AbstrakPenggunaan serum otologus merupakan pendekatan terapeutik yang direkomendasikan untuk mengobati beberapa jenis penyakit yang bersifat kronis, menahun bahkan dapat bertahan seumur hidup. Serum otologus mengandung epidermal growth factor (EGF) yang dapat menstimulasi proses migrasi dan proliferasi keratinosit pada proses re-epitelisasi dalam penyembuhan luka. Penambahan hEGF pada serum otologus dilakukan sebagai upaya untuk mempercepat proses penyembuhan luka. Pengujian aktivitas dilakukan dengan mengukur proliferasi dan migrasi sel keratinosit menggunakan cell line HaCaT. Proliferasi sel diukur dengan metode Water Soluble Tetrazolium-8 (WST-8) dan migrasi sel diukur dengan metode Scratch Assay. Variasi konsentrasi hEGF yang ditambahkan adalah 0,5; 1; 5; 10; 25 dan 50 ng/mL. Terdapat perbedaan yang bermakna pada hasil proliferasi sel HaCaT yang ditambahkan hEGF dan konsentrasi optimal pada penambahan hEGF 25 ng/mL (p<0,05), sedangkan penambahan variasi konsentrasi hEGF pada serum otologus tidak menunjukkan perbedaan yang bermakna terhadap proliferasi sel HaCaT (p>0,05 Autologous Serum and human Epidermal Growth Factor (hEGF) Accelerate Keratinocyte Proliferation and Migration during Re-epithelialization Process AbstractAutologous serum is used as a therapeutic approach recommended to treat certain types of chronic diseases that can even last a lifetime. Autologous serum contains Epidermal Growth Factor (EGF), which can stimulate the migration and proliferation of keratinocytes in the re-epithelialization process in wound healing. The addition of hEGF on autologous serum is a part of efforts to accelerate the wound healing process. Tests were performed by measuring proliferation and migration activities of keratinocytes cells using HaCaT cell line. The cell proliferation was measured using Water Soluble Tetrazolium-8 (WST-8) method while the cell migration was measured using Scratch Assay method. Variations in the concentration of hEGF added were 0.5, 1, 5, 10, 25, and 50 ng / mL. There were significant differences in the results of cell proliferation in hEGF-added HaCaTcells and the optimum concentration was seen in 25 ng/mL hEGF group (p <0.05). On the contrary, the addition of various concentrations of hEGF in autologous serum resulted in no significant difference in HaCaT cell proliferation (p>0 , 05). The addition of hEGF and autologous serum showed a visible role in accelerating the pace of cell migration. Within 18 hours, the percentage of cell migration in HaCaT cells added by hEGF and autologous serum reaches 55-70% and then 65-80% within 24 hours while HaCaT cells that receive hEGF only only reaches 25-45% cell migration which increases to 35-70% within 24 hours. [MKB. 2016;48(4):205-10]

The autologous serum is widely used as therapy for epithelial cell treatment and dry eye syndrome due to its epitheliotropic properties, which are similar to tears. The serum contains human epidermal growth factor (hEGF) and retinol, which play important roles in cell proliferation and differentiation. Freeze-drying improves the stability of therapeutic proteins for long-term storage. Sucrose is known to stabilize protein during freeze-drying and storage. Also, antioxidants such as vitamin E can maintain unstable retinol levels. This study aims to investigate the effect of the addition of sucrose to preserve dried autologous serum during freeze-drying and storage, as well as the effects of the serum on keratinocyte proliferation and migration. The addition of sucrose to autologous serum could preserve the dried autologous serum components above the minimum requirement for substitute tears after 6 months of storage. The highest cell proliferation was achieved by the addition of 25 ng/ml hEGF and 1% vitamin E, while the highest migration activity was achieved by the addition of 25 ng/ml hEGF. Freeze-drying autologous serum with the addition of sucrose and storage at low temperatures can extend its shelf life. Also, the addition of exogenous hEGF and vitamin E to autologous serum can increase the proliferation and migration of keratinocyte cells.