Multipotent stem cells from umbilical cord: Cord is richer than blood!

The identification of mesenchymal stem cell ( MSC) sources that are easily obtainable is of utmost importance. Several studies have shown that MSCs could be isolated from umbilical cord (UC) units. However, the presence of MSCs in umbilical cord blood (UCB) is controversial. A possible explanation for the low efficiency of MSCs from UCB is the use of different culture conditions by independent studies. Here, we compared the efficiency in obtaining MSCs from unrelated paired UCB and UC samples harvested from the same donors. Samples were processed simultaneously, under the same culture conditions. Although MSCs from blood were obtained from only 1 of the 10 samples, we were able to isolate large amounts of multi-potent MSCs from all UC samples, which were able to originate different cell lineages. Since the routine procedure in UC banks has been to store the blood and discard other tissues, such as the cord and/or placenta, we believe our results are of immediate clinical value. Furthermore, the possibility of originating different cell lines from the UC of neonates born with genetic defects may provide new cellular research models for understanding human malformations and genetic disorders, as well as the possibility of testing the effects of different therapeutic drugs.

Gene Expression Profile of Mesenchymal Stem Cells from Paired Umbilical Cord Units: Cord is Different from Blood

Mesenchymal stem cells (MSC) are multipotent cells which can be obtained from several adult and fetal tissues including human umbilical cord units. We have recently shown that umbilical cord tissue (UC) is richer in MSC than umbilical cord blood (UCB) but their origin and characteristics in blood as compared to the cord remains unknown. Here we compared, for the first time, the exonic protein-coding and intronic noncoding RNA (ncRNA) expression profiles of MSC from match-paired UC and UCB samples, harvested from the same donors, processed simultaneously and under the same culture conditions. The patterns of intronic ncRNA expression in MSC from UC and UCB paired units were highly similar, indicative of their common donor origin. The respective exonic protein-coding transcript expression profiles, however, were significantly different. Hierarchical clustering based on protein-coding expression similarities grouped MSC according to their tissue location rather than original donor. Genes related to systems development, osteogenesis and immune system were expressed at higher levels in UCB, whereas genes related to cell adhesion, morphogenesis, secretion, angiogenesis and neurogenesis were more expressed in UC cells. These molecular differences verified in tissue-specific MSC gene expression may reflect functional activities influenced by distinct niches and should be considered when developing clinical protocols involving MSC from different sources. In addition, these findings reinforce our previous suggestion on the importance of banking the whole umbilical cord unit for research or future therapeutic use.

In vitro cultivation of canine multipotent mesenchymal stromal cells on collagen membranes treated with hyaluronic acid for cell therapy and tissue regeneration

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Periodontal Tissue Engineering After Tooth Replantation

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Comparison of Single-Standing or Connected Implants on Stress Distribution in Bone of Mandibular Overdentures: A Two-Dimensional Finite Element Analysis

This study aimed to compare the influence of single-standing or connected implants on stress distribution in bone of mandibular overdentures by means of two-dimensional finite element analysis. Two finite element models were designed using software (ANSYS) for 2 situations: bar-clip (BC) group-model of an edentulous mandible supporting an overdenture over 2 connected implants with BC system, and o'ring (OR) group-model of an edentulous mandible supporting an overdenture over 2 single-standing implants with OR abutments. Axial loads (100 N) were applied on either central (L1) or lateral (L2) regions of the models. Stress distribution was concentrated mostly in the cortical bone surrounding the implants. When comparing the groups, BC (L1, 52.0 MPa and L2, 74.2 MPa) showed lower first principal stress values on supporting tissue than OR (L1, 78.4 MPa and L2, 76.7 MPa). Connected implants with BC attachment were more favorable on stress distribution over peri-implant-supporting tissue for both loading conditions.

