RESUMEN
BACKGROUND:Bushen Yiqi Huayu granule has the effect of replenishing qi and eliminating blood stasis,tonifying kidney and smoothing collages,and is often used in the treatment of osteoporosis.At present,there are few studies on the effect of Bushen Yiqi Huayu granule on the callus angiogenesis of osteoporotic fractures. OBJECTIVE:To explore the mechanism of Bushen Yiqi Huayu granule by up-regulating extracellular signal-regulated kinase/mitogen-activated protein kinases(ERK/MAPK)signaling pathway to improve callus angiogenesis after fracturing in osteoporotic rats. METHODS:(1)Bone marrow mesenchymal stem cells from osteoporotic SD rats and bone marrow monocytes from C57BL/6 mice were collected.MTT assay was used to detect different doses of Bushen Yiqi Huayu granule on bone mesenchymal stem cell toxicity.Bone mesenchymal stem cells and bone marrow monocytes were cultured in the medium supplemented with 0 and 1.5 mg/mL Bushen Yiqi Huayu granules,respectively,and osteogenic differentiation and osteoclast differentiation were carried out in vitro.(2)144 SD rats were randomly divided into sham operation group,model group,granule group and granule+PD98059 group with 36 rats in each group.The osteoporotic model was established in the model group,granule group and granule+PD98059 group,and only part of the adipose tissue near the ovary was removed in the sham operation group.8 weeks later,all rats received left tibial osteotomy.In the granule+PD98059 group,5 g/kg Bushen Yiqi Huayu granule was given intragastrically,and 0.3 mg/kg PD98059 was injected into the tail vein.The granule group was given 5 g/kg Bushen Yiqi Huayu granule,and the same volume of normal saline was injected into the tail vein.The sham operation group and model group were given an equal volume of normal saline,and the caudal vein was injected with an equal volume of normal saline.Drug administration was conducted once a day for 8 weeks.Fracture healing and callus angiogenesis were observed by X-ray,micro-computed tomography and microvascular perfusion angiography.Bone mineral density and mechanical strength of the callus were measured.The tibia was observed by hematoxylin-eosin staining and Masson staining.Western blot assay and immunohistochemistry were used to examine the expression of ERK/MAPK signaling pathway-related molecules. RESULTS AND CONCLUSION:(1)Compared with the 0 mg/mL group,the alkaline phosphatase activity,mineralization level,p-ERK1/2 and p-p38 MAPK expression levels of osteocytes increased(P<0.05),while the density and volume of osteoclasts decreased in the 1.5 mg/mL group(P<0.05).(2)Compared with the sham operation group,the fracture healing degree of the granule group was similar at 4 and 8 weeks,but there was no significant difference in Lane-Sandhu score,total callus volume,mineralized callus volume,mineralized callus volume/total callus volume,trabecular thickness,vascular number,spacing,thickness,vascular volume/total volume,bone mineral density,ultimate load of callus,stiffness,p-ERK1/2,p-p38 MAPK,p-IκB-α,and p-p65 expression levels(P>0.05).In the model group and granule+PD98059 group,fracture healing was slow,Lane-Sandhu score,total callus volume,mineralized callus volume,mineralized callus volume/total callus volume,trabecular thickness,vascular number,thickness,vascular volume/total volume,bone mineral density,ultimate load of callus,stiffness,p-ERK1/2,p-p38 MAPK,p-IκB-α,and p-p65 expression levels decreased(P<0.05),and vascular spacing increased(P<0.05),compared with the sham operation group.(3)It is indicated that Bushen Yiqi Huayu granule can improve fracture healing,promote callus angiogenesis and alleviate the symptoms of osteoporosis by enhancing the expression of the ERK/MAPK signaling pathway-related molecules in osteoporotic rats.
RESUMEN
BACKGROUND:In recent years, studies on the application of traditional Chinese medicine in bone tissue engineering are stil at the initial stage in China. Reports about psoralen antibiotic biphasic ceramic bone have not been seen in bone tissue engineering research. OBJECTIVE:To fabricate the psoralen antibiotic biphasic ceramic bone by vacuum evaporator and to observe the physicochemical properties, antibiotic activity and biocompatibility of the material. METHODS:Biphasic ceramic bone was prepared by twice low-temperature calcining pig vertebrae combined with sodium pyrophosphate, and then the ceramic bone was immersed in chitosan/psoralen compound solution to prepare psoralen antibiotic biphasic ceramic bone. The surface morphology was observed by scanning electron microscope, and the compressive strength was tested. Antibiotic activity of the psoralen antibiotic biphasic ceramic bone on Escherichia coli and Staphylococcus aureus was tested by microbiological methods. Mouse osteoblasts MC3T3-E1 were co-cultured with the psoralen antibiotic biphasic ceramic bone for 4 and 8 days, and the cel adhesion and proliferation on the scaffold surface were observed under the scanning electron microscope. RESULTS AND CONCLUSION:Psoralen antibiotic biphasic ceramic bone had natural pore structure with the trabecular bone, trabecular space and lumen system and exerted great antibiotic effect on Escherichia coli and Staphylococcus aureus. The compressive strength was (4.69±0.50) MPa. Mouse osteoblasts MC3T3-E1M could be adherent to the psoralen antibiotic biphasic ceramic bone and proliferate rapidly, suggesting the psoralen antibiotic biphasic ceramic bone has good cytocompatibility.
