Increasing the osteogenic differentiation ability and decreasing the adipogenic differentiation ability of bone marrow mesenchymal stem cells(BMSCs)is a potential strategy for the treatment of osteoporosis(OP).Natural...Increasing the osteogenic differentiation ability and decreasing the adipogenic differentiation ability of bone marrow mesenchymal stem cells(BMSCs)is a potential strategy for the treatment of osteoporosis(OP).Naturally derived oligosaccharides have shown significant anti-osteoporotic effects.Nystose(NST),an oligosaccharide,was isolated from the roots of Morinda officinalis How.(MO).The aim of the present study was to investigate the effects of NST on bone loss in ovariectomized mice,and explore the underlying mechanism of NST in promoting differentiation of BMSCs to osteoblasts.Administration of NST(40,80 and 160 mg/kg)and the positive control of estradiol valerate(0.2 mg/kg)for 8 weeks significantly prevented bone loss induced by ovariectomy(OVX),increased the bone mass density(BMD),improved the bone microarchitecture and reduced urine calcium and deoxypyridinoline(DPD)in ovariectomized mice,while inhibited the increase of body weight without significantly affecting the uterus weight.Furthermore,we found that NST increased osteogenic differentiation,inhibited adipogenic differentiation of BMSCs in vitro,and upregulated the expression of the key proteins of BMP and Wnt/β-catenin pathways.In addition,Noggin and Dickkopf-related protein-1(DKK-1)reversed the effect of NST on osteogenic differentiation and expression of the key proteins in BMP and Wnt/β-catenin pathway.The luciferase activities and the molecular docking analysis further supported the mechanism of NST.In conclusion,these results indicating that NST can be clinically used as a potential alternative medicine for the prevention and treatment of postmenopausal osteoporosis.展开更多
Areca nut is used worldwide as a hallucinogenic addicting drug along the tropical belt.Arecoline,a toxic compound,is the most important alkaloid in areca nuts.The adverse effects of oral uptake and chewing of areca nu...Areca nut is used worldwide as a hallucinogenic addicting drug along the tropical belt.Arecoline,a toxic compound,is the most important alkaloid in areca nuts.The adverse effects of oral uptake and chewing of areca nut are well known.For example,the possibility of cancer caused by chewing areca nuts is widely discussed.Chewing areca nut has other adverse effects on other organs,including abnormal cell differentiation,oral cancer,and several other diseases.The use of areca nut is also associated with low birthweight.Skeletal musculature is the largest organ in the body and is attached to the bones.During embryo development,the differentiation of bone and muscle cells is critical.In this article,we reviewed the effects of areca nut and arecoline on embryonic cell differentiation,particularly osteoblasts,myoblasts,and fibroblasts.展开更多
Bone formation is a highly energy-demanding process that can be impacted by metabolic disorders.Glucose has been considered the principal substrate for osteoblasts,although fatty acids are also important for osteoblas...Bone formation is a highly energy-demanding process that can be impacted by metabolic disorders.Glucose has been considered the principal substrate for osteoblasts,although fatty acids are also important for osteoblast function.Here,we report that osteoblasts can derive energy from endogenous fatty acids stored in lipid droplets via lipolysis and that this process is critical for bone formation.As such,we demonstrate that osteoblasts accumulate lipid droplets that are highly dynamic and provide the molecular mechanism by which they serve as a fuel source for energy generation during osteoblast maturation.Inhibiting cytoplasmic lipolysis leads to both an increase in lipid droplet size in osteoblasts and an impairment in osteoblast function.The fatty acids released by lipolysis from these lipid droplets become critical for cellular energy production as cellular energetics shifts towards oxidative phosphorylation during nutrient-depleted conditions.In vivo,conditional deletion of the ATGL-encoding gene Pnpla2 in osteoblast progenitor cells reduces cortical and trabecular bone parameters and alters skeletal lipid metabolism.Collectively,our data demonstrate that osteoblasts store fatty acids in the form of lipid droplets,which are released via lipolysis to support cellular bioenergetic status when nutrients are limited.Perturbations in this process result in impairment of bone formation,specifically reducing ATP production and overall osteoblast function.展开更多
Objective:To investigate whether acupotomy could inhibit subchondral bone remodeling in knee osteoarthritis(KOA)rabbits by regulating the activity of osteoblasts and osteoclasts.Methods:KOA rabbits were prepared by im...Objective:To investigate whether acupotomy could inhibit subchondral bone remodeling in knee osteoarthritis(KOA)rabbits by regulating the activity of osteoblasts and osteoclasts.Methods:KOA rabbits were prepared by immobilization for 6 and 9 weeks by Videman method.Nine groups of rabbits(control,6 weeks and 9 weeks model,6 weeks and 9 weeks acupotomy,6 weeks and 9 weeks electroacupuncture,and 6 weeks and 9 weeks drug groups)received acupotomy,electroacupuncture and risedronate sodium intervention,respectively,for 3 weeks.Results:Acupotomy can inhibit the activity of osteoclasts and osteoblasts in subchondral bone by reducing the proteins expression of cathepsin K(CK)and tartrate-resistant acid phosphatase(TRAP)and decreasing the proteins expression of osteocalcin(OCN)and alkaline phosphatase(ALP),to intercept the abnormal bone resorption and bone formation of subchondral bone in 6-week and 9-week immobilization-induced KOA rabbits.Conclusion:These findings indicated that acupotomy may be more advantageous than risedronate sodium intervention in modulating subchondral bone remodeling in KOA rabbits,especially in 9-week immobilization-induced KOA rabbits.展开更多
Summary: Rat transforming growth factor β1 (rTGFβ1) cDNA from rat lymphocytes was cloned by RT-PCR and inserted into pcDNA3 to construct an eukaryotic expression vector, which was named pcDNA3-TGFβ1. The cloned gen...Summary: Rat transforming growth factor β1 (rTGFβ1) cDNA from rat lymphocytes was cloned by RT-PCR and inserted into pcDNA3 to construct an eukaryotic expression vector, which was named pcDNA3-TGFβ1. The cloned gene was confirmed to code rat TGFβ1 by restriction enzyme analysis. pcDNA3-TGFβ1 plasmid was transfected into rat osteoblasts by using liposome-mediated gene transfer technique and the expression of TGFβ1 was detected by using irnmunohistochemical staining assay. It was found that the rat TGFβ1 expression product was obviously detectable in the transfected osteoblasts in 48 h. High expression of TGFβ1 was obtained in the rat osteoblasts in which the constructed TGFβ1 expression vector was transfected.展开更多
