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1.
PLoS One ; 9(9): e106120, 2014.
Article in English | MEDLINE | ID: mdl-25180783

ABSTRACT

A recent study showed that ergometry increased circulating hematopoietic stem and progenitor cell (CPC) numbers, but reduced hematopoietic colony forming capacity/functionality under normoxia and normobaric hypoxia. Herein we investigated whether an exercise-induced elevated plasma free/bound norepinephrine (NE) concentration could be responsible for directly influencing CPC functionality. Venous blood was taken from ten healthy male subjects (25.3+/-4.4 yrs) before and 4 times after ergometry under normoxia and normobaric hypoxia (FiO2<0.15). The circulating hematopoietic stem and progenitor cell numbers were correlated with free/bound NE, free/bound epinephrine (EPI), cortisol (Co) and interleukin-6 (IL-6). Additionally, the influence of exercise-induced NE and blood lactate (La) on CPC functionality was analyzed in a randomly selected group of subjects (n = 6) in vitro under normoxia by secondary colony-forming unit granulocyte macrophage assays. Concentrations of free NE, EPI, Co and IL-6 were significantly increased post-exercise under normoxia/hypoxia. Ergometry-induced free NE concentrations found in vivo showed a significant impairment of CPC functionality in vitro under normoxia. Thus, ergometry-induced free NE was thought to trigger CPC mobilization 10 minutes post-exercise, but as previously shown impairs CPC proliferative capacity/functionality at the same time. The obtained results suggest that an ergometry-induced free NE concentration has a direct negative effect on CPC functionality. Cortisol may further influence CPC dynamics and functionality.


Subject(s)
Colony-Forming Units Assay , Exercise , Hematopoietic Stem Cells/cytology , Norepinephrine/blood , Adult , Antigens, CD34/metabolism , Blood Cell Count , Cell Hypoxia , Cell Proliferation , Epinephrine/blood , Ergometry , Humans , Hydrocortisone/blood , Interleukin-6/blood , Lactates/blood , Leukocyte Common Antigens/metabolism , Male
2.
Histochem Cell Biol ; 140(6): 611-21, 2013 Dec.
Article in English | MEDLINE | ID: mdl-23996194

ABSTRACT

The aim of the present study was to evaluate the potential of intraoral harvested alveolar bone as an alternative source of multipotent mesenchymal stromal cells for future applications in oral and maxillofacial tissue engineering. Explant cultures were established from 20 alveolar bone samples harvested from the oblique line immediately before wisdom tooth removal. Morphology and proliferation characteristics of the in vitro expanded cells, referred to as human alveolar bone-derived cells (hABDCs), were studied using phase-contrast microscopy. Immunocytochemical analysis of their surface marker expression was conducted using monoclonal antibodies defining mesenchymal stromal cells. To evaluate their multilineage differentiation potential, hABDCs were induced to differentiate along the osteogenic, adipogenic, and chondrogenic lineage and compared to bone marrow mesenchymal stromal cells (hBMSCs) on mRNA and protein levels applying RT-PCR and cytochemical staining methods. hABDCs showed typical morphological characteristics comparable to those of hBMSCs such as being mononuclear, fibroblast-like, spindle-shaped, and plastic adherent. Immunophenotypically, cells were positive for CD105, CD90, and CD73 while negative for CD45, CD34, CD14, CD79α, and HLA-DR surface molecules, indicating an antigen expression pattern considered typical for multipotent mesenchymal stromal cells. As evidenced by RT-PCR and cytochemistry, hABDCs showed multilineage differentiation and similar chondrogenic and osteogenic differentiation potentials when compared to hBMSCs. Our findings demonstrate that human alveolar bone contains mesenchymal progenitor cells that can be isolated and expanded in vitro and are capable of trilineage differentiation, providing a reservoir of multipotent mesenchymal cells from an easily accessible tissue source.


Subject(s)
Alveolar Process/cytology , Bone Marrow Cells/cytology , Mesenchymal Stem Cells/cytology , Stromal Cells/cytology , Cell Proliferation , Humans
3.
J Craniomaxillofac Surg ; 41(2): 110-8, 2013 Mar.
Article in English | MEDLINE | ID: mdl-22898339

ABSTRACT

The purpose of this study was to analyse the potential of intraoral tissues as a source of mesenchymal stromal and progenitor cells (MSPCs) for usage in future cell-based therapy models. Cells were isolated from four different tissues harvested during oral surgery intervention: (1) bone explants from the posterior maxilla, (2) bone explants from the oblique line, (3) from the mandibular periosteum, and (4) from the dental pulp. Donor sites and tissues were evaluated in terms of their accessibility, donor-site morbidity and average time period until appearance of MSPC colonies. Cell characterization was performed by flow cytometry and evaluation of in vitro osteogenic, adipogenic and chondrogenic differentiation potential. Adherent cell colonies were isolated from tissues from all sites after 4-8 days. The cells showed characteristics of MSPCs, so they were expanded up to clinical scales and demonstrated multipotency. The lowest donor-site morbidity was observed in the posterior maxilla harvests, while the highest donor-site morbidity was associated with harvests from mandibular sites. All sites seem to be potential sources of mesenchymal stromal and progenitor cells for tissue engineering approaches. Therefore, harvest morbidity and patient acceptance should affect the choice of the appropriate site.


