| Part one Experimental study on the inhibitory effect of zoledronic acid on the function of osteoclast in vitroObjectives:Bisphosphonates are newly developed in recent years in Switzerland and are gradually applied to a new class of drugs to regulate bone metabolism.The chemical structure of zoledronic acid is similar to that of pyrophosphoric acid,and can resist the hydrolysis of some enzymes.Zoledronic acid can inhibit bone metabolism and absorption mediated by osteoclasts.As early as 1980,the first generation of bisphosphonates began to be used in the prevention and treatment of senile osteoporosis.However,some scholars have found that zoledronic acid can prevent mineralization of normal bone tissue and even cause fractures in long or large doses of zoledronic acid.Zoledronic acid is a imidazole heterocyclic bisphosphonate,developed by Novartis,Switzerland.Zoledronic acid belongs to the third generation of bisphosphonates.The first clinical application of zoledronic acid was first reported in Canada.Zoledronic acid was subsequently approved in more than80 countries and regions around the world.Zoledronic acid for treatment of malignant hypercalcemia satisfaction.Zoledronic acid and zoledronic acid have good curative effect in the treatment of bone metastases of multiple myeloma and solid tumor.The drug zoledronic acid can be effective in the treatment of patients with hypercalcemia.At the same time,zoledronic acid for advanced cancer bone metastasis and Paget,can reduce the incidence of bone related events.Zoledronic acid can significantly alleviate the symptoms of patients and improve the quality of life of the patients.The biological activity of zoledronic acid has been confirmed by preclinical trials.Zoledronic acid is the strongest anti bone resorption in bisphosphonates that have been found so far.Zoledronic acid is about 100 to 850 times the effect of the previous generations of bisphosphonates.Zoledronic acid is also the only effective bisphosphonate drug that is currently approved by FDA for solid bone metastases.We have found zoledronic acid in the treatment of osteoclast tumor of the spine,and it has been found to be effective in the treatment of osteoclast.However,we have not yet fully understood the characteristics of zoledronic acid in the inhibition of osteoclast function.Therefore,we have designed this experimental study to determine the characteristics of zoledronic acid in inhibiting the function of osteoclast in vitro.Methods:The bone marrow mononuclear cells of 6 weeks old C57 mice were cultured in vitro,and the bone marrow mononuclear cells were induced and differentiated to produce osteoclasts.In the experimental group,the concentration of zoledronic acid was different according to the culture medium in vitro.It is divided into 1×10-8mol/L,1×10-7mol/L,1×10-6mol/L,1×10-5mol/L and 1 x 10-4mol/L.And the experiment set the buffer solution that did not contain any bisphosphonates as the control group.The anti tartaric acid phosphatase staining?TRAP?was used to count the multinucleated cells.The calculation was calculated through the cell count.Are attached to the wall of cell count and bone absorption lacuna.The inhibitory effects of zoledronic acid on cultured osteoclasts in vitro were observed.Results:The early growth of OCLs?60min?was the growth of wall attachment.After 5 days of cultivation,it was found that with the increase of zoledronic acid,the number of multinucleated cells in the experimental group decreased with the increase of concentration.The minimum effective inhibitory concentration of zoledronic acid in inhibiting the formation of osteoclast in vitro is 1×10-6mol/L.The inhibitory effect on osteoclast migration and bone resorption was more significant at 1×10-5mol/L concentration,and the effect was further enhanced at 1×10-4mol/L concentration,but the correlation curve between concentration and inhibition ability gradually levelled off.Summary:Different concentrations of zoledronic acid have different effects on the differentiation,migration and bone resorption of osteoclasts.The 1×10-6mol/L concentration of zoledronic acid has obvious anti-osteoclast action.The inhibition ability curve of high concentration gradually tends to be slow,which suggests that the suitable concentration and dose of zoledronic acid should be selected in the clinical application.Part two Experimental study on zoledronic acid