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1.
Semin Thromb Hemost ; 50(2): 236-252, 2024 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-37611623

RESUMO

There is a lack of evidence-based reviews on the effects of laser irradiation on blood coagulation in the literature, despite a large number of clinical trials. We therefore evaluated the available evidence on laser irradiation parameters used in coagulation of blood to optimize physical parameters. We performed a literature search for recent scientific studies indexed between 2017 and 2023 using the databases of PubMed and ScienceDirect. Articles were selected based on defined inclusion and exclusion criteria, and 78 publications in total were eventually included in this scoping review. The following were found to produce significant benefits in blood coagulation for surgical application: (1) dentistry and oral surgeries: 980 nm, 27 s, 2 W, 1502.7 W/cm2, 26.5 J, 622 J/cm2, 400 µm; (2) urogenital disorders: 532 nm, 4 s, 40 W, 10600 W/cm2, 1.3 J, 424 J/cm2, 600 µm; (3) ophthalmic disorders: 810 nm, 1 s, 1 W, 3540 W/cm2, 0.75 J, 1326 J/cm2, 100 µm; (4) embryological surgeries: 1064 nm, 10 s, 25 W, 35400 W/cm2, 262.5 J, 371000 J/cm2, 332.5 µm; (5) dermatological disorders: 1064 nm, 20 W, 2440 W/cm2, 0.1 J, 24 J/cm2, 670 µm; (6) gastrointestinal disorders: 532 nm, 3 s, 20 W, 1051 W/cm2, 120 J, 26500 J/cm2, 760 µm; (7) neurological surgeries: 2.5 s, 1.5 W, 1035 W/cm2, 2 J, 1584 J/cm2, 385 µm; (8) pulmonary disorders: 1320 nm, 5s, 35 W, 12450 W/cm2, 250 J, 65000 J/cm2, 700 µm (9) cardiovascular disorders: 1064 nm, 16.5 s, 5 W, 1980.5 W/cm2, 900 J, 760 J/cm2, 400 µm. In conclusion, our scoping review identifies that combining data from all clinically heterogeneous studies suggests that laser irradiation reflects an effective method for inducing blood coagulation in several medical fields.


Assuntos
Terapia a Laser , Humanos , Terapia a Laser/métodos , Lasers , Coagulação Sanguínea
2.
J Mater Sci Mater Med ; 34(4): 15, 2023 Apr 01.
Artigo em Inglês | MEDLINE | ID: mdl-37004591

RESUMO

Based on the concept of tissue engineering (Cells-Scaffold-Bioactive molecules), regenerative endodontics appeared as a new notion for dental endodontic treatment. Its approaches aim to preserve dental pulp vitality (pulp capping) or to regenerate a vascularized pulp-like tissue inside necrotic root canals by cell homing. To improve the methods of tissue engineering for pulp regeneration, numerous studies using in vitro, ex vivo, and in vivo models have been performed. This review explores the evolution of laboratory models used in such studies and classifies them according to different criteria. It starts from the initial two-dimensional in vitro models that allowed characterization of stem cell behavior, through 3D culture matrices combined with dental tissue and finally arrives at the more challenging ex vivo and in vivo models. The travel which follows the elaboration of such models reveals the difficulty in establishing reproducible laboratory models for dental pulp regeneration. The development of well-established protocols and new laboratory ex vivo and in vivo models in the field of pulp regeneration would lead to consistent results, reduction of animal experimentation, and facilitation of the translation to clinical practice.


Assuntos
Polpa Dentária , Regeneração , Animais , Polpa Dentária/fisiologia , Células-Tronco , Engenharia Tecidual/métodos , Alternativas aos Testes com Animais/métodos
3.
Int J Mol Sci ; 24(16)2023 Aug 09.
Artigo em Inglês | MEDLINE | ID: mdl-37628774

