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1.
Polymers (Basel) ; 15(10)2023 May 16.
Article in English | MEDLINE | ID: mdl-37242899

ABSTRACT

Articular cartilage is a specialized tissue that provides a smooth surface for joint movement and load transmission. Unfortunately, it has limited regenerative capacity. Tissue engineering, combining different cell types, scaffolds, growth factors, and physical stimulation has become an alternative for repairing and regenerating articular cartilage. Dental Follicle Mesenchymal Stem Cells (DFMSCs) are attractive candidates for cartilage tissue engineering because of their ability to differentiate into chondrocytes, on the other hand, the polymers blend like Polycaprolactone (PCL) and Poly Lactic-co-Glycolic Acid (PLGA) have shown promise given their mechanical properties and biocompatibility. In this work, the physicochemical properties of polymer blends were evaluated by Fourier Transform Infrared Spectroscopy (FTIR) and Scanning Electron Microscope (SEM) and were positive for both techniques. The DFMSCs demonstrated stemness by flow cytometry. The scaffold showed to be a non-toxic effect when we evaluated it with Alamar blue, and the samples were analyzed using SEM and phalloidin staining to evaluate cell adhesion to the scaffold. The synthesis of glycosaminoglycans was positive on the construct in vitro. Finally, the PCL/PLGA scaffold showed a better repair capacity than two commercial compounds, when tested in a chondral defect rat model. These results suggest that the PCL/PLGA (80:20) scaffold may be suitable for applications in the tissue engineering of articular hyaline cartilage.

2.
J Biomater Appl ; 37(10): 1776-1788, 2023 05.
Article in English | MEDLINE | ID: mdl-37025081

ABSTRACT

Today, regenerative osteogenesis represents a clinical need, due to the incidence of bone defects that involve groups of pathologies ranging from congenital anomalies to traumatic injuries, as well as problems presented surgically. This is why the design of a polymeric biomaterial (scaffold) of chitosan, carboxymethylcellulose, zinc oxide, and calcium carbonate with similar characteristics in terms of composition and bone structure offers high potential to help address this health problem. The technique for obtaining the scaffolds of this research was to develop a physical hydrogel to have the biofunctionality of the active groups of the polymer chains used, then make use of the lyophilization process to obtain three-dimensional (3D) porous scaffolds. The physicochemical and biological properties of the scaffolds were evaluated. The scaffolds presented morphology with pore size and interconnectivity that favor the need for cell proliferation and viability. The biocompatibility tests confirm that the designed scaffolds do not present cytotoxicity and the analyzes with alizarin red staining show calcium deposits in the materials with CaCO3 and ZnO. Osteoinduction assays to osteogenic lineage using runt-related transcription factor type 2 (RUNX2) and collagen type 1 (COL-1) antibodies allowed expression in differentiated cells. Therefore, the calcium carbonate-containing scaffolds stabilized by physical bonds have characteristics of being non-cytotoxic, bioactive, and osteoinductive, which motivate their use in future tests to evaluate their demeanor with rat models for bone engineering studies.


Subject(s)
Chitosan , Zinc Oxide , Rats , Animals , Tissue Engineering/methods , Chitosan/chemistry , Tissue Scaffolds/chemistry , Carboxymethylcellulose Sodium , Calcium Carbonate , Osteogenesis , Cell Differentiation , Cell Proliferation
3.
Article in English | MEDLINE | ID: mdl-36497984

ABSTRACT

Jarosite-type compounds precipitated in the zinc industry for iron control can also incorporate arsenic and can be used for wastewater treatment for As elimination. According with the last, this work is related to arsenic incorporation at room temperature in decomposed potassium jarosite. The work began with the synthesis of the compound at 75 °C for 9 h using Fe2(SO4)3 and K2SO4 at a pH of 1.1. Once jarosite was obtained, solids were subjected to an alkaline decomposition using NaOH at pH 10 for 30 min, and then As was added to the solution as HAsNaO4 and the pH modified by adding HNO3 until it reached a value of 1.1. The initial, intermediate, and final products were wholly characterized by scanning electron microscopy (SEM) in conjunction with energy dispersive spectrometry (EDS), X-ray diffraction (XRD), Fourier transform infrared spectroscopy (FTIR), Raman spectroscopy (RS), and X-ray photoelectron spectrometry (XPS). The obtained results show that As(V) can be adsorbed by ionic exchange in the amorphous FeOH structure of decomposed jarosite and when pH decreased to 1.1, the compound recrystallized, incorporating up to 6% As on average, which is indicative that this process can be used to reduce As in contaminated waters.


