mouse, were classified into 4 groups: a control group, a hydrogel-only group, a PRP-only group, and a combined-treatment group. All mice were assessed for changes in wound size and photographed on scheduled dates. The number of blood vessels was measured in all specimens. Immunohistochemical staining was used for the analysis of vascular endothelial growth factor (VEGF) expression. Results Differences in the decrease and change in wound size in the combined-treatment group were more significant than those in the single-treatment groups on days 3, 5, 7, and 10. Analysis of the number of blood vessels through histological examination showed a pattern of increase over time that occurred in all groups, but the combined-treatment group exhibited the greatest increase on days 7 and 14. Immunohistochemical staining showed that VEGF expression in the combined-treatment group exhibited its highest value on day 7. Conclusions This experiment demonstrated improved wound healing using a PRP-hydrogel combined treatment compared to either treatment individually, resulting in a decrease in wound size and a shortening of the healing period.
Background Platelet-rich plasma (PRP) contains high concentrations of growth factors involved in wound healing. Hydrogel is a 3-dimensional, hydrophilic, high-molecular, reticular substance generally used as a dressing formulation to accelerate wound healing, and also used as a bio-applicable scaffold or vehicle. This study aimed to investigate the effects of PRP and hydrogel on wound healing, in combination and separately, in an animal wound model. Methods A total of 64 wounds, with 2 wounds on the back of each nude mouse, were classified into 4 groups: a control group, a hydrogel-only group, a PRP-only group, and a combined-treatment group. All mice were assessed for changes in wound size and photographed on scheduled dates. The number of blood vessels was measured in all specimens. Immunohistochemical staining was used for the analysis of vascular endothelial growth factor (VEGF) expression. Results Differences in the decrease and change in wound size in the combined-treatment group were more significant than those in the single-treatment groups on days 3, 5, 7, and 10. Analysis of the number of blood vessels through histological examination showed a pattern of increase over time that occurred in all groups, but the combined-treatment group exhibited the greatest increase on days 7 and 14. Immunohistochemical staining showed that VEGF expression in the combined-treatment group exhibited its highest value on day 7. Conclusions This experiment demonstrated improved wound healing using a PRP-hydrogel combined treatment compared to either treatment individually, resulting in a decrease in wound size and a shortening of the healing period.
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제안 방법
On this basis, an experiment was carried out using an animal model under the assumption that in a combined treatment of hydrogel and PRP, hydrogel would act as an effective scaffold for growth factors and help in their maintenance and expression, thereby accelerating wound healing.
For each wound, occlusive film dressing was applied using Opsite (Smith & Nephew, London, UK) and Surgifix (Panamedic, Seoul, Korea). The dressing was replaced daily, and the wounds were photographed on days 3, 5, 7, 10, and 14 to assess their size.
To determine the number of blood vessels, structures with the shape of the inner diameter of a blood vessel were observed with a microscope in each of the different fields of 3 sites for each slide, at × 100 magnification.
The collected samples were stored by freezing them at −70°C and fixed in 10% formalin for 1 day in order to produce histopathology slides.
Thereafter, they were passed through a nitrocellulose membrane, and the first and second antibodies were reacted. Immune protein analysis was performed through chemical fluorescence analysis of the X-ray film, and the size of the expressed band was determined by measuring the area using the software program ImageJ. The values compared to the reference value of beta-actin expression were used for relative comparison of the groups,
, the concentration) may not be consistent. To overcome this limitation, a study was conducted on increasing the action time and effect by utilizing a scaffold or vehicle along with a bioactive substance such as PRP [13]; in particular, this study aimed to investigate the effect by using hydrogel as a scaffold.
However, the distance between these 2 areas was within 10 mm, with wounds largely located in the dorsum and central portions, and we believe this limitation is unlikely to have significantly affected the results. Further work including quantitative analysis and measurement is required to accurately assess the effects of VEGF expression. Although H&E staining was used in our study, another important way of identifying the number of blood vessels generated would be through the use of CD31 staining, which would provide a potential area for further research.
Therefore, a total of 64 wounds (n = 16 per group) were created. They were divided into 4 treatment method groups: the control group, to which neither PRP nor hydrogel was applied, the hydrogel-only group, the PRP-only group, and the combined-treatment group, to which both hydrogel and PRP treatment were applied. For each wound, occlusive film dressing was applied using Opsite (Smith & Nephew, London, UK) and Surgifix (Panamedic, Seoul, Korea).
After the creation of the wound, the dressing was replaced daily, and photos of the wound were taken on days 3, 5, 7, 10, and 14 at the same resolution and distance using a digital single-lens reflex camera (Nikon D90, macro-lens 50 mm, Tokyo, Japan), to reduce errors. The photos were taken with a ruler placed next to the wound to show the wound size.
대상 데이터
For this experiment, 32 9-week-old female BALB/c nude mice with a weight of 20–30 g (26 g on average) and without a thymus gland (Orientbio Inc., Sungnam, South Korea) were used.
The production of PRP required the collection of 54 mL of blood from the vein of a donor, a 30-year-old adult male. The collected blood was mixed with 6 mL of anticoagulant citrate dextrose solution.
The hydrogel that was used in this experiment was Purilon gel (Coloplast, Humlebæk, Denmark), which has an amorphous property.
Immunohistochemistry was performed to analyze vascular endothelial growth factor (VEGF) expression. The VEGF signal protein (1:1,000; Santa Cruz Biotechnology, Dallas, TX, USA) was used as a mouse monoclonal antibody, with beta-actin (1:1,000, Abcam, Cambridge, UK) used as a reference value antibody. After dissolving the tissue samples in a sodium dodecyl sulfate (SDS) sample and buffer solution, they were passed through 10% polyacrylamide gel.
데이터처리
To compare the wound sizes, change rates, and numbers of blood vessels at the different time points per group, 2-way repeated-measures analysis of variance (ANOVA) was conducted. To determine if the difference between the groups at each time point was significant, ANOVA tests were performed at each time point, and post hoc tests were conducted using the Tukey honest significant difference test. All statistical analysis was conducted using SPSS ver.
These results were derived through 2-way repeated-measures ANOVA.
At each time point, a post-hoc comparison was made using the Tukey HSD test.
To compare the wound sizes, change rates, and numbers of blood vessels at the different time points per group, 2-way repeated-measures analysis of variance (ANOVA) was conducted. To determine if the difference between the groups at each time point was significant, ANOVA tests were performed at each time point, and post hoc tests were conducted using the Tukey honest significant difference test.
성능/효과
After treatment, the differences in the average number of blood vessels in the 4 groups were found to be significant (P < 0.05), and the increase in the number of blood vessels over time was found to be significant in all groups (P < 0.05) (Table 2).
In this study, the application of PRP–hydrogel combination therapy resulted in more effective wound healing acceleration.
This study showed that wound healing was significantly accelerated in the combined-treatment (PRP and hydrogel) group compared to the PRP- and hydrogel-only groups. The degree of wound healing acceleration in the PRP-only group was significant compared to the control group, but there was no significant difference between the PRP- and hydrogel-only groups in this regard.
참고문헌 (17)
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