RESUMO
We compared qualitative characteristics of the connective tissue at the site of implantation of polypropylene or a biodegradable polymer synthesized by electrospinning and consisting of 65% polycaprolactone and 35% polytrimethylene carbonate. Synthetic materials were implanted into the interfascial space of the muscles on the back of Wistar rats. The parameters of cellular and non-cellular structures of the forming connective tissue were studied in 1, 2, 3, 6, 9, and 12 months after implantation. It was found that the number of fibrocytes and fibroblasts around biopolymer and polypropylene increased during the period from 1 to 9 months, with a significant lag in case of biopolymer implantation; by 12 months, the number of fibrocytic cells did not significantly differ. The fibroplastic and angioplastic processes developed in the thickness of both materials, but with a one-month lag in case of biopolymer implantation due to degradation properties and nonporous structure of this polymer. Collagen fibers were actively synthesized around both materials, but in case of polypropylene, this process was more pronounced. In 6 months after implantation of polypropylene, dense regular connective tissue was formed; in 12 months after implantation of the biopolymer, the formation of dense irregular connective tissue was observed at the site of implantation. The biopolymer can be used to strengthen the core muscles.
Assuntos
Tecido Conjuntivo/química , Polipropilenos/química , Animais , Biodegradação Ambiental , Biopolímeros/química , Dioxanos/química , Masculino , Polímeros/química , Ratos , Ratos WistarRESUMO
BACKGROUND: Protein glycation by Maillard reaction is commonly used to improve the functional and bioactive properties of food proteins. It is also known that this glycation method can be accelerated by heat without the need for chemical reagents that could be harmful to health. In this study, glycoconjugates were obtained from a mixture of connective tissue proteins (CTP) from jumbo squid (Dosidicus gigas) and two different sugars, dextran (DEX; 10 kDa) and glucose (GLU), using protein-to-carbohydrate ratios of 1:2 and 1:3, in solution at 50 °C for 6 h. The glycation products were characterized by means of their physicochemical properties and cytotoxic effect. RESULTS: The intensity of the browning measured at A420nm and A294nm in glycoconjugates showed no significant difference (P < 0.05). CTP-DEX (1:2) and CTP-DEX (1:3) were those products with the greatest fluorescence related to the intermediate stage in the Maillard reaction, and also with the highest degree of glycation, which was confirmed using o-phthaldialdehyde assay and Fourier transform infrared analysis. The values of cellular viability for CTP-GLU (1:3), CTP-DEX (1:2, 1:3) as well as CTP (0, 6 h) were around 92-103%. CONCLUSIONS: The operational parameters used in the glycation process achieved the formation of glycoconjugates from proteins of D. gigas, showing no cytotoxic effect on the HaCaT cell line. This research proposes an alternative for the modification of proteins and opens the way to future investigations regarding the bioactivity of these macromolecules to have applications for the use of byproducts in food science and technology. © 2020 Society of Chemical Industry.
Assuntos
Decapodiformes/química , Glicoconjugados/química , Resíduos/análise , Animais , Tecido Conjuntivo/química , Dextranos/química , Glucose/química , Glicosilação , Reação de Maillard , Proteínas/químicaRESUMO
Collagen accounts for the major extracellular matrix (ECM) component in many tissues and provides mechanical support for cells. Magnetic Resonance (MR) Imaging, MR based diffusion measurements and MR Elastography (MRE) are considered sensitive to the microstructure of tissues including collagen networks of the ECM. However, little is known whether water diffusion interacts with viscoelastic properties of tissues. This study combines highfield MR based diffusion measurements, novel compact tabletop MRE and confocal microscopy in collagen networks of different cross-linking states (untreated collagen gels versus additional treatment with glutaraldehyde). The consistency of bulk rheology and MRE within a wide dynamic range is demonstrated in heparin gels, a viscoelastic standard for MRE. Additional crosslinking of collagen led to an 8-fold increased storage modulus, a 4-fold increased loss modulus and a significantly decreased power law exponent, describing multi-relaxational behavior, corresponding to a pronounced transition from viscous-soft to elastic-rigid properties. Collagen network changes were not detectable by MR based diffusion measurements and microscopy which are sensitive to the micrometer scale. The MRE-measured shear modulus is sensitive to collagen fiber interactions which take place on the intrafiber level such as fiber stiffness. The insensitivity of MR based diffusion measurements to collagen hydrogels of different cross-linking states alludes that congeneric collagen structures in connective tissues do not hinder extracellular diffusive water transport. Furthermore, the glutaraldehyde induced rigorous changes in viscoelastic properties indicate that intrafibrillar dissipation is the dominant mode of viscous dissipation in collagen-dominated connective tissue.