Subject(s)
Arsenic , Water Pollutants, Chemical , Potassium , Water Pollutants, Chemical/analysis , Ferric Compounds/chemistry , Adsorption , Arsenic/analysis , Water , Spectroscopy, Fourier Transform Infrared
4.
Polymers (Basel) ; 14(3)2022 Jan 28.
Article in English | MEDLINE | ID: mdl-35160534

ABSTRACT

Cleft palate (CP) is one of the most common birth defects, presenting a multitude of negative impacts on the health of the patient. It also leads to increased mortality at all stages of life, economic costs and psychosocial effects. The embryological development of CP has been outlined thanks to the advances made in recent years due to biomolecular successions. The etiology is broad and combines certain environmental and genetic factors. Currently, all surgical interventions work off the principle of restoring the area of the fissure and aesthetics of the patient, making use of bone substitutes. These can involve biological products, such as a demineralized bone matrix, as well as natural-synthetic polymers, and can be supplemented with nutrients or growth factors. For this reason, the following review analyzes different biomaterials in which nutrients or biomolecules have been added to improve the bioactive properties of the tissue construct to regenerate new bone, taking into account the greatest limitations of this approach, which are its use for bone substitutes for large areas exclusively and the lack of vascularity. Bone tissue engineering is a promising field, since it favors the development of porous synthetic substitutes with the ability to promote rapid and extensive vascularization within their structures for the regeneration of the CP area.

5.
Medicine (Baltimore) ; 100(25): e26435, 2021 06 25.
Article in English | MEDLINE | ID: mdl-34160434

ABSTRACT

ABSTRACT: The aim of this study was to evaluate the association between sociodemographic factors and noncavitated and cavitated caries lesions in Mexican schoolchildren.This cross-sectional study was conducted in 2020 on 8-to-12-year-old schoolchildren of different socioeconomic status (SES). The caries was evaluated using ICDAS II, SES was evaluated using three categories---a high, middle, or low-income level---of the CONAPO. Multinomial logistic regression analyses were performed in order to ascertain the associations between socioeconomic factors and noncavitated and cavitated caries lesions.The prevalence of noncavitated lesions was 38.0% and cavitated lesions was 43.4% in permanent dentition. In all the samples, 50.6% of schoolchildren had poor oral hygiene. About 52.5% of the mothers and 64.7% of the fathers had less than 9 years of education. Schoolchildren with a low-income level have more cavitated lesions (ICDAS II 4-6) than schoolchildren with high-income level (56.3% vs 15.8%, P = .009). The multinomial logistic regression models showed that mother's level of education <9 years and low-income level were significantly associated with cavitated caries lesions (ICDAS II 4-6), [odds ratio = 1.79 (1.17 - 2.75); P = .007], [OR = 2.21 (1.23 - 3.97); P = .008], respectively. The socioeconomic level was not associated with noncavitated caries lesions (ICDAS II 1-3).An association was found between the presence of cavitated caries lesions and the subject's mother's level of education and a low-income level. Socioeconomic factors were found to be associated with inequalities in caries distribution in the age group studied.


Subject(s)
Dental Caries/epidemiology , Dental Pulp Exposure/epidemiology , Child , Cross-Sectional Studies , Dental Caries/complications , Dental Caries/diagnosis , Dental Pulp Exposure/diagnosis , Dental Pulp Exposure/etiology , Educational Status , Female , Humans , Income , Male , Mexico/epidemiology , Mothers/education , Mothers/statistics & numerical data , Prevalence , Risk Assessment/statistics & numerical data , Risk Factors , Social Class
6.
Polymers (Basel) ; 13(7)2021 Apr 04.
Article in English | MEDLINE | ID: mdl-33916576