RESUMO

Mesenchymal stem cell secretome or conditioned medium (MSC-CM) is a combination of biomolecules and growth factors in cell culture growth medium, secreted by mesenchymal stem cells (MSCs), and the starting point of several derived products. MSC-CM and its derivatives could be applied after injuries and could mediate most of the beneficial regenerative effects of MSCs without the possible side effects of using MSCs themselves. However, before the clinical application of these promising biopharmaceuticals, several issues such as manufacturing protocols and quality control must be addressed. This review aims to underline the influence of the procedure for conditioned medium production on the quality of the secretome and its derivatives and highlights the questions considering cell sources and donors, cell expansion, cell passage number and confluency, conditioning period, cell culture medium, microenvironment cues, and secretome-derived product purification. A high degree of variability in MSC secretomes is revealed based on these parameters, confirming the need to standardize and optimize protocols. Understanding how bioprocessing and manufacturing conditions interact to determine the quantity, quality, and profile of MSC-CM is essential to the development of good manufacturing practice (GMP)-compliant procedures suitable for replacing mesenchymal stem cells in regenerative medicine.


Assuntos
Células-Tronco Mesenquimais , Secretoma , Meios de Cultivo Condicionados/farmacologia , Controle de Qualidade , Padrões de Referência
4.
Lasers Med Sci ; 36(2): 437-445, 2021 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-32621128

RESUMO

Photobiomodulation (PBM) has been shown to improve cell proliferation and cell migration. Many cell types have been investigated, with most studies using deep penetrating red light irradiation. Considering the interest of surface biostimulation of oral mesenchymal cells after surgical wound, the present study aimed to assess green light irradiation effects on Dental Pulp Stem Cells' (DPSC) proliferation and migration. To understand the mechanisms underlying these effects, we investigated cytoskeleton organization and subsequent cell shape and stiffness. A 532-nm wavelength Nd:YAG laser (30 mW) was applied between 30 and 600 s on DPSC in vitro. Cell proliferation was analyzed at 24, 48, and 72 h after irradiation, by cell counting and enzymatic activity quantification (paranitrophenylphosphate phosphatase (pNPP) test). A wound healing assay was used to study cell migration after irradiation. Effects of PBM on cytoskeleton organization and cell shape were assessed by actin filaments staining. Elasticity changes after irradiation were quantified in terms of Young's modulus measured using Atomic Force Microscopy (AFM) force spectroscopy. Green light significantly improved DPSC proliferation with a maximal effect obtained after 300-s irradiation (energy fluence 5 J/cm2). This irradiation had a significant impact on cell migration, improving wound healing after 24 h. These results were concomitant with a decrease of cells' Young's modulus after irradiation. This cell softening was explained by actin cytoskeleton reorganization, with diminution of cell circularity and more abundant pseudopodia. This study highlights the interest of green laser PMB for the proliferation and migration of mesenchymal stem cells, with encouraging results for clinical application, especially for surgical wound healing procedures.


Assuntos
Citoesqueleto/efeitos da radiação , Polpa Dentária/citologia , Terapia com Luz de Baixa Intensidade , Células-Tronco Mesenquimais/citologia , Células-Tronco Mesenquimais/efeitos da radiação , Cicatrização/efeitos da radiação , Adolescente , Adulto , Fenômenos Biomecânicos/efeitos da radiação , Movimento Celular/efeitos da radiação , Proliferação de Células/efeitos da radiação , Forma Celular/efeitos da radiação , Células Cultivadas , Humanos , Adulto Jovem
5.
Anal Chem ; 92(24): 15745-15756, 2020 12 15.
Artigo em Inglês | MEDLINE | ID: mdl-33225709

RESUMO

The variable configuration of Raman spectroscopic platforms is one of the major obstacles in establishing Raman spectroscopy as a valuable physicochemical method within real-world scenarios such as clinical diagnostics. For such real world applications like diagnostic classification, the models should ideally be usable to predict data from different setups. Whether it is done by training a rugged model with data from many setups or by a primary-replica strategy where models are developed on a 'primary' setup and the test data are generated on 'replicate' setups, this is only possible if the Raman spectra from different setups are consistent, reproducible, and comparable. However, Raman spectra can be highly sensitive to the measurement conditions, and they change from setup to setup even if the same samples are measured. Although increasingly recognized as an issue, the dependence of the Raman spectra on the instrumental configuration is far from being fully understood and great effort is needed to address the resulting spectral variations and to correct for them. To make the severity of the situation clear, we present a round robin experiment investigating the comparability of 35 Raman spectroscopic devices with different configurations in 15 institutes within seven European countries from the COST (European Cooperation in Science and Technology) action Raman4clinics. The experiment was developed in a fashion that allows various instrumental configurations ranging from highly confocal setups to fibre-optic based systems with different excitation wavelengths. We illustrate the spectral variations caused by the instrumental configurations from the perspectives of peak shifts, intensity variations, peak widths, and noise levels. We conclude this contribution with recommendations that may help to improve the inter-laboratory studies.