Objective:To study the changes of gene expression profiles associated with osteoblasts differentiated from rat bone marrow stromal cells in vitro by gene chip technique.Methods:rat Rone marrow stromal cells were isola...Objective:To study the changes of gene expression profiles associated with osteoblasts differentiated from rat bone marrow stromal cells in vitro by gene chip technique.Methods:rat Rone marrow stromal cells were isolated and cultured,and differentiation was induced by dexamethasone,β-glycerol phosphate and vitamin C.Cellular mRNA was extracted and reverse transcribed into cDNA,thus related genes expression differences were detected by gene expression profile chip.Results:Calcifying nodules were visible in the induced cells.There were27.7%genes expressed differentially,three times more than the normal and induced cells,and some genes were related to transcription,translation,glycosylation modification.Extracellular matrix,signal molecules and metabolism were up—regulated.Conclusions:The gene chip technique can be used to detect the multi-gene different expression in the differentiationinduceed rat BMSCs,and these differentially expressed genes are necessary genes related to rat BMSCs proliferation and induction of osteoblastic differentiation.展开更多
The effects of cerium ion(Ce3+) on the proliferation,differentiation,adipocytic transdifferentiation and mineralization function of primary mouse osteoblasts(OBs) were investigated.The results indicated that Ce3+ at a...The effects of cerium ion(Ce3+) on the proliferation,differentiation,adipocytic transdifferentiation and mineralization function of primary mouse osteoblasts(OBs) were investigated.The results indicated that Ce3+ at all concentrations(1×10-9,1×10-8,1×10-7,1×10-6,1×10-5,and 1×10-4 mol/L) promoted the proliferation of osteoblasts(OBs).On day 1 and 3,Ce3+ promoted the differentiation of OBs at concentrations of 1×10-9,1×10-7,and 1×10-6 mol/L,but inhibited the differentiation of OBs at higher concentrations.On day 2,Ce3+ inhibited the differentiation of OBs at tested concentrations.On day 9 and 12,Ce3+ inhibited the adipocytic transdifferentiation of OBs at most concentrations.On day 15,Ce3+ promoted the adipocytic transdifferentiation of OBs at concentrations of 1×10-9,1×10-6,1×10-5,and 1×10-4 mol/L,but had no effects at other concentrations.Ce3+ inhibited the formation of mineralized matrix nodules of OBs at concentrations of 1×10-9,1×10-8 and 1×10-7 mol/L,and promoted the formation of mineralized matrix nodules of OBs at other concentrations.These findings suggested that the effects of Ce3+ on the proliferation,differentiation,adipocytic transdifferentiation and mineralization function of primary OBs depended on the concentration and culture time;moreover,they were pivotal factors for switching the biological effects of Ce3+ from toxicity to activity,from damage to protection,or from down-regulation to up-regulation.展开更多
Cigarette consumption increases oxidative stress in many organs. Increased oxidative stress harms bone cells, which negatively affects bone-matter and -stability. This leads to an increased fracture risk and delayed f...Cigarette consumption increases oxidative stress in many organs. Increased oxidative stress harms bone cells, which negatively affects bone-matter and -stability. This leads to an increased fracture risk and delayed fracture healing in smokers. A supporting therapy with antioxidants could be of great benefit for surgeons dealing with delayed fracture healing due to increased oxidative stress. In this article we complement and compare our published data with hitherto unpublished data and show the protective effect of 15 different antioxidants on cigarette smoke induced damage in primary human osteoblasts. Exposure to cigarette smoke medium (CSM) rapidly induces formation of ROS in osteoblasts in a concentration- and time-dependent manner. Massive cell damage is seen already after 4 h (EC50 ≈ 0.75 OD320). Pre-, co- and post-incubation with the different antioxidants reduces the formation of ROS and consequently improves the viability of the CSM exposed osteoblasts. Small compounds, e.g. N-acetylcysteine, proved highly effective if pre- or co-incubated before exposure to the CSM. Thus, they are good candidates for acute therapy support as they can be administered in high doses. However, our data suggest that a balanced daily diet could lead to an accumulation of various natural antioxidants (flavonoids) that effectively protect osteoblasts from oxidative stress-induced damage in all three settings investigated. Together with their partly phytoestrogenic properties this may even abate alterations in bone and thus reduce fracture risk on the long run.展开更多
Summary: In order to investigate the effect of TGFβ 1 gene transfer on the biological characteristics, the effects of gene transfer and supernatant of transfected osteoblasts on the proliferation and ALP activity of ...Summary: In order to investigate the effect of TGFβ 1 gene transfer on the biological characteristics, the effects of gene transfer and supernatant of transfected osteoblasts on the proliferation and ALP activity of osteoblasts were detected by 3H-TdR and MTT. Our results showed that TGFβ 1 gene transfer had no effect on the biological characteristics and the activated supernatant of transfected osteoblasts stimulated proliferation and inhibited ALP activity of osteoblasts. TGFβ 1 gene transfer could promote the expression of TGFβ 1 and the biological characteristics of transfected osteoblasts were stable, which might be helpful for gene therapy of bone defects in vivo.展开更多
Objective:To verify whether icariin has drug toxicity to osteoblasts and its pre protective effect on inflammatory osteoblasts induced by lipopolysaccharide.Methods:(1)The osteoblasts of newborn SD rats were extracted...Objective:To verify whether icariin has drug toxicity to osteoblasts and its pre protective effect on inflammatory osteoblasts induced by lipopolysaccharide.Methods:(1)The osteoblasts of newborn SD rats were extracted,cultured and passaged.By observing the morphology of osteoblasts and ALP staining,the activity and proliferation of osteoblasts were determined.(2)CCK-8 method was used to observe the effect of Icariin on osteoblast activity.(3)Different concentrations of lipopolysaccharide(LPS)were used to induce inflammatory osteoblasts in fetal rats.CCK-8 method was used to select the best concentration for induction.(4)This experiment was divided into control group,low concentration group,middle concentration group and high concentration group.CCK-8 method was used to observe whether icariin could protect osteoblasts from inflammatory reaction.Results:(1)The number of osteoblasts in the third generation increased,the shape of osteoblasts overlapped like tiles,most of osteoblasts grew in the center,the center was dense and the specific shape was difficult to see.After ALP staining,the positive cells showed gray black granules in cytoplasm and irregular cell body shape under inverted microscope;(2)DWhen the concentration of icariin was lower than 1μg/ml,it had no significant effect on osteoblasts(P<0.05);when it was higher than 10μg/ml,icariin had no significant effect on osteoblasts(P<0.05);(3)When the concentration of lipopolysaccharide was higher than 80μg/ml,there was a significant trend of inflammatory damage to osteoblasts,which was statistically significant(P<0.01).Therefore,80μg/ml was selected as the best injury concentration in this experiment;(4)When the concentration of icariin was lower than 1μg/ml,there was no significant pre protective effect on the inflammatory response of osteoblasts(P<0.05);when it was higher than 10μg/ml,there was significant pre protective effect on the inflammatory response of osteoblasts(P<0.05).Conclusion:When the concentration of icariin reaches a certain level,it can promote the proliferation of osteoblasts and has a certain pre protective effect on inflammatory osteoblasts.展开更多