ABSTRACT

The search of suitable combinations of stem cells, biomaterials and scaffolds manufacturing methods have become a major focus of research for bone engineering. The aim of this study was to test the potential of dental pulp stem cells to attach, proliferate, mineralize and differentiate on 3D printed polycaprolactone (PCL) scaffolds. A 100% pure Mw: 84,500 ± 1000 PCL was selected. 5 × 10 × 5 mm3 parallelepiped scaffolds were designed as a wood-pilled structure composed of 20 layers of 250 µm in height, in a non-alternate order ([0,0,0,90,90,90°]). 3D printing was made at 170 °C. Swine dental pulp stem cells (DPSCs) were extracted from lower lateral incisors of swine and cultivated until the cells reached 80% confluence. The third passage was used for seeding on the scaffolds. Phenotype of cells was determined by flow Cytometry. Live and dead, Alamar blue™, von Kossa and alizarin red staining assays were performed. Scaffolds with 290 + 30 µm strand diameter, 938 ± 80 µm pores in the axial direction and 689 ± 13 µm pores in the lateral direction were manufactured. Together, cell viability tests, von Kossa and Alizarin red staining indicate the ability of the printed scaffolds to support DPSCs attachment, proliferation and enable differentiation followed by mineralization. The selected material-processing technique-cell line (PCL-3D printing-DPSCs) triplet can be though to be used for further modelling and preclinical experiments in bone engineering studies.

7.
J Mech Behav Biomed Mater ; 119: 104510, 2021 07.
Article in English | MEDLINE | ID: mdl-33872923

ABSTRACT

The present study elucidates the impact of detergent-based chemical decellularization on the micro-mechanical properties of porcine and rabbit corneas for the purpose of extracellular matrix (ECM) derived scaffolds. Aiming to optimize the decellularization process, different concentrations of Sodium Dodecyl Sulfate (SDS), Triton X-100 and CHAPS detergents were assessed on their ability to decellularize corneas from both bio-models at incubation periods of 12 and 24h. We evaluated the effect of decellularization on corneal ECM Young's Modulus and various area's roughness parameters (topography features) at a microscale by using Atomic Force Microscopy (AFM). Only SDS presented adequate decellularization properties at the selected concentrations (0.2, 0.5 and 1%) and incubation periods. All topography features displayed by native corneas were preserved after SDS treatments, while no statistically significant differences were identified for the average value of Young's Modulus between the control samples and those treated with 0.2% SDS (rabbit corneas) and 0.5% SDS (porcine corneas) after 12h. In this sense, cornea decellularization procedures can be improved by simultaneously reducing SDS concentration and incubation period. AFM is a useful tool to perform biomechanical analysis of the effect of decellularization on scaffold micro-mechanics. Evaluation of the scaffold mechanical behavior at a microscale could help in understanding cell-scaffold interactions in terms of mechanotransduction, complementing macroscale techniques (e.g. tensile tests) relevant for tissue engineering quality control and decision-making.


Subject(s)
Mechanotransduction, Cellular , Tissue Scaffolds , Animals , Cornea , Extracellular Matrix , Rabbits , Swine , Tissue Engineering
8.
Materials (Basel) ; 14(1)2021 Jan 05.
Article in English | MEDLINE | ID: mdl-33466418

ABSTRACT

This work shows the synthesis of a polyvinylpyrrolidone (PVP) hydrogel by heat-activated polymerization and explores the production of hydrogels with an open porous network by lyophilisation to allow the three-dimensional culture of human oral mucosa stem cells (hOMSCs). The swollen hydrogel showed a storage modulus similar to oral mucosa and elastic solid rheological behaviour without sol transition. A comprehensive characterization of porosity by scanning electron microscopy, mercury intrusion porosimetry and nano-computed tomography (with spatial resolution below 1 µm) showed that lyophilisation resulted in the heterogeneous incorporation of closed oval-like pores in the hydrogel with broad size distribution (5 to 180 µm, d50 = 65 µm). Human oral mucosa biopsies were used to isolate hOMSCs, expressing typical markers of mesenchymal stem cells in more than 95% of the cell population. Direct contact cytotoxicity assay demonstrated that PVP hydrogel have no negative effect on cell metabolic activity, allowing the culture of hOMSCs with normal fusiform morphology. Pore connectivity should be improved in future to allow cell growth in the bulk of the PVP hydrogel.

