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1.
J Contemp Dent Pract ; 21(7): 776-780, 2020 Jul 01.
Artigo em Inglês | MEDLINE | ID: mdl-33020362

RESUMO

AIM: To evaluate the ability of osteogenic culture media in comparison with regular growth culture media in enhancing the osteoblastic cell differentiation of human periodontal ligament stem cells (hPDLSCs). MATERIALS AND METHODS: In vitro cultures of commercially obtained hPDLSCs were seeded onto xenograft bone blocks in both regular and osteogenic media. Confocal laser microscope images were obtained for cellular differentiation and adhesion, and scanning electron microscopy (SEM) images were obtained to validate the osteogenic differentiation by showing the morphological characteristics of the newly formed cells. RESULTS: Confocal laser microscope analysis showed positive staining for new bone cells with an increased signal intensity when samples were cultured in osteogenic culture media compared with regular culture media. These findings indicate the effect of the active ingredients of the osteogenic culture media in enhancing the osteogenic differentiation hPDLSC. Scanning electron microscopy images validated the osteogenic differentiation showing a flattened, polygonal morphology with multiple extending cytoplasmic processes of new cells. CONCLUSION: Xenograft bone blocks are biocompatible scaffold for the osteogenic differentiation of seeded hPDLSCs. Osteogenic culture media enhances and increases the osteogenic differentiation of hPDLSCs into new bone cells more than regular growth culture media. Periodontal ligament stem cells are a predictable biological input as a cell-based tissue-engineered construct and biologically acceptable when it is cultured in a suitable growth media that mimics the intended environment. CLINICAL SIGNIFICANCE: Consideration of the clinical use of equine bone blocks and periodontal ligament stem cells in a suitable biological environment as a potential new option for bone regeneration techniques.


Assuntos
Osteogênese , Ligamento Periodontal , Animais , Células Cultivadas , Meios de Cultura , Cavalos , Humanos , Microscopia Eletrônica de Varredura , Células-Tronco
2.
Beijing Da Xue Xue Bao Yi Xue Ban ; 52(5): 952-958, 2020 Oct 18.
Artigo em Chinês | MEDLINE | ID: mdl-33047736

RESUMO

OBJECTIVE: To prepare and evaluate the basic properties in vitro of a novel small intestinal submucosa (SIS) sponge, and to describe the bone formation ability of the SIS sponge in vivo. METHODS: The SIS sponge was prepared by freeze-drying method. To evaluate the physicochemical properties of the sponge, electron microscope observation, porosity test, water absorption ability and mechanical property were conducted in vitro. The cytotoxicity of the SIS sponge was performed by cell counting kit-8 method. In vivo experiments, eighteen extraction sockets of premolar of three Beagle dogs were randomly divided into three groups: SIS sponge group (SIS sponge), positive control group (Bio-Oss granules and Bio-Gide membrane) and control group(no treatment). The animals were sacrificed 4 weeks and 12 weeks after operation, and micro computed tomography (Micro-CT) was applied to measure the bone volume fraction (BV/TV) and bone mineralized density (BMD). The data were analyzed with one-way ANOVA. RESULTS: The average pore diameter of the SIS sponge was (194.90±30.39) µm, the porosity was 92.31%±0.24%, the water absorption rate was 771.50%±40.90%, and the compressive elastic modulus was (2.20±0.19) kPa. There was no significant difference in cell proliferation ability between SIS sponge and control group (P>0.05). Micro-CT quantitative results showed that BV/TV of SIS sponge group (52.81%±3.21%) and positive control group (58.30%±9.36%) were significantly higher than that of control group (38.65%±4.80%) 4 weeks after operation (P < 0.05). The BMD of SIS sponge group [(887.09±61.02) mg/cm3], positive control group [(952.05±132.78) mg/cm3] and control group [(879.29±74.27) mg/cm3] showed no statistical difference 4 weeks after operation (P>0.05). The BV/TV of positive control group (60.57%± 6.56%) was significantly higher than that of SIS sponge group (47.89%±3.59%) and control group (42.99%±2.54%) 12 weeks after operation (P < 0.05). BMD of SIS sponge group [(1047±89.95) mg/cm3] and positive control group [(1101.37±98.85) mg/cm3] were significantly higher than that of control group [(890.36±79.79) mg/cm3] 12 weeks after operation (P < 0.05). CONCLUSION: The SIS sponge has satisfying physicochemical properties and biocompatibility. The SIS sponge significantly increased bone volume fraction in the early stage of bone formation (4 weeks) and bone mineralized density in the late stage of bone formation (12 weeks).


Assuntos
Osteogênese , Animais , Cães , Microtomografia por Raio-X
3.
Shanghai Kou Qiang Yi Xue ; 29(3): 225-230, 2020 Jun.
Artigo em Chinês | MEDLINE | ID: mdl-33043336

RESUMO

PURPOSE: To investigate the effects of exendin-4(EX-4) on proliferation, migration and osteogenic differentiation of human periodontal ligament stem cells(PDLSCs). METHODS: PDLSCs were isolated and cultured using limited dilution method in vitro. Colony formation assay, osteogenic and adipogenic differentiation were applied to identify the stem cells. Immunofluorescence staining was used to detect the expression of EX-4 receptor glucagon-like peptide-1 receptor (GLP-1R) on the surface of PDLSCs. PDLSCs were stimulated with 5, 10, 20 or 50 nmol/L EX-4 in vitro. CCK-8, Transwell assay and alkaline phosphatase(ALP) activity assay were used to determine the effects of EX-4 on PDLSCs proliferation, migration and osteogenic differentiation. Quantitative real-time polymerase chain reaction was used to determine the expression of osteogenic related genes ALP, runt-related transcription factor 2(Runx2) and osteocalcin (OCN). The data were analyzed by Graphpad Prims 6.0 software package. RESULTS: PDLSCs were successfully isolated and cultivated. GLP-1R positively expressed on the surface of PDLSCs. EX-4 exerted no significant effect on PDLSCs proliferation(P>0.05). EX-4 significantly promoted migration, ALP activity and osteogenic related genes expression of PDLSCs (P<0.05). CONCLUSIONS: 10 nmol/L EX-4 could promote migration and osteogenic differentiation of PDLSCs.