and ibandronate inhibit osteoclast characteristic comparisonObjective:Zoledronic acid and ibandronate bisphosphonate are commonly used in clinical work.Found in the process of treatment of osteoclastoma in when the suppression of ibandronic acid,osteoclastoma recurrence,osteoclastoma application of zoledronic acid in the treatment of recurrence is still valid.At the same time,in clinical application,it is found that the effect of 4mg zoledronic acid is stronger,but the incidence of adverse drug reaction is high.Such as:influenza like symptoms,mandibular necrosis,abnormal renal function,fever,and so on.The effect of medicine application4mg ibandronic acid intravenous zoledronic acid although relatively weak,but the incidence of adverse reactions were mild and.Our research is through the method of in vitro studies,found that zoledronic acid and ibandronate inhibit osteoclast characteristics.Through the comparison of zoledronic acid and ibandronic acid inhibition curves,found that zoledronic acid and ibandronate minimal effective concentration of phosphonic acid,in order to guide the clinical application.Methods:The bone marrow mononuclear cells of 6 weeks old C57 mice were cultured in vitro,and the bone marrow mononuclear cells were induced and differentiated to produce osteoclasts.Zoledronic acid was added to the zoledronic acid group according to the concentration of zoledronic acid in the culture medium.The further step by step is divided into 1×10-8mol/L,1×10-7mol/L,1×10-6mol/L,1×10-5mol/L,1×10-4mol/L,×10-3mol/L.Ibandronic acid group according to the concentration of adding zoledronic acid in vitro.It is divided into 1×10-8mol/L,1×10-7mol/L,1×10-6mol/L,1×10-5mol/L,1×10-4mol/L,1×10-3mol/L.And the experiment set the buffer solution that did not contain any bisphosphonates as the control group.The anti tartaric acid phosphatase staining?TRAP?was used to count the multinucleated cells.The calculation was calculated through the cell count.Are attached to the wall of cell count and bone absorption lacuna.Were observed in different concentrations of zoledronic acid and ibandronic acid culture characteristics of inhibitory effect on osteoclasts in vitro,and to compareResults:OCLs training for 5 days,with the discovery of sequential phosphonic acid and ibandronic acid concentration increases,the number of phosphonic acid group and ibandronic acid group coenocyte decrease with increasing concentration.The minimum effective inhibitory concentration of zoledronic acid in inhibiting the formation of osteoclast in vitro is1×10-6mol/L.The inhibitory effect on osteoclast migration and bone resorption was more significant at 1×10-5mol/L concentration,and the effect was further enhanced at 1×10-4mol/L and 1×10-3mol/L concentration,but the correlation curve between concentration and inhibition ability gradually levelled off.However,the minimum effective concentration of ibandronate inhibit osteoclast formation is 1×10-5mol/L.The inhibitory effect on osteoclast migration and bone resorption was more significant at 1×10-4mol/L concentration,and the effect was further enhanced at 1×10-3mol/L concentration,but the correlation curve between concentration and inhibition ability gradually levelled off.Summary:At the concentration of 1×10-6mol/L,zoledronic acid has obvious anti-osteoclast action.However,the lowest effective anti-osteoclast concentration of ibandronate is 1×10-5mol/L.Part three Clinical study of bisphosphonates adjuvant treatment of spinal giant cell tumor after surgeryObjectives:Giant cell tumor is a benign invasive tumor.The treatment of giant cell tumor of the extremities is varied and the therapeutic effect is good.But the spinal giant cell tumor of the bone can cause serious consequences.The operation space is small,the bleeding is more in the operation,the spinal cord may cause the nerve symptom after the operation.Moreover,the treatment of giant cell tumors of the spine,especially the sacrosacral giant cell tumor,is particularly difficult.More bleeding during the operation,the surgical field fuzzy nerve distribution and shape,causing confusion in operation.At the same time,the side effect of enlarged excision is very large,bleeding can cause shock,and nerve injury can cause incontinence and paralysis.However,the spinal osteoclast tumor is still difficult to be curettaged,and the recurrence rate is very high,and the literature is reported