Assuntos
Colágeno/química , Colágeno/metabolismo , Tecido Conjuntivo/química , Tecido Conjuntivo/metabolismo , Elasticidade , Solventes/química , Água/química , Animais , Bovinos , Tecido Conjuntivo/diagnóstico por imagem , Difusão , Imageamento por Ressonância Magnética , ViscosidadeRESUMO
Natural teeth are supported by connective tissue collagen fibers that insert perpendicularly in the tooth cementum. Perpendicular insertion plays an important role in the maintenance of the junction between the oral epithelium and the periodontal connective tissue. Most titanium dental implant surfaces have no micro or macro structure to support perpendicularly oriented collagen attachment. Without this tight biologic seal to resist bacterial invasion and epithelial downgrowth, progressive bone loss in peri-implantitis is seen around dental implants. The purpose of this study was to establish the perpendicularly oriented collagen attachment to titanium oxide nanotube (TNT), and to assess its binding stability. TNT was prepared on the titanium-surface by anodization. Scanning electron microscopy (SEM) showed a regularly aligned TNT with an average 67 nm-diameter when anodized at 30 V for 3 h. Subsequently, collagen type I (CoI) was electrophoretically fused to anodic TNT in native polyacrylamide gel system where negatively charged CoI-C term was perpendicularly navigated to TNT. SEM and atomic force microscopy (AFM) were used to analyze CoI on the TiO2 and TNT surface. Several tens of nanometers of CoI protrusion were recorded by AFM. These protrusions may be long enough to be priming sites for cell-secreted CoI. CoI laid parallel to the titanium surface when fused by a chemical linker. Binding resistance of CoI against drastic ultrasonication was measured by Fourier-transform infrared spectroscopy attenuated total reflection (FTIR-ATR). The electrophoretically fused CoI in the titanium nanotube (TNT-CoIEPF) showed the significantly greatest binding resistance than the other groups (P < 0.01, a 1-way ANOVA and Tukey HSD post hoc test). Furthermore, TNT-CoIEPF surface rejected epithelial cell stretching and epithelial sheet formation. Chemically linked horizontal CoI on titanium oxide (TiO2) facilitated epithelial cell stretching and sheet formation.
Assuntos
Colágeno Tipo I/química , Tecido Conjuntivo/química , Implantes Dentários , Nanotubos/química , Titânio/química , Materiais Biocompatíveis/química , Adesão Celular/efeitos dos fármacos , Técnicas Eletroquímicas , Células Epiteliais/efeitos dos fármacos , Humanos , Tamanho da Partícula , Propriedades de SuperfícieRESUMO
OBJECTIVE: To define the indication for preventive mesh implantation in patients scheduled for laparotomy. MATERIAL AND METHODS: There were 108 patients divided into the control (52 patients without anterior abdominal wall hernia) and the main (56 patients with ventral hernia) groups. RESULTS: Predominance of collagen type III over type I was observed in patients older 60 years in the main group. The maximum ratio of collagen types I and III was 1.4 in the main group. The minimum number of inter-fiber spaces (n=5) was noted in patients aged 30-40 years. The maximum density of connective tissue was 250 pixels per inch. CONCLUSION: Analysis of connective tissue structures revealed some important age-related features and markers of connective tissue dysplasia in the main group. An indication for preventive mesh implantation in patients scheduled for laparotomy was determined.