Assuntos
Exenatida , Ligamento Periodontal , Diferenciação Celular , Proliferação de Células , Células Cultivadas , Exenatida/farmacologia , Humanos , Osteogênese , Células-Tronco
4.
Shanghai Kou Qiang Yi Xue ; 29(3): 242-249, 2020 Jun.
Artigo em Chinês | MEDLINE | ID: mdl-33043339

RESUMO

PURPOSE: This study was aimed to investigate the mesenchymal stem cell (MSC) phenotypes of immortalized odontoblasts(iODs) and bone morphogenetic protein -2, -4, -6, -7, and -9 (BMPs) differentially regulate the mineralization of iODs. METHODS: ODs were immortalized by SV40 T antigen to establish iOD lineages, and the endogenous expression of BMPs was successively examined. Recombinant adenoviruses expressing BMPs and GFP were generated using Ad-Easy technology. The proliferation capability of iODs was examined using an MTT kit. MSC markers of iODs were examined by immunofluorescence. In vitro, semiquantitative RT-PCR, alkaline phosphatase(ALP) activity assay, matrix mineralization assay and oil red O staining assay were used to examine the osteogenic and adipogenic differentiation capabilities of iODs. Statistical significance among groups was analyzed by one-way analysis of variance and Scheffe's multiple comparison test was SPSS 21.0 software package. Finally, the volume and density of ectopic mineralized tissues formed in vivo were assessed by micro-CT and histological analysis. RESULTS: ODs can be efficiently immortalized by SV40 T antigen, and the resulting iODs maintained an excellent proliferative activity, expressed certain MSC markers and possessed multiple differentiation capabilities. BMP-2 and BMP-9 regulated iODs osteogenic differentiation better than BMP-4, -6, and -7. CONCLUSIONS: Our findings suggest that ODs and osteogenic growth factors such as BMP-2 and BMP-9 can be used as an efficacious strategy for bone tissue engineering.


Assuntos
Células-Tronco Mesenquimais , Osteogênese , Proteínas Morfogenéticas Ósseas , Diferenciação Celular , Odontoblastos
5.
Int J Oral Implantol (Berl) ; 13(3): 213-232, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32879927

RESUMO

PURPOSE: The evidence pertaining to the contribution of the sinus membrane to new bone formation following maxillary sinus augmentation procedures is equivocal. The purpose of this study was to analyse the evidence currently available on the osteogenic capacity of the sinus membrane following maxillary sinus augmentation procedures, and the effect of local delivery of recombinant human bone morphogenic proteins (rhBMPs) on the bone-forming potential of the sinus membrane. MATERIALS AND METHODS: An electronic search was conducted using six different databases to identify controlled trials, prospective and retrospective cohort studies, case series and case reports, as well as preclinical (animal) studies reporting on new bone formation in close proximity with the sinus membrane after maxillary sinus augmentation procedures, assessed through histological and/or histomorphometrical evaluation, on the basis of pre-established eligibility criteria. RESULTS: No clinical studies were identified. Twenty-six preclinical studies were included in the review. Nine of them supported the osteogenic potential of the sinus membrane, while eight reported no evidence of osteogenicity from the sinus membrane. The nine remaining studies reported on the local effect of rhBMPs. The majority of these nine studies reported enhanced new bone formation in the sinus membrane region. CONCLUSIONS: The sinus membrane contains pluripotent mesenchymal cells with the capacity to differentiate and participate in the process of new bone formation. However, the findings from the studies selected in this systematic review do not consistently support that the sinus membrane significantly contributes to new bone formation following maxillary sinus augmentation procedures.


Assuntos
Seio Maxilar , Osteogênese , Animais , Humanos , Maxila , Estudos Prospectivos , Estudos Retrospectivos
6.
Zhongguo Zhong Yao Za Zhi ; 45(15): 3603-3607, 2020 Aug.
Artigo em Chinês | MEDLINE | ID: mdl-32893549

RESUMO

Osteoporosis fracture with high disability and mortality is a difficult problem that seriously affects the life quality of individuals. At present, there is still a lack of anti-osteoporosis drugs with clear target and significant efficacy in the clinical practice. Rehmanniae Radix and its prescriptions have significant clinical effects. In this regard, more and more studies have reported the effects and mechanisms of Rehmanniae Radix and its active components, and the certain research outputs have been achieved. In this article, the PubMed, Web of science, China National Knowledge Infrastructure, and Wanfang database were searched to collect and organize the latest research progress of Rehmanniae Radix treatment of osteoporosis in the recent 10 years. We summarized the research dynamics as well as the function indexes and mechanisms of the raw and processed Rehmanniae Radix, active ingredients such as catalpol, aucubin, acteoside and Rehmanniae Radix polysaccharide, and their formulating prescriptions, and then excavated the potential active ingredients, targets and signaling pathways, including the effect on bone marrow mesenchymal stem cells, promoting the osteoblast proliferation and promoting osteogenesis differentiation(increasing alkaline phosphatase, typeⅠ collagen, osteoprotegerin, and osteocalcin and promoting calcium deposits), increasing the bone density, inhibiting the osteoclast quantity and differentiation, promoting the osteoclast apoptosis, and reducing tartrate resistant acid phosphatase and bone resorption pit area to provide the reference and develop new ideas for developing Rehmanniae Radix prescriptions for treatment of osteoporosis and exploring its mechanism.