to be as high as fifty percent.However,the application of chemotherapy to spinal osteoclast curettage is not effective.The traditional adjuvant therapy is radiation therapy,but radiation therapy may cause the wound to be difficult to heal,and the sarcomatosis of the osteoclast tumor.Therefore,this study retrospectively analyzed the clinical data of different types of adjuvant therapy after spinal osteoclastoma,aiming to further improve the adjuvant treatment of spinal osteoclastoma after curettage.Methods:From July 2008 to July 2016,128 cases of spinal osteoclastoma were retrospectively analyzed,and the clinical data of six cases of spinal osteoclastoma after operation were analyzed statistically.Zoledronic acid 4mg?zoledronate 4mg?was used every month for 24 months in the zoledronic acid group.Ibandronic acid group of 32 patients,spinal resection and interbody fusion with internal fixation per month after application of ibandronic acid 4mg treatment,treatment for 24 months.31 patients were treated with conventional radiotherapy,and routine radiotherapy was used after operation.That is,the X ray of a linear accelerator is used to irradiate.The irradiated field is a osteoclast seen in the naked eye;and the edge of the tumor bed extends outward 2cm.The absorbed dose of the irradiated tumor was 30 to 50Gy,2Gy each time,5 times a week.Denosumab patients 32 cases,spinal resection and interbody fusion with internal fixation after application of denosumab monthly subcutaneous injection of 120mg treatment,treatment for24 months.All patients with osteoclast tumor must be examined by MRI before operation.At the same time,all the patients were followed up with the X-ray of the spinal column,the CT scan of the vertebral body and the two dimensional reconstruction.The four groups of patients with the actual operation time,intraoperative bleeding,postoperative wound healing,wound flow rate and other indexes were compared statistically.At the same time,the clinical treatment effect score,the recurrence rate of osteoclast tumor and the rate of ossification of the lesion were compared between the four groups.The adverse reaction of postoperative adjuvant therapy was observed.Results:The clinical study was followed up for 26 months to 86 months,with an average of 58 months after the spinal fusion and internal fixation.The four groups of patients with the actual operation time,intraoperative blood loss,postoperative wound healing,wound drainage and other indicators were no significant significance?P>0.05?.The VAS score and ODI score of the four groups of patients were statistically significant?P<0.01?.Zoledronic acid group the recurrence rate was 6%,ibandronic acid group was 12.5%,conventional radiotherapy group was 32.2%and 3.1%in the denosumab group.The recurrence rate of tumor in the four groups was statistically significant?P<0.01?.Zoledronic acid group vertebral reconstruction rate was86.6%,ibandronic acid group was 76.2%,conventional radiotherapy group was 58.5%and 92.8%in the denosumab group.The rate of reconstruction and reconstruction of vertebral body after operation in four groups was statistically significant?P<0.01?.Zoledronic acid group the incidence of adverse reaction was 51.5%,ibandronic acid group was 31.2%,conventional radiotherapy group and poor wound healing of skin irritation and other adverse reaction rate was 51.6%,12.5%in the denosumab group.There was a significant difference in the incidence of adverse reactions between the four groups?P<0.01?.Summary:The recurrence rate of tumor with conventional radiotherapy alone is still high after the resection of bone giant cell tumor of the spine.Zoledronic acid,Ibandronic acid,Denosumab were significantly decreased the giant cell tumor of the spine resection recurrence rate after resection of bone graft.The three kinds of drugs have significant clinical significance for the repair,reconstruction and reossification of the spinal vertebra structure.Effect of zoledronic acid is stronger than ibandronic acid.But a slightly higher incidence of adverse reactions was found.Part four A clinical study of zoledronic acid on the treatment of giant cell tumor of thoracic vertebraObjectives:Although the thoracic spinal giant cell tumor is mostly an invasive benign tumor.However,because of the special anatomical structure of thoracic vertebrae,the thoracic spinal cord injury after thoracoslac osteoclastoma