9.
Article in English | MEDLINE | ID: mdl-33153121

ABSTRACT

This cross-sectional study aimed to establish a relationship between the Normative Need for Orthodontic Treatment (NNOT) and oral health among Mexican adolescents aged 13-15 years old. A convenience sample of 424 subjects in Mexico City participated in the study. The dependent variable used was NNOT, which was determined via the dental health component (grades 4 and 5) of the Index of Orthodontic Treatment Need (IOTN). The variables for oral health were as follows: caries experience, oral hygiene, self-reported temporomandibular joint pain, and self-reported bruxism. Logistic regression models were fitted to determine the association between NNOT and oral health. The prevalence of NNOT was 66.0% (280/424), and the crowding was the most prevalent occlusal anomaly with 36.1% (n = 135). Multivariate models showed that subjects with NNOT were more than twice as likely to present poor hygiene (OR = 2.56; p = 0.001) as subjects presenting crowding (>4 mm) (OR = 1.99; p = 0.004) and increased overjet (>6 mm) (OR = 1.74; p = 0.046). Those schoolchildren who presented anterior guidance were 72% less likely to present NNOT (OR = 0.28; p < 0.001). In conclusion, the risk of presenting NNOT in Mexican adolescents is high, with a prevalence of over 50% of which the most prevalent occlusal anomaly was crowding. On the other hand, poor oral hygiene was associated with crowding and increased overjet.


Subject(s)
Malocclusion , Oral Health , Adolescent , Cross-Sectional Studies , Female , Humans , Index of Orthodontic Treatment Need , Male , Malocclusion/epidemiology , Mexico/epidemiology , Orthodontics
10.
J Biomater Appl ; 35(4-5): 485-499, 2020.
Article in English | MEDLINE | ID: mdl-32659135

ABSTRACT

Polyglycerol sebacate (PGS) scaffolds obtained using a leaching technique were modified with iodine-doped polypyrrole (PPy-I) in a plasma reactor in order to study the effect of exposure time on the cell viability of hDPSCs. SEM analysis showed the formation and growth of PPy-I particles as the exposure time was increased, while FTIR and XPS analysis revealed the presence of -NH- and N+ groups in the chemical composition of the surfaces, relating to the increase in the amount of PPY-I particles. The water contact angle measurements showed an increase in the scaffold's hydrophilicity with greater exposure times which was also attributed to the rising of PPy-I particles. It was also observed that PPy-I promotes the rigidity of the treated PGS scaffolds. when in direct contact with treated PGS scaffolds, cell viability improved with respect to non-treated scaffolds, however only at shorter time exposures. Extracts of plasma-treated PGS scaffolds showed high cytotoxicity as the time exposure to plasma treatment was increased.


Subject(s)
Biocompatible Materials/chemistry , Decanoates/chemistry , Glycerol/analogs & derivatives , Iodine/chemistry , Plasma Gases/chemistry , Polymers/chemistry , Pyrroles/chemistry , Tissue Scaffolds/chemistry , Biocompatible Materials/metabolism , Cell Proliferation , Cell Survival , Cross-Linking Reagents/chemistry , Dental Pulp/cytology , Glycerol/chemistry , Humans , Mechanical Tests , Stem Cells/cytology , Surface Properties , Time Factors , Tissue Engineering
12.
J Biomater Sci Polym Ed ; 30(15): 1415-1432, 2019 10.
Article in English | MEDLINE | ID: mdl-31233380

ABSTRACT

Polyurethanes (PU) foams with titanium particles (Ti) were prepared with castor oil (CO) and isophorone diisocyanate (IPDI) as polymeric matrix, and 1, 3 and 5 wt.% of Ti. Composites were physicochemically and mechanically characterized and their biocompatibility assessed using human dental pulp stem cells (HDPSC). PU synthesis was confirmed by FTIR, but the presence of Ti was detected by RAMAN, X-ray diffraction (peak at 2θ = 40.2°) and by EDX-mapping. Materials showed three decomposition temperatures between 300 °C and 500 °C and their decomposition were not catalyzed by Ti particles. Compressive modulus (164-846 kPa), compressive strength (12.9-116.7 kPa) and density (128-240 kg/m3) tend to increase with Ti concentration but porosity was reduced (87% to 80%). Composites' foams were fully degraded in acid and oxidative media while remained stable in distilled water. HDPSC viability on all composites was higher than 80% up to 14 days while proliferation dropped up to 60% at 21 days. Overall, these results suggest that these foams can be used as scaffolds for bone tissue regeneration.