RESUMEN
Objective To investigate the changes and significance of plasma interleukin (IL)-6,IL-8,tumor necrosis factor-alpha (TNF-α) and nitric oxide synthase (NOS) in acute blood loss patients.Methods The levels of plasma IL-6,IL-8,TNF-α and NOS were determined in 25 patients with acute blood loss(blood loss group) and 25 healthy controls(control group) by enzyme linked immunosorbent assay.Result The levels of plasma IL-6,IL-8,TNF-α and NOS in blood loss group were higher than those in control group (0.284 ± 0.027 vs.0.204 ± 0.016,0.059 ± 0.079 vs.0.037 ± 0.039,0.460 ± 0.024 vs.0.372 ±0.018,0.637 ±0.054 vs.0.443 ±0.040,P＜0.05 or ＜0.01).Conclusions First,the expressions of IL-6,IL-8,TNF-α and NOS are strongly induced at the circumstances of acute early stage blood loss which can cause inflammatory reaction,aggravate the damage of soft tissues and organs,and easy to lead the blood loss to uncontrolled hemorrhagic shock.Second,the expression of NOS is strongly induced at the circumstances of acute blood loss which should reduce the damage of soft tissue,and alleviate the hemorrhagic shock.Therefore,the levels of plasma IL-6,IL-8,TNF-α and NOS can estimate the illness prognosis and curative effect,which have important clinical instruction value for effective treatment of hemorrhagic shock.
RESUMEN
BACKGROUND:Natural biologic bone-derived products have been prepared by raw ox bone but not by swine vertebra. Previous research has indicated that biphasic ceramic-like biologic active bone made by swine vertebra has osteoinductive ability. OBJECTIVE:To investigate the feasibility of biphasic ceramic-like biologic active bone and biphasic ceramic-like biologic bone (BCBB) to repair the segmental defect in radius of rabbits. DESIGN,TIME AND SETTING:A randomized controlled animal experiment was performed at Institute of Orthopedics,the Fourth Military Medical University of Chinese PLA and Central Laboratory of Qingdao Medical College from October 2004 to October 2005. MATERIALS:Biphasic ceramic biologic bone was made of swine vertebra and sodium pyrophosphate after two calcinations at low temperature. The mixture which consisted of hydroxyapatite and tricalcium phosphate,sizing 0.3 cm ? 0.3 cm ? 1.5 cm was combined with type I collagen to make BCBB. Bone morphogenetic protein was mixed with type I collagen,and then the mixture was combined with biphasic ceramic biologic bone to make biphasic ceramic-like biologic active bone. METHODS:A total of 48 healthy Japanese rabbits were randomly divided into biologic active bone group,biologic bone group,and blank control group,with 16 rabbits per group. A segmental defect which was 15 mm along bilateral radius was established. Biphasic ceramic-like biologic active bone and BCBB were implanted into defect region,respectively,but implantation was not treated in the blank control group. MAIN OUTCOME MEASURES:Materials were harvested at weeks 2,4,8,12 for the gross observation under microscope. Tissue sections were selected for hematoxylin-eosin (HE) staining and Masson staining. RESULTS:① Gross observation:At 2 weeks,there were no differences between the two groups,and the materials were connected to bone bed with fibrous tissues. At 4 and 8 weeks,there was more new callus in biologic active bone group than in biologic bone group. At 12 weeks,some bionic biphasic ceramic-like biologic active bone were absorbed and new bone regenerated in biologic active bone group,being similar to appearance of host bone,but there were few callus in biologic bone group. In blank control group,bone defect could not be repaired and there were fibrous tissue in bone defect. ② Staining results:At 2 weeks after operation,there were no differences between the two groups,and the materials were connected to bone bed with fibrous tissues. At 4 and 8 weeks,more vessels and fibrous tissues grew into BCBB,new bone was formed,and bone connect was found between BCBB and bone bed in the biologic active bone group. At 12 weeks,in the biologic active bone group,more materials degraded,new bone was integrated with bone bed,and bone marrow-like structure was formed. At 12 weeks,the broken ends were sclerosis and blocked in the blank control group. CONCLUSION:Bionic biphasic ceramic-like biologic active bone characterizing by good bone induction and biocompatibility can remarkably repair the segmental defect in radius of rabbits. BCBB is perspective for repairing the segmental bone defect.