6.
Caries Res ; 54(2): 144-153, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32107346

RESUMO

The development of new diagnostic technologies based on the light scattering and autofluorescence properties of dental tissues is required to improve the diagnostic ability of initial caries lesions earlier than previously done and promoting the potential of treatment without surgical intervention. The aim of this study is to correlate fluorescence-based results provided by multiphoton microscopy (MPM) with confocal Raman microscopy records using phosphate level at 960 cm-1 and the organic matrix at ∼2,931 cm-1 in healthy and demineralized human enamel. Measurements on 14 teeth were made using two incident lights of different wavelengths, released by confocal Raman microscopy and MPM. Raman phosphate peak intensity at 960 cm-1 along with organic to mineral ratio at (2,931/430 cm-1) and nonlinear optical signals (second harmonic generation [SHG] and intrinsic two-photon excited fluorescence [I2PEF]) were recorded from the demineralized and healthy enamel sites. Raman spectral maps showed that the higher the organic/mineral ratio in the demineralized enamel, the lower the intensity of mineral component in the same zone. MPM revealed new optical indicators of carious lesion as shown by the presence of a red-shifted fluorescence peak in the 650- to 750-nm area of the fluorescence spectrum of demineralized enamel. Moreover, on sample regions with insignificant autofluorescence, the emergence of the SHG signal could be noted. By comparing I2PEF images with the structural motifs observed by the confocal Raman imaging system, the morphological similarity of the acquired images was quite evident. Any change in the I2PEF spectra reflects alterations in the chemical composition of enamel. These findings may provide an important basis for potentially valuable applications of photonic tools in the clinical diagnosis of tooth pathological conditions, besides exposing the fundamental role of organic matrix in enamel integrity and reparation.


Assuntos
Cárie Dentária , Dente , Cárie Dentária/diagnóstico por imagem , Esmalte Dentário/diagnóstico por imagem , Humanos , Fosfatos , Desmineralização do Dente/diagnóstico por imagem
7.
Clin Oral Investig ; 23(7): 2959-2973, 2019 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-30374828

RESUMO

OBJECTIVES: Functional tooth replacement and bone regeneration are parts of the daily practice in modern dentistry, but well-reproducible and relatively inexpensive experimental models are still missing. We aimed to develop a new small animal model to monitor osseointegration utilizing the combination of multiple evaluation protocols. MATERIAL AND METHODS: After cutting the tail between the C4 and C5 vertebrae in Wistar rats, costume made, parallel walled, non-threaded implants were placed into the center of the tail parallel with its longitudinal axis using a surgical guide. Osseointegration of the titanium implants was followed between 4 and 16 weeks after surgery applying axial extraction force, and resonance frequency analysis as functional tests, and histomorphometry and micro-CT as structural evaluations. RESULTS: In functional tests, we observed that both methods are suitable for the detection of the time-dependent increase in osseointegration, but the sensitivity of the pull-out technique (an approximately five times increase with rather low standard error) was much higher than that of the resonance frequency analysis. In structural evaluations, changes in the detected bone implant contact values measured by histomorphometry (yielding 1.5 times increase, with low variations of data) were more reliable than micro-CT based evaluations to screen the developments of contact between bone and implant. CONCLUSION: Our results provide evidence that the caudal vertebrae osseointegration model is useful for the preclinical evaluation of implant integration into the bone. CLINICAL RELEVANCE: The combination of the biomechanical and structural tests offers a well-reproducible small animal system that can be suitable for studying the integration of various implant materials and surface treatments.