Subject(s)
Bone and Bones/cytology , Castor Oil/chemistry , Polyurethanes/chemistry , Polyurethanes/pharmacology , Tissue Engineering , Titanium/chemistry , Biocompatible Materials/chemistry , Biocompatible Materials/pharmacology , Cell Proliferation/drug effects , Chemical Phenomena , Dental Pulp/cytology , Humans , Mechanical Phenomena , Stem Cells/cytology , Stem Cells/drug effects , Temperature , Tissue Scaffolds/chemistry
13.
Int J Mol Sci ; 20(9)2019 May 02.
Article in English | MEDLINE | ID: mdl-31052503

ABSTRACT

Craniofacial bone defect anomalies affect both soft and hard tissues and can be caused by trauma, bone recessions from tumors and cysts, or even from congenital disorders. On this note, cleft/lip palate is the most prevalent congenital craniofacial defect caused by disturbed embryonic development of soft and hard tissues around the oral cavity and face area, resulting in most cases, of severe limitations with chewing, swallowing, and talking as well as problems of insufficient space for teeth, proper breathing, and self-esteem problems as a consequence of facial appearance. Spectacular advances in regenerative medicine have arrived, giving new hope to patients that can benefit from new tissue engineering therapies based on the supportive action of 3D biomaterials together with the synergic action of osteo-inductive molecules and recruited stem cells that can be driven to the process of bone regeneration. However, few studies have focused on the application of tissue engineering to the regeneration of the cleft/lip and only a few have reported significant advances to offer real clinical solutions. This review provides an updated and deep analysis of the studies that have reported on the use of advanced biomaterials and cell therapies for the regeneration of cleft lip and palate regeneration.


Subject(s)
Biocompatible Materials/therapeutic use , Cleft Lip/therapy , Cleft Palate/therapy , Regenerative Medicine/methods , Animals , Cleft Lip/epidemiology , Cleft Lip/pathology , Cleft Lip/physiopathology , Cleft Palate/epidemiology , Cleft Palate/pathology , Cleft Palate/physiopathology , Folic Acid/analogs & derivatives , Folic Acid/therapeutic use , Humans , Intercellular Signaling Peptides and Proteins/therapeutic use , Osteogenesis/drug effects , Stem Cell Transplantation/methods , Tissue Engineering/methods
14.
Molecules ; 24(9)2019 Apr 27.
Article in English | MEDLINE | ID: mdl-31035627

ABSTRACT

Nowadays, regenerative medicine has paid special attention to research (in vitro and in vivo) related to bone regeneration, specifically in the treatment of bone fractures or skeletal defects, which is rising worldwide and is continually demanding new developments in the use of stem cells, growth factors, membranes and scaffolds based on novel nanomaterials, and their applications in patients by using advanced tools from molecular biology and tissue engineering. Strontium (Sr) is an element that has been investigated in recent years for its participation in the process of remodeling and bone formation. Based on these antecedents, this is a review about the Strontium Folate (SrFO), a recently developed non-protein based bone-promoting agent with interest in medical and pharmaceutical fields due to its improved features in comparison to current therapies for bone diseases.


Subject(s)
Bone Regeneration , Folic Acid/metabolism , Strontium/metabolism , Tissue Scaffolds , Animals , Biocompatible Materials/chemical synthesis , Biocompatible Materials/chemistry , Bone and Bones/drug effects , Bone and Bones/metabolism , Dental Pulp/cytology , Folic Acid/chemistry , Humans , Osteogenesis/drug effects , Osteogenesis/physiology , Stem Cells/cytology , Stem Cells/drug effects , Stem Cells/metabolism , Strontium/chemistry , Tissue Engineering , Tissue Scaffolds/chemistry , Vitamin B 12/chemistry , Vitamin B 12/metabolism , Vitamin B 6/chemistry , Vitamin B 6/metabolism
15.
J Biomater Appl ; 33(1): 11-22, 2018 07.
Article in English | MEDLINE | ID: mdl-29726734

