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1.
Cell Tissue Res ; 383(3): 1061-1075, 2021 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-33242173

RESUMO

Adipose-derived stem cells (ASC) have been used as an alternative to bone marrow mesenchymal stem cells (BMSC) for bone tissue engineering. However, the efficacy of ASC in bone regeneration in comparison with BMSC remains debatable, since inconsistent results have been reported. Comparing ASC with BMSC obtained from different individuals might contribute to this inconsistency in results. Therefore, this study aimed to compare the bone regenerative capacity of donor-matched human ASC and BMSC seeded onto poly(L-lactide-co-ε-caprolactone) scaffolds using calvarial bone defects in nude rats. First, donor-matched ASC and BMSC were seeded onto the co-polymer scaffolds to evaluate their in vitro osteogenic differentiation. Seeded scaffolds and scaffolds without cells (control) were then implanted in calvarial defects in nude rats. The expression of osteogenesis-related genes was examined after 4 weeks. Cellular activity was investigated after 4 and 12 weeks. Bone formation was evaluated radiographically and histologically after 4, 12, and 24 weeks. In vitro, ASC and BMSC demonstrated mineralization. However, BMSC showed higher alkaline phosphatase activity than ASC. In vivo, human osteogenesis-related genes Runx2 and collagen type I were expressed in defects with scaffold/cells. Defects with scaffold/BMSC had higher cellular activity than defects with scaffold/ASC. Moreover, bone formation in defects with scaffold/BMSC was greater than in defects with scaffold/ASC, especially at the early time-point. These results suggest that although ASC have the potential to regenerate bone, the rate of bone regeneration with ASC may be slower than with BMSC. Accordingly, BMSC are more suitable for bone regenerative applications.


Assuntos
Células da Medula Óssea/citologia , Regeneração Óssea , Células-Tronco Mesenquimais/citologia , Osteogênese , Engenharia Tecidual/métodos , Alicerces Teciduais , Animais , Diferenciação Celular , Células Cultivadas , Criança , Feminino , Humanos , Masculino , Ratos
2.
Stem Cell Res Ther ; 9(1): 168, 2018 06 19.
Artigo em Inglês | MEDLINE | ID: mdl-29921311

RESUMO

BACKGROUND: Adipose-derived stem cells (ASCs) have been introduced as an alternative to bone marrow mesenchymal stem cells (BMSCs) for cell-based therapy. However, different studies comparing ASCs and BMSCs have shown conflicting results. In fact, harvesting ASCs and BMSCs from different individuals might influence the results, making comparison difficult. Therefore, this study aimed to characterize donor-matched ASCs and BMSCs in order to investigate proliferation, differentiation potential and possible effects of donor variation on these mesenchymal stem cells (MSCs). METHODS: Human bone marrow and adipose tissue samples were obtained from nine donors aged 8-14. ASCs and BMSCs were isolated and characterized based on expression of surface markers using flow cytometry. The proliferation up to 21 days was investigated. Multi-lineage differentiation was induced using osteogenic, chondrogenic and adipogenic differentiation media. Alkaline phosphatase (ALP) activity was monitored and collagen type I formation was evaluated by immunofluorescence staining. In vitro multi-potency was studied using tissue-specific stains and lineage-specific gene expression. In addition, the osteogenic lineage was evaluated at protein level. RESULTS: Isolated ASCs and BMSCs from all donors demonstrated morphologic and immunophenotypic characteristics of MSCs, with expression of MSCs markers and negative expression of hematopoietic markers. Unlike BMSCs, ASCs showed high expression of CD49d and low expression of Stro-1. In general, ASCs showed significantly higher proliferation and adipogenic capacity with more lipid vesicle formation and expression of the adipogenesis-related genes than BMSCs. In contrast, BMSCs showed significantly higher osteogenic and chondrogenic capacity compared to ASCs. BMSCs had earlier and higher ALP activity, calcium deposition, and expression of the osteogenesis- and chondrogenesis-related genes and the osteogenesis-related protein osteopontin. Proliferation and differentiation capacity of ASCs and BMSCs varied significantly among the donors. CONCLUSIONS: ASCs and BMSCs showed tissue-specific differentiation abilities, but with significant variation between donors. The similarities and differences in the properties of ASCs and BMSCs should be taken into consideration when planning stem cell-based therapy.


