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1.
Life (Basel) ; 12(6)2022 Jun 15.
Artigo em Inglês | MEDLINE | ID: mdl-35743928

RESUMO

Human skeletal stem cells (hSSCs) were recently identified as podoplanin (PDPN)/CD73/CD164-positive and CD146-negative cells that decline with age, and play a role in the pathogenesis of osteoarthritis (OA). The aim of this study was to identify the hSSC-like properties of bone-derived mesenchymal stem/stromal cells (MSCs) of patients with late and early OA. Methods: First, we performed gene expression profiling for the hSSC markers in 32 patients with late and early OA, and donors without OA. Having identified the low expression of hSSC markers in late OA patients, we further performed trilineage differentiation and immunophenotyping for hSSC makers in the selected subsets from each donor group. Results: Our results show no differences in osteogenesis, chondrogenesis, and adipogenesis between the MSCs from the three groups. However, the immunophenotyping shows lower CD164 in MSCs from early OA patients in comparison with late and no OA subjects (p = 0.002 and p = 0.017). Conclusions: Our study shows that the in vitro hSSC-like properties of bone-derived MSCs are similar in patients with early and late OA, and in donors without OA. However, the lower percentage of CD164-positive MSCs in early OA patients indicates the potential of CD164 as a marker of the onset of OA.

2.
Int J Mol Sci ; 23(6)2022 Mar 15.
Artigo em Inglês | MEDLINE | ID: mdl-35328593

RESUMO

Tissues of post mortem donors represent valuable alternative sources for the isolation of primary cells with mesenchymal stem/stromal cell (MSC)-like properties. However, the properties of primary cells derived from different tissues and at different post mortem times are poorly recognized. Here, we aim to identify the optimal tissue source between three knee and peri-knee tissues for the isolation of primary cells with MSC-like properties, and to define the influence of the time post mortem on the properties of these cells. We harvested tissues from subchondral bone marrow, synovium and periosteum from 32 donors at various post mortem times. Primary cells were evaluated using detailed in vitro analyses, including colony formation, trilineage differentiation, immunophenotyping and skeletal stem cell marker-gene expression profiling. These data show that the primary cells with MSC-like properties isolated from these three tissues show no differences in their properties, except for higher expression of CD146 in bone-marrow cells. The success rate of the primary cell isolation is dependent on the post mortem time. However, synovium and periosteum cells isolated more than 48 h post mortem show improved osteogenic and chondrogenic potential. This study suggests that knee and peri-knee tissues from donors even 3 days post mortem are strategic sources of MSCs for regenerative procedures.


Assuntos
Células-Tronco Mesenquimais , Células da Medula Óssea , Diferenciação Celular , Células Cultivadas , Condrogênese , Células-Tronco Mesenquimais/metabolismo , Osteogênese
3.
Int J Legal Med ; 135(4): 1245-1256, 2021 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-33624158

RESUMO

DNA analysis of Second World War skeletal remains is challenging because of the limited yield of DNA that is usually recovered. Recent forensic research has focused on determining which skeletal elements are superior in their preservation of DNA, and little focus has been placed on measuring intra-bone variability. Metatarsals and metacarpals outperformed all the other bones in DNA yield when analyzing all representative skeletal elements of three Second World War victims, and intra-bone variability was not studied. Soft-tissue remnants were found to contribute to higher DNA yield in trabecular bone tissue. Because metatarsals and metacarpals are composed of trabecular epiphyses and a dense diaphysis, the goal of this study was to explore intra-bone variability in DNA content by measuring nuclear DNA quantity and quality using the PowerQuant System (Promega). A total of 193 bones from a single Second World War mass grave were examined. From each bone, DNA was extracted from the compact diaphysis and from both spongy epiphyses combined. This study confirms higher DNA quantity in epiphyses than diaphyses among all the bones analyzed, and more DNA was obtained from metacarpal epiphyses than from metatarsal epiphyses. Therefore, whenever the possibility for sampling both metacarpals and metatarsals from skeletal remains exists, collecting metacarpals is recommended. In cases in which the hands are missing, metatarsals should be sampled. In any case, epiphyses are a richer source of DNA than diaphyses.


Assuntos
DNA/análise , Diáfises/química , Epífises/química , Ossos Metacarpais , Ossos do Metatarso , Restos Mortais , Humanos , II Guerra Mundial
4.
Forensic Sci Res ; 5(1): 74-84, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32490313

RESUMO

Examination of a person who has been a victim of a physical or sexual assault may be very important for upcoming legal proceedings. In the context of a clinical forensic examination, physical findings are recorded and biological trace material is gathered and secured. Ideally, all forensic findings are documented in a detailed report combined with photographic documentation, which employs a forensic scale to depict the size of the injuries. However, the integrity of such forensic findings depends particularly on two factors. First, the examination needs to be conducted professionally to ensure that the findings are properly admissible as court evidence. Second, the examination should take place as soon as possible because the opportunity to successfully secure biological samples declines rapidly with time. Access to low-threshold clinical forensic examinations is not evenly provided in all member states of the European Union (EU); in some states, they are not available at all. As part of the JUSTeU! (Juridical standards for clinical forensic examinations of victims of violence in Europe) project, the Ludwig Boltzmann Institute for Clinical Forensic Imaging in Graz, Austria created (in cooperation with its international partner consortium) a questionnaire: the purpose was to collect information about support for victims of physical and/or sexual assault in obtaining a low-threshold clinical forensic examination in various countries of the EU. Our paper provides a summary of the responses and an overview of the current situation concerning provided clinical forensic services.

