| It is well known that bFGF is of great significance in the field of wound repair.Because it can effectively promote wound healing and inhibit scar formation,it has good prospects for development in medical and cosmetic fields.However,most of the bFGF on the market are expressed in prokaryotic and yeast systems,which leads to high cost of separation and purification of bFGF and unstable protein.Once dissolved in solution,it must be stored at low temperature(4?)and used within 2 weeks,which is not easy to preserve.Therefore,it is of great significance to develop a new expression and purification system of bFGF,improve the drug formulation of bFGF,and make it more stable in storage at room temperature.Vegetable oil was selected as the expression vector of bFGF in our laboratory.The method of separation and purification was simple and easy to operate.The freeze-dried powder as a drug formulation can effectively prolong the storage time of the product and make the transportation and storage of the product more convenient,thus effectively reducing the production and transportation costs.To further determine the stability and dermal toxicity of linseed mustard oil lyophilized powder(oleosin-bFGF)fused with bFGF protein,as well as the development,quality and safety standard system of oleosin-bFGF lyophilized powder.In this study,the influence factors,accelerated stability and long-term stability of oleosin-bFGF freeze-dried powder were investigated to determine its storage conditions and storage time.Secondly,the acute and subchronic dermal toxicity of oleosin-bFGF lyophilized powder was preliminarily studied,and the drug safety was evaluated.The main results are as follows:1.The indexes of oleosin-bFGF lyophilized powder placed at 60?,high temperature,high humidity and strong light were detected.The appearance,microscopic examination,pH value,protein content and cell proliferation promoting activity of oleosin-bFGF freeze-dried powder decreased obviously under high temperature and humidity conditions.There was no significant change in the above indicators under illumination.The results show that oleosin-bFGF lyophilized powder is mainly unstable under high temperature and humidity.Therefore,oleosin-bFGF lyophilized powder should avoid high temperature above 40? during storage and transportation,and be packed in vacuum sealed package.2.After accelerated stability test(6 months)and long-term stability test(12 months),the appearance,microscopic examination,pH value,protein content,effect of promoting cell proliferation and aseptic test results of oleosin-bFGF freeze-dried powder did not change significantly.The results show that the freeze-dried bFGF powder has good stability at room temperature and can only be preserved at low temperature.3.It was found that even the high dose of oleosin-bFGF freeze-dried powder had no significant effect on rats after one-time skin administration.All rats showed no signs of poisoning.The activity of rats in the experimental group was normal,with almost no detectable erythema and edema.There was no significant difference in body weight and visceral ratio between the experimental group and the control group.The maximum tolerable dose of oleosin-bFGF lyophilized powder was 2000 mg/kg.4.It was found that the oleosin-bFGF freeze-dried powder had no significant effect on the general state,weight change,blood routine and biochemical parameters,organ coefficient and histopathological examination of rats even under the condition of continuous administration for 28 days.It is concluded that oleosin-bFGF freeze-dried powder is safe for skin administration and meets the requirements of OECD criteria.It is worth further development and promotion. |
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