Assuntos
Medicamentos de Ervas Chinesas , Osteoporose , Rehmannia , China , Humanos , Osteogênese
7.
Zhongguo Xiu Fu Chong Jian Wai Ke Za Zhi ; 34(9): 1170-1176, 2020 Sep 15.
Artigo em Chinês | MEDLINE | ID: mdl-32929912

RESUMO

Objective: To investigate the effects of three-dimensional (3D) printed Ti6Al4V-4Cu alloy on inflammation and osteogenic gene expression in mouse bone marrow mesenchymal stem cells (BMSCs) and mouse mononuclear macrophage line RAW264.7. Methods: Ti6Al4V and Ti6Al4V-4Cu alloys were prepared by selective laser melting, and the extracts of the two materials were prepared according to the biological evaluation standard of medical devices. The effects of two kinds of extracts on the proliferation of mouse BMSCs and mouse RAW264.7 cells were detected by cell counting kit 8 method. After co-cultured with mouse BMSCs for 3 days, the expression of osteogenesis- related genes [collagen type Ⅰ (Col-Ⅰ), alkaline phosphatase (ALP), Runx family transcription factor 2 (Runx-2), osteoprotegerin (OPG), and osteopontin (OPN)] were detected by real-time fluorescence quantitative PCR. After co-cultured with mouse RAW264.7 cells for 1 day, the expressions of inflammation-related genes [interleukin 4 (IL-4) and nitric oxide synthase 2 (iNOS)] were detected by real-time fluorescence quantitative PCR, and the supernatants of the two groups were collected to detect the secretion of vascular endothelial growth factor a (VEGF-a) and bone morphogenetic protein 2 (BMP-2) by ELISA. The osteogenic conditioned medium were prepared with the supernatants of the two groups and co-cultured with BMSCs for 3 days. The expressions of osteogenesis-related genes (Col-Ⅰ, ALP, Runx-2, OPG, and OPN) were detected by real-time fluorescence quantitative PCR. Results: Compared with Ti6Al4V alloy extract, Ti6Al4V-4Cu alloy extract had no obvious effect on the proliferation of BMSCs and RAW264.7 cells, but it could promote the expression of OPG mRNA in BMSCs, reduce the expression of iNOS mRNA in RAW264.7 cells, and promote the expression of IL-4 mRNA. It could also promote the secretions of VEGF-a and BMP-2 in RAW264.7 cells. Ti6Al4V-4Cu osteogenic conditioned medium could promote the expressions of Col-Ⅰ, ALP, Runx-2, OPG, and OPN mRNAs in BMSCs. The differences were all significant ( P<0.05). Conclusion: 3D printed Ti6Al4V-4Cu alloy can promote RAW264.7 cells to secret VEGF-a and BMP-2 by releasing copper ions, thus promoting osteogenesis through bone immune regulation, which lays a theoretical foundation for the application of metal prosthesis.


Assuntos
Ligas , Osteogênese , Animais , Células da Medula Óssea , Proteína Morfogenética Óssea 2/metabolismo , Diferenciação Celular , Células Cultivadas , Regulação da Expressão Gênica , Camundongos , Titânio , Fator A de Crescimento do Endotélio Vascular/metabolismo
8.
Int J Oral Maxillofac Implants ; 35(5): 910-916, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32991640

RESUMO

PURPOSE: This study evaluated the bone-forming potential of the demineralized human dentin matrix by performing histologic and morphometric analyses. The immunolabeling of osteopontin, a determinant protein for bone repair, was also evaluated. MATERIALS AND METHODS: Wistar rats were selected and submitted to the extraction of the right and left second molars. Tooth sockets were separated into two groups: the control group (right), which was filled with the blood clot, and the experimental group (left), which was filled with demineralized human dentin matrix. Animals were sacrificed at 5, 10, and 21 days. Histologic and histoquantitative analyses (analyses of variance [ANOVA] and Tukey's test) were performed, as well as immunostaining for osteopontin as an osteogenesis indicator. RESULTS: After 5 days, demineralized human dentin matrix was incorporated by new trabeculae. After 10 days, connective tissue organization and new trabeculae were observed in the experimental group, and intense staining for osteopontin close to demineralized human dentin matrix was observed in the experimental group. After 21 days, the experimental group was showing mature trabeculae. A statistical difference was observed (P < .05). There was a higher number of trabeculae in the experimental groups in all periods of analysis. The presence of osteopontin was observed more intensely at 10 days close to demineralized human dentin matrix. CONCLUSION: This study indicates that demineralized human dentin matrix implanted in tooth sockets induces the acceleration of osteogenesis.