ABSTRACT

Segmented polyurethanes were prepared with polycaprolactone diol as soft segment and 4,4-methylene-bis cyclohexyl diisocyanate and l-glutamine as the rigid segment. These polyurethanes were filled with 1 wt.% to 5 wt.% titanium particles (Ti), physicochemically characterized and their biocompatibility assessed using human dental pulp stem cells and mice osteoblasts. Physicochemical characterization showed that composites retained the properties of the semicrystalline polyurethane as they exhibited a glass transition temperature (Tg) between -35°C and -45°C, melting temperature (Tm) at 52°C and crystallinity close to 40% as determined by differential scanning calorimetry. In agreement with this, X-ray diffraction showed reflections at 21.3° and 23.6° for polycaprolactone diol and reflections at 35.1°, 38.4°, and 40.2° for Ti particles suggesting that these particles are not acting as nucleating sites. The addition of up to 5 wt.% of Ti reduced both, tensile strength and maximum strain from 1.9 MPa to 1.2 MPa, and from 670% to 172% for pristine and filled polyurethane, respectively. Although there were differences between composites at low strain rates, no significant differences in mechanical behavior were observed at higher strain rate where a tensile stress of 8.5 MPa and strain of 223% were observed for 5 wt.% composites. The addition to titanium particles had a beneficial effect on both human dental pulp stem cells and osteoblasts viability, as it increased with the amount of titanium in composites up to 10 days of incubation.


Subject(s)
Biocompatible Materials/chemistry , Bone and Bones/chemistry , Polyurethanes/chemistry , Tissue Scaffolds/chemistry , Titanium/chemistry , Adolescent , Animals , Cell Line , Cell Proliferation , Cell Survival , Dental Pulp/cytology , Female , Humans , Mice , Osteoblasts/cytology , Phase Transition , Stem Cells/cytology , Stress, Mechanical , Temperature , Tensile Strength , Tissue Engineering , Young Adult
16.
Polymers (Basel) ; 10(3)2018 Mar 07.
Article in English | MEDLINE | ID: mdl-30966314

ABSTRACT

In craniofacial tissue regeneration, the current gold standard treatment is autologous bone grafting, however, it presents some disadvantages. Although new alternatives have emerged there is still an urgent demand of biodegradable scaffolds to act as extracellular matrix in the regeneration process. A potentially useful element in bone regeneration is strontium. It is known to promote stimulation of osteoblasts while inhibiting osteoclasts resorption, leading to neoformed bone. The present paper reports the preparation and characterization of strontium (Sr) containing hybrid scaffolds formed by a matrix of ionically cross-linked chitosan and microparticles of poly(ε-caprolactone) (PCL). These scaffolds of relatively facile fabrication were seeded with osteoblast-like cells (MG-63) and human bone marrow mesenchymal stem cells (hBMSCs) for application in craniofacial tissue regeneration. Membrane scaffolds were prepared using chitosan:PCL ratios of 1:2 and 1:1 and 5 wt % Sr salts. Characterization was performed addressing physico-chemical properties, swelling behavior, in vitro biological performance and in vivo biocompatibility. Overall, the composition, microstructure and swelling degree (≈245%) of scaffolds combine with the adequate dimensional stability, lack of toxicity, osteogenic activity in MG-63 cells and hBMSCs, along with the in vivo biocompatibility in rats allow considering this system as a promising biomaterial for the treatment of craniofacial tissue regeneration.

17.
J Appl Oral Sci ; 25(2): 186-195, 2017.
Article in English | MEDLINE | ID: mdl-28403359

ABSTRACT

OBJECTIVE: The aim of this study was to obtain autologous dermal-epidermal skin substitutes from oral mucosa from diabetic subjects as a first step towards a possible clinical application for cases of diabetic foot. MATERIAL AND METHODS: Oral mucosa was obtained from diabetic and healthy subjects (n=20 per group). Epidermal cells were isolated and cultured using autologous fibrin to develop dermal-epidermal in vitro substitutes by the air-liquid technique with autologous human serum as a supplement media. Substitutes were immunocharacterized with collagen IV and cytokeratin 5-14 as specific markers. A Student´s t- test was performed to assess the differences between both groups. RESULTS: It was possible to isolate epidermal cells from the oral mucosa of diabetic and healthy subjects and develop autologous dermal-epidermal skin substitutes using autologous serum as a supplement. Differences in the expression of specific markers were observed and the cytokeratin 5-14 expression was lower in the diabetic substitutes, and the collagen IV expression was higher in the diabetic substitutes when compared with the healthy group, showing a significant difference. CONCLUSION: Cells from oral mucosa could be an alternative and less invasive source for skin substitutes and wound healing. A difference in collagen production of diabetic cells suggests diabetic substitutes could improve diabetic wound healing. More research is needed to determine the crosstalk between components of these skin substitutes and damaged tissues.