Assuntos
Implantes Dentários , Osseointegração , Animais , Implantes Experimentais , Masculino , Ratos , Ratos Wistar , Propriedades de Superfície , Titânio
8.
J Contemp Dent Pract ; 20(10): 1126-1131, 2019 Oct 01.
Artigo em Inglês | MEDLINE | ID: mdl-31883244

RESUMO

AIM: Modern clinical caries management involves early stage caries diagnosis and should fit with dental health policy. The objective of this study was to achieve early caries detection in enamel and dentine with a laser-based system (DIAGNOdent™ pen) first and secondary with a new fluorescence intra-oral camera (Soprolife®). A visual inspection with a loupe was used as control. MATERIALS AND METHODS: Following the consolidated standards of reporting trials recommendations, 628 occlusal fissures were included for analysis. RESULTS: The sensitivity and specificity of both devices varied depending on the cutoff threshold of the caries score, and the ROC curve showed higher values for the Soprolife® than for DIAGNOdent™ pen. The values of the area under the curve decreased from 0.81 (Soprolife® in daylight) to 0.79 (Soprolife® in fluorescent mode) and 0.67 for DIAGNOdent™ pen. DIAGNOdent™ pen reproducibility (intra and inter-investigator) showed a wide dispersion, with many values scattered beyond the confidence limits (±2 SD), and the weighted kappa coefficient, which was quite low (0.58), confirmed this tendency. CONCLUSION: Caries prevalence in terms of public health policy is of interest and caries detection increased significantly when using an fluorescence-based intra-oral camera. CLINICAL SIGNIFICANCE: The clinical significance of these findings is that fluorescence could help improve caries diagnosis, reduce clinical misinterpretations, and finally benefit the patients. How to cite this article: Terrer E, Slimani A, Giraudeau N, et al. Performance of Fluorescence-based Systems in Early Caries Detection: A Public Health Issue. J Contemp Dent Pract 2019;20(10):1126-1132.


Assuntos
Cárie Dentária , Saúde Pública , Esmalte Dentário , Dentina , Fluorescência , Humanos , Reprodutibilidade dos Testes , Sensibilidade e Especificidade
9.
Caries Res ; 52(5): 359-366, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-29510396

RESUMO

Dentin carious lesion is a dynamic process that involves demineralization and collagen denaturation. Collagen type I is the major protein in dentin and it has been investigated based on its optical properties. Multiphoton microscopy (MPM) is a nonlinear imaging technique that reveals the caries process using the collagen two-photon excitation fluorescence (2PEF) and its second-harmonic generation (SHG). Combining the histological and the International Caries Detection and Assessment System (ICDAS) classifications with nonlinear optical spectroscopy (NLOS), 2PEF and SHG intensities of enamel and dentin were highly altered during the caries process. It has been proven that the ratio SHG/2PEF is a relevant indicator of the organic matrix denaturation [Terrer et al.: J Dent Res 2016; 96: 574-579]. In the present study, a series of measurable signals is made to detect early stages of carious lesion according to the ICDAS classification and to explore the relationship between these measures and the ICDAS scale. Comparison of the efficiency of nonlinear optical signals for caries detection with the ICDAS classification is essential to evaluate their potential for clinical application. In our study, the use of the NLOS measured by MPM allowed us to monitor a quantitative parameter (SHG/2PEF ratio) according to the dentin carious lesion state (ICDAS and histological examination). Three coherent new groups were defined (ICDAS 0/1; ICDAS 2/3; ICDAS 4/5/6), where the carious process can be clearly described with a statistically significant decrease of the SHG/2PEF ratio.


Assuntos
Cárie Dentária/diagnóstico , Adulto , Cárie Dentária/classificação , Cárie Dentária/diagnóstico por imagem , Cárie Dentária/patologia , Esmalte Dentário/diagnóstico por imagem , Esmalte Dentário/patologia , Progressão da Doença , Humanos , Microscopia de Fluorescência por Excitação Multifotônica
10.
J Mater Sci Mater Med ; 28(10): 145, 2017 Aug 19.
Artigo em Inglês | MEDLINE | ID: mdl-28823063

RESUMO

For many practitioners, longevity of full glass ceramic crowns in the posterior area, molars and premolars, remains a real challenge. The purpose of this article is to identify and evaluate the parameters that can significantly influence their resistance when preparing a tooth. The analysis proposed in this article relies on interrelated studies conducted at three levels: in vitro (mechanical tests), in silico (finite elements simulations) and in vivo (clinical survival rates). The in vitro and the in silico studies proved that an appropriate variation of the geometric design of the preparations enables to increase up to 80% the mechanical strength of ceramic reconstructions. The in vivo clinical study of CAD/CAM full ceramic crowns was performed in accordance with the principles stated within the in vitro and the in silico studies and provided a 98.97% success rate over a 6 years period. The variations of geometric design parameters for dental preparation allows for reconstructions with a mechanical breaking up to 80% higher than that of a non-appropriate combination. These results are confirmed in clinical practice.