Assuntos
Osteogênese , Alvéolo Dental/cirurgia , Animais , Dentina , Humanos , Osteopontina , Ratos , Ratos Wistar
9.
Int J Oral Maxillofac Implants ; 35(5): 939-947, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32991644

RESUMO

PURPOSE: This study investigated the effects of bisphosphonates, namely, alendronate and zoledronate, on the osteogenic activity of osteoprogenitor cells cultured on titanium surfaces at therapeutic doses in order to assess if altered osteoblastogenesis could compromise osseointegration and contribute to etiopathogenesis of painful disorders such as bisphosphonates-related osteonecrosis of the jaw (BRONJ) following implant placement. MATERIALS AND METHODS: MC3T3-E1 Subclone 4 cells were utilized in this study. Therapeutic doses of alendronate and zoledronate were calculated based on reported peak plasma concentrations. The viability, proliferation, adhesion, and mineralization potential of cells was assessed using a LIVE/DEAD stain, alamarBlue assay, immunofluorescence microscopy, and Alizarin Red S staining, respectively. RESULTS: Therapeutic doses of zoledronate negatively affected cell viability, whereas therapeutic doses of alendronate significantly enhanced cell differentiation and the amount of bone formation compared with the control. CONCLUSION: The findings of this study may provide some insight into the pathogenesis of BRONJ development following implant placement in patients treated with zoledronate and may have promising implications toward improved wound healing and osseointegration in patients treated with alendronate.


Assuntos
Difosfonatos/efeitos adversos , Osteogênese , Alendronato , Células Cultivadas , Humanos , Titânio
10.
Pediatrics ; 146(3)2020 09.
Artigo em Inglês | MEDLINE | ID: mdl-32868470

RESUMO

Pediatric care providers, pediatricians, pediatric subspecialty physicians, and other health care providers should be able to recognize children with abnormal head shapes that occur as a result of both synostotic and deformational processes. The purpose of this clinical report is to review the characteristic head shape changes, as well as secondary craniofacial characteristics, that occur in the setting of the various primary craniosynostoses and deformations. As an introduction, the physiology and genetics of skull growth as well as the pathophysiology underlying craniosynostosis are reviewed. This is followed by a description of each type of primary craniosynostosis (metopic, unicoronal, bicoronal, sagittal, lambdoid, and frontosphenoidal) and their resultant head shape changes, with an emphasis on differentiating conditions that require surgical correction from those (bathrocephaly, deformational plagiocephaly/brachycephaly, and neonatal intensive care unit-associated skill deformation, known as NICUcephaly) that do not. The report ends with a brief discussion of microcephaly as it relates to craniosynostosis as well as fontanelle closure. The intent is to improve pediatric care providers' recognition and timely referral for craniosynostosis and their differentiation of synostotic from deformational and other nonoperative head shape changes.


Assuntos
Craniossinostoses/diagnóstico , Acrocefalossindactilia/genética , Fenótipo de Síndrome de Antley-Bixler/genética , Suturas Cranianas/anatomia & histologia , Disostose Craniofacial , Craniossinostoses/classificação , Craniossinostoses/etiologia , Craniossinostoses/cirurgia , Cabeça/anormalidades , Humanos , Lactente , Hipertensão Intracraniana/etiologia , Ilustração Médica , Microcefalia/etiologia , Osteogênese/fisiologia , Fenótipo , Fotografação , Polidactilia/genética , Receptores de Fatores de Crescimento de Fibroblastos/metabolismo , Procedimentos Cirúrgicos Reconstrutivos , Crânio/anatomia & histologia , Crânio/diagnóstico por imagem , Crânio/crescimento & desenvolvimento , Sinostose/complicações , Sinostose/diagnóstico por imagem
11.
Zhong Nan Da Xue Xue Bao Yi Xue Ban ; 45(6): 678-683, 2020 Jun 28.
Artigo em Inglês, Chinês | MEDLINE | ID: mdl-32879125

RESUMO

OBJECTIVES: To explore the difference in odontoblast differentiation capacity between stem cells from human exfoliated deciduous teeth (SHED) and dental pulp stem cells (DPSCs), and to examine the expression level of ephrinB1 in odontoblast differentiation of these stem cells. METHODS: The stems cells were divided into a SHED group and a DPSCs group. After odontoblast differentiation induction, the above 2 groups were also randomly divided into a 3 d group and a 7 d group, respectively.The calcium deposition was detected by alkaline phosphatase (ALP) staining and alizarin red staining.The mRNA and protein expressions of ephrinB1, dentin matrix protein-1 (DMP-1) and dentin sialophosphoprotein (DSPP) were detected by real-time PCR and Western blotting. RESULTS: ALP staining and alizarin red staining showed that there was stronger mineralization capacity in the SHED group than that in the DPSCs group. The relative mRNA and protein expressions of DMP-1, DSPP, and ephrinB1 in the SHED group were higher than those in the DPSCs group except for the protein expression of DMP-1 in the SHED 3 d group (all P<0.05). CONCLUSIONS: SHED has stronger odontoblast differentiation capacity than DPSCs. In addition, ephrinB1 may be involved in the processes of odontoblast differentiation in the SHED and DPSCs.