Objectives: This study explores feasibility of tissue-engineered osteogenesis using sterile coral implants loaded with homologous osteoblasts to repair bone defects. Study Design: A unilateral 4 mm transverse dis- con...Objectives: This study explores feasibility of tissue-engineered osteogenesis using sterile coral implants loaded with homologous osteoblasts to repair bone defects. Study Design: A unilateral 4 mm transverse dis- continuity defect was produced approximately mid-way along left radius of young female rabbits using ro- tary diamond disc under continuous saline irrigation and stabilised with autoclaved steel miniplate and screws. The defect was then fitted with sterile bioresorbable coral implant loaded with homologous neonatal calvarial osteoblasts or control implants without osteoblasts. All animals underwent radiography immedi- ately post-operative, at weekly intervals for four weeks and at fortnightly intervals thereafter. Operated bones were histologically evaluated for osteogenesis at 12 weeks. Results: Findings demonstrate osteogenesis and complete repair of bioresorbable coral implant by homologous osteoblasts loaded on coral scaffold. Conclu- sions: Single stage surgery using this technique to induce osteogenesis and closure of discontinuity bone de- fects including palatal clefts and peripheral reduction of large craniofacial defects might prove better thera- peutic modality than autologous bone grafting or tissue distraction osteogenesis.展开更多
Vascular calcification, which causes occlusion and rupture of the vascular, is often observed in patients in the advanced stages of arteriosclerosis. One of the best procedures for inhibiting the accumulation of vascu...Vascular calcification, which causes occlusion and rupture of the vascular, is often observed in patients in the advanced stages of arteriosclerosis. One of the best procedures for inhibiting the accumulation of vascular calcification is to obstruct the differentiation of mesenchymal stem cells (MSCs) and/or vascular endothelial cells (VECs) in the vascular to osteoblasts. In this study, we evaluated the biochemical and genetic characteristics of the process of differentiation of MSCs and VECs to osteoblasts. C3H10T1/2 MSCs, TKD2 VECs and MC3T3-E1 preosteoblasts (POBs) were cultured in medium containing both hydrocortisone and glycerophosphate. These compounds showed strong effects promoting the differentiation of VECs as well as POBs, although the effect was weak in the MSCs. Moreover, C3H10T1/2 MSCs and TKD2 VECs were cultured in medium containing 10 mM retinol, after which the alkali phosphatase (ALP) activity of the MSCs and production of calcified nodules of TKD2 were significantly increased, whereas the marker genes for the osteoblasts were not. These results suggest that retinol does not have an effect in inducing the differentiation of VECs to osteoblasts, but rather exhibits a strong promoting effect on differentiation.展开更多
PURPOSE: This study was designed to investigate and compare in vitro survivin activity in normal human osteoblast and MG-63 osteosarcoma cell cultures with and without vitamin D3. METHODS: Normal human alveolar bone e...PURPOSE: This study was designed to investigate and compare in vitro survivin activity in normal human osteoblast and MG-63 osteosarcoma cell cultures with and without vitamin D3. METHODS: Normal human alveolar bone explants were recovered from extraction sites of non-carious teeth of 9 healthy donors and cultured to the 2nd passage. MG-63 osteosarcoma cells were obtained from ATCC. To compare the survivin activities in these two types of cells and to determine the effect of vitamin D3 on survivin expression and associated activities of cell proliferation and differentiation, levels of survivin, osteocalcin and alkaline phosphatase were measured at 7-20 day cultures with and without vitamin D3 in both cultures of normal osteoblasts and osteosarcoma cells. Matched pair t-test and two sample independent t-test were applied for statistical analysis of the data. P values of 0.05 or less were considered statistically significant. RESULTS: A significantly higher level of survivin was detected in normal osteoblasts compared to osteosarcoma cells at 7 days without vitamin D3 treatment (P<0.01). Survivin expression in normal osteoblasts significantly decreased after vitamin D3 treatment at 7 days (P<0.05), but not at 20 days of culture (P=NS), compared to that in normal osteoblast culture without vitamin D3 treatment. Vitamin D3 had no effect on survivin expression in osteosarcoma cells at 7 or 20 days (P=NS). CONCLUSIONS: Expression of survivin in cultured normal human osteoblasts and osteosarcoma cells was positively identified. There is a positive correlation between higher expression of survivin and less differentiated osteoblasts that still retain their proliferative ability. Vitamin D3 has significant negative effect on expression of survivin in normal human osteoblasts but not on that in osteosarcoma cells.展开更多
Bone-resorbing osteoclasts are formed from a monocyte/macrophage lineage under the strict control o bone-forming osteoblasts. So far,macrophage colonystimulating factor(M-CSF),receptor activator o nuclear factor-κB l...Bone-resorbing osteoclasts are formed from a monocyte/macrophage lineage under the strict control o bone-forming osteoblasts. So far,macrophage colonystimulating factor(M-CSF),receptor activator o nuclear factor-κB ligand(RANKL),and osteoprotegerin(OPG) produced by osteoblasts play major roles in the regulation of osteoclast differentiation. Recent studies have shown that osteoblasts regulate osteoclastogenesis through several mechanisms independent o M-CSF,RANKL,and OPG production. Identification o osteoclast-committed precursors in vivo demonstrated that osteoblasts are involved in the distribution o osteoclast precursors in bone. Interleukin 34(IL-34)a novel ligand for c-Fms,plays a pivotal role in maintaining the splenic reservoir of osteoclast-committed precursors in M-CSF deficient mice. IL-34 is also able to act as a substitute for osteoblast-producing M-CSF in osteoclastogenesis. Wnt5 a,produced by osteoblasts,enhances osteoclast differentiation by upregulating RANK expression through activation of the noncanonical Wnt pathway. Semaphorin 3A produced by osteoblasts inhibits RANKL-induced osteoclast differentiation through the suppression of immunoreceptortyrosine-based activation motif signals. Thus,recent findings show that osteoclast differentiation is tightly regulated by osteoblasts through several different mechanisms. These newly identified molecules are expected to be promising targets of therapeutic agents in bone-related diseases.展开更多