Assuntos
Dente Pré-Molar , Cerâmica , Desenho Assistido por Computador , Dente Molar , Coroa do Dente , Cimentação/métodos , Materiais Dentários , Falha de Restauração Dentária , Humanos , Teste de Materiais , Mecânica
11.
Fogorv Sz ; 110(1): 7-14, 2017 Mar.
Artigo em Inglês, Húngaro | MEDLINE | ID: mdl-29847062

RESUMO

During that last decade a large number of experiments showed the successful application of stem cells in achieving large bone volume regeneration. On the contrary, our knowledge about the promotion of implant osseointegration by stem cell is sporadic. Recently, our research group has carried out an array of studies aiming the characterization of postnatal stem cells of dental origin. In addition, we have developed a novel quantitative model for implant osseointegration in rat tail vertebrae. In the present work we aimed to study how the implant osseointegration process is affected by mesenchymal stem cells of rat dental pulp origin (DPSC) when cells are undifferentiated or predifferentiated into osteogenic direction. Our results show that undifferentiated pulp cells inserted between the implant and the bone slow down the osseointegration process. On the other hand, pre-differentiated DPSCs do not have a similar adverse effect any more. Our data suggest that the success of mesenchymal stem cell application to promote implant osseointegration is highly dependent on the applied conditions, particularly on the parallel applicatioh of scaffolds and osteogenic components.


Assuntos
Implantes Dentários , Polpa Dentária/citologia , Células-Tronco Mesenquimais , Osseointegração , Titânio , Animais , Modelos Animais , Ratos , Coluna Vertebral
12.
J Mater Sci Mater Med ; 27(7): 118, 2016 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-27259708

RESUMO

Polyetheretherketone (PEEK) is a polyaromatic semi-crystalline thermoplastic polymer with mechanical properties favorable for bio-medical applications. Polyetheretherketone forms: PEEK-LT1, PEEK-LT2, and PEEK-LT3 have already been applied in different surgical fields: spine surgery, orthopedic surgery, maxillo-facial surgery etc. Synthesis of PEEK composites broadens the physicochemical and mechanical properties of PEEK materials. To improve their osteoinductive and antimicrobial capabilities, different types of functionalization of PEEK surfaces and changes in PEEK structure were proposed. PEEK based materials are becoming an important group of biomaterials used for bone and cartilage replacement as well as in a large number of diverse medical fields. The current paper describes the structural changes and the surface functionalization of PEEK materials and their most common biomedical applications. The possibility to use these materials in 3D printing process could increase the scientific interest and their future development as well.


Assuntos
Materiais Biocompatíveis/química , Cetonas/química , Teste de Materiais/métodos , Polietilenoglicóis/química , Animais , Benzofenonas , Osso e Ossos/metabolismo , Cartilagem/metabolismo , Humanos , Ortopedia/métodos , Polímeros , Ratos , Propriedades de Superfície , Procedimentos Cirúrgicos Torácicos , Engenharia Tecidual/instrumentação , Engenharia Tecidual/métodos
13.
Folia Med (Plovdiv) ; 57(1): 11-8, 2015.
Artigo em Inglês | MEDLINE | ID: mdl-26431090

RESUMO

Numerous approaches have been used to improve the tissue-implant interface of titanium (Ti) and titanium alloy (Ti6Al4V). They all aim at increasing cell migration and attachment to the metal, preventing unspecific protein adsorption and improving post-implantation healing process. Promising methods for titanium and titanium alloy surface modification are based on the immobilization of biologically active organic molecules. New and interesting biochemical approaches to such surface modification include layer-by-layer deposition of polyelectrolyte films, phage display-selected surface binding peptides and self-assembled DNA monolayer systems. The present review summarizes the scientific information about these methods, which are at in vitro or in vivo development stages, and hopes to promote their future application in dental implantology and in oral and maxillofacial surgery.