Characterization and in vitro evaluation of bacterial cellulose membranes functionalized with osteogenic growth peptide for bone tissue engineering

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Bacterial cellulose-collagen nanocomposite for bone tissue engineering

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

In vitro evaluation of three different biomaterials as scaffolds for canine mesenchymal stem cells

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Tissue-engineered blood vessel substitute by reconstruction of endothelium using mesenchymal stem cells induced by platelet growth factors

Background: Cardiovascular diseases remain leaders as the major causes of mortality in Western society. Restoration of the circulation through construction of bypass surgical treatment is regarded as the gold standard treatment of peripheral vascular diseases, and grafts are necessary for this purpose. The great saphenous vein is often not available and synthetic grafts have their limitations. Therefore, new techniques to produce alternative grafts have been developed and, in this sense, tissue engineering is a promising alternative to provide biocompatible grafts. This study objective was to reconstruct the endothelium layer of decellularized vein scaffolds, using mesenchymal stem cells (MSCs) and growth factors obtained from platelets. Methods: Fifteen nonpregnant female adult rabbits were used for all experiments. Adipose tissue and vena cava were obtained and subjected to MSCs isolation and tissue decellularization, respectively. MSCs were subjected to differentiation using endothelial inductor growth factor (EIGF) obtained from human platelet lysates. Immunofluorescence, histological and immunohistochemical analyses were employed for the final characterization of the obtained blood vessel substitute. Results: The scaffolds were successfully decellularized with sodium dodecyl sulfate. MSCs actively adhered at the scaffolds, and through stimulation with EIGF were differentiated into functional endothelial cells, secreting significantly higher quantities of von Willebrand factor (0.85 μg/mL; P < .05) than cells cultivated under the same conditions, without EIGF (0.085 μg/mL). Cells with evident morphologic characteristics of endothelium were seen at the lumen of the scaffolds. These cells also stained positive for fascin protein, which is highly expressed by differentiated endothelial cells. Conclusions: Taken together, the use of decellularized bioscaffold and subcutaneous MSCs seems to be a potential approach to obtain bioengineered blood vessels, in the presence of EIGF supplementation. © 2013 Society for Vascular Surgery.

Characterization of mesenchymal stem cells derived from equine adipose tissue

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Bone stress and strain after use of a miniplate for molar protraction and uprighting: a 3-dimensional finite element analysis

The aim of this study was to use the finite element method to evaluate the distribution of stresses and strains on the local bone tissue adjacent to the miniplate used for anchorage of orthodontic forces. Methods: A 3-dimensional model composed of a hemimandible and teeth was constructed using dental computed tomographic images, in which we assembled a miniplate with fixation screws. The uprighting and mesial movements of the mandibular second molar that was anchored with the miniplate were simulated. The miniplate was loaded with horizontal forces of 2, 5, and 15 N. A moment of 11.77 N.mm was also applied. The stress and strain distributions were analyzed, and their correlations with the bone remodeling criteria and miniplate stability were assessed. Results: When orthodontic loads were applied, peak bone strain remained within the range of bone homeostasis (100-1500 mu m strain) with a balance between bone formation and resorption. The maximum deformation was found to be 1035 mu m strain with a force of 5 N. At a force of 15 N, bone resorption was observed in the region of the screws. Conclusions: We observed more stress concentration around the screws than in the cancellous bone. The levels of stress and strain increased when the force was increased but remained within physiologic levels. The anchorage system of miniplate and screws could withstand the orthodontic forces, which did not affect the stability of the miniplate.