Persistent generalized low bone mineral density (BMD) has been reported in patients with adolescent idiopathic scoliosis (AIS).However,the exact mechanisms and causes of the low BMD in AIS patients are largely unknown...Persistent generalized low bone mineral density (BMD) has been reported in patients with adolescent idiopathic scoliosis (AIS).However,the exact mechanisms and causes of the low BMD in AIS patients are largely unknown.The purpose of this study was to examine the relationship between the receptor activator of NF-κB ligand (RANKL)/osteoprotegerin (OPG) levels in osteoblasts (OBs) from AIS patients with low BMD and with comparison made between the patients and controls.Twenty AIS patients and eight age-matched controls were included in the present study.The BMD of lumbar spine and proximal femur was measured in all subjects.OBs from the cancellous bone of each subject was harvested and primarily cultured.The mRNA and protein expression of RANKL and OPG in OBs was detected by RT-PCR and Western blotting.The results showed BMD was lower in AIS patients than in controls.A significantly higher mRNA and protein expression of RANKL was observed in OBs from AIS patients,while no significant difference was found in the expression of OPG between AIS patients and controls.As a result,RANKL/OPG ratio in patients with AIS was remarkably higher than controls.Our study preliminarily demonstrated expression of RANKL was higher in OBs from AIS patients with low BMD as compared with controls,suggesting the unbalanced RANKL/OPG ratio caused by an over-expression of RANKL in OBs may be responsible for the low BMD in AIS patients.展开更多
BACKGROUND Cigarette smoking(CS)is the most common method of consuming tobacco.Deleterious effects on bone integrity,increased incidence of fractures,and delayed fracture healing are all associated with CS.Over 150 of...BACKGROUND Cigarette smoking(CS)is the most common method of consuming tobacco.Deleterious effects on bone integrity,increased incidence of fractures,and delayed fracture healing are all associated with CS.Over 150 of the 6500 molecular species contained in cigarette smoke and identified as toxic compounds are inhaled by CS and,via the bloodstream,reach the skeletal system.New technologies designed to develop a reduced-risk alternative for smokers are based on electronic nicotine delivery systems,such as e-cigarettes and tobacco heating systems(THS).THS are designed to heat tobacco instead of burning it,thereby reducing the levels of harmful toxic compounds released.AIM To examine the effects of THS on osteoprogenitor cell viability and function compared to conventional CS.METHODS Human immortalized mesenchymal stem cells(n=3)and primary human preosteoblasts isolated from cancellous bone samples from BG Unfall Klinik Tübingen(n=5)were osteogenically differentiated in vitro with aqueous extracts generated from either the THS 2.4“IQOS”or conventional“Marlboro”cigarettes for up to 21 d.Cell viability was analyzed using resazurin conversion assay(mitochondrial activity)and calcein-AM staining(esterase activity).Osteogenic differentiation and bone cell function were evaluated using alkaline phosphatase(AP)activity,while matrix formation was analyzed through alizarin red staining.Primary cilia structure was examined by acetylatedα-tubulin immunofluorescent staining.Free radical production was evaluated with 2’,7’-dichlorofluoresceindiacetate assay.RESULTS Our data clearly show that THS is significantly less toxic to bone cells than CS when analyzed by mitochondrial and esterase activity(P<0.001).No significant differences in cytotoxicity between the diverse flavors of THS were observed.Harmful effects from THS on bone cell function were observed only at very high,non-physiological concentrations.In contrast,extracts from conventional cigarettes significantly reduced the AP activity(by two-fold)and matrix mineralization(four-fold)at low concentrations.Additionally,morphologic analysis of primary cilia revealed no significant changes in the length of the organelle involved in osteogenesis of osteoprogenitor cells,nor in the number of ciliated cells following THS treatment.Assessment of free radical production demonstrated that THS induced significantly less oxidative stress than conventional CS in osteoprogenitor cells.CONCLUSIONTHS was significantly less harmful to osteoprogenitor cells during osteogenesisthan conventional CS. Additional studies are required to confirm whether THS isa better alternative for smokers to improve delays in bone healing followingfracture.展开更多
Since 3D printed hard materials could match the shape of bone,cell survival and fate determination towards osteoblasts in such materials have become a popular research target.In this study,a scaffold of hardmaterial f...Since 3D printed hard materials could match the shape of bone,cell survival and fate determination towards osteoblasts in such materials have become a popular research target.In this study,a scaffold of hardmaterial for 3D fabrication was designed to regulate developmental signal(Notch)transduction guiding osteoblast differentiation.We established a polycaprolactone(PCL)and cell-integrated 3D printing system(PCI3D)to reciprocally print the beams of PCL and cell-laden hydrogel for a module.This PCI3D module holds good cell viability of over 87%,whereas cells show about sixfold proliferation in a 7-day culture.The osteocytic MLO-Y4 was engineered to overexpress Notch ligand Dll4,making up 25%after mixing with 75%stromal cells in the PCI3D module.Osteocytic Dll4,unlike other delta-like family members such as Dll1 or Dll3,promotes osteoblast differentiation and themineralization of primary mouse and a cell line of bone marrow stromal cells when cultured in a PCI3D module for up to 28 days.Mechanistically,osteocytic Dll4 could not promote osteogenic differentiation of the primary bone marrow stromal cells(BMSCs)after conditional deletion of the Notch transcription factor RBPjκby Cre recombinase.These data indicate that osteocytic Dll4 activates RBPjκ-dependent canonical Notch signaling in BMSCs for their oriented differentiation towards osteoblasts.Additionally,osteocytic Dll4 holds a great potential for angiogenesis in human umbilical vein endothelial cells within modules.Our study reveals that osteocytic Dll4 could be the osteogenic niche determining cell fate towards osteoblasts.This will open a new avenue to overcome the current limitation of poor cell viability and low bioactivity of traditional orthopedic implants.展开更多