Subject(s)
Cell Transplantation/methods , Diabetes Mellitus, Type 2 , Epidermal Cells , Epithelial Cells/transplantation , Mouth Mucosa/cytology , Skin, Artificial , Adult , Aged , Biocompatible Materials , Case-Control Studies , Cell Culture Techniques , Cell Proliferation , Cells, Cultured , Collagen/analysis , Diabetes Mellitus, Type 2/therapy , Female , Fibroblasts , Humans , Keratinocytes/cytology , Male , Middle Aged , Reproducibility of Results , Skin Ulcer/therapy , Time Factors , Transplantation, Autologous , Wound Healing
18.
J. appl. oral sci ; 25(2): 186-195, Mar.-Apr. 2017. tab, graf
Article in English | LILACS, BBO - Dentistry | ID: biblio-841186

ABSTRACT

Abstract Oral mucosa has been highlighted as a suitable source of epidermal cells due to its intrinsic characteristics such as its higher proliferation rate and its obtainability. Diabetic ulcers have a worldwide prevalence that is variable (1%-11%), meanwhile treatment of this has been proven ineffective. Tissue-engineered skin plays an important role in wound care focusing on strategies such autologous dermal-epidermal substitutes. Objective The aim of this study was to obtain autologous dermal-epidermal skin substitutes from oral mucosa from diabetic subjects as a first step towards a possible clinical application for cases of diabetic foot. Material and Methods Oral mucosa was obtained from diabetic and healthy subjects (n=20 per group). Epidermal cells were isolated and cultured using autologous fibrin to develop dermal-epidermal in vitro substitutes by the air-liquid technique with autologous human serum as a supplement media. Substitutes were immunocharacterized with collagen IV and cytokeratin 5-14 as specific markers. A Student´s t- test was performed to assess the differences between both groups. Results It was possible to isolate epidermal cells from the oral mucosa of diabetic and healthy subjects and develop autologous dermal-epidermal skin substitutes using autologous serum as a supplement. Differences in the expression of specific markers were observed and the cytokeratin 5-14 expression was lower in the diabetic substitutes, and the collagen IV expression was higher in the diabetic substitutes when compared with the healthy group, showing a significant difference. Conclusion Cells from oral mucosa could be an alternative and less invasive source for skin substitutes and wound healing. A difference in collagen production of diabetic cells suggests diabetic substitutes could improve diabetic wound healing. More research is needed to determine the crosstalk between components of these skin substitutes and damaged tissues.


Subject(s)
Humans , Male , Female , Adult , Middle Aged , Aged , Skin, Artificial , Cell Transplantation/methods , Diabetes Mellitus, Type 2 , Epidermis/cytology , Epithelial Cells/transplantation , Mouth Mucosa/cytology , Skin Ulcer/therapy , Time Factors , Transplantation, Autologous , Wound Healing , Biocompatible Materials , Case-Control Studies , Keratinocytes/cytology , Cells, Cultured , Reproducibility of Results , Collagen/analysis , Cell Culture Techniques , Cell Proliferation , Diabetes Mellitus, Type 2/therapy , Fibroblasts
19.
J Nanobiotechnology ; 15(1): 10, 2017 Jan 31.
Article in English | MEDLINE | ID: mdl-28143540