Assuntos
Odontoblastos , Osteogênese , Diferenciação Celular , Proliferação de Células , Células Cultivadas , Polpa Dentária , Humanos , Células-Tronco , Dente Decíduo
12.
Zhong Nan Da Xue Xue Bao Yi Xue Ban ; 45(6): 684-692, 2020 Jun 28.
Artigo em Inglês, Chinês | MEDLINE | ID: mdl-32879126

RESUMO

OBJECTIVES: To evaluate the repairing ability of nano-pearl powder bone substitute in rabbit with defect of distal femur bone. METHODS: Thirty-two New Zealand rabbits were randomly divided into four groups: a nano-pearl powder/recombinant human bone morphogenetic protein 2 (rhBMP-2)/hyaluronic acid group, a nano-pearl powder/hyaluronic acid group, a nano-pearl powder group and a blank control group (n=8 in each group). A defect with the diameter of 7 mm and height of 10 mm was prepared at the distal femoral metaphysis line of the rabbit.Different bone substitutes were planted, and the effect of repair was evaluated by macroscopic observation, imaging examination, and histopathological examination. RESULTS: The results of imageology showed that: the bone repairing effect in the nano-pearl powder/rhBMP-2/hyaluronic acid group was better than that in the pure pearl powder group and the nano-pearl powder/hyaluronic acid group, and which in the 3 experimental groups was better than that in the blank control group; The results of histology showed that: at the 4th, 8th and 12th weeks after the modeling operation, the speed of bone repair in the nano-pearl powder/rhBMP-2/hyaluronic acid group was faster than that in the pure pearl powder group and the nano-pearl powder/hyaluronic acid group, and which in the blank control group was far slower than that in the 3 experimental groups. The results of immunohistochemistry staining for osteocalcin antibody showed that: the osteogenic effect in the nano-pearl powder/rhBMP-2/hyaluronic acid group was better than that in the pure pearl powder group and the nano-pearl powder/hyaluronic acid group (both P<0.05); there was no significant difference between the nano-pearl powder/hyaluronic acid group and the pure pearl powder group (P>0.05); however, there was significant difference between the pure pearl powder group and the blank control group (P<0.05). According to the staining results of Type I collagen antibody, there was no significant difference in the osteogenic effect between the nano-pearl powder/rhBMP-2/hyaluronic acid group and the nano-pearl powder/hyaluronic acid group (P>0.05), but the osteogenic effect in the nano-pearl powder/hyaluronic acid group was better than that in the pure pearl powder group and the blank control group (both P<0.05). CONCLUSIONS: Nano-pearl powder and its bone substitute can promote the repair of bone defect, and the nano-pearl powder which contains rhBMP-2 has better osteogenic and repairing effect on defect.


Assuntos
Substitutos Ósseos , Animais , Proteína Morfogenética Óssea 2 , Colágeno , Fêmur , Humanos , Osteogênese , Pós , Coelhos , Proteínas Recombinantes , Fator de Crescimento Transformador beta
13.
Acta Odontol Latinoam ; 33(2): 125, 2020 Sep 01.
Artigo em Inglês | MEDLINE | ID: mdl-32920615

RESUMO

Melatonin (MLT) is a potential signaling molecule in the homeostasis of bone metabolism and may be an important mediator of bone formation and stimulation. The aim of this in vitro study was to evaluate the effect of MLT on the viability, mRNA/protein expression and mineralization of pre-osteoblastic cells. The concentrations 5, 2.5, 1, 0.1 and 0.01 mM MLT were tested on pre-osteoblastic cells (MC3T3) compared to control (no MLT), evaluating proliferation and cell viability (C50), gene expression (RT-PCR) and secretion (ELISA) of COL-I and OPN at 24h, 48h and 72h, and the formation of mineral nodules (alizarin red and fast red) after 10 days of treatment. MLT at 5 and 2.5 mM proved to be cytotoxic (C50), so only 0.01, 0.1 and 1 mM were used for the subsequent analyses. OPN mRNA expression increased with MLT at 0.1 mM - 1 mM, which was followed by increased secretion of OPN both at 24h and 72h compared to the remaining groups (p <0.05). COL-I mRNA and COL-1 secretion followed the same pattern as OPN at 0.1 mM MLT at 72h of treatment (p <0.05). Regarding mineralization, all MLT doses (except 1mM) caused an increase (p <0.05) in the formation of mineral nodules compared to the control. Melatonin at 0.01mM - 1mM had a stimulatory effect on osteoblasts by upregulating COL-I and OPN expression/ secretion and mineralization, thereby fostering osteogenesis.


Assuntos
Diferenciação Celular/efeitos dos fármacos , Diferenciação Celular/genética , Metaloproteinase 2 da Matriz/metabolismo , Melatonina/farmacologia , Osteoblastos/efeitos dos fármacos , Osteogênese/efeitos dos fármacos , Osteogênese/genética , Osteopontina/metabolismo , Fragmentos de Peptídeos/metabolismo , Proliferação de Células/efeitos dos fármacos , Proliferação de Células/genética , Ensaio de Imunoadsorção Enzimática , Expressão Gênica , Regulação da Expressão Gênica no Desenvolvimento/efeitos dos fármacos , Humanos , Metaloproteinase 2 da Matriz/genética , Osteoblastos/metabolismo , Osteopontina/genética , Fragmentos de Peptídeos/genética , RNA Mensageiro/genética , Reação em Cadeia da Polimerase em Tempo Real
14.
Lancet ; 396(10252): 684-692, 2020 09 05.
Artigo em Inglês | MEDLINE | ID: mdl-32891212