Summary: 17β-estradiol (E2), progesterone (P) and testosterone (T) were investigated for their effects on the proliferation and differentiation of primary rat calvarial osteoblasts in vitro. Rat calvarial osteoblasts...Summary: 17β-estradiol (E2), progesterone (P) and testosterone (T) were investigated for their effects on the proliferation and differentiation of primary rat calvarial osteoblasts in vitro. Rat calvarial osteoblasts were cultured with 10-10 mol/L E2, 10-9-10-6 mol/L P and 10-10 mol/L T for 20 days. Cell proliferation was determined by 3H-thymidine incorporation and cell counting. Cell differentiation was examined by measuring alkaline phosphatase (ALP) activity, osteocalcin gene expression and production, bone nodule formation in different periods of culture. Our results showed that no effect of three sex hormones was observed on cell proliferation, but, the responses of cell differentiation to the hormones were more or less different. Among these three hormones used in this study, P appeared to have multi-stimulating effect on cell differentiation. Effect of T seemed not so significant as that of P on cell differentiation. Although ALP activity and osteocalcin production were increased significantly by T, it had slight effect on osteocalcin mRNA and bone nodule formation. Besides, E2 did not demonstrate any effect on cell differentiation. It is concluded that the proliferation of rat calvarial osteoblasts was independent of the presence of sex hormones. However, the differentiation of these cells were stimulated in different levels and to different extent either by P or T. P appeared to be a multi-stimulator on differentiation of such cells.展开更多
基金support from the Public Platform of Medical Research Center,Academy of Chinese Medical Science,Zhejiang Chinese Medical Universitysponsored by the National Natural Science Foundation of China(81973534,U1505226)。
文摘Increasing the osteogenic differentiation ability and decreasing the adipogenic differentiation ability of bone marrow mesenchymal stem cells(BMSCs)is a potential strategy for the treatment of osteoporosis(OP).Naturally derived oligosaccharides have shown significant anti-osteoporotic effects.Nystose(NST),an oligosaccharide,was isolated from the roots of Morinda officinalis How.(MO).The aim of the present study was to investigate the effects of NST on bone loss in ovariectomized mice,and explore the underlying mechanism of NST in promoting differentiation of BMSCs to osteoblasts.Administration of NST(40,80 and 160 mg/kg)and the positive control of estradiol valerate(0.2 mg/kg)for 8 weeks significantly prevented bone loss induced by ovariectomy(OVX),increased the bone mass density(BMD),improved the bone microarchitecture and reduced urine calcium and deoxypyridinoline(DPD)in ovariectomized mice,while inhibited the increase of body weight without significantly affecting the uterus weight.Furthermore,we found that NST increased osteogenic differentiation,inhibited adipogenic differentiation of BMSCs in vitro,and upregulated the expression of the key proteins of BMP and Wnt/β-catenin pathways.In addition,Noggin and Dickkopf-related protein-1(DKK-1)reversed the effect of NST on osteogenic differentiation and expression of the key proteins in BMP and Wnt/β-catenin pathway.The luciferase activities and the molecular docking analysis further supported the mechanism of NST.In conclusion,these results indicating that NST can be clinically used as a potential alternative medicine for the prevention and treatment of postmenopausal osteoporosis.
基金the funding provided by the Ministry of Science and Technology,Taiwan(108-2314-B-037-075)the Kaohsiung Medical University Research Foundation(KMU-M103001,KMU-M104003,KMU-TP104PR16).
文摘Areca nut is used worldwide as a hallucinogenic addicting drug along the tropical belt.Arecoline,a toxic compound,is the most important alkaloid in areca nuts.The adverse effects of oral uptake and chewing of areca nut are well known.For example,the possibility of cancer caused by chewing areca nuts is widely discussed.Chewing areca nut has other adverse effects on other organs,including abnormal cell differentiation,oral cancer,and several other diseases.The use of areca nut is also associated with low birthweight.Skeletal musculature is the largest organ in the body and is attached to the bones.During embryo development,the differentiation of bone and muscle cells is critical.In this article,we reviewed the effects of areca nut and arecoline on embryonic cell differentiation,particularly osteoblasts,myoblasts,and fibroblasts.
基金supported by National Institutes of Health(NIH),the National Institute of Arthritis and Musculoskeletal and Skin Diseases(NIAMS)Grant K01AR072123(ER-R)the American Society of Bone and Mineral Research(ASBMR)Rising Star awardprovided by NIH-National Institute of Diabetes and Digestive and Kidney Disease(NIDDK)Grant DK116056(MPC)。
文摘Bone formation is a highly energy-demanding process that can be impacted by metabolic disorders.Glucose has been considered the principal substrate for osteoblasts,although fatty acids are also important for osteoblast function.Here,we report that osteoblasts can derive energy from endogenous fatty acids stored in lipid droplets via lipolysis and that this process is critical for bone formation.As such,we demonstrate that osteoblasts accumulate lipid droplets that are highly dynamic and provide the molecular mechanism by which they serve as a fuel source for energy generation during osteoblast maturation.Inhibiting cytoplasmic lipolysis leads to both an increase in lipid droplet size in osteoblasts and an impairment in osteoblast function.The fatty acids released by lipolysis from these lipid droplets become critical for cellular energy production as cellular energetics shifts towards oxidative phosphorylation during nutrient-depleted conditions.In vivo,conditional deletion of the ATGL-encoding gene Pnpla2 in osteoblast progenitor cells reduces cortical and trabecular bone parameters and alters skeletal lipid metabolism.Collectively,our data demonstrate that osteoblasts store fatty acids in the form of lipid droplets,which are released via lipolysis to support cellular bioenergetic status when nutrients are limited.Perturbations in this process result in impairment of bone formation,specifically reducing ATP production and overall osteoblast function.
基金supported by the Beijing Municipal Natural Science Foundation(7192110)。
文摘Objective:To investigate whether acupotomy could inhibit subchondral bone remodeling in knee osteoarthritis(KOA)rabbits by regulating the activity of osteoblasts and osteoclasts.Methods:KOA rabbits were prepared by immobilization for 6 and 9 weeks by Videman method.Nine groups of rabbits(control,6 weeks and 9 weeks model,6 weeks and 9 weeks acupotomy,6 weeks and 9 weeks electroacupuncture,and 6 weeks and 9 weeks drug groups)received acupotomy,electroacupuncture and risedronate sodium intervention,respectively,for 3 weeks.Results:Acupotomy can inhibit the activity of osteoclasts and osteoblasts in subchondral bone by reducing the proteins expression of cathepsin K(CK)and tartrate-resistant acid phosphatase(TRAP)and decreasing the proteins expression of osteocalcin(OCN)and alkaline phosphatase(ALP),to intercept the abnormal bone resorption and bone formation of subchondral bone in 6-week and 9-week immobilization-induced KOA rabbits.Conclusion:These findings indicated that acupotomy may be more advantageous than risedronate sodium intervention in modulating subchondral bone remodeling in KOA rabbits,especially in 9-week immobilization-induced KOA rabbits.