Assuntos
Adipogenia/fisiologia , Tecido Adiposo/metabolismo , Células-Tronco Mesenquimais/metabolismo , Diferenciação Celular , Proliferação de Células , Humanos , Doadores de Tecidos
3.
BMC Oral Health ; 15: 86, 2015 Jul 27.
Artigo em Inglês | MEDLINE | ID: mdl-26211001

RESUMO

BACKGROUND: Pathological changes in periodontal tissues are mediated by the interaction between microorganisms and the host immune-inflammatory response. Hyperglycemia may interfere with this process. The aim of this study was to compare the levels of 27 inflammatory molecules in the gingival crevicular fluid (GCF) of patients with type 2 diabetes, with and without chronic periodontitis, and of chronic periodontitis subjects without diabetes. A putative correlation between glycated haemoglobin (HbA1c) and levels of the inflammatory molecules was also investigated. METHODS: The study population comprised a total of 108 individuals, stratified into: 54 with type 2 diabetes and chronic periodontitis (DM + CP), 30 with chronic periodontitis (CP) and 24 with type 2 diabetes (DM). Participants were interviewed with the aid of structured questionnaire. Periodontal parameters (dental plaque, bleeding on probing and periodontal pocket depth) were recorded. The GCF levels of the 27 inflammatory molecules were measured using multiplex micro-bead immunoassay. A glycated haemoglobin (HbA1c) test was performed for patients with diabetes by boronate affinity chromatography. RESULTS: After adjustment for potential confounders, the DM + CP group had higher levels of IL-8 and MIP-1ß, and lower levels of TNF-α, IL-4, INF-γ, RANTES and IL-7 compared to the CP group. Moreover, the DM + CP group had lower levels of IL-6, IL-7 and G-CSF compared to the DM group. The DM group had higher levels of IL-10, VEGF, and G-CSF compared to the CP group. The levels of MIP-1α and FGF were lower in diabetes patients (regardless of their periodontal status) than in chronic periodontitis subjects without diabetes. Diabetes patients (DM + CP and DM) had higher Th-2/Th-1 ratio compared to the CP group. HbA1c correlated positively with the pro-inflammatory cytokines (Pearson correlation coefficient = 0.27, P value: 0.02). CONCLUSION: Type 2 diabetes and chronic periodontitis may influence the GCF levels of inflammatory molecules synergistically as well as independently. Type 2 diabetes was associated with high Th-2/Th-1 ratio, and modulated the local expression of molecules involved in the anti-inflammatory and healing processes.


Assuntos
Periodontite Crônica/imunologia , Diabetes Mellitus Tipo 2/imunologia , Líquido do Sulco Gengival/imunologia , Mediadores da Inflamação/análise , Adulto , Idoso , Quimiocina CCL3/análise , Quimiocina CCL4/análise , Quimiocina CCL5/análise , Periodontite Crônica/sangue , Estudos Transversais , Índice de Placa Dentária , Diabetes Mellitus Tipo 2/sangue , Feminino , Fatores de Crescimento de Fibroblastos/análise , Hemoglobinas Glicadas/análise , Fator Estimulador de Colônias de Granulócitos/análise , Humanos , Mediadores da Inflamação/sangue , Interferon gama/análise , Interleucina-10/análise , Interleucina-4/análise , Interleucina-6/análise , Interleucina-7/análise , Interleucina-8/análise , Masculino , Pessoa de Meia-Idade , Índice Periodontal , Células Th1/imunologia , Células Th2/imunologia , Fator de Necrose Tumoral alfa/análise , Fator A de Crescimento do Endotélio Vascular/análise , Adulto Jovem
4.
PLoS One ; 10(5): e0127660, 2015.
Artigo em Inglês | MEDLINE | ID: mdl-25993052

RESUMO

The relationship between diabetes and periodontal disease is bidirectional, but information about the effect of chronic periodontitis on the levels of the glucoregulatory biomarkers locally in gingival crevicular fluid (GCF) is limited. The aim of this study was to compare the levels of 10 glucoregulatory biomarkers in GCF, firstly in subjects with type 2 diabetes (T2DM) presenting with and without chronic periodontitis and secondly, in subjects without diabetes, with and without chronic periodontitis. The material comprised a total of 152 subjects, stratified as: 54 with T2DM and chronic periodontitis (G1), 24 with T2DM (G2), 30 with chronic periodontitis (G3) and 44 without T2DM or periodontitis (G4). The levels of the biomarkers were measured using multiplex biometric immunoassays. Periodontal pocket depths were recorded in mm. Subsets G1 and G2 and subsets G3 and G4 were compared independently. Among T2DM subjects, GIP, GLP-1 and glucagon were significantly up-regulated in G1 compared to G2. Moreover, there were no statistical differences between the two groups regarding C-peptide, insulin, ghrelin, leptin and PAI-1. Comparisons among individuals without T2DM revealed significantly lower amounts of C-peptide and ghrelin in G3 than in G4. The number of sites with pocket depth ≥ 4mm correlated negatively with C-peptide (Spearman's correlation co-efficient: -0.240, P < 0.01) and positively with GIP and visfatin (Spearman's correlation co-efficient: 0.255 and 0.241, respectively, P < 0.01). The results demonstrate that chronic periodontitis adversely influences the GCF levels of glucoregulatory biomarkers, as it is associated with disturbed levels of biomarkers related to the onset of T2DM and its medical complications.


