ObjectiveTo evaluate the effect of leucocyte- and platelet-rich plasma (L-PRP) on the osteogenic differentiation of bone marrow mesenchymal stem cells (BMSCs) in treating avascular necrosis of the femoral head (ANFH) in rabbits. MethodsTwenty-four New Zealand white rabbits (4-6 months old, both genders, weighing 2.0-3.0 kg) were used for the establishment of bilateral ANFH models and divided into 4 groups (n=6). BMSCs were isolated from the bone marrow of iliac crest, cultured and identified. L-PRP was prepared by Landesberg method. Core decompression only (group A), core decompression and L-PRP implantation (group B), core decompression and BMSCs implantation (group C), and core decompression and implantation of BMSCs and L-PRP were performed in 4 groups. To evaluate bone formation and remodeling of the defects, X-ray photography was taken at 2, 4, and 8 weeks postoperatively. The modified Lane-Sandhu scoring system was used to evaluate the bone formation. Two rabbits were sacrificed at 2, 4, 8 weeks after operation to harvest the specimens for histological observation, new blood vessel count and new bone area ratio. ResultsThe observations of radiology and histology displayed different degrees of bone regeneration at bone defect sites in each group. At 2, 4, and 8 weeks postoperatively, the results of Lane-Sandhu X-ray photography scoring, new blood vessel count, and new bone area ratio showed that groups C and D were significantly better than groups A and B, group D was significantly better than group C. and group B was significantly better than group A (P<0.05). ConclusionThese findings demonstrate that L-PRP can promote osteogenic differentiation of BMSCs in treating ANFH in rabbits, and core decompression associated with BMSCs and L-PRP is an effective and feasible method to treat ANFH.
ObjectiveTo explore the effect of fetal bovine serum (FBS) of different concentrations in the culture medium on osteogenic growth peptide (OGP) promoting bone marrow mesenchymal stem cells (BMSCs) proliferation and differentiation. MethodsBMSCs were separated from limb bones of 8 Sprague Dawley rats (5 weeks old) and purified by adherence method, and BMSCs at passage 3 were divided into 4 groups according to OGP concentration: OGP 1×10-10 mol/L group, OGP 1×10-9mol/L group, OGP 1×10-8 mol/L group, and control group without OGP; and 0, 2%, 5%, 8%, and 10%FBS concentration gradient was used in each group. The cell proliferation rate was detected by MTT method at 1, 3, 5, 7, 9, and 12 days after culture, and the activity of intracellular alkaline phosphatase (ALP) was determined by the method of p-nitrophenyl phosphate disodium at 9 days after culture. ResultsBMSCs showed adherent growth, rapid proliferation, long fiber vortex, and typical morphology. MTT analysis showed that cells could not sustain proliferation when FBS concentration was less than 5% in each group; when FBS concentration was above 8%, cells proliferated continually. Proliferation promoting effect of OGP 1×10-8 mol/L and 1×10-9 mol/L groups was significantly higher than that of the control group in all serum concentrations (P<0.05); when FBS concentration was lower than 10%, the proliferation promoting effect of OGP 1×10-8 mol/L group was significantly higher than that of the other 2 OGP groups (P<0.05), but when FBS concentration was 10%, OGP 1×10-8 mol/L group had no advantage of promoting proliferation. ALP test results showed that as the FBS concentration increased, ALP activity of all groups also significantly increased (P<0.05). Under the condition of 5%FBS and 8%FBS, the ALP activity of each OGP group was significantly greater than that of the control group, and it was the highest in OGP 1×10-8 mol/L group (P<0.05). Under the condition of 10%FBS, the ALP activity of each OGP group was still greater than that of the control group (P<0.05), but no significant difference was found between the OGP 1×10-8 mol/L group and OGP 1×10-9 mol/L group (P>0.05). ConclusionThe concentration of 8%FBS is the best concentration of serum for OGP promoting the proliferation and differentiation of BMSCs, and the most suitable concentration of promoting the proliferation and differentiation of BMSCs is OGP 1×10-8 mol/L.
