| Basic Fibroblast Growth Factor [bFGF] is a peptide factor that strongly promotes cell growth, and can exert extensive effects in various cell growth, development and function. BFGF can promote the mitosis of most mesoderm-derived and ectoderm-derived cells, and change the non-mitosis function of a large amount of cells. Its biological effects are mainly expressed in: (1) promoting the formation of blood vessels: FGF is a strong proliferation factor in the walls of blood vessels, which facilitates the chemotaxis and mitosis of the vascular endothelium cells, and induces the formation of capillary-alike cavity for new capillary. (2) taking part in the repairing of cardiac muscles damage: FGF is concerned with the repairing process of myocardial ischemia and infarction. (3) lowering the blood pressure: FGF can lead to the increasing of the intracellular calcium ion concentration and endothelium derived relaxing factor [EDRF] synthesis, and result in vasodilatation. (4) affecting the nervous system: as a neuro-nutritional factor, FGF can facilitate neuron viability, neuroganglion proliferation and neuron dendrites and axon growing; it can also prevent neuron cell body from dying. (5) promoting the healing of wound and tissue repair: including the repair of soft tissue, cartilage tissue, and bone tissue damage, and the regeneration of limbs. (6) taking part in the embryonic development and differentiation. (7)regulating endocrine. (8) relating to cell proliferation diseases. Because of the same biologic functions of the recombinant FGF and natural FGF, the succese of recombinant bFGF can solve a industrial equation of bFGF essentially. It provided a good condition for clinical application. At present, domestic productions of bFGF are all in aqueous solution (pressurized cintainer); they have some disadvantages, such as short period of validity, inconvenient in transportation, getting polluted easily, poor patient compliance, etc. Contrapose to these problems, the bFGF gelling agent is studied. The advantages of the bFGF gelling agent are: (1)the barrier action: after daubing it on the surface ofwound, a protective film will formate, which prevents the bacillosis; (2) the slow release effect: the gelling agent can stay much longer than the aqueous solution; (3) the appeasing effect: it can alleviate the direct stimulation of the dressing to the wounded area; (4) the humification: the gel matrix can keep humidity and lubrication, which can lessen the conglutination and ease the main. Thus, the bFGF gelling agent has some advantage in therapeutic action than the aqueous solution, and it has a better applying foreground. This study is divided into four sections: 1 The study of fermentation process for bFGF We studied bacterium growing and bFGF expression in different glucose, dissolved oxygen and inductor (IPTG), and set up the fermentative technology for industrialization. 2 The study of purification technology for bFGF The bFGF is a kind of intracellular expressed soluble protein that has a good affinity with heparin. Therefore, we homogenizing comminuted and centrifuged the bacterial strain, and got the extracted pure protein, without denaturalization and renaturation. After exchanging through CM positive ion and heparin affinity chromatography, we got the recombination FGF pure product. The comprehensive detection of the pure product is done with MTT method for the titer, and the average specific activity is 2.8×106AU/mg (which equals to 4.9×106IU/mg). SDS-PAGE electrophoresis and HPLC showed that the purity of the product is 99.3%. The solid phase spot crossbreed test showed that the amount of extraneous sourced DNA is less than 10ng. The ELIZA test showed the remaining bacteril protein had an amount that is less than 0.1%. The hydrogen bromide cracking test showed that there were two strips in the peptide picture. The immunoblotting differentiation test result is positive.The purification techniques is stable, and each index of the bFGF undiluted liquid is equal or higher than the national standard. 3 The study of the bFGF gelling agent matrix We use the sodium carboxymethyl cellulose, Carbomer940 and sodium alginate, etc. as the macromolecule matrix. After autoclaving them, we compare the results of the characteristics, viscosity, cold-resistant and hot-resistant test, and find that the Carbomer940 predominating matrix has the best stabilization. We mixed the bFGF and the Carbomer940 matrix with different protective agent, and made them into different bFGF gelling agent. The temperature test showed that the most stable bFGF gelling agent matrix has a prescription as following: 0.5% human serum albumin, 2.5% mannitol and 1% HP-β-CD; mixed even with bFGF and stored for 24 months under 4 ℃, 18 months under 25 ℃, and 6 months under 40℃. This illuminates the choice and the proportioning of the gelling agent are appropriate. 4 The study of toxicology and pharmacology for bFGF gelling agent We compared the results of the local excitation test, the acute toxicity test and different forms of bFGF used for burned surface of wound, and found the bFGF gelling agent had no stimulation; it could be used externally on skin. The wounded area daubed with bFGF gelling agent healed two to three days earlier than that daubed with bFGF aqueous solution; the result was obvious. To sum up, bFGF gelling agent preparation technology is stable; the product is eligible. The animal test shows that it has good security, validity and practicability. Different forms of its matrix used on burned surface of wound indicate that the necessary times are fewer, and the patient compliance is better. Nonetheless, the period of validity of bFGF gelling agent is longer than the aqueous solution, and it is much easier in storing and transportation, less easily gets polluted, and more convenient to...
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