Recombinant Human Epidermal Growth Factor Derivative For External Use,Liquid(Shenzhen Watsin Genetech Ltd.)

To determine the role of the intralesional recombinant epidermal growth factor (rEGF) in the healing and prevention of extremity amputation in advanced diabetic foot ulcer patients.

Observational study.

Department of Cardiovascular Surgery, Duzce State Hospital, Duzce, Turkey, between November 2018 and September 2019.

A total of 58 patients with diabetic foot ulcers that were treated at the study place were enrolled. The lesions were graded with Wagner Classification System. EGF (75 microg of Heberprot-P) vials were stored at +4°C and cold-chain requirements were followed. EGF 5 mL was dissolved with 0.09% saline solution; and 0.5-1 ml of the solution was injected into the tissues and edge of the lesions regularly. The data was evaluated at the end of two years of the treatment period. The primary objective was wound healing, formation of granulation tissue; and the secondary objective was the prevention of lower extremity amputation.

Diabetic foot ulcers wound healing was achieved in 93.1% (n=54) of patients with the formation of granulation tissue. The complete recovery was observed in 94.1% (n=32) of the patients who had Grade III and IV lesions. Lower extremity amputation was performed in two (3.4%) subjects. The lesions of two patients required flap surgery. The most common adverse events were tremor and syncope.

Recombinant epidermal growth factor is highly effective for the treatment of diabetic foot ulcers and prevention of extremity amputation. Intralesional rEGf provides efficient and safe wound healing/closure in patients with diabetic foot ulcers. Key Words: Amputation, Epidermal growth factor, Diabetic foot, Wound healing.

Vero cells have been widely used in the viral vaccine production due to the recommendation of the World Health Organization regarding its safety and non-tumorigenicity. The aim of this study was to describe the development a modified serum-free medium for Vero cell cultures. Two protein hydrolysates (Bacto™ soytone and Bacto™ yeast extract), vitamin C, vitamin B12, SITE liquid media supplement, and recombinant human epidermal growth factor (rEGF) were investigated as serum substitutes. A sequential experiment of fractional factorial and central composite design was applied. A modified serum-free medium obtained (named as SFM01-M) was verified. Contrary to P0, the cell yields obtained at P1, P2, and P3 decreased continuously during the verification experiments indicating that Vero cells could not adapt to SFM01-M as expected according to the empirical mathematical model. To improve cell growth after P0, protein hydrolysates, l-glutamine, and SITE liquid media supplement were further investigated. The results showed that cell yields gradually decreased from P1 to P3 when a fixed concentration of Bacto™ yeast extract (7.0 g/L) combined with various concentrations of Bacto™ soytone (0.1-7.0 g/L) in SFM01-M were used. Similarly, cell yields also gradually decreased from P1 to P3 when a fixed concentration of Bacto™ soytone (7.0 g/L) combined with various concentrations of Bacto™ yeast extract (0.1-7.0 g/L) in SFM01-M were used. However, the combination of Bacto™ soytone at 0.1 g/L and Bacto™ yeast extract at 7.0 g/L or Bacto™ soytone at 7.0 g/L and Bacto™ yeast extract at 0.1 g/L in SFM01-M could give the maximum cell yield at P3 when compared with other combinations. In addition, the addition of SITE liquid media supplement (0.1-2.0% v/v) in SFM01-M in which the concentrations of Bacto™ soytone, Bacto™ yeast extract, and l-glutamine were fixed at 0.1 g/L, 0.1 g/L, and 4.0 mM, respectively, the results showed that the cell yields obtained at P3 were not significantly different. From this study, the optimum concentrations of SFM01-M components were as follows: Bacto™ soytone (0.1 g/L), Bacto™ yeast extract (0.1 g/L), vitamin C (9.719 mg/L), vitamin B12 (0.1725 mg/L), SITE liquid media supplement (0.1-2.0% v/v), rEGF (0.05756 mg/L), l-glutamine (4.0 mM), MEM non-essential amino acids (1.0% v/v), sodium pyruvate (1.0 mM), MEM (9.4 g/L), and sodium hydrogen carbonate (2.2 g/L). However, to evaluate SFM01-M in the long-term subculture of Vero cells, the efficiency of SFM01-M will be further investigated.