Assuntos
Implantes Dentários , Proteínas Imobilizadas/química , Titânio , Adsorção , Ligas , Sequência de Aminoácidos , Proteínas Sanguíneas/química , Dados de Sequência Molecular , Propriedades de Superfície
14.
J Chem Inf Model ; 53(12): 3273-9, 2013 Dec 23.
Artigo em Inglês | MEDLINE | ID: mdl-24289530

RESUMO

Engineering shape-controlled bionanomaterials requires comprehensive understanding of interactions between biomolecules and inorganic surfaces. We explore the origin of facet-selective binding of peptides adsorbed onto Pt(100) and Pt(111) crystallographic planes. Using molecular dynamics simulations, we show that upon adsorption the peptides adopt a predictable conformation. We compute the binding energies of the amino acids constituting two adhesion peptides for Pt, S7, and T7 and demonstrate that peptides' surface recognition behavior that makes them unique among populations originates from differential adsorption of their building blocks. We find that the degree of peptide binding is mainly due to polar amino acids and the molecular architecture of the peptides close to the Pt facets. Our analysis is a first step in the prediction of enhanced affinity between inorganic materials and a peptides, toward the synthesis of novel nanomaterials with programmable shape, structure, and properties.


Assuntos
Aminoácidos/química , Peptídeos/química , Platina/química , Adsorção , Cristalização , Simulação de Dinâmica Molecular , Nanoestruturas , Ligação Proteica , Conformação Proteica , Propriedades de Superfície , Termodinâmica
15.
Clin Oral Investig ; 17(3): 757-63, 2013 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-22855265

RESUMO

OBJECTIVES: Our aim was to determine the origin of the red fluorescence of carious dentine observed with the Soprolife® camera. METHODS: We conducted in vitro studies to evaluate the origin of the red fluorescence using acids and matrix metalloproteinase (MMP) to mimic caries and methylglycoxal (MGO) to evaluate the effect of glycation reactions on the red fluorescence. In every step of these models, we detected the changes of dentin photonic response with Soprolife® in daylight mode and in treatment mode. A Raman spectroscopy analysis was performed to determine the variations of the dentin organic during the in vitro caries processes. Raman microscopy was performed to identify change in the collagen matrix of dentine. RESULTS: The red fluorescence observed in carious dentine using a Soprolife® camera corresponds to the brownish color observed using daylight. Demineralization using nitric acid induces a loss of the green fluorescence of dentine. The red fluorescence of carious dentine is resistant to acid treatment. Immersion of demineralized dentine in MGO induces a change of color from white to orange-red. This indicates that the Maillard reaction contributes to lesion coloration. Immersion of demineralized dentine in an MMP-1 solution followed by MGO treatment results in a similar red fluorescence. Raman microspectroscopy analysis reveals accumulation of AGE's product in red-colored dentine. CONCLUSIONS: Our results provide important information on the origin of the fluorescence variation of dentine observed with the Soprolife® camera. We demonstrate that the red fluorescence of carious dentine is linked to the accumulation of Advanced Glycation End products (AGE). CLINICAL RELEVANCE: The study provides a new biological basis for the red fluorescence of carious dentine and reinforces the importance of the Soprolife® camera in caries diagnostics.


Assuntos
Cárie Dentária/patologia , Dentina/patologia , Fotografia Dentária , Colágeno/química , Cárie Dentária/diagnóstico , Dentina/química , Fluorescência , Produtos Finais de Glicação Avançada/análise , Humanos , Reação de Maillard , Metaloproteinase 1 da Matriz , Fotografia Dentária/instrumentação , Análise Espectral Raman
16.
Biomedicines ; 11(8)2023 Jul 30.
Artigo em Inglês | MEDLINE | ID: mdl-37626649

