The effect of Gu-Sui-Bu (Drynaria fortunei J. Sm) immobilized modified calcium hydrogenphosphate on bone cell activities

In our previous study,we have validated the ef ficacy and the safety of Gu-Sui-Bu [Drynaria fortunei (Kunze)J.Sm.]by the bone cells culture.However,a satisfactory delivery system for Gu-Sui-Bu must be developed before it can be used in clinical medicine.In this study,we try to use modi fied calcium hydrogenphosphate (MCHP)bioceramic as a carrier to transport Gu-Sui-Bu into the bone cell culture system.Toward this goal,we evaluated the effect of a Gu-Sui-Bu-immobilized modi fied calcium hydrogenphosphate (GI-MCHP)on the bone cells activities. The Chinese medicine:Gu-Sui-Bu [Drynaria fortunei (kunze)J.Sm ]was extractedandthen immobilizedon the surface of MCHP. The rat osteoblasts –osteoclasts co-culture system was usedas the experimental model.After the cells grew to 80%con fluence, different sizes of GI-MCHP particles were tested.The mitochondria activity of the bone cells after exposure was determined by colorimetric assay.Biochemical markers such as lactate dehydrogenase (LDH),alkaline phosphatase (ALP),acid phosphatase (ACP)andprostaglandin E2 titer were analyzedto evaluate the bone cells activities.Histomorphometric study of osteoclasts activities andthe phenotype expression of osteoblasts were also evaluated. There is no detectable titer of LDH secretion into the medium and no signi ficant change in the intracellular ALP content.The ALP titer in the culture medium did increase signi ficantly at 3 days ’culture,while there is a signi ficant decrease in the intracellular ACP content andsigni ficant increase in the ACP titer in the medium.The concentrations of PGE2 in testedmedium are always signi ficantly higher than that of control medium during the 7 days ’culture.At the end of 7 days ’culture,the PGE2 concentrations in the tested medium were still 4.74 times that of the control medium.After GI-MCHP treatment on bone cells,the size of the osteoclasts seems decreased and their cell integrity seems lost,while the osteoblasts phenotype expression was relatively preserved. From this study,we demonstrated that Gu-Sui-Bu [Drynaria fortunei (Kunze)J.Sm.]immobilizedMCHP has well preservedthe potential bene ficial effects of Gu-Sui-Bu on the bone cells culture