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1.
J Clin Periodontol ; 47(11): 1391-1402, 2020 11.
Article in English | MEDLINE | ID: mdl-32946590

ABSTRACT

AIM: To evaluate the safety and efficacy of autologous periodontal ligament-derived mesenchymal stem cells (PDL-MSCs) embedded in a xenogeneic bone substitute (XBS) for the regenerative treatment of intra-bony periodontal defects. MATERIAL AND METHODS: This quasi-randomized controlled pilot phase II clinical trial included patients requiring a tooth extraction and presence of one intra-bony lesion (1-2 walls). Patients were allocated to either the experimental (XBS + 10 × 106 PDL-MSCs/100 mg) or the control group (XBS). Clinical and radiographical parameters were recorded at baseline, 6, 9 and 12 months. The presence of adverse events was also evaluated. Chi-square, Student's t test, Mann-Whitney U, repeated-measures ANOVA and regression models were used. RESULTS: Twenty patients were included. No serious adverse events were reported. Patients in the experimental group (n = 9) showed greater clinical attachment level (CAL) gain (1.44, standard deviation [SD] = 1.87) and probing pocket depth (PPD) reduction (2.33, SD = 1.32) than the control group (n = 10; CAL gain = 0.88, SD = 1.68, and PPD reduction = 2.10, SD = 2.46), without statistically significant differences. CONCLUSION: The application of PDL-MSCs to XBS for the treatment of one- to two-wall intra-bony lesions was safe and resulted in low postoperative morbidity and appropriate healing, although its additional benefit, when compared with the XBS alone, was not demonstrated.


Subject(s)
Alveolar Bone Loss , Bone Substitutes , Mesenchymal Stem Cells , Alveolar Bone Loss/diagnostic imaging , Alveolar Bone Loss/surgery , Bone Regeneration , Bone Substitutes/therapeutic use , Guided Tissue Regeneration, Periodontal , Humans , Periodontal Attachment Loss/surgery , Periodontal Ligament
2.
Am J Physiol Heart Circ Physiol ; 309(11): H1964-73, 2015 Dec 01.
Article in English | MEDLINE | ID: mdl-26408535

ABSTRACT

The objective of this article is to present an in vitro model of atrial cardiac tissue that could serve to study the mechanisms of remodeling related to atrial fibrillation (AF). We analyze the modification on gene expression and modifications on rotor dynamics following tissue remodeling. Atrial murine cells (HL-1 myocytes) were maintained in culture after the spontaneous initiation of AF and analyzed at two time points: 3.1 ± 1.3 and 9.7 ± 0.5 days after AF initiation. The degree of electrophysiological remodeling (i.e., relative gene expression of key ion channels) and structural inhomogeneity was compared between early and late cell culture times both in nonfibrillating and fibrillating cell cultures. In addition, the electrophysiological characteristics of in vitro fibrillation [e.g., density of phase singularities (PS/cm(2)), dominant frequency, and rotor meandering] analyzed by means of optical mapping were compared with the degree of electrophysiological remodeling. Fibrillating cell cultures showed a differential ion channel gene expression associated with atrial tissue remodeling (i.e., decreased SCN5A, CACN1C, KCND3, and GJA1 and increased KCNJ2) not present in nonfibrillating cell cultures. Also, fibrillatory complexity was increased in late- vs. early stage cultures (1.12 ± 0.14 vs. 0.43 ± 0.19 PS/cm(2), P < 0.01), which was associated with changes in the electrical reentrant patterns (i.e., decrease in rotor tip meandering and increase in wavefront curvature). HL-1 cells can reproduce AF features such as electrophysiological remodeling and an increased complexity of the electrophysiological behavior associated with the fibrillation time that resembles those occurring in patients with chronic AF.


Subject(s)
Atrial Fibrillation/physiopathology , Atrial Remodeling , Action Potentials , Animals , Anti-Arrhythmia Agents/pharmacology , Atrial Fibrillation/drug therapy , Atrial Fibrillation/genetics , Atrial Fibrillation/metabolism , Atrial Remodeling/drug effects , Calcium Channels, L-Type/genetics , Calcium Channels, L-Type/metabolism , Cell Line , Connexin 43/genetics , Connexin 43/metabolism , Gene Expression Regulation , Heart Atria/drug effects , Heart Atria/metabolism , Heart Atria/physiopathology , Mice , Models, Cardiovascular , NAV1.5 Voltage-Gated Sodium Channel/genetics , NAV1.5 Voltage-Gated Sodium Channel/metabolism , Potassium Channels, Inwardly Rectifying/genetics , Potassium Channels, Inwardly Rectifying/metabolism , Shal Potassium Channels/genetics , Shal Potassium Channels/metabolism , Time Factors , Voltage-Sensitive Dye Imaging
3.
Injury ; 45 Suppl 4: S42-7, 2014 Oct.
Article in English | MEDLINE | ID: mdl-25384474

ABSTRACT

INTRODUCTION: The most common method to obtain human mesenchymal stem cells (MSCs) is bone marrow aspiration from the iliac crest, but MSCs have also been isolated from different bones. The main purpose of this study was to compare bone marrow MSCs aspirated from the metaphysis of the distal femur and the proximal tibia with those obtained from the iliac crest, and to determine whether these locations represent potential alternative sources of MSCs for research and clinical application. MATERIALS AND METHODS: Bone marrow was aspirated from the iliac crest and the metaphysis of the distal femur and the proximal tibia during total knee arthroplasty in 20 patients. The aspirates were centrifuged by density gradient, then mononucleated cell (MNC) concentration in the different aspirates was determined using a Coulter counter. MSCs were isolated, cultivated and characterised by their immunophenotype and by their in vitro potential for differentiation into osteoblasts, chondroblasts and adipocytes in specific media. Expansion and cell viability were quantified using trypan blue staining and cell counting with a haemocytometer (Neubauer chamber). The three sources were compared in terms of MNC concentration, viability of the cultures and presence of MSC using the Wilcoxon test. RESULTS: MNC concentration was significantly higher in the iliac crest (10.05 Millions/ml) compared with the femur (0.67 Millions/ml) and tibia (1.7 Millions/ml). Culture success rates were 90%, 71% and 47% for MSCs from the iliac crest, femur and tibia, respectively. Flow cytometry analysis showed the presence of CD90+, CD105+, CD73+, VEGF+, CD71+, HLA-DR-, CD45-, CD34-, CD19-, and CD14- cells. The immunophenotype pattern of MSCs was similar for the three locations. Trilineage differentiation was achieved with all samples. CONCLUSIONS: MSCs can be found in bone marrow from the metaphysis of both the distal femur and the proximal tibia. The phenotype and differentiation potential of these cells are similar to those of bone marrow MSCs from the iliac crest. Bone marrow aspiration from these locations is a relatively easy and safe alternative to that from the iliac crest for obtaining MSCs. Further study is required to assess whether the concentrations of MSCs obtained from these sources are sufficient for one-step therapeutic purposes.


Subject(s)
Bone Marrow Cells/cytology , Cell Culture Techniques/methods , Ilium/cytology , Knee Joint/cytology , Mesenchymal Stem Cells/cytology , Aged , Bone Marrow Cells/physiology , Female , Humans , Male , Mesenchymal Stem Cells/physiology , Middle Aged
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