Scientific use of the finite element method in Orthodontics

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Effect of crown-to-implant ratio on peri-implant stress: a finite element analysis

The aim of this study was to evaluate stress distribution in the fixation screws and bone tissue around implants in single-implant supported prostheses with crowns of different heights (10,12.5, 15 mm crown-to-implant ratio 1:1, 1.25:1, 1.5:1, respectively). It was designed using three 3-Dmodels. Each model was developed with a mandibular segment of bone block including an internal hexagon implant supporting a screw-retained, single metalceramic crown. The crown height was set at 10, 12.5, and 15 mm with crown-to-implant ratio of 1:1, 1.25:1, 1.5:1, respectively. The applied forces were 200 N (axial) and 100 N (oblique). The increase of crown height showed differences with the oblique load in some situations. By von Mises'criterion, a high stress area was concentrated at the implant/fixation screw and abutment/implant interfaces at crown-to-implant ratio of 1:1, 1.25:1, 1.5:1, respectively. Using the maxiinum principal criteria, the buccal regions showed higher traction stress intensity, whereas the distal regions showed the largest compressive stress in all models. The increase of C/I ratio must be carefully evaluated by the dentist since the increase of this C/I ratio is proportional to the increase of average stress for both screw fixation (C/I 1:1 to 1:1.25 ratio = 30.1% and C/I 1:1 to 1 :1.5 ratio = 46.3%) and bone tissue (C/I 1:1 to 1:1.25 ratio = 30% and C/I 1:1 to 1:1.5 ratio = 51.5%). (C) 2014 Elsevier B.V. All rights reserved.

Straight and angulated abutments in platform switching: influence of loading on bone stress by three-dimensional finite element analysis

PURPOSE: In view of reports in the literature on the benefits achieved with the use of platform switching, described as the use of an implant with a larger diameter than the abutment diameter, the goal being to prevent the (previously) normal bone loss down to the first thread that occurs around most implants, thus enhancing soft tissue aesthetics and stability and the need for implant inclination due to bone anatomy in some cases, the aim of this study was to evaluate bone stress distribution on peri-implant bone, by using three-dimensional finite element analysis to simulate the influence of implants with different abutment angulations (0 and 15 degrees) in platform switching. METHODS: Four mathematical models of an implant-supported central incisor were created with varying abutment angulations: straight abutment (S1 and S2) and angulated abutment at 15 degrees (A1 and A2), submitted to 2 loading conditions (100 N): S1 and A1-oblique loading (45 degrees) and S2 and A2-axial loading, parallel to the long axis of the implant. Maximum (σmax) and minimum (σmin) principal stress values were obtained for cortical and trabecular bone. RESULTS: Models S1 and A1 showed higher σmax in cortical and trabecular bone when compared with S2 and A2. The highest σmax values (in MPa) in the cortical bone were found in S1 (28.5), followed by A1 (25.7), S2 (11.6), and A2 (5.15). For the trabecular bone, the highest σmax values were found in S1 (7.53), followed by A1 (2.87), S2 (2.85), and A2 (1.47). CONCLUSIONS: Implants with straight abutments generated the highest stress values in bone. In addition, this effect was potentiated when the load was applied obliquely.

Characterization and in vitro evaluation of bacterial cellulose membranes functionalized with osteogenic growth peptide for bone tissue engineering

The aim of this study was to characterize the physicochemical properties of bacterial cellulose (BC) membranes functionalized with osteogenic growth peptide (OGP) and its C-terminal pentapeptide OGP[10-14], and to evaluate in vitro osteoinductive potential in early osteogenesis, besides, to evaluate cytotoxic, genotoxic and/or mutagenic effects. Peptide incorporation into the BC membranes did not change the morphology of BC nanofibers and BC crystallinity pattern. The characterization was complemented by Raman scattering, swelling ratio and mechanical tests. In vitro assays demonstrated no cytotoxic, genotoxic or mutagenic effects for any of the studied BC membranes. Culture with osteogenic cells revealed no difference in cell morphology among all the membranes tested. Cell viability/proliferation, total protein content, alkaline phosphatase activity and mineralization assays indicated that BC-OGP membranes enabled the highest development of the osteoblastic phenotype in vitro. In conclusion, the negative results of cytotoxicity, genotoxicity and mutagenicity indicated that all the membranes can be employed for medical supplies, mainly in bone tissue engineering/regeneration, due to their osteoinductive properties.