ABSTRACT

BACKGROUND: Neovascularization over dental implants is an imperative requisite to achieve successful osseointegration onto implanted materials. The aim of this study was to investigate the effects on in vitro angiogenesis of anodized 70 nm diameter TiO2 nanotubes (NTs) on Ti6Al4V alloy synthesized and disinfected by means of a novel, facile, antibacterial and cost-effective method using super oxidized water (SOW). We also evaluated the role of the surface roughness and chemical composition of materials of materials on angiogenesis. METHODS: The Ti6Al4V alloy and a commercially pure Ti were anodized using a solution constituted by SOW and fluoride as electrolyte. An acid-etched Ti6Al4V was evaluated to compare the effect of micro-surface roughness. Mirror-polished materials were used as control. Morphology, roughness, chemistry and wettability were assessed by field emission scanning electron microscopy (FE-SEM), transmission electron microscopy, atomic force microscopy, energy dispersive X-ray spectroscopy (EDX) and using a professional digital camera. Bovine coronary artery endothelial cells (BCAECs) were seeded over the experimental surfaces for several incubation times. Cellular adhesion, proliferation and monolayer formation were evaluated by means of SEM. BCAEC viability, actin stress fibers and vinculin cellular organization, as well as the angiogenic receptors vascular endothelial growth factor 2 (VEGFR2) and endothelial nitric oxide synthase (eNOS) were measured using fluorescence microscopy. RESULTS: The anodization process significantly increased the roughness, wettability and thickness of the oxidized coating. EDX analysis demonstrated an increased oxygen (O) and decreased carbon (C) content on the NTs of both materials. Endothelial behavior was solidly supported and improved by the NTs (without significant differences between Ti and alloy), showing that endothelial viability, adhesion, proliferation, actin arrangement with vinculin expression and monolayer development were evidently stimulated on the nanostructured surface, also leading to increased activation of VEGFR2 and eNOS on Ti6Al4V-NTs compared to the control Ti6Al4V alloy. Although the rougher alloy promoted BCAECs viability and proliferation, filopodia formation was poor. CONCLUSION: The in vitro results suggest that 70 nm diameter NTs manufactured by anodization and cleaned using SOW promotes in vitro endothelial activity, which may improve in vivo angiogenesis supporting a faster clinical osseointegration process.


Subject(s)
Angiogenesis Inducing Agents/pharmacology , Nanotubes/chemistry , Neovascularization, Physiologic/drug effects , Titanium/chemistry , Alloys , Animals , Cattle , Cell Adhesion , Cell Proliferation , Cells, Cultured , Coronary Vessels/cytology , Dental Implants , Endothelial Cells/drug effects , Microscopy, Atomic Force , Microscopy, Electron, Scanning , Nitric Oxide Synthase Type III/metabolism , Particle Size , Spectrometry, X-Ray Emission , Surface Properties , Vascular Endothelial Growth Factor Receptor-2/metabolism , Wettability
20.
J Biomed Mater Res B Appl Biomater ; 105(4): 785-794, 2017 05.
Article in English | MEDLINE | ID: mdl-26750608

ABSTRACT

Fiber scaffolds are attractive materials for mimicking, within a 3D in vitro system, any living environment in which animal cells can adhere and proliferate. In three dimensions, cells have the ability to communicate and organize into complex architectures similar to those found in their natural environments. The aim of this study was to evaluate, in terms of cell reactivity, a new in vitro cell model: dental pulp stem cells (DPSCs) in a 3D polymeric textile. Scaffolds were knitted from polyglycolic acid (PGA) or polydioxanone (PDO) fibers differing in surface roughness. To promote cell adhesion, these hydrophobic fabrics were also functionalized with either chitosan or the peptide arginine-glycine-aspartic acid (RGD). Cell behavior was examined 1, 10, and 21 days post-seeding with a LIVE/DEAD® Kit. Confocal laser scanning microscopy (CLSM) highlighted the biocompatibility of these materials (cell survival rate: 94% to 100%). Fiber roughness was found to influence cell adhesion and viability significantly and favorably. A clear benefit of polymeric textile functionalization with chitosan or RGD was demonstrated in terms of cell adhesion and viability. © 2016 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 105B: 785-794, 2017.


Subject(s)
Dental Pulp/metabolism , Polydioxanone/chemistry , Polyglycolic Acid/chemistry , Stem Cells/metabolism , Textiles , Tissue Scaffolds/chemistry , Adolescent , Adult , Cell Adhesion , Cell Survival , Chitosan/chemistry , Dental Pulp/cytology , Female , Humans , Hydrophobic and Hydrophilic Interactions , Male , Oligopeptides/chemistry , Stem Cells/cytology
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