文摘Summary: Rat transforming growth factor β1 (rTGFβ1) cDNA from rat lymphocytes was cloned by RT-PCR and inserted into pcDNA3 to construct an eukaryotic expression vector, which was named pcDNA3-TGFβ1. The cloned gene was confirmed to code rat TGFβ1 by restriction enzyme analysis. pcDNA3-TGFβ1 plasmid was transfected into rat osteoblasts by using liposome-mediated gene transfer technique and the expression of TGFβ1 was detected by using irnmunohistochemical staining assay. It was found that the rat TGFβ1 expression product was obviously detectable in the transfected osteoblasts in 48 h. High expression of TGFβ1 was obtained in the rat osteoblasts in which the constructed TGFβ1 expression vector was transfected.
文摘Objective:To study the changes of gene expression profiles associated with osteoblasts differentiated from rat bone marrow stromal cells in vitro by gene chip technique.Methods:rat Rone marrow stromal cells were isolated and cultured,and differentiation was induced by dexamethasone,β-glycerol phosphate and vitamin C.Cellular mRNA was extracted and reverse transcribed into cDNA,thus related genes expression differences were detected by gene expression profile chip.Results:Calcifying nodules were visible in the induced cells.There were27.7%genes expressed differentially,three times more than the normal and induced cells,and some genes were related to transcription,translation,glycosylation modification.Extracellular matrix,signal molecules and metabolism were up—regulated.Conclusions:The gene chip technique can be used to detect the multi-gene different expression in the differentiationinduceed rat BMSCs,and these differentially expressed genes are necessary genes related to rat BMSCs proliferation and induction of osteoblastic differentiation.
基金supported by the Natural Science Foundation of China (20971034)Foundation for Key Program of Ministry of Education of China (208018)+2 种基金Returned Scholars of Hebei Province (207041)Natural Science Foundation of Hebei Province (B2009000161)Natural Science Foundation of Hebei University
文摘The effects of cerium ion(Ce3+) on the proliferation,differentiation,adipocytic transdifferentiation and mineralization function of primary mouse osteoblasts(OBs) were investigated.The results indicated that Ce3+ at all concentrations(1×10-9,1×10-8,1×10-7,1×10-6,1×10-5,and 1×10-4 mol/L) promoted the proliferation of osteoblasts(OBs).On day 1 and 3,Ce3+ promoted the differentiation of OBs at concentrations of 1×10-9,1×10-7,and 1×10-6 mol/L,but inhibited the differentiation of OBs at higher concentrations.On day 2,Ce3+ inhibited the differentiation of OBs at tested concentrations.On day 9 and 12,Ce3+ inhibited the adipocytic transdifferentiation of OBs at most concentrations.On day 15,Ce3+ promoted the adipocytic transdifferentiation of OBs at concentrations of 1×10-9,1×10-6,1×10-5,and 1×10-4 mol/L,but had no effects at other concentrations.Ce3+ inhibited the formation of mineralized matrix nodules of OBs at concentrations of 1×10-9,1×10-8 and 1×10-7 mol/L,and promoted the formation of mineralized matrix nodules of OBs at other concentrations.These findings suggested that the effects of Ce3+ on the proliferation,differentiation,adipocytic transdifferentiation and mineralization function of primary OBs depended on the concentration and culture time;moreover,they were pivotal factors for switching the biological effects of Ce3+ from toxicity to activity,from damage to protection,or from down-regulation to up-regulation.
文摘Cigarette consumption increases oxidative stress in many organs. Increased oxidative stress harms bone cells, which negatively affects bone-matter and -stability. This leads to an increased fracture risk and delayed fracture healing in smokers. A supporting therapy with antioxidants could be of great benefit for surgeons dealing with delayed fracture healing due to increased oxidative stress. In this article we complement and compare our published data with hitherto unpublished data and show the protective effect of 15 different antioxidants on cigarette smoke induced damage in primary human osteoblasts. Exposure to cigarette smoke medium (CSM) rapidly induces formation of ROS in osteoblasts in a concentration- and time-dependent manner. Massive cell damage is seen already after 4 h (EC50 ≈ 0.75 OD320). Pre-, co- and post-incubation with the different antioxidants reduces the formation of ROS and consequently improves the viability of the CSM exposed osteoblasts. Small compounds, e.g. N-acetylcysteine, proved highly effective if pre- or co-incubated before exposure to the CSM. Thus, they are good candidates for acute therapy support as they can be administered in high doses. However, our data suggest that a balanced daily diet could lead to an accumulation of various natural antioxidants (flavonoids) that effectively protect osteoblasts from oxidative stress-induced damage in all three settings investigated. Together with their partly phytoestrogenic properties this may even abate alterations in bone and thus reduce fracture risk on the long run.
文摘Summary: In order to investigate the effect of TGFβ 1 gene transfer on the biological characteristics, the effects of gene transfer and supernatant of transfected osteoblasts on the proliferation and ALP activity of osteoblasts were detected by 3H-TdR and MTT. Our results showed that TGFβ 1 gene transfer had no effect on the biological characteristics and the activated supernatant of transfected osteoblasts stimulated proliferation and inhibited ALP activity of osteoblasts. TGFβ 1 gene transfer could promote the expression of TGFβ 1 and the biological characteristics of transfected osteoblasts were stable, which might be helpful for gene therapy of bone defects in vivo.
基金The 61th Batch of China Postdoctoral Science Foundation(No.2017M611884)General topic of medical science and Technology development in Nanjing,Jiangsu Province(No.YKK17148)。
文摘Objective:To verify whether icariin has drug toxicity to osteoblasts and its pre protective effect on inflammatory osteoblasts induced by lipopolysaccharide.Methods:(1)The osteoblasts of newborn SD rats were extracted,cultured and passaged.By observing the morphology of osteoblasts and ALP staining,the activity and proliferation of osteoblasts were determined.(2)CCK-8 method was used to observe the effect of Icariin on osteoblast activity.(3)Different concentrations of lipopolysaccharide(LPS)were used to induce inflammatory osteoblasts in fetal rats.CCK-8 method was used to select the best concentration for induction.(4)This experiment was divided into control group,low concentration group,middle concentration group and high concentration group.CCK-8 method was used to observe whether icariin could protect osteoblasts from inflammatory reaction.Results:(1)The number of osteoblasts in the third generation increased,the shape of osteoblasts overlapped like tiles,most of osteoblasts grew in the center,the center was dense and the specific shape was difficult to see.After ALP staining,the positive cells showed gray black granules in cytoplasm and irregular cell body shape under inverted microscope;(2)DWhen the concentration of icariin was lower than 1μg/ml,it had no significant effect on osteoblasts(P<0.05);when it was higher than 10μg/ml,icariin had no significant effect on osteoblasts(P<0.05);(3)When the concentration of lipopolysaccharide was higher than 80μg/ml,there was a significant trend of inflammatory damage to osteoblasts,which was statistically significant(P<0.01).Therefore,80μg/ml was selected as the best injury concentration in this experiment;(4)When the concentration of icariin was lower than 1μg/ml,there was no significant pre protective effect on the inflammatory response of osteoblasts(P<0.05);when it was higher than 10μg/ml,there was significant pre protective effect on the inflammatory response of osteoblasts(P<0.05).Conclusion:When the concentration of icariin reaches a certain level,it can promote the proliferation of osteoblasts and has a certain pre protective effect on inflammatory osteoblasts.