Assuntos
Colágeno/análise , Doenças do Tecido Conjuntivo/complicações , Hérnia Ventral/prevenção & controle , Hérnia Incisional/prevenção & controle , Laparotomia/efeitos adversos , Telas Cirúrgicas , Parede Abdominal/patologia , Parede Abdominal/cirurgia , Adulto , Tecido Conjuntivo/química , Tecido Conjuntivo/patologia , Doenças do Tecido Conjuntivo/patologia , Hérnia Ventral/etiologia , Hérnia Ventral/patologia , Herniorrafia/métodos , Humanos , Hérnia Incisional/etiologia , Hérnia Incisional/patologia , Pessoa de Meia-IdadeRESUMO
The mammalian lung´s structural design is optimized to serve its main function: gas exchange. It takes place in the alveolar region (parenchyma) where air and blood are brought in close proximity over a large surface. Air reaches the alveolar lumen via a conducting airway tree. Blood flows in a capillary network embedded in inter-alveolar septa. The barrier between air and blood consists of a continuous alveolar epithelium (a mosaic of type I and type II alveolar epithelial cells), a continuous capillary endothelium and the connective tissue layer in-between. By virtue of its respiratory movements, the lung has to withstand mechanical challenges throughout life. Alveoli must be protected from over-distension as well as from collapse by inherent stabilizing factors. The mechanical stability of the parenchyma is ensured by two components: a connective tissue fiber network and the surfactant system. The connective tissue fibers form a continuous tensegrity (tension + integrity) backbone consisting of axial, peripheral and septal fibers. Surfactant (surface active agent) is the secretory product of type II alveolar epithelial cells and covers the alveolar epithelium as a biophysically active thin and continuous film. Here, we briefly review the structural components relevant for gas exchange. Then we describe our current understanding of how these components function under normal conditions and how lung injury results in dysfunction of alveolar micromechanics finally leading to lung fibrosis.
Assuntos
Tecido Conjuntivo/metabolismo , Alvéolos Pulmonares/metabolismo , Surfactantes Pulmonares/metabolismo , Animais , Tecido Conjuntivo/química , Humanos , Alvéolos Pulmonares/química , Surfactantes Pulmonares/químicaRESUMO
Despite their soft body and slow motion, sea cucumbers present a low predation rate, reflecting the presence of efficient defence systems. For instance, members of the family Holothuriidae rely on Cuvierian tubules for their defence. These tubules are normally stored in the posterior coelomic cavity of the animal, but when the sea cucumber is threatened by a potential predator, they are expelled through the cloacal aperture, elongate, become sticky and entangle and immobilise the predator in a matter of seconds. The mechanical properties (extensibility, tensile strength, stiffness and toughness) of quiescent (i.e. in the body cavity) and elongated (i.e. after expulsion) Cuvierian tubules were investigated in the species Holothuria forskali using traction tests. Important mechanical differences were measured between the two types of tubules, reflecting adaptability to their operating mode: to ease elongation, quiescent tubules present a low resistance to extension, while elongated tubules present a high toughness to resist tractions generated by the predator. We demonstrate that a mutable collagenous tissue (MCT) is involved in the functioning of these organs: (1) some mechanical properties of Cuvierian tubules are modified by incubation in a cell-disrupting solution; (2) the connective tissue layer encloses juxtaligamental-like cells, a cell type present in all MCTs; and (3) tensilin, a MCT stiffening protein, was localised inside these cells. Cuvierian tubules thus appear to enclose a new type of MCT which shows irreversible stiffening.
Assuntos
Colágeno/fisiologia , Tecido Conjuntivo/química , Holothuria/fisiologia , Sequência de Aminoácidos , Animais , Fenômenos Biomecânicos , Proteínas de Transporte , Colágeno/efeitos dos fármacos , Tecido Conjuntivo/ultraestrutura , Octoxinol , Resistência à Tração/fisiologiaRESUMO
Connective tissues such as tendon, ligament and cartilage are mostly composed of extracellular matrix (ECM). These tissues are insoluble, mainly due to the highly cross-linked ECM proteins such as collagens. Difficulties obtaining suitable samples for mass spectrometric analysis render the application of modern proteomic technologies difficult. Complete solubilization of them would not only elucidate protein composition of normal tissues but also reveal pathophysiology of pathological tissues. Here we report complete solubilization of human Achilles tendon and yellow ligament, which is achieved by chemical digestion combined with successive protease treatment including elastase. The digestion mixture was subjected to liquid chromatography-mass spectrometry. The low specificity of elastase was overcome by accurate mass analysis achieved using FT-ICR-MS. In addition to the detailed proteome of both tissues, we also quantitatively determine the major protein composition of samples, by measuring peak area of some characteristic peptides detected in tissue samples and in purified proteins. As a result, differences between human Achilles tendon and yellow ligament were elucidated at molecular level.
Assuntos
Tendão do Calcâneo/química , Tecido Conjuntivo/química , Matriz Extracelular/química , Ligamentos/química , Proteoma/análise , Cromatografia Líquida , Humanos , Espectrometria de Massas , Peptídeo Hidrolases/metabolismo , Proteômica/métodos , SolubilidadeRESUMO
BACKGROUND: The camel is an excellent source of high quality meat and camel meat might be a potential alternative for beef. This study aimed to manipulate the raw camel meat for the production of stable and acceptable emulsion sausage, as well as to study the effect of cooking at different core temperatures on the tenderness, sensory quality and microstructure of produced sausage. RESULTS: Increasing the cooking temperature of sausages resulted in reduction of the shear force values from 2.67 kgf after cooking at 85 °C to 1.57 kgf after cooking at 105 °C. The sensory scores of sausages have been improved by increasing the cooking core temperature of meat batter. The light and scanning electron microscope micrographs revealed solubilisation of the high quantity of connective tissue of camel meat. High emulsion stability values for the camel meat batter associated with high values of water-holding capacity for raw camel meat and meat batter have been recorded. CONCLUSION: Stable and acceptable camel meat emulsion can be developed from camel meat. Increasing the cooking core temperature of meat batter improved the quality of produced sausages. Therefore, camel meat emulsion sausages might be a potential alternative for beef particularly in Asian and African countries. © 2015 Society of Chemical Industry.
Assuntos
Camelus , Culinária , Qualidade dos Alimentos , Produtos da Carne/análise , Carne/análise , Animais , Tecido Conjuntivo/química , Tecido Conjuntivo/ultraestrutura , Egito , Emulsões , Fermentação , Preferências Alimentares/etnologia , Armazenamento de Alimentos , Temperatura Alta/efeitos adversos , Humanos , Carne/microbiologia , Produtos da Carne/microbiologia , Fenômenos Mecânicos , Micrococcus/crescimento & desenvolvimento , Micrococcus/metabolismo , Microscopia Eletrônica de Varredura , Músculo Esquelético/química , Músculo Esquelético/ultraestrutura , Refrigeração , Sensação , Resistência ao Cisalhamento , Água/análiseRESUMO
Computer simulation of the electrical activity in sinoatrial node cells interacting via gap junctions with fibroblasts revealed that interaction with fibroblasts results in greater oscillation frequency of sinoatrial node cells. We have found out that fibroblasts also decrease the oscillation amplitude of the intrinsic central cells or completely suppress their spontaneous activity, while weakly affect the oscillation amplitude of peripheral cells.
Assuntos
Simulação por Computador , Fibroblastos/química , Modelos Teóricos , Nó Sinