Fracture healing is a process that involves the proliferation and differentiation of cells and tissues and can be stimulated by drugs. Traditional medicine has a reputation in the therapy on bone fracture. It is believed that herbal formulae promotes bone repair through multiple functions of “anti-inflammation”, “promoting blood circulation” and “stimulating bone formation”. This study aims to engineer a herbal formula under the guidance of Chinese medicine theories, then define the effective dosage ratio between the herbs and demonstrate the biological effects of the formulae on different parameters of healing using relevant in vitro assays. Six herbs which are frequently used for fractures were combined into five formulae according to the “Uniform Design” method. Assays of nitric oxide (NO) inhibition in mouse macrophages, human umbilical vein endothelial cells (HUVEC) and rat osteoblasts proliferation were used to reflect the “anti-inflammation inflammatory”, “circulation promotion” and “bone formation stimulation” functions. The results showed that all the formulae expressed significant activities of NO inhibition, and HUVEC cell and osteoblasts proliferation. Formula 5 with the largest doses (containing 30g Radix et Rhizoma Rhei, 9g Fructus Gardeniae, 9g Radix et Rhizoma Notoginseng, 9g Flos Carthami, 30g Ramulus Sambucus Willamsii, 15g Radix Dipsaci) demonstrated the best results in all the three assays.
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