文摘Objectives: This study explores feasibility of tissue-engineered osteogenesis using sterile coral implants loaded with homologous osteoblasts to repair bone defects. Study Design: A unilateral 4 mm transverse dis- continuity defect was produced approximately mid-way along left radius of young female rabbits using ro- tary diamond disc under continuous saline irrigation and stabilised with autoclaved steel miniplate and screws. The defect was then fitted with sterile bioresorbable coral implant loaded with homologous neonatal calvarial osteoblasts or control implants without osteoblasts. All animals underwent radiography immedi- ately post-operative, at weekly intervals for four weeks and at fortnightly intervals thereafter. Operated bones were histologically evaluated for osteogenesis at 12 weeks. Results: Findings demonstrate osteogenesis and complete repair of bioresorbable coral implant by homologous osteoblasts loaded on coral scaffold. Conclu- sions: Single stage surgery using this technique to induce osteogenesis and closure of discontinuity bone de- fects including palatal clefts and peripheral reduction of large craniofacial defects might prove better thera- peutic modality than autologous bone grafting or tissue distraction osteogenesis.
文摘Vascular calcification, which causes occlusion and rupture of the vascular, is often observed in patients in the advanced stages of arteriosclerosis. One of the best procedures for inhibiting the accumulation of vascular calcification is to obstruct the differentiation of mesenchymal stem cells (MSCs) and/or vascular endothelial cells (VECs) in the vascular to osteoblasts. In this study, we evaluated the biochemical and genetic characteristics of the process of differentiation of MSCs and VECs to osteoblasts. C3H10T1/2 MSCs, TKD2 VECs and MC3T3-E1 preosteoblasts (POBs) were cultured in medium containing both hydrocortisone and glycerophosphate. These compounds showed strong effects promoting the differentiation of VECs as well as POBs, although the effect was weak in the MSCs. Moreover, C3H10T1/2 MSCs and TKD2 VECs were cultured in medium containing 10 mM retinol, after which the alkali phosphatase (ALP) activity of the MSCs and production of calcified nodules of TKD2 were significantly increased, whereas the marker genes for the osteoblasts were not. These results suggest that retinol does not have an effect in inducing the differentiation of VECs to osteoblasts, but rather exhibits a strong promoting effect on differentiation.
文摘PURPOSE: This study was designed to investigate and compare in vitro survivin activity in normal human osteoblast and MG-63 osteosarcoma cell cultures with and without vitamin D3. METHODS: Normal human alveolar bone explants were recovered from extraction sites of non-carious teeth of 9 healthy donors and cultured to the 2nd passage. MG-63 osteosarcoma cells were obtained from ATCC. To compare the survivin activities in these two types of cells and to determine the effect of vitamin D3 on survivin expression and associated activities of cell proliferation and differentiation, levels of survivin, osteocalcin and alkaline phosphatase were measured at 7-20 day cultures with and without vitamin D3 in both cultures of normal osteoblasts and osteosarcoma cells. Matched pair t-test and two sample independent t-test were applied for statistical analysis of the data. P values of 0.05 or less were considered statistically significant. RESULTS: A significantly higher level of survivin was detected in normal osteoblasts compared to osteosarcoma cells at 7 days without vitamin D3 treatment (P<0.01). Survivin expression in normal osteoblasts significantly decreased after vitamin D3 treatment at 7 days (P<0.05), but not at 20 days of culture (P=NS), compared to that in normal osteoblast culture without vitamin D3 treatment. Vitamin D3 had no effect on survivin expression in osteosarcoma cells at 7 or 20 days (P=NS). CONCLUSIONS: Expression of survivin in cultured normal human osteoblasts and osteosarcoma cells was positively identified. There is a positive correlation between higher expression of survivin and less differentiated osteoblasts that still retain their proliferative ability. Vitamin D3 has significant negative effect on expression of survivin in normal human osteoblasts but not on that in osteosarcoma cells.
基金This work was supported by grantsfromThe Int .Cooperation Projectfor National &Abroad Lab.of the National Natural Sciences Foundation ofChina(2002008) and The Science &Technology Foundation of Liaoningprovince (20022140)
文摘Bone-resorbing osteoclasts are formed from a monocyte/macrophage lineage under the strict control o bone-forming osteoblasts. So far,macrophage colonystimulating factor(M-CSF),receptor activator o nuclear factor-κB ligand(RANKL),and osteoprotegerin(OPG) produced by osteoblasts play major roles in the regulation of osteoclast differentiation. Recent studies have shown that osteoblasts regulate osteoclastogenesis through several mechanisms independent o M-CSF,RANKL,and OPG production. Identification o osteoclast-committed precursors in vivo demonstrated that osteoblasts are involved in the distribution o osteoclast precursors in bone. Interleukin 34(IL-34)a novel ligand for c-Fms,plays a pivotal role in maintaining the splenic reservoir of osteoclast-committed precursors in M-CSF deficient mice. IL-34 is also able to act as a substitute for osteoblast-producing M-CSF in osteoclastogenesis. Wnt5 a,produced by osteoblasts,enhances osteoclast differentiation by upregulating RANK expression through activation of the noncanonical Wnt pathway. Semaphorin 3A produced by osteoblasts inhibits RANKL-induced osteoclast differentiation through the suppression of immunoreceptortyrosine-based activation motif signals. Thus,recent findings show that osteoclast differentiation is tightly regulated by osteoblasts through several different mechanisms. These newly identified molecules are expected to be promising targets of therapeutic agents in bone-related diseases.