5.
Stem Cell Res Ther ; 11(1): 146, 2020 04 03.
Artigo em Inglês | MEDLINE | ID: mdl-32245507

RESUMO

BACKGROUND: Mesenchymal stem/stromal cells (MSCs) can replenish the aged cells of the musculoskeletal system in adult life. Stem cell exhaustion and decrease in their regenerative potential have been suggested to be hallmarks of aging. Here, we investigated whether muscle- and bone-derived MSCs of patients with osteoarthritis and osteoporosis are affected by this exhaustion, compared to healthy donors. METHODS: Patients with primary osteoarthritis, femoral neck fractures due to osteoporosis, and healthy donors (controls) were included. MSCs were isolated from the skeletal muscle and subchondral bone from each patient and compared using ex vivo and in vitro analyses, including immunophenotyping, colony-forming unit fibroblast assays, growth kinetics, cell senescence, multilineage potential, and MSC marker gene expression profiling. RESULTS: Freshly isolated cells from muscle from patients with osteoarthritis showed a lower proportion of CD45/CD19/CD14/CD34-negative cells compared to patients with osteoporosis and healthy donors. Freshly isolated muscle cells from patients with osteoarthritis and osteoporosis also showed higher clonogenicity compared to healthy donors. MSCs from both tissues of osteoarthritis patients showed significantly reduced osteogenesis and MSCs from the bone also reduced adipogenesis. Chondrogenic pellet diameter was reduced in bone-derived MSCs from both patient groups compared to healthy donors. A significant positive correlation was observed between adipogenesis and CD271 expression in muscle-derived MSCs. CD73 was significantly lower in bone-derived MSCs from osteoarthritis patients, compared to osteoporosis patients. Gene expression profiling showed significantly lower expression of MSC marker gene leptin receptor, LEPR, previously identified as the major source of the bone and adipocytes in the adult bone marrow, in bone-derived MSCs from patients with osteoarthritis in comparison with osteoporotic patients and healthy donors. CONCLUSIONS: Our results show deficient ex vivo and in vitro properties of both skeletal muscle- and bone-derived MSCs in osteoarthritis and osteoporosis patients, compared to healthy donors. In bone-derived MSCs from patients with osteoarthritis, we also identified a lower expression of the leptin receptor, a marker of MSCs that present a major source of MSCs in the adult bone marrow. This suggests that exhaustion of skeletal muscle- and bone-derived MSCs is a hallmark of osteoarthritis and osteoporosis, which defines the need for further clinical trials of stem cell transplantation in these patients.


Assuntos
Fraturas do Colo Femoral , Células-Tronco Mesenquimais , Osteoartrite , Adulto , Idoso , Células da Medula Óssea , Diferenciação Celular , Células Cultivadas , Humanos , Músculo Esquelético , Osteogênese
6.
Adv Exp Med Biol ; 1288: 33-46, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32036570

RESUMO

Stem cells provide for all of the tissues in our body during embryogenesis. In adult organisms, they can be found as rare populations of tissue-specific stem cells in quiescent states, although they can still regenerate damaged tissues. Astonishingly, these cells are retained in tissues even post-mortem. There have been several reports that have provided evidence that cells with stem-like capabilities can be isolated, expanded, and differentiated in vitro from various tissues several hours, or even several days, post-mortem. Moreover, some post-mortem-tissue-derived stem cells can successfully engraft and regenerate injured host tissues. Here, we review in-vitro and in-vivo studies that provide evidence of isolation and characterization of stem cells from different tissues post-mortem, with a focus on the musculoskeletal and neural systems. Finally, we discuss their potential for use in regenerative medicine, and what needs to be done in further research toward their better exploitation.


Assuntos
Autopsia , Separação Celular/métodos , Medicina Regenerativa/métodos , Células-Tronco/citologia , Diferenciação Celular , Humanos
7.
Growth Factors ; 36(5-6): 263-273, 2018 12.
Artigo em Inglês | MEDLINE | ID: mdl-30764675

RESUMO

Paired cartilage and subchondral bone of subjects with no clinical history of joint disorders were analyzed to determine whether antioxidant enzymes, inflammatory cytokines and growth factors can be linked to a pre-osteoarthritis. Tissue explants were phenotyped according to Osteoarthritis Research Society International grading and micro-computed tomography, and also screened for the expression of several markers using quantitative polymerase chain reaction. The expression of these same genes was measured in SW1353 cells treated with hydrogen peroxide, to gain insight into the pathways involved with oxidative stress responses. Vascular endothelial growth factor A (VEGF-A) was up-regulated in the cartilage samples that showed early cartilage or bone degeneration. Oxidative stress in chondrocytes provoked up-regulation of interleukin-1ß, interleukin-6, aggrecan, and SRY-box containing gene 9. Our results confirm the hitherto evidence of the deteriorating effects of the oxidative stress on cartilage and suggest the link between VEGF-A and pre-osteoarthritis.