RESUMO

Amyotrophic lateral sclerosis (ALS) is a fatal and incurable paralytic disorder caused by the progressive death of upper and lower motoneurons. Although numerous strategies have been developed to slow disease progression and improve life quality, to date only a few therapeutic treatments are available with still unsatisfactory therapeutic benefits. The secretome of dental pulp stem cells (DPSCs) contains numerous neurotrophic factors that could promote motoneuron survival. Accordingly, DPSCs confer neuroprotective benefits to the SOD1G93A mouse model of ALS. However, the mode of action of DPSC secretome on motoneurons remains largely unknown. Here, we used conditioned medium of human DPSCs (DPSCs-CM) and assessed its effect on survival, axonal length, and electrical activity of cultured wildtype and SOD1G93A motoneurons. To further understand the role of individual factors secreted by DPSCs and to circumvent the secretome variability bias, we focused on GDF15 and HB-EGF whose neuroprotective properties remain elusive in the ALS pathogenic context. DPSCs-CM rescues motoneurons from trophic factor deprivation-induced death, promotes axon outgrowth of wildtype but not SOD1G93A mutant motoneurons, and has no impact on the spontaneous electrical activity of wildtype or mutant motoneurons. Both GDF15 and HB-EGF protect SOD1G93A motoneurons against nitric oxide-induced death, but not against death induced by trophic factor deprivation. GDF15 and HB-EGF receptors were found to be expressed in the spinal cord, with a two-fold increase in expression for the GDF15 low-affinity receptor in SOD1G93A mice. Therefore, the secretome of DPSCs appears as a new potential therapeutic candidate for ALS.

17.
Bioengineering (Basel) ; 10(7)2023 Jul 19.
Artigo em Inglês | MEDLINE | ID: mdl-37508879

RESUMO

To date, bone regeneration techniques use many biomaterials for bone grafting with limited efficiencies. For this purpose, tissue engineering combining biomaterials and stem cells is an important avenue of development to improve bone regeneration. Among potentially usable non-toxic and bioresorbable scaffolds, porous silicon (pSi) is an interesting biomaterial for bone engineering. The possibility of modifying its surface can allow a better cellular adhesion as well as a control of its rate of resorption. Moreover, release of silicic acid upon resorption of its nanostructure has been previously proved to enhance stem cell osteodifferentiation by inducing calcium phosphate formation. In the present study, we used a rat tail model to experiment bone tissue engineering with a critical size defect. Two groups with five rats per group of male Wistar rats were used. In each rat, four vertebrae were used for biomaterial implantation. Randomized bone defects were filled with pSi particles alone or pSi particles carrying dental pulp stem cells (DPSC). Regeneration was evaluated in comparison to empty defect and defects filled with xenogenic bone substitute (Bio-Oss®). Fluorescence microscopy and SEM evaluations showed adhesion of DPSCs on pSi particles with cells exhibiting distribution throughout the biomaterial. Histological analyzes revealed the formation of a collagen network when the defects were filled with pSi, unlike the positive control using Bio-Oss®. Overall bone formation was objectivated with µCT analysis and showed a higher bone mineral density with pSi particles combining DPSC. Immunohistochemical assays confirmed the increased expression of bone markers (osteocalcin) when pSi particles carried DPSC. Surprisingly, no grafted cells remained in the regenerated area after one month of healing, even though the grafting of DPSC clearly increased bone regeneration for both bone marker expression and overall bone formation objectivated with µCT. In conclusion, our results show that the association of pSi with DPSCs in vivo leads to greater bone formation, compared to a pSi graft without DPSCs. Our results highlight the paracrine role of grafted stem cells by recruitment and stimulation of endogenous cells.

18.
Biomater Sci ; 11(20): 6919-6930, 2023 Oct 10.
Artigo em Inglês | MEDLINE | ID: mdl-37655620

RESUMO

In dental practice, Regenerative Endodontic Treatment (RET) is applied as an alternative to classical endodontic treatments of immature necrotic teeth. This procedure, also known as dental pulp revitalization, relies on the formation of a blood clot inside the root canal leading to the formation of a reparative vascularized tissue similar to dental pulp, which would provide vitality to the affected tooth. Despite the benefit of this technique, it lacks reproducibility due to the fast degradation and poor mechanical properties of blood clots. This work presents a method for constructing a fibrinogen-blood hydrogel that mimics the viscoelastic properties of human dental pulp while preserving the biological properties of blood for application in RET. By varying the blood and fibrinogen concentrations, gels with different biomechanical and biological properties were obtained. Rheology and atomic force microscopy (AFM) were combined to study the viscoelastic properties. AFM was used to evaluate the elasticity of human dental pulp. The degradation and swelling rates were assessed by measuring weight changes. The biomimetic properties of the gels were demonstrated by studying the cell survival and proliferation of dental pulp cells (DPCs) for 14 days. The formation of an extracellular matrix (ECM) was assessed by multiphoton microscopy (MPM). The angiogenic potential was evaluated by an ex vivo aortic ring assay, in which the endothelial cells were observed by histological staining after migration. The results show that the Fbg-blood gel prepared with 9 mg ml-1 fibrinogen and 50% blood of the Fbg solution volume has similar elasticity to human dental pulp and adequate degradation and swelling rates. It also allows cell survival and ECM secretion and enhances endothelial cell migration and formation of neovessel-like structures.