Assuntos
Biomarcadores/metabolismo , Periodontite Crônica/complicações , Periodontite Crônica/metabolismo , Diabetes Mellitus Tipo 2/complicações , Diabetes Mellitus Tipo 2/metabolismo , Líquido do Sulco Gengival/metabolismo , Glucose/metabolismo , Adulto , Demografia , Feminino , Humanos , Modelos Logísticos , Masculino , Pessoa de Meia-Idade
5.
PLoS One ; 8(12): e82158, 2013.
Artigo em Inglês | MEDLINE | ID: mdl-24349205

RESUMO

AIM: The purpose of this study was to compare the clinical and subjective oral health indicators of type 2 diabetic patients (T2DM) with age and gender matched non-diabetic controls. A second aim was to identify clinical and subjective oral health indicators that discriminate between well-controlled and poorly controlled T2DM patients as well as between patients with long and short duration of the disease. METHODS: A total of 457 individuals participated in the study (154 T2DM cases and 303 non-diabetic controls). The T2DM group was sub-divided according to metabolic control [(well-controlled: glycosylated haemoglobin test 8%), (poorly controlled: glycosylated haemoglobin test > 8%)] and according to duration of T2DM [(long duration: >10 years), (short duration: 10 years)]. Participants were interviewed using a structured questionnaire including socio-demographics, lifestyle and oral health related quality of life factors. The clinical examination comprised full mouth probing depths, plaque index, tooth mobility index, furcation involvement and coronal and root surface caries. RESULTS: The T2DM patients presented with more probing depths 4 mm, furcation involvement, tooth mobility, missing teeth, and oral impacts on daily performance (OIDP). The corresponding adjusted odds ratios and their 95% confidence intervals were 4.07 (1.74-9.49), 2.96 (1.36-6.45), 5.90 (2.26-15.39), 0.23 (0.08-0.63) and 3.46 (1.61-7.42), respectively. Moreover, the odds ratio was 2.60 (1.21-5.55) for the poorly controlled T2DM patients to have high levels of mobility index and 2.94 (1.24-6.94) for those with long duration of T2DM to have high decayed, missed and filled teeth (DMFT) values. CONCLUSION: This study revealed that chronic periodontitis, tooth mobility, furcation involvement and OIDP were more prevalent among T2DM patients compared to their non-diabetic controls.


Assuntos
Diabetes Mellitus Tipo 2/patologia , Nível de Saúde , Saúde Bucal , Adulto , Estudos de Casos e Controles , Fatores de Confusão Epidemiológicos , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Análise de Regressão , Autorrelato , Sudão
6.
Tissue Eng Part A ; 17(21-22): 2817-31, 2011 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-21905880

RESUMO

One of the principal goals in tissue engineering is to produce scaffold materials that will guide cells to differentiate and regenerate functional replacement tissue at the site of injury. Poly(l-lactide-co-1,5-dioxepan-2-one) [Poly(LLA-co-DXO)], a potential scaffolding material for bone tissue engineering, has high hydrophilicity. Previous in vitro studies using human osteoblast-like cells (HOBs) demonstrated greater cytocompatibility and enhanced osteogenic differentiation when HOBs were seeded onto Poly(LLA-co-DXO) compared to Poly(l-lactide) [P(LLA)] scaffolds. The aim of the study was to identify the gene expression profiles of HOBs obtained from alveolar bone and grown on Poly(LLA-co-DXO) biodegradable polymer scaffolds compared to P(LLA) one. Illumina HumanWG-6 v3.0 Expression BeadChips were used for the gene expression analysis. Several genes were found as differentially expressed at 24 h and at 21 days. Expression of genes related to cell adhesion, cytoskeleton, antiapoptosis, proliferation, and bone mineralization was influenced by adding the monomer 1,5-dioxepan-2-one to the L-lactide. Genes related to three biological pathways involving Integrin, Notch, and Ras were found to be upregulated. For selected genes, results were confirmed by quantitative reverse transcriptase-polymerase chain reaction. Further, calcium content analysis revealed a significant enhancement of calcium deposition on both tested scaffolds. This observation was confirmed by Von Kossa and Alizarin Red S staining. Findings of this study are relevant to a better understanding of the molecular mechanisms underlying the behavior of HOBs in bone regenerative procedure.


Assuntos
Perfilação da Expressão Gênica/métodos , Osteoblastos/citologia , Osteoblastos/metabolismo , Poliésteres/química , Polímeros/química , Engenharia Tecidual/métodos , Alicerces Teciduais/química , Células Cultivadas , Humanos , Análise de Sequência com Séries de Oligonucleotídeos , Reação em Cadeia da Polimerase Via Transcriptase Reversa
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