Objective To systematically evaluate the effectiveness and safety of zoledronic acid combined with radiotherapy in treating bone metastasis of malignant tumor. Methods Such databases as PubMed, EMbase, The Cochrane Library (Issue 10, 2012), CBM, CNKI, VIP and WanFang Data were searched to collect randomized clinical trials (RCTs) on bone metastasis of malignant tumor from inception to October, 2012. References of included studies were also retrieved. Two reviewers independently screened studies according to exclusion and inclusion criteria, extracted data, and assessed the methodological quality. Then, meta-analysis was performed using RevMan 5.1 software. Results Twenty nine trials were included involving 2 021 patients. The results of meta-analysis showed that, compared with the radiotherapy alone group, zoledronic acid combined with radiotherapy improved the effectiveness rate of pain relieving at the end of treatment (OR=3.08, 95%CI 2.30 to 4.12, Plt;0.000 01), the effectiveness rate of pain relieving two weeks after treatment (OR=3.39, 95%CI 2.52 to 4.56, Plt;0.000 01), the quality of life (OR=2.74, 95%CI 1.66 to 4.52, Plt;0.000 01) and the ability of movement (OR=2.96, 95%CI 2.16 to 4.05, Plt;0.000 01). Zoledronic acid combined with radiotherapy also reduced the incidence of new bone metastasis (OR=0.21, 95%CI 0.10 to 0.45, Plt;0.000 1) and the incidence rate of bone-related events (OR=0.17, 95%CI 0.03 to 0.92, P=0.04). The adverse reactions of zoledronic acid combined with radiotherapy such as fever (OR=11.92, 95%CI 6.31 to 22.48, Plt;0.000 01) and hypocalcaemia (OR=8.82, 95%CI 1.61 to 48.36, P=0.01), significantly increased. Conclusion Compared with radiotherapy alone, zoledronic acid combined with radiotherapy can relieve bone metastatic pain, effectively enhance patients’ ability of movement, improve quality of life, and decrease new bone metastasis and the occurrence of bone-related events.
Objective To determine whether statins has some effects on the treatment of cardio-cerebral vascular diseases or hyperlipdemia increases bone mineral density （BMD）. Methods One hundred and sixty-two patients aged over 60 were identified in the outpatient-department of Geriatrics of West China Hospital from Jan. 1998 to Aug. 2003. Seventy cases were exposed to statins with follow-up for 5 years. BMD of the spine, femoral neck, femoral wards triangle and femoral trochanter was measured by dual-energy X-ray absorptiometry. The multiple regression analysis was used to exclude potential confounders, e.g. age, BMI, comorbidity,etc. Results Those elderly patients with a history of taking statins had higher BMD, such as femoral neck with t =-2. 466 （P =0. 015）, femoral wards triangle with t =-2. 483 （P = 0. 014 ）and femoral trochanter with t =-2. 743 （P =0. 007 ）than the control elderly at the end of follow-up. Conclusions It has been found that HMG-CoA reductase inhibitors （statins ） may prevent bone loss in elderly patients by increasing BMD. Further prospective studies of statins are needed to confirm these observatioris.
Objective To explore repair role of allogeneic bone marrow mesenchymal stem cells （BM-MSCs） transplantation on treating hepatic ischemia reperfusion injury （HIRI） in rats. Methods Ten rats were executed to get BM-MSCs， then BM-MSCs were cultured in vitro and dyed by 4，6-diamidino-2-phenylindole （DAPI）. Models of 70% hepatic ischemia reperfusion injury were eatablished. Thirty two rats were randomly divided into sham operation group （Sham group）， ischemia reperfusion group （I/R group）， Vitamin C group （VC group）， and BM-MSCs group. Serum samples were analyzed for ALT and AST， and hepatic tissue were for superoxide dismutase （SOD） and malondialdehyde （MDA）. Liver sections were stain with hematoxylin and eosin （HE） for histological analysis， TUNEL staining was applied to detect hepatic apoptosis. Serum and tissues were both collected at 24 h after reperfusion. Results The isolated BM-MSCs maintained vigorous growth in vitro. Specific markers for MSCs antigens CD29 and CD44 were detected by flow cytometry， but antigens CD34 and CD45 were not be detected. Models of HIRI were stable， and BM-MSCs were detected around the periportal area by DAPI staining. Compared with I/R group， levels of ALT， AST， MDA， and AI in the VC group and BM-MSCs group decreased at 24 h after reperfusion （P＜0.05）， meanwhile SOD level increased （P＜0.05）. Compared with VC group， levels of ALT， AST， MDA， and AI in the BM-MSC group decreased at 24 h after reperfusion （P＜0.05）， meanwhile SOD level increased （P＜0.05）. Conclusion BM-MSCs could protect HIRI by alleviating oxidative stress and inhibiting cellular apoptosis.