Assuntos
Polpa Dentária , Regeneração , Humanos , Células Endoteliais , Fibrinogênio , Hidrogéis/farmacologia , Reprodutibilidade dos Testes
19.
Expert Rev Med Devices ; 20(8): 633-642, 2023.
Artigo em Inglês | MEDLINE | ID: mdl-37294872

RESUMO

INTRODUCTION: This review summarizes the available recent literature on different mechanisms and parameters of pulsed ultrasound (US) that have been used during orthodontic treatments to prevent and repair root resorption. AREAS COVERED: A literature search was conducted between January (2002) and September (2022) in the following databases: PubMed, Google-Scholar, Embase, and The-Cochrane-Library. After exclusions, a total of 19 papers were included in the present review. The most used US parameters with positive outcomes were frequency of 1.5 MHz, pulse repetition frequency of 1000 Hz, output intensity of 30 mW/cm2, duration of application of 20 min and total number sessions were 14 with a repetition interval of 1 day. The suggested mechanisms induced by US were alteration of cementoblasts, osteoblasts, osteoclasts, alkaline-phosphatase (ALP), runt-related-gene-2 (Runx2), osteoprotegerin (OPG), type-I-collagen (Col-I), C-telopeptide-type-I-collagen (CTX-I), hepatocyte-growth-factor (HGF), bone morphogenetic protein-2 (BMP-2), cyclooxygenase-2 (Cox-2), calcium (Ca2+), receptor activator of nuclear factor-kappa-B ligand (RANKL), and receptor activator of nuclear factor-kappa-B (RANK). EXPERT OPINION: Understanding mechanisms and deciding which parameters of US that can be used during orthodontic treatment to prevent and repair root resorption is a great challenge. This work summarizes all the available data that can aid this process and suggest that US is an effective noninvasive method not only in prevention and repairing of orthodontic induced root resorption but also in accelerating teeth movement.


Assuntos
Reabsorção da Raiz , Humanos , Reabsorção da Raiz/diagnóstico por imagem , Reabsorção da Raiz/prevenção & controle , Ultrassonografia , Técnicas de Movimentação Dentária , Osteoclastos , Colágeno
20.
Arch Oral Biol ; 152: 105733, 2023 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-37247560

RESUMO

OBJECTIVE: Dentin, enamel and the transition zone, called the dentin-enamel junction (DEJ), have an organization and properties that play a critical role in tooth resilience and in stopping the propagation of cracks. Understanding their chemical and micro-biomechanical properties is then of foremost importance. The aim of this study is to apply Brillouin microscopy on a complex biological structure, that is, the DEJ, and to compare these results with those obtained with Raman microscopy. DESIGN: Both techniques allow noncontact measurements at the microscopic scale. Brillouin microscopy is based on the interaction between acoustic phonons and laser photons and gives a relation between the frequency shift of the scattered light and the stiffness of the sample. Raman spectra contain peaks related to specific chemical bonds. RESULTS: Comparison of the Brillouin and Raman cartographies reveals correlations between mechanical and chemical properties. Indeed, the shapes of the phosphate content and stiffness curves are similar. The two spectroscopies give compatible values for the mean distance between two tubules, i.e., 4-6 µm. Moreover, for the first time, the daily cross striations of enamel could be studied, indicating a relationship between the variation in the phosphate concentration and the variation in the rigidity within the enamel prisms. CONCLUSIONS: We demonstrate here the possibility of using Brillouin scattering microscopy to both study complex biological materials such as the enamel-dentin junction and visualize secondary structures. Correlations between the chemical composition and mechanical properties could help in better understanding the tissue histology.


Assuntos
Dentina , Dente , Dentina/química , Microscopia , Esmalte Dentário/química
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