RESUMO

BACKGROUND: There are no effective therapies for achondroplasia. An open-label study suggested that vosoritide administration might increase growth velocity in children with achondroplasia. This phase 3 trial was designed to further assess these preliminary findings. METHODS: This randomised, double-blind, phase 3, placebo-controlled, multicentre trial compared once-daily subcutaneous administration of vosoritide with placebo in children with achondroplasia. The trial was done in hospitals at 24 sites in seven countries (Australia, Germany, Japan, Spain, Turkey, the USA, and the UK). Eligible patients had a clinical diagnosis of achondroplasia, were ambulatory, had participated for 6 months in a baseline growth study and were aged 5 to less than 18 years at enrolment. Randomisation was done by means of a voice or web-response system, stratified according to sex and Tanner stage. Participants, investigators, and trial sponsor were masked to group assignment. Participants received either vosoritide 15·0 µg/kg or placebo, as allocated, for the duration of the 52-week treatment period administered by daily subcutaneous injections in their homes by trained caregivers. The primary endpoint was change from baseline in mean annualised growth velocity at 52 weeks in treated patients as compared with controls. All randomly assigned patients were included in the efficacy analyses (n=121). All patients who received one dose of vosoritide or placebo (n=121) were included in the safety analyses. The trial is complete and is registered, with EudraCT, number, 2015-003836-11. FINDINGS: All participants were recruited from Dec 12, 2016, to Nov 7, 2018, with 60 assigned to receive vosoritide and 61 to receive placebo. Of 124 patients screened for eligibility, 121 patients were randomly assigned, and 119 patients completed the 52-week trial. The adjusted mean difference in annualised growth velocity between patients in the vosoritide group and placebo group was 1·57 cm/year in favour of vosoritide (95% CI [1·22-1·93]; two-sided p<0·0001). A total of 119 patients had at least one adverse event; vosoritide group, 59 (98%), and placebo group, 60 (98%). None of the serious adverse events were considered to be treatment related and no deaths occurred. INTERPRETATION: Vosoritide is an effective treatment to increase growth in children with achondroplasia. It is not known whether final adult height will be increased, or what the harms of long-term therapy might be. FUNDING: BioMarin Pharmaceutical.


Assuntos
Acondroplasia/tratamento farmacológico , Peptídeo Natriurético Tipo C/análogos & derivados , Osteogênese , Absorciometria de Fóton , Acondroplasia/sangue , Adolescente , Biomarcadores/sangue , Estatura , Densidade Óssea , Criança , Pré-Escolar , Colágeno Tipo X/sangue , Método Duplo-Cego , Feminino , Humanos , Reação no Local da Injeção , Injeções Subcutâneas , Masculino , Peptídeo Natriurético Tipo C/uso terapêutico
15.
Chin J Dent Res ; 23(3): 169-176, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32974616

RESUMO

OBJECTIVE: To explore the effects of Sirtuin 7 (SIRT7) on the gene expression profile of stem cells from the apical papilla (SCAPs). METHODS: SCAPs were isolated and cultured. SIRT7 short hairpin ribonucleic acid (shRNA) was used to knock down the expression of SIRT7 in SCAPs. After library construction and RNA sequencing (RNA-seq), differentially expressed genes were identified using Cuffdiff with a false discovery rate (FDR) ≤ 0.05 and fold change ≥ 2. Pathway and Gene Ontology (GO) analyses were conducted to elucidate the changes in important functions and pathways after SIRT7 gene knockdown. Gene set enrichment analysis (GSEA) was performed and enrichment of a gene set with an FDR lower than 0.25 was considered significant. RESULTS: The most striking GO terms related to SIRT7sh SCAPs and Consh SCAPs were response to nucleus, nucleolus, cytoplasm, protein binding and intrinsic apoptotic signalling pathway. Signalling pathway analysis revealed the top five pathways to be metabolic, pyrimidine metabolism, protein processing in endoplasmic reticulum, phosphatidylinositol 3-kinase/protein kinase B (PI3K/Akt) signalling and p53 signalling. The results of GSEA showed that genes were mainly enriched in cell cycle, cell proliferation, transforming growth factor beta (TGF-ß) signalling and cytokine-cytokine receptor interaction pathways. CONCLUSION: SIRT7 may affect the functions of SCAPs through cell cycle, cell proliferation and apoptosis pathways.


Assuntos
Papila Dentária , RNA Longo não Codificante , Diferenciação Celular , Proliferação de Células/genética , Células Cultivadas , Osteogênese , Fosfatidilinositol 3-Quinases , Sirtuínas , Células-Tronco
16.
Nat Commun ; 11(1): 4465, 2020 09 08.
Artigo em Inglês | MEDLINE | ID: mdl-32901012

RESUMO

Titanium implants have been widely used in bone tissue engineering for decades. However, orthopedic implant-associated infections increase the risk of implant failure and even lead to amputation in severe cases. Although TiO2 has photocatalytic activity to produce reactive oxygen species (ROS), the recombination of generated electrons and holes limits its antibacterial ability. Here, we describe a graphdiyne (GDY) composite TiO2 nanofiber that combats implant infections through enhanced photocatalysis and prolonged antibacterial ability. In addition, GDY-modified TiO2 nanofibers exert superior biocompatibility and osteoinductive abilities for cell adhesion and differentiation, thus contributing to the bone tissue regeneration process in drug-resistant bacteria-induced implant infection.