基金supported by the National Natural ScienceFoundation of China (No.81101335)
文摘Persistent generalized low bone mineral density (BMD) has been reported in patients with adolescent idiopathic scoliosis (AIS).However,the exact mechanisms and causes of the low BMD in AIS patients are largely unknown.The purpose of this study was to examine the relationship between the receptor activator of NF-κB ligand (RANKL)/osteoprotegerin (OPG) levels in osteoblasts (OBs) from AIS patients with low BMD and with comparison made between the patients and controls.Twenty AIS patients and eight age-matched controls were included in the present study.The BMD of lumbar spine and proximal femur was measured in all subjects.OBs from the cancellous bone of each subject was harvested and primarily cultured.The mRNA and protein expression of RANKL and OPG in OBs was detected by RT-PCR and Western blotting.The results showed BMD was lower in AIS patients than in controls.A significantly higher mRNA and protein expression of RANKL was observed in OBs from AIS patients,while no significant difference was found in the expression of OPG between AIS patients and controls.As a result,RANKL/OPG ratio in patients with AIS was remarkably higher than controls.Our study preliminarily demonstrated expression of RANKL was higher in OBs from AIS patients with low BMD as compared with controls,suggesting the unbalanced RANKL/OPG ratio caused by an over-expression of RANKL in OBs may be responsible for the low BMD in AIS patients.
文摘BACKGROUND Cigarette smoking(CS)is the most common method of consuming tobacco.Deleterious effects on bone integrity,increased incidence of fractures,and delayed fracture healing are all associated with CS.Over 150 of the 6500 molecular species contained in cigarette smoke and identified as toxic compounds are inhaled by CS and,via the bloodstream,reach the skeletal system.New technologies designed to develop a reduced-risk alternative for smokers are based on electronic nicotine delivery systems,such as e-cigarettes and tobacco heating systems(THS).THS are designed to heat tobacco instead of burning it,thereby reducing the levels of harmful toxic compounds released.AIM To examine the effects of THS on osteoprogenitor cell viability and function compared to conventional CS.METHODS Human immortalized mesenchymal stem cells(n=3)and primary human preosteoblasts isolated from cancellous bone samples from BG Unfall Klinik Tübingen(n=5)were osteogenically differentiated in vitro with aqueous extracts generated from either the THS 2.4“IQOS”or conventional“Marlboro”cigarettes for up to 21 d.Cell viability was analyzed using resazurin conversion assay(mitochondrial activity)and calcein-AM staining(esterase activity).Osteogenic differentiation and bone cell function were evaluated using alkaline phosphatase(AP)activity,while matrix formation was analyzed through alizarin red staining.Primary cilia structure was examined by acetylatedα-tubulin immunofluorescent staining.Free radical production was evaluated with 2’,7’-dichlorofluoresceindiacetate assay.RESULTS Our data clearly show that THS is significantly less toxic to bone cells than CS when analyzed by mitochondrial and esterase activity(P<0.001).No significant differences in cytotoxicity between the diverse flavors of THS were observed.Harmful effects from THS on bone cell function were observed only at very high,non-physiological concentrations.In contrast,extracts from conventional cigarettes significantly reduced the AP activity(by two-fold)and matrix mineralization(four-fold)at low concentrations.Additionally,morphologic analysis of primary cilia revealed no significant changes in the length of the organelle involved in osteogenesis of osteoprogenitor cells,nor in the number of ciliated cells following THS treatment.Assessment of free radical production demonstrated that THS induced significantly less oxidative stress than conventional CS in osteoprogenitor cells.CONCLUSIONTHS was significantly less harmful to osteoprogenitor cells during osteogenesisthan conventional CS. Additional studies are required to confirm whether THS isa better alternative for smokers to improve delays in bone healing followingfracture.
基金the National Natural Science Foundation of China(Nos.U1601220,82072450,and 81672118)Chongqing Science and Technology Commission-Basic Science and Frontier Technology Key Project(No.cstc2015jcyjBX0119)Chongqing Medical University Intelligent Medicine Research Project(No.ZHYX202115).
文摘Since 3D printed hard materials could match the shape of bone,cell survival and fate determination towards osteoblasts in such materials have become a popular research target.In this study,a scaffold of hardmaterial for 3D fabrication was designed to regulate developmental signal(Notch)transduction guiding osteoblast differentiation.We established a polycaprolactone(PCL)and cell-integrated 3D printing system(PCI3D)to reciprocally print the beams of PCL and cell-laden hydrogel for a module.This PCI3D module holds good cell viability of over 87%,whereas cells show about sixfold proliferation in a 7-day culture.The osteocytic MLO-Y4 was engineered to overexpress Notch ligand Dll4,making up 25%after mixing with 75%stromal cells in the PCI3D module.Osteocytic Dll4,unlike other delta-like family members such as Dll1 or Dll3,promotes osteoblast differentiation and themineralization of primary mouse and a cell line of bone marrow stromal cells when cultured in a PCI3D module for up to 28 days.Mechanistically,osteocytic Dll4 could not promote osteogenic differentiation of the primary bone marrow stromal cells(BMSCs)after conditional deletion of the Notch transcription factor RBPjκby Cre recombinase.These data indicate that osteocytic Dll4 activates RBPjκ-dependent canonical Notch signaling in BMSCs for their oriented differentiation towards osteoblasts.Additionally,osteocytic Dll4 holds a great potential for angiogenesis in human umbilical vein endothelial cells within modules.Our study reveals that osteocytic Dll4 could be the osteogenic niche determining cell fate towards osteoblasts.This will open a new avenue to overcome the current limitation of poor cell viability and low bioactivity of traditional orthopedic implants.
文摘Summary: 17β-estradiol (E2), progesterone (P) and testosterone (T) were investigated for their effects on the proliferation and differentiation of primary rat calvarial osteoblasts in vitro. Rat calvarial osteoblasts were cultured with 10-10 mol/L E2, 10-9-10-6 mol/L P and 10-10 mol/L T for 20 days. Cell proliferation was determined by 3H-thymidine incorporation and cell counting. Cell differentiation was examined by measuring alkaline phosphatase (ALP) activity, osteocalcin gene expression and production, bone nodule formation in different periods of culture. Our results showed that no effect of three sex hormones was observed on cell proliferation, but, the responses of cell differentiation to the hormones were more or less different. Among these three hormones used in this study, P appeared to have multi-stimulating effect on cell differentiation. Effect of T seemed not so significant as that of P on cell differentiation. Although ALP activity and osteocalcin production were increased significantly by T, it had slight effect on osteocalcin mRNA and bone nodule formation. Besides, E2 did not demonstrate any effect on cell differentiation. It is concluded that the proliferation of rat calvarial osteoblasts was independent of the presence of sex hormones. However, the differentiation of these cells were stimulated in different levels and to different extent either by P or T. P appeared to be a multi-stimulator on differentiation of such cells.