Objective To elucidate whether glucose transporters-4 （GLUT-4） takes part in glucose uptake of mesenchymal stem cells （MSCs） and whether Akt gene improves translocation and expression of GLUT-4 in MSCs under hypoxic environment ex vivo. Methods MSCs, transfected by Akt gene and no, were cultured with normoxia (5% CO2) or hypoxia (94%N2, 1%O2 and 5% CO2) at 37 ℃ for 8 h. Glucose uptake was assayed by using radiation isotope 2-［3H］-deoxy-Dglucose (3H-G) and the expression of GLUT-4 protein and mRNA was assayed by immunocytochemistry, Western blot and RT-PCR, respectively. Results ①3 H-G intake of MSCs was significantly increased in hypoxiatransfection group than that in hypoxia-non-transfection 〔（1.39±0.13） fold, P＜0.05〕, but which was lower than that in normoxia-non-transfection group, P＜0.05. ②GLUT-4 was expressed by MSCs under any conditions. Compared with normoxia-non-transfection group, hypoxia decreased the expressions of GLUT-4 mRNA and protein significantly (P＜0.05). ③Compared with hypoxianontransfection group, the expression of GLUT-4 〔mRNA（1.756±0.152） fold, total protein in cell （1.653±0.312） fold, protein in plasma membrane （2.041±0.258） fold〕 was increased in hypoxia-transfection group significantly (P＜0.05), but which was lower than that in normoxianontransfection group (P＜0.05). ④There was significantly positive relation between 3H-G intake and GLUT-4 protein expression in plasma membrane (r=0.415, P=0.001).Conclusion GLUT-4 may take part in glucose uptake of MSCs, and the capability of Akt gene to improve MSCs anti-hypoxia may be finished by its role in increasing the expression and translocation of GLUT-4.
Objective To introduce the current studies on bone biochemical markers in breast cancer with bone metastasis. Methods The papers in recent 8 years about the application of bone biochemical markers in the diagnosis and treatment of breast cancer with bone metastasis were reviewed. Results NTX had the best relation with bone metastasis. ICTP was much more worthy than NTX in diagnosis of breast cancer with bone metastasis. Osteogenesis markers were little worthy in diagnosis of breast cancer with bone metastasis. Conclusion Bone biochemical markers can not replace the image exams and biopsy in diagnosing the bone metastasis of breast cancer,but may be one of the factors to get the early diagnosis.
【Abstract】Objective To introduce three methods of creating the animal model of bone metastasis from breast cancer and the advances in the application of these models. Methods The related literatures were collected and reviewed.Results In summary, breast cancer cells injected through left ventricles was commonly used. Breast cancer cells injected into medullary cavity of shaft of femur was simple and effective, but it was very different from the real condition of bone metastasis of patients. The development of animal model created by surgical orthotopic implantation gives the researchs an ideal instrument similar with the condition of patients to research the mechanism of bone metastasis and the treatment. Conclusion Each animal model of bone metastasis from breast cancer has itself usefulness. Our destination is to create the real model of bone metastasis from breast cancer that is very similar with the patients.
With immunohistochemical technique, epithelial membrane antigen monoclonal antibody (EMA) has been used to detect the micrometastatic focus in bone marrow of patients with primary gastric cancer since 1992. In the exmamination of 65 patients, the positive rate of bone metastasis was 58.46%. After comprehensive treatment to these patients, comparative observation showed that there were marked differences between pre-therapeutic (9.45) and post-therapeutic (2.19). The result demonstrates that this technique provides identification of blood micrometastases and has insttructive signnificance for clinical comprehensive treatment.
Objective To investigate the therapeutic effects of strontium-89 to prevent bone metastases of lung neoplasms.Methods Thirty patients with bone metastases of lung neoplasms received strontium-89 treatment (89SrCl2) at a dose of 148 MBq through intravenous injection.The analgesic effect was assessed by VAS method and doses or frequency of using analgesic drugs.Other efficacy parameters included changes in the number of osseous lesions and urinary levels of pyridinoline and deoxypyridinoline on the day 28 after therapy.Results The total pain relief rate was 73.3%(22/30)，among which 5(16.6%) cases with pain vanished，suggesting significant alleviation of the pain intensity by the treatment(Plt;0.001) on the day 28 after therapy.The number of lesions decreased in 16 cases with effective rate of 53.3%，showing the bone metastases significantly decreased after the therapy (Plt;0.001).The urinary levels of pyridinoline and deoxypyridinoline on the day 28 after therapy were (62.48±37.25)nmol/mmol Cr and (13.94±8.66)nmol/mmol Cr，respectively，which were decreased significantly compared to the levels before treatment which were (100.15±48.65)nmol/mmol Cr and (31.25±15.32)nmol/mmol Cr，respectively (both Plt;0.001).Conclusion Strontium-89 is effective to relieve pain and prevent bone lesions in patients with bone metastases of the lung neoplasms.