Assuntos
Antibacterianos/química , Grafite , Nanofibras/química , Próteses e Implantes , Infecções Relacionadas à Prótese/prevenção & controle , Titânio , Células 3T3 , Animais , Materiais Biocompatíveis/química , Regeneração Óssea , Sobrevivência Celular , Modelos Animais de Doenças , Feminino , Teste de Materiais , Staphylococcus aureus Resistente à Meticilina/efeitos dos fármacos , Camundongos , Nanocompostos/química , Osteogênese , Processos Fotoquímicos , Infecções Estafilocócicas/prevenção & controle
17.
Int J Nanomedicine ; 15: 5825-5838, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32821104

RESUMO

Background and Purpose: The extracellular matrix (ECM) derived from bone marrow mesenchymal stem cells (BMSCs) has been used in regenerative medicine because of its good biological activity; however, its poor mechanical properties limit its application in bone regeneration. The purpose of this study is to construct a three dimensional-printed hydroxyapatite (3D-HA)/BMSC-ECM composite scaffold that not only has biological activity but also sufficient mechanical strength and reasonably distributed spatial structure. Methods: A BMSC-ECM was first extracted and formed into micron-sized particles, and then the ECM particles were modified onto the surface of 3D-HA scaffolds using an innovative linking method to generate composite 3D-HA/BMSC-ECM scaffolds. The 3D-HA scaffolds were used as the control group. The basic properties, biocompatibility and osteogenesis ability of both scaffolds were tested in vitro. Finally, a critical skull defect rat model was created and the osteogenesis effect of the scaffolds was evaluated in vivo. Results: The compressive modulus of the composite scaffolds reached 9.45±0.32 MPa, which was similar to that of the 3D-HA scaffolds (p>0.05). The pore size of the two scaffolds was 305±47 um and 315±34 um (p>0.05), respectively. A CCK-8 assay indicated that the scaffolds did not have cytotoxicity. The composite scaffolds had good cell adhesion ability, with a cell adhesion rate of up to 76.00±6.17% after culturing for 7 hours, while that of the 3D-HA scaffolds was 51.85±4.77% (p<0.01). In addition, the composite scaffold displayed higher alkaline phosphatase (ALP) activity, osteogenesis-related mRNA expression, and calcium nodule formation, thus confirming that the composite scaffolds had good osteogenic activity. The composite scaffolds exhibited good bone repair in vivo and were superior to the 3D-HA scaffolds. Conclusion: We conclude that BMSC-ECM is a good osteogenic material and that the composite scaffolds have good osteogenic ability, which provides a new method and concept for the repair of bone defects.


Assuntos
Durapatita/farmacologia , Matriz Extracelular/metabolismo , Células-Tronco Mesenquimais/citologia , Tecidos Suporte/química , Animais , Regeneração Óssea/efeitos dos fármacos , Osso e Ossos/diagnóstico por imagem , Osso e Ossos/efeitos dos fármacos , Osso e Ossos/patologia , Adesão Celular/efeitos dos fármacos , Morte Celular/efeitos dos fármacos , Diferenciação Celular/efeitos dos fármacos , Matriz Extracelular/efeitos dos fármacos , Hidrodinâmica , Masculino , Células-Tronco Mesenquimais/efeitos dos fármacos , Células-Tronco Mesenquimais/ultraestrutura , Osteogênese/efeitos dos fármacos , Osteogênese/genética , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Ratos Sprague-Dawley , Cicatrização/efeitos dos fármacos
18.
Environ Pollut ; 265(Pt A): 114734, 2020 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-32806408

RESUMO

Excess fluoride in drinking water is an environmental issue of increasing worldwide concern, because of its adverse effect on human health. Skeletal fluorosis caused by chronic exposure to excessive fluoride is a metabolic bone disease characterized by accelerated bone turnover accompanied by aberrant activation of osteoblasts. It is not clear whether Wnt/ß-catenin signaling, an important signaling pathway regulating the function of osteoblasts, mediates the pathogenesis of skeletal fluorosis. A cross-sectional case-control study was conducted in Tongyu County, Jilin Province, China showed that fluoride stimulated the levels of OCN and OPG, resulting in accelerated bone turnover in patients with skeletal fluorosis. To investigate the influence of fluoride on Wnt/ß-catenin signaling pathway, 64 male BALB/c mice were allotted randomly to four groups and treated with deionized water containing 0, 55, 110 and 221 mg/L NaF for 3 months, respectively. The results demonstrated that fluoride significantly increased mouse cancellous bone formation and the protein expression of Wnt3a, phospho-GSK3ß (ser 9) and Runx2. Moreover, partial correlation analysis indicated that there was no significant correlation between fluoride exposure and Runx2 protein levels, after adjusting for ß-catenin, suggesting that ß-catenin might play a crucial role in fluoride-induced aberrant osteogenesis. In vivo, viability of SaoS2 cells was significantly facilitated by 4 mg/L NaF, and fluoride could induce the abnormal activation of Wnt/ß-catenin signaling, the expression of its target gene Runx2 and significantly increased Tcf/Lef reporter activity. Importantly, inhibition of ß-catenin suppressed fluoride-induced Runx2 protein expression and the osteogenic phenotypes. Taken together, the present study provided in vivo and in vitro evidence reveals a potential mechanism for fluoride-induced aberrant osteoblast activation and indicates that ß-catenin is the pivot molecule mediating viability and differentiation of osteoblasts and might be a therapeutic target for skeletal fluorosis.