is very easy to cause paraplegia and fecal incontinence.Therefore,the treatment of the osteoclast tumor of the thoracic vertebra is very difficult.The incision and curettage of the thoracic osteoclast tumor is difficult.The surgeon must try to cut the tumor as far as possible in the blurred vision of the bleeding.At the same time,try to avoid the injury of the spinal cord as much as possible.Therefore,the traditional thoracic osteoclast tumor curettage is very difficult to clean the tumor.On the basis of the intractable osteoclast tumor of the thoracic vertebra,the scholars have carried out repeated exploration in the clinical work.Experience in the treatment of thoracic osteoclast tumor.Some scholars improved the operation mode,completely removed and subtotal resection of the affected vertebral body.Some scholars reduced postoperative complications and prevented postoperative recurrence through adjuvant therapy.Therefore,through the analysis of osteoclastoma modified thoracic surgery and postoperative adjuvant therapy of the method of actual operation time,intraoperative blood loss,postoperative wound healing,wound drainage,recurrence rate,disease rate,ossification of adverse events and other indicators,to further discuss the clinical effect of two different ways of thinking under the guidance of treatment.Methods:From July 2011 to July 2016,62 patients with osteoclast tumor of the thoracic vertebra were analyzed retrospectively.The total resection group of 32 patients underwent integral resection of the thoracic vertebra osteoclast with bone graft fusion and internal fixation.30 patients with curettage group were treated with zoledronic acid 4mg every month for 24months after the thoracic osteoclast curettage and fusion.All patients with osteoclast tumor of the thoracic vertebra must be examined by MRI before operation.At the same time,all the patients were followed up with the X-ray of the spinal column,the CT scan of the vertebral body and the two dimensional reconstruction.The two groups of patients with the actual operation time,intraoperative bleeding,postoperative wound healing,wound flow rate and other indexes were compared statistically.At the same time,the clinical curative effect score of two groups of patients,the recurrence rate of osteoclast tumor and the rate of ossification of the lesion were compared.The occurrence of adverse events in the perioperative period and after the operation.Results:The clinical study was followed up for 25 months to 58 months,with an average of 36 months after the spinal fusion and internal fixation.The two groups of patients with the actual operation time,intraoperative blood loss,postoperative wound healing,wound drainage and other indicators were statistically significant?P<0.01?.The VAS score and ODI score of the two groups of patients were statistically significant?P<0.01?.The tumor recurrence rate in the total resection group was 6.3%,and the lesion group was6.7%.The recurrence rate of tumor in the two groups was not statistically significant?P>0.05?.The rate of ossification was 89.6%in the total resection group and 86.5%in the curettage group.There was no significant difference in the incidence of adverse reactions between the two groups?P>0.05?.The incidence of adverse events was 53.3%in the total resection group and 12.5%in the curettage group.There was a significant difference in the incidence of adverse reactions between the two groups?P<0.01?.Summary:Clinical study found that the resection of thoracic vertebra,fusion and internal fixation was an effective way to reduce the recurrence rate of giant cell tumor.And the application of bisphosphonates in the treatment of giant cell tumor of thoracic vertebra also has a good clinical effect.Conclusions:1.Bisphosphonates can inhibit osteoclasts formation,migration,absorption and bone resorption in vitro.2.Clinical studies have found that bisphosphonates have a clear inhibitory effect on recurrent giant cell tumor after operation.It is suitable for the treatment of primary and recurrent giant cell tumor,and it can be used gradually in clinical application.3.Zoledronic acid is the most potent bisphosphonate and most powerful in inhibiting osteoclasts formation,migration,adsorption,bone absorption and so on.It plays the lowest effective drug concentration in inhibiting osteoclasts. |
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