Computational protocol: The Effects of Aloe vera Cream on the Expression of CD4+ and CD8+ Lymphocytes in Skin Wound Healing

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Protocol publication

[…] This study was taken in Laboratory of Pathology, Faculty of Veterinary Medicine, Gadjah Mada University. All animal procedures in this study were approved by ethical clearance committee of Gadjah Mada University, with license numbers 00052/04/LPPT/VII/2016. Aloe vera from Kalimantan, Indonesia, was used in this study. Aloe vera leafs were peeled, cut, put in a blender mix, and extracted by alcohol 70%. Aloe vera extract was made into 1% and 2% cream concentration. The cream was made from the mixture of Aloe vera extract, stearic acid, potassium hydroxide (KOH), glycerin, methyl paraben, propylparaben, and water [].Thirty-six male Sprague-Dawley rats weighing 150–200 grams were divided into four groups. All groups were shaved and exposed to round 4 mm full-thickness punch biopsy on the back under ketamine 50 mg/kg BW and xylazine 4 mg/kg BW anesthesia. Next, group I was the control group; group II was treated with topical application of 1% Aloe vera cream; group III was treated with 2% Aloe vera cream; and group IV was treated with madecassol. The treatments were given once a day, for fifteen days. Three rats from each group were euthanized with cervical dislocation on days 5, 10, and 15 to collect skin specimen. The collected specimen was stored in neutral buffer formalin 10%, dehydrated, blocked in paraffin, and cut using microtome for H&E and IHC stain. IHC stain used monoclonal antibodies for CD4+ (anti-rat CD4+, Novocastra, RTU-CD4-1F6, Cat. number PA0427) and CD8+ (anti-rat CD8+, Novocastra, RTU-CD8-295, Cat. number PA0183).The data was divided into macroscopical and microscopical data. Macroscopical data was the percentage of the wound area. Microscopical data was epidermal thickness, neutrophils infiltration, fibroblast, angiogenesis, and CD4+ and CD8+ lymphocytes. Microscopical examination was performed by ImageJ software. All data were analyzed by SPSS 16. Epidermal thickness, neutrophils infiltration, fibroblast, angiogenesis, and CD8+ lymphocytes data were analyzed with Kruskal-Wallis and Mann–Whitney U Test. CD4+ lymphocytes were analyzed with two-way ANOVA and Post hoc test. […]

Pipeline specifications

Software tools ImageJ, SPSS
Applications Miscellaneous, Microscopic phenotype analysis
Organisms Homo sapiens, Rattus norvegicus