Assuntos
Osteogênese , beta Catenina , Animais , Estudos de Casos e Controles , China , Estudos Transversais , Fluoretos , Humanos , Camundongos , Camundongos Endogâmicos BALB C , Osteoblastos
19.
Zhongguo Xiu Fu Chong Jian Wai Ke Za Zhi ; 34(8): 1044-1051, 2020 Aug 15.
Artigo em Chinês | MEDLINE | ID: mdl-32794677

RESUMO

Objective: To investigate the effect of graphene oxide (GO)-carboxymethyl chitosan (CMC) hydrogel loaded with interleukin 4 (IL-4) and bone morphogenetic protein 2 (BMP-2) on macrophages M2 type differentiation and osteogenic differentiation of bone marrow mesenchymal stem cells (BMSCs). Methods: GO solution was mixed with CMC, then the phosphate buffered saline (PBS), IL-4, BMP-2, or IL-4+BMP-2 were added to prepare different GO-CMC hydrogel scaffolds with or without different cytokines under crosslinking agents. The characteristics of pure GO-CMC hydrogel were characterized by gross observation, scanning electron microscope (SEM), and Fourier transform infrared spectroscopy (FTIR), and the CMC hydrogel was used as control. The sustained release of GO-CMC hydrogels with different cytokines was also tested. Macrophages were isolated and cultured from female Sprague Dawley rats aged 4-5 weeks, and then cultured with GO-CMC hydrogels with and without different cytokines, respectively. CD206 immunofluorescence staining was used to detect the differentiation of macrophages after 24 hours. The 3rd generation of rats BMSCs were cultured with GO-CMC hydrogels with and without different cytokines respectively for osteogenic induction. The early osteogenesis was observed by alkaline phosphatase (ALP) staining after 10 days, and the late osteogenesis was observed by alizarin red staining after 21 days. Results: Generally, GO-CMC hydrogel was brown and translucent. SEM showed that the pore diameter and wall thickness of GO-CMC hydrogel were similar to that of CMC hydrogel, but the inner wall roughness increased. FTIR test showed that CMC polymerized to form hydrogel. In vitro, the sustained release experiments showed that the properties of GO-CMC hydrogels loaded with different cytokines were similar. CD206 immunofluorescence detection showed that GO-CMC hydrogels could induce macrophages differentiation into M2-type. ALP and alizarin red staining showed that GO-CMC hydrogels could induce BMSCs osteogenic differentiation, in which GO-CMC hydrogel loaded with IL-4+BMP-2 showed the most significant effect ( P<0.05). Conclusion: The GO-CMC hydrogel loaded with IL-4 and BMP-2 can induce macrophages differentiation into M2-type and enhance the ability of BMSCs with osteogenic differentiation in vitro, which provide a new strategy for bone defect repair and immune regulation.


Assuntos
Proteína Morfogenética Óssea 2 , Quitosana , Animais , Diferenciação Celular , Células Cultivadas , Feminino , Grafite , Hidrogéis , Interleucina-4 , Osteogênese , Ratos , Ratos Sprague-Dawley
20.
Zhongguo Xiu Fu Chong Jian Wai Ke Za Zhi ; 34(8): 1052-1058, 2020 Aug 15.
Artigo em Chinês | MEDLINE | ID: mdl-32794678

RESUMO

Objective: To investigate the effect of small interfering RNA (siRNA) lentivirus-mediated silencing of P75 neurotrophin receptor (P75NTR) gene on osteogenic differentiation of bone marrow mesenchymal stem cells (BMSCs) in rats. Methods: Three lentivirus-mediated P75NTR gene siRNA sequences (P75NTR-siRNA-1, 2, 3) and negative control (NC)-siRNA were designed and transfected into the 3rd generation Sprague Dawley (SD) rat BMSCs. The cells morphological changes were observed under an inverted microscope, and the expressions of P75NTR gene and protein in cells were detected by real-time fluorescence quantitative PCR and Western blot. Then the best silencing P75NTR-siRNA for subsequent osteogenic differentiation experiments was screened out. The 3rd generation SD rat BMSCs were randomly divided into experimental group, negative control group, and blank control group (normal BMSCs). The BMSCs of negative control group and experimental group were transfected with NC-siRNA and the selected P75NTR-siRNA lentiviral vector, respectively. The cells of each group were cultured by osteogenic induction. The expressions of osteogenic related proteins [osteocalcin (OCN) and Runx related transcription factor 2 (Runx2)] were detected by Western blot; the collagen type Ⅰ expression was observed by immunohistochemical staining; the osteogenesis of BMSCs was observed by alkaline phosphatase (ALP) detection and alizarin red staining. Results: After lentivirus-mediated P75NTR transfected into BMSCs, the expressions of P75NTR mRNA and protein significantly reduced ( P<0.05), and the best silencing P75NTR-siRNA was P75NTR-siRNA-3. After P75NTR gene was silenced, MTT test showed that the cell proliferation in the experimental group was significantly faster than those in the two control groups ( P<0.05). After osteogenic induction, the relative expressions of OCN and Runx2 proteins, collagen type Ⅰ expression, and ALP activity were significantly higher in the experimental group than in the two control groups, the differences were significant ( P<0.05). With the prolongation of osteogenic induction, the mineralized nodules in the experimental group gradually increased. Conclusion: Silencing the P75NTR gene with siRNA lentivirus can promote the osteogenic differentiation of rat BMSCs and provide a new idea for the treatment of bone defects.


Assuntos
Células-Tronco Mesenquimais , Osteogênese , Animais , Células da Medula Óssea , Diferenciação Celular , Células Cultivadas , Lentivirus , Ratos , Ratos Sprague-Dawley , Receptor de Fator de Crescimento Neural
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