In vitro & in vivo assessment of a herbal formula used topically for bone fracture treatment.
OBJECTIVE
A novel topical paste used for fracture healing (FH), consisting of the extracts of six herbs, Radix Dipsaci, Ramulus Sambucus Williamsii, Rhizoma Notoginseng, Flos Carthami, Rhizoma Rhei and Fructus Gardeniae, was developed according to the classical theory of traditional Chinese medicine. This study aimed to determine the effectiveness of this formula, and some of its important chemical components in the promotion of fracture healing. The transdermal transport of FH was also examined.


METHODS
The osteogenic, angiogenic and nitric oxide suppressing effects of FH and its important chemical marker components were assessed by using osteoblastosacroma UMR-106 cells, human umbilical vein endothelial cells (HUVEC) and murine macrophage RAW264.7 cells, respectively. The bone healing effects of the FH paste and its transdermal absorption were determined using a rabbit fracture model. The callus sizes, bone specific alkaline phosphatase levels and biomechanical properties of the healed bone were assessed.


RESULTS
FH significantly increased the cell proliferation in UMR-106 and HUVEC cells and inhibited the nitric oxide production in murine macrophage in dose-dependent manner. Its important chemical components asperosaponin VI, ginsenoside Rg1 and emodin were shown to be acting positively in the respective in vitro studies. FH paste significantly improved the bone healing in the rabbit fracture model, as was indicated by the increases in callus size at weeks 2-5, and the elevations in bone specific alkaline phosphatase activities at weeks 5-6. The analysis using LC/MS/MS also showed the presence of important chemical marker components of the FH formula in the plasma after 8 weeks of topical treatment.


CONCLUSIONS
This study presents the first scientific evidence of the efficacy of a herbal paste in the promotion of fracture healing. There were evidences of transdermal transport of the chemical components, control the inflammation through nitric oxide inhibition, promotion of angiogenesis, and bone healing in the in vitro tests, as well as in the experimental animal.
Peng
Li Hua
LH
College of Pharmaceutical Sciences, ZheJiang University, Hangzhou City, ZheJiang Province, PR China.
Ko
Chun Hay
CH
Siu
Sum Wing
SW
Koon
Chi Man
CM
Yue
Gar Lee
GL
Cheng
Wai Hing
WH
Lau
Tai Wai
TW
Han
Quan Bin
QB
Ng
Ka Ming
KM
Fung
Kwok Pui
KP
Lau
Clara Bik San
CB
Leung
Ping Chung
PC
D000279:Administration, Cutaneous; D000469:Alkaline Phosphatase / Q000097:blood; D000818:Animals; D002146:Bony Callus / Q000187:drug effects; D002460:Cell Line; D049109:Cell Proliferation / Q000187:drug effects; D004195:Disease Models, Animal; D004305:Dose-Response Relationship, Drug; D004365:Drugs, Chinese Herbal / Q000493:pharmacokinetics / Q000494:pharmacology / Q000627:therapeutic use*; D004642:Emodin / Q000493:pharmacokinetics / Q000494:pharmacology / Q000627:therapeutic use; D042783:Endothelial Cells / Q000187:drug effects / Q000502:physiology; D004730:Endothelium, Vascular / Q000187:drug effects; D017102:Fracture Healing / Q000187:drug effects*; D036145:Ginsenosides / Q000493:pharmacokinetics / Q000494:pharmacology / Q000627:therapeutic use; D006801:Humans; D008264:Macrophages / Q000187:drug effects; D019684:Magnoliopsida / Q000737:chemistry*; D008297:Male; D051379:Mice; D009569:Nitric Oxide / Q000096:biosynthesis; D010012:Osteogenesis / Q000187:drug effects*; D008517:Phytotherapy; D011817:Rabbits; D012503:Saponins / Q000493:pharmacokinetics / Q000494:pharmacology / Q000627:therapeutic use; D012869:Skin Absorption / Q000187:drug effects; D013978:Tibial Fractures / Q000188:drug therapy / Q000378:metabolism; D014471:Umbilical Veins
D004365:Drugs, Chinese Herbal; D036145:Ginsenosides; D012503:Saponins; C534004:akebia saponin D; D009569:Nitric Oxide; D000469:Alkaline Phosphatase; D004642:Emodin; C035054:ginsenoside Rg1