| Dental implant has been accepted to use by more and more people due to its incomparable advantages. But the lack of bone quality and bone quantity in patients caused by various reasons, not only greatly increase time and cost of the treatment, also seriously restrict the dental implant indications in clinical. Traditional Chinese medicine can increase bone mass, and improve bone structure, promote the synthesis of bone mineralization and sedimentary, mature Chinese herbal has been used for many years. In recent years, some people found that it can accelerate osseointegration after dental implant operation during experimental research, to a certain extent. This study intends to explore effects and underlying mechanisms of a kidney-reinforcing Chinese herbal mixture on the proliferation and function of osteoblasts through test in vivo and in vitro, to observe the differentiation of bone marrow stem cells, the role of osteoblast and osteoclast proliferation, as well as the protein expression and signaling pathway, etc., and micro-ct methods such as qualitative and quantitative detection of traditional Chinese medicine mixture how to accelerate osseointegration after dental implant operation.Part one:the test in vitroTest one:kidney-reinforcing Chinese herbal mixture on proliferation of osteoblasts in vitroObjective:To examine the effects of the kidney-reinforcing Chinese herbal medicated serum on the proliferation of human osteoblastic cells (hFOB1.19) cultured in vitro. Methods:The MTT assay was used to evaluate cell viability (under low and high doses), BrdU labeling was used to assess the effects of the medicated sera (48h incubation) on cell proliferation. Results:Exposure of hFOB1.19cells to the prepared medicated sera for24h significantly increased cell viability in comparison with the control group (p<0.05); extension of the incubation time to48h further augmented cell viability (p<0.05) and growth appearance. Conclusion: The medicated sera of the kidney-reinforcing Chinese herbal mixture could significantly promote the proliferation of osteoblasts. Test two:The mechanism research of kidney-reinforcing Chinese herbal mixture promote proliferation and function of osteoblast in vitroObjective:To examine the effects of the medicated serum of a kidney-reinforcing Chinese herbal mixture on the proliferation and function of human osteoblastic cells (hFOB1.19) cultured in vitro and explored the relevant molecular mechanisms. Methods:We prepared medicated sera using different doses of a kidney-reinforcing Chinese herbal mixture to treat hFOB1.19cells cultured in vitro. A kit was used to determine the dynamics of cellular alkaline phosphatase (ALP) activity. ELISA was used to examine the influence of the medicated sera on osteocalcin secretion, and Western blot analysis was used to elucidate the effects of the medicated sera on the protein levels of osteoprotegerin (OPG), osteopontin (OPN), and receptor activator of NF-kB ligand (RANKL). Results:The medicated sera were found to enhance ALP activity in hFOB1.19cells in a dose-dependent manner. Both treatments, at the low dose (p<0.05) and high dose (p<0.01), clearly increased the cellular ALP activity compared with the cells in the mock control. Exposing hFOB1.19cells to the medicated sera for48h did not result in any statistical difference in osteocalcin secretion among the experimental groups; however, a96h exposure to the medicated sera promoted secretion, which was dose dependent (compared to the mock control, low-dose serum, p<0.05, and high-dose serum, p<0.01). Finally, the medicated sera were shown to boost markedly the expression of OPG and OPN (p<0.05) but suppress that of RANKL (p<0.01) in hFOB1.19cells. Conclusion:The medicated sera of the kidney-reinforcing Chinese herbal mixture could significantly promote the proliferation and differentiation of osteoblasts. The underlying molecular mechanism is associated with singnal regulating system of osteoprotegerin, receptor activator of nuclear factor kappa B and its ligand.Part two:the test in vivoTest three:Histopathological assessment of osseointegration of dental implant after kidney-reinforcing Chinese herbal mixtureObjective:Histopathological method was used to assess osseointegration of dental implant after kidney-reinforcing Chinese herbal mixture;Methods:12rabbits were divided into two groups at random, both at the top of the the tibial inserted implant pure titanium implant, a group kidney-reinforcing Chinese herbal mixture were added to the feed, another group didn't join any drugs, after30days,60days and90days put to death, with histopathologic evaluation of osseointegration of dental implant and bone tissue morphology metrology method;Results:The results of experimental pathology and bone tissue morphology metrology parameters was better than control group;Conclusion:Kidney-reinforcing Chinese herbal mixture may promote osseointegration of dental implant.Test four:Imageology assessment of osseointegration of dental implant after kidney-reinforcing Chinese herbal mixtureobjective:Imageology method was used to assess osseointegration of dental implant after kidney-reinforcing Chinese herbal mixture;Methods:12rabbits were divided into two groups at random, both at the top of the the tibial inserted implant pure titanium implant, a group kidney-reinforcing Chinese herbal mixture were added to the feed, another group didn't join any drugs, after30days,60days put to death, with Imageology evaluation of osseointegration of dental implant; Results: the experimental group of radiographic outcomes, such as x-rays, CT showed the osteogenesis effect was obviously better than control group; Conclusions:kidney in traditional Chinese medicine mixture may promote dental implant speed of osseointegration.Test five:Immunohistochemical assessment of osseointegration of dental implant after kidney-reinforcing Chinese herbal mixtureobjective:Immunohistochemical method was used to assess osseointegration of dental implant after kidney-reinforcing Chinese herbal mixture;Methods:12rabbits were divided into two groups at random, both at the top of the the tibial inserted implant pure titanium implant, a group kidney-reinforcing Chinese herbal mixture were added to the feed, another group didn't join any drugs, after30days,60days,90days put to death, with Immunohistochemical evaluation of osseointegration of dental implant; Results:Immunohistochemical results show that OPG and OPGL, cox-2can be seen moderate positive expression in both the control group and experimental group near dental implant after30days; After60days OPG and OPGL can be seen moderately positive expression, but cox-2weakly positive expression in both control group and experimental group; After90days OPG and OPGL, cox-2can be seen weakly positive expression in the both control group, and experimental group; Conclusions:kidney-reinforcing chinese herbal mixture may promote dental implant speed of osseointegration. |
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