Pharmacodynamic and Pharmacology Research

OsteoKing has been granted China National Approval Certificate in year 2002 by completing all the very strict researches and tests according to China nation standards. It has been proven by clinical applications that OsteoKing has remarkable effectiveness and very safe in treating bone diseases, especially for femoral head necrosis, prolapse of lumbar intervertebral disc, osteoarthritis, osteoporosis and fractures.

In order to explore its mechanism of the therapeutic effects, the company has taken the lead and completed the researches in foremost level of biochemistry, histomorphology, molecular biology for OsteoKing. The research results show that OsteoKing can reduce blood viscosity, improve hemodynamics, reduce internal pressure of bone, improve blood supply, reduce blood fat concentration, reduce the formation of fat embolism and promote the genetic expression of endogenous VEGF(Vascular endothelial growth factor), cbf ?1(core binding factor ?1) and BMP-2(Bone morphogenetic proteins-2). It can also strengthen the activity of bone cells, inhibit osteoclast cells and promote the repair of bone.

OsteoKing has solved the core problems of bone disease by decreasing blood fat, anti-coagulation, fibrinolysis and anti-inflammation so that patients can speed up their recoveries effectively. OsteoKing provides scientific evidence for non-surgery treatment for bone diseases and fractures and it is the best solution for clinical treatment. It has been listed as the research project founded by China National Natural Science.

The results of pharmacodynamic show that OsteoKing has the following functions:

Anti-inflammation and detumescence
Effective Pain Relief
Inhibiting thrombosis, improving blood circulation and microcirculation.
Activating the cells of periost osteogenesis, promoting the formation of new bone cells and the repair of necrotic bone.
Promoting the regeneration and matureness of bone trabecula and increasing the bone density
Decreasing blood fat, anti-coagulation, and fibrinolysis
Preventing the deep vein thrombosis (DVT) after bone diseases operation and protecting and repairing the venous endothelial cell injured by mechanical force.
Promoting the formation of blood vessel by promoting the genetic expression of endogenous VEGF, effectively improve the activity of osteoblasts and speed the formation of bone cells.
Promoting the development, differentiation, regulation of osteoblasts and bone calcification by boosting the genetic of cbf ?1.
Inducting ossification and stimulating the formation of bone cells by boosting the genetic of BMP-2.

Femoral head necrosis is a difficult and complex bone disease. In recent years, femoral head necrosis has become the key problem that medical experts in the world are working on. After extensive studies done by medical experts worldwide on the mechanism of necrosis of femoral head, the results point out that the principle for treating necrosis of femoral head is to decrease blood fat, anti-coagulation, fibrinolysis and anti-inflammation. Those concepts have now been widely accepted by the medical field.

Based on the above principle, western medicine uses compound medicines to treat necrosis of femoral head. Long term usage of them has caused lots of side effects to patients and also high treatment costs but the effectiveness is not long lasting. Therefore non-operation treatment by using herbs is becoming the new focus all over the world.

A large of clinical research have approved that OsteoKing is the unique herbal medicine with practical value. It is one product, but has four functions of decreasing blood fat, anti-coagulation, fibrinolysis and anti-inflammation, which solves the core problems of bone diseases - blood supply and thrombosis. It is one of the very unique herb products.

OsteoKing not only has unique and remarkable curative effects to femoral head necrosis, but also to prolapse of lumbar intervertebral disc, osteoarthritis, osteoporosis and fractures. OsteoKing greatly reduces the disable rate and facilitates the patient’s early recovery.

The researches on the functions:
Anti-inflammation and detumescence
Test: OsteoKing was applied to rat metatarsal swelling caused by egg whites.
The results showed that the product significantly inhibits rat metatarsal swelling. (compared with control group, p <0.05)
Test: OsteoKing was applied to mice injected 0.7% acetic acid into their abdomen cavities.

The results indicated that OsteoKing possesses significant property in inhibiting the raising of capillary permeability of mice caused by injected acetic acid into their abdominal cavities. (compared with control group, p<0.01)

Pain Relief
OsteoKing was applied to mice whose pain threshold stimulated by a hot plate. The results indicate that the product can relieve the pain caused by a hot plate. (compared with control group, p<0.05)

OsteoKing was given to a mouse with convulsion caused by an acetic acid injection into its abdominal cavity.

The results showed that the product produces a significant analgesic effect on pain caused by chemical substances. (compared with control group, p<0.01)

Inhibiting thrombosis, improving blood circulation and microcirculation
OsteoKing was given to rats with thrombosis. The results showed that the product could inhibit rat thrombosis in vivo and improve blood circulation and microcirculation. (compared with control group, p<0.01)

Decreasing blood fat
To perform the test of blood fat of necrotic bone, rabbits were divided into three groups: no-treatment control group, model group, and OsteoKing group.

The test measured the serum cholesterol (CHOL) and triglyceride (TG) and high density lipoprotein (HDL) and the results were statistically evaluated by using SPSS 10.0 s software, t examination and multivariate variance analysis for the enumeration data. The difference has statistics significance (compared with control group, p<0.01).

The results showed that necrotic bone is closely related to disturbance of lipid metabolism. OsteoKing can decrease the serum cholesterol (CHOL) and triglyceride (TG) significantly and increase HDL significantly. OsteoKing possesses significant function in decreasing blood fat.