Assuntos
Osso e Ossos/metabolismo , Cartilagem/metabolismo , Osteoartrite/metabolismo , Fator A de Crescimento do Endotélio Vascular/metabolismo , Idoso , Agrecanas/genética , Agrecanas/metabolismo , Osso e Ossos/patologia , Cartilagem/patologia , Linhagem Celular Tumoral , Células Cultivadas , Feminino , Humanos , Interleucina-1beta/genética , Interleucina-1beta/metabolismo , Masculino , Pessoa de Meia-Idade , Estresse Oxidativo , Fatores de Transcrição SOX9/genética , Fatores de Transcrição SOX9/metabolismo , Regulação para Cima , Fator A de Crescimento do Endotélio Vascular/genética
8.
J Bone Miner Metab ; 31(5): 512-9, 2013 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-23588618

RESUMO

The role of bone marrow adipocytes in bone tissue is not yet understood. Adipocytes express enzymes for metabolism of free fatty acids and adipokines such as adiponectin, which have been shown to exert different effects on bone cells. Our aim was to find out whether triglyceride (TG) metabolism in bone tissue is associated with osteoblast and osteoclast differentiation by gene expression analysis of lipoprotein lipase (LPL), hormone sensitive lipase (HSL), fatty acid synthase (FASN), adiponectin, RUNX2, RANK, RANKL and OPG. Bone tissue was obtained from patients undergoing hip arthroplasty due to osteoporosis (OP) (50) or osteoarthritis (OA) (48) or from healthy autopsy controls (14). Lower bone mineral density and microstructural parameters were observed in OP compared to OA. The FASN expression did not differ between groups suggesting similar de novo lipogenesis. Lower LPL and HSL in OP suggest lower FFA release and uptake in OP bone tissue. Adiponectin expression was lower in OP than in OA and a trend was seen for controls. These results suggest OP bone has lower TG metabolism than OA and normal bone. In OP bone, lower osteoblastogenesis and higher osteoclast formation were observed and correlation analysis suggests adiponectin, LPL and HSL are associated with higher osteoblastogenesis and lower osteoclastogenesis. This study gives insights into TG metabolism in the human bone microenvironment. We conclude that OP bone tissue exhibits lower osteoblastogenesis, higher osteoclastogenesis and lower TG metabolism compared to OA or healthy controls.


Assuntos
Osso e Ossos/citologia , Osso e Ossos/metabolismo , Osteoblastos/citologia , Osteoblastos/metabolismo , Osteoclastos/citologia , Osteoclastos/metabolismo , Triglicerídeos/metabolismo , Idoso , Feminino , Humanos , Masculino , Osteoartrite/metabolismo , Osteoporose/metabolismo
9.
J Orthop Res ; 31(7): 1059-66, 2013 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-23362137

RESUMO

Several studies have shown that in contrast to osteoporosis (OP), osteoarthritis (OA) is characterized by high bone mineral density (BMD). Bone strength not only depends on mineral content as determined by dual X-ray absorptiometry (DXA), but also on bone microarchitecture. We studied intertrochanteric bone from normal controls and OA and OP patients by bone histomorphometry (BHM) and microcomputed tomography (µCT) as well as DXA in order to first, test the differences between OA and OP comparing both groups to healthy controls, second, to assess variations between three different skeletal sites in controls and third, to determine the level of agreement between µCT, BHM, and DXA. Analysis was performed on 115 samples from OA and OP patients, and controls. We found significant differences between OA and OP samples in structural parameters and in the osteoid fraction (p < 0.05). The majority of the intra-skeletal differences were shown between lumbar spine and femoral head samples (p < 0.05). Significant agreements were found between µCT and BHM and DXA (r = 0.32-0.45, p < 0.05). Our findings suggest differences in intertrochanteric bone between OA and OP, the age-related intra-skeletal variations and a correlation between microscopic and macroscopic bone evaluation methods.


Assuntos
Absorciometria de Fóton , Osso e Ossos/diagnóstico por imagem , Osteoartrite/diagnóstico por imagem , Osteoporose/diagnóstico por imagem , Microtomografia por Raio-X , Idoso , Idoso de 80 Anos ou mais , Densidade Óssea , Osso e Ossos/patologia , Estudos de Casos e Controles , Feminino , Cabeça do Fêmur/diagnóstico por imagem , Cabeça do Fêmur/patologia , Humanos , Vértebras Lombares/diagnóstico por imagem , Vértebras Lombares/patologia , Masculino , Pessoa de Meia-Idade , Osteoartrite/patologia , Osteoporose/patologia
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