Subject(s)
Dental Pulp/cytology , Mandible/cytology , Maxilla/cytology , Mesenchymal Stem Cells/physiology , Periosteum/cytology , Adipogenesis/physiology , Adult , Aged , Aggrecans/analysis , Alkaline Phosphatase/analysis , Calcium/analysis , Cell Culture Techniques , Cell Differentiation/physiology , Cell Lineage/physiology , Chondrogenesis/physiology , Humans , Lipids/analysis , Middle Aged , Multipotent Stem Cells/physiology , Osteogenesis/physiology , Time Factors , Tissue and Organ Harvesting/methods , Transplant Donor Site , Young Adult
4.
Stem Cells Dev ; 21(16): 2915-25, 2012 Nov 01.
Article in English | MEDLINE | ID: mdl-22616638

ABSTRACT

Circulating hematopoietic progenitor cells (CPCs) may be triggered by physical exercise and/or normobaric hypoxia from the bone marrow. The aim of the study was to investigate the influence of physical exercise and normobaric hypoxia on CPC number and functionality in the peripheral blood as well as the involvement of oxidative stress parameters as possibly active agents. Ten healthy male subjects (25.3±4.4 years) underwent a standardized cycle incremental exercise test protocol (40 W+20 W/min) under either normoxic (FiO2 ∼0.21) or hypoxic conditions (FiO2<0.15, equals 3,500 m, 3 h xposure) within a time span of at least 1 week. Blood was drawn from the cubital vein before and 10, 30, 60, and 120 min after exercise. The number of CPCs in the peripheral blood was analyzed by flow cytometry (CD34/CD45-positive cells). The functionality of cells present was addressed by secondary colony-forming unit-granulocyte macrophage (CFU-GM) assays. To determine a possible correlation between the mobilization of CPCs and reactive oxygen species, parameters for oxidative stress such as malondialdehyde (MDA) and myeloperoxidase (MPO) were obtained. Data showed a significant increase of CPC release under normoxic as well as hypoxic conditions after 10 min of recovery (P<0.01). Most interestingly, although CD34+/CD45dim cells increased in number, the proliferative capacity of CPCs decreased significantly 10 min after cessation of exercise (P<0.05). A positive correlation between CPCs and MDA/MPO levels turned out to be significant for both normoxic and hypoxic conditions (P<0.05/P<0.01). Hypoxia did not provoke an additional effect. Although the CPC frequency increased, the functionality of CPCs decreased significantly after exercise, possibly due to the influence of increased oxidative stress levels.


Subject(s)
Cell Movement , Colony-Forming Units Assay , Exercise/physiology , Hematopoiesis , Hematopoietic Stem Cells/cytology , Adult , Antigens, CD34/metabolism , Blood Cell Count , Cell Proliferation , Erythropoietin/metabolism , Flow Cytometry , Humans , Hypoxia/blood , Kinetics , Leukocyte Common Antigens/metabolism , Male , Malondialdehyde/metabolism , Oxidative Stress , Peroxidase/metabolism
5.
J Oral Maxillofac Surg ; 70(1): 154-62, 2012 Jan.
Article in English | MEDLINE | ID: mdl-22014939

ABSTRACT

PURPOSE: The aim of the present study was to compare the influence of 2 different bone scrapers with respect to graft quality. MATERIALS AND METHODS: The study was conducted as a prospective, controlled experimental study of patients selected from the outpatient unit of the Department of Oral Surgery and Radiology (Dental Clinic, Medical University, Graz, Austria). Bone samples were obtained during routine lower third molar removal. Both a manual bone scraper (MS) and a piezoelectric device (PD) were used in directly adjacent regions in each case. As variables, the chip morphology, cell viability, and osteogenic differentiation were investigated. For statistical analysis, the Student t test and Fisher's exact test (P < .05) were applied. RESULTS: A total of 20 patients (12 women and 8 men, mean age 28.15 ± 5.8 years) were included in the study. A series of 40 bone samples was obtained during lower third molar removal. MS and PD enabled similar intraoral harvest of bone chips. In vitro outgrowth of adherent cells was found in 90% of the MS and 80% of the PD samples after 7 to 18 days, without statistical significance (P = .67). Similar cell viability of outgrowing cells in both groups was observed (94.7% ± 2.2% in the MS group and 94.1% ± 1.6% in the PD group). Reverse transcriptase-polymerase chain reaction analysis and the staining pattern verified osteopotent cells in both groups. CONCLUSIONS: Both manual and piezoelectric techniques are adequate harvesting technologies for limited intraoral augmentations. Our results did not show an advantage for the piezoelectric device.


Subject(s)
Bone Transplantation/pathology , Osteotomy/instrumentation , Piezosurgery/instrumentation , Tissue and Organ Harvesting/instrumentation , Adult , Alkaline Phosphatase/analysis , Cell Culture Techniques , Cell Differentiation/physiology , Cell Line , Cell Survival/physiology , Collagen Type I/analysis , Female , Guided Tissue Regeneration , Humans , Immunohistochemistry , Male , Microscopy, Electron, Scanning , Minimally Invasive Surgical Procedures , Osteoblasts/physiology , Osteocalcin/analysis , Osteogenesis/physiology , Osteonectin/analysis , Osteopontin/analysis , Prospective Studies , Tooth Socket/surgery , Young Adult
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