Promoting the repair of necrotic bone.
Observation with optical microscope, transmission electron microscope and

scanning electron microscope, the results showed

The general observation:
No-treatment control group: normal femoral head
Model group: modeling cartilage has become thin; the bone trabecula has disordered and appeared oily condition.
OsteoKing group: the thickness of cartilage is normal and fat is little.
The results observed with optical microscope:
No-treatment control group: normal femoral head
Model group: the bone trabecula has disordered and become thinner, sparse. The medullar cavity is full of empty lipid cells.
OsteoKing group: more new bone is formed on the surface of bone trabecula.
The results observed with transmission electron microscope:
Model group: necrosis occurred, the medullar cavity bled, collagen was disturbance
Model group: necrosis occurred, the medullar cavity bled seriously,
Model group: bone tissues have become degenerately necrosis and bleeding
OsteoKing group: new bone trabecula and original bone trabecula
OsteoKing group: more blood capillary appeared
OsteoKing group: more osteoblasts and new bone cells appeared
The results observed with scanning electron microscope:
No-treatment control group: the normal membranous structure on the surface of bone trabecula
Model group: the membranous structure was destroyed
OsteoKing group: the membranous structure was normal and the formation of bone was active
The results indicate that OsteoKing can promote the repair of necrotic bone. Necrotic bone cells are less in the OsteoKing group, more osteoblasts and osteoclasts, new bone cells and bone matrix are appeared as well as calcium deposit occurs. The results observed with scanning electron microscope indicate that more osteoblasts appeared on the surface of bone trabecula. OsteoKing group has remarkable superiority compares with model group.

Promoting the formation of blood vessel by promoting the genetic expression of endogenous VEGF, effectively improve the activity of osteoblasts and speed the formation of bone cells.

VEGF (Vascular endothelial growth factor) is very closely related to bone repair and plays a very important role in the formation and repair of bone. (Lieberman JR, et al. Instr Course Lect, 2003; 52:337-55. ?Carano RA, et al. Drug Discov Today. 2003, 8(21): 980-989.?Yancopoulos GD,et al. Nature. 2000, 407(6801):242-248. Review.)

Tested through the methods of immunohistochemistry, hybridization in situ, real-time fluorescent fixed quantity and PCR and verified each other then draw the same conclusion that OsteoKing can promote the expression of endogenous VEGF in the necrotic femoral head and increase the expression volume significantly. Compared with Model group, the difference has statistics significance (p<0.05).

Promoting the development, differentiation, regulation of osteoblasts and bone calcification by boosting the genetic of cbf ?1.

cbf ?1 (core binding factor ?1) gene is osteogenesis specificity transcription activation factor, It plays a very important role during the development and differentiation of the osteoblast and the bone formation.(Ducy P, et al. Cell, 1997, 89(5):747~754.?Komori T, et al. Cell, 1997, 89(5):755-764.?He J, et al. Oral Surg Oral Med Oral Pathol Oral Radiol Endod, 2004, 97(2):239-245.)

Tested through the methods of immunohistochemistry, hybridization in situ, real-time fluorescent fixed quantity and PCR and verified each other then draw the same conclusion that OsteoKing can promote the repair of necrotic bone through promoting the expression of cbf ?1 in the necrotic femoral head and increase the expression volume significantly. Compared with the model group, the difference has statistics significance (p<0.05).

Inducting ossification and stimulating the formation of bone cells by boosting the genetic of BMP-2.

BMP-2 (Bone morphogenetic proteins-2) has very powerful activity in inducting ossification (Wozney JM, et al. Science, 1988, 242(4885): 1528-1534.?Croteau S,et al. Orthopedics, 1999, 22(7):686-695.?Shore EM, et al. Clin Orthop, 1995, (311):199-209.)

Tested through the methods of immunohistochemistry, hybridization in situ, real-time fluorescent fixed quantity and PCR and verified each other then draw the same conclusion that OsteoKing can promote the repair of necrotic bone through promoting the expression of BMP-2 in the necrotic femoral head and increase the expression volume significantly. Compared with the model group, the difference has statistics significance (p<0.05).

Promoting healing of fractures
Test of the effect of OsteoKing on fractures: The radius diaphysis of rabbits was totally fractured with a double blade saw. The gap between the broken parts is 3mm.The models were separated into: a no-treatment control group, a drug-taking group (taking another bone medicine) and an OsteoKing group. X-rays were taken, fracture-resistant strength tests were administered, and a histopathological examination was done.

The results showed that OsteoKing can promote the healing of fractures and speed of recovery.

The researches of OsteoKing are performed with the latest modern methods and receive the scientific data, from many angles and aspects to analysis the mechanism of the therapeutic effects of OsteoKing deeply to the genetic level. Those provide solid scientific data for the clinical application.

The results of pharmacology research showed:
OsteoKing strengthens the function of the pituitary -- adrenal cortex system, inhibits the function of hyaluronidase, chrondroitin enzyme, PG synthetase, 5_HTp and histamine. All those develop stronger anti-inflammatory and analgesic effects as well as reduction of swelling.
OsteoKing enlarges blood vessels, reduces peripheral resistance and strengthens general blood circulation.
OsteoKing decreases the synthesis of TXA2, increases the content of cAMP by inhibiting thrombus alkane A2 (TXA2) synthetase, activates adenyl cyclase in blood platelets, and inhibits the function of phosphodiesterase so it can develop the functions of anti-platelet aggregation and inhibiting thrombosis.
OsteoKing activates periost osteoblast, the growth and reconstruction of callus tissues, and the formation of new bone cells that improve the healing of fractures and promote the repair of femoral head.
Some ingredients in OsteoKing can block the path of receptive calcium ions from blood vessel's smooth muscle, block the internal flow of Ca2+ and the release of Ca2+ within cells, and therefore enlarge the external blood vessels, especially for the spastic small vessels. These factors improve microcirculation and the supply of blood and oxygen to the cells of injured tissues.
OsteoKing contains several amino acids, ossein, and several microelements that play important roles in the treatment of bone injuries and diseases, therefore it promotes the healing of factures and the recovery of bone diseases. It shortens the period of treatment.
All these pharmacology functions create a favorable condition for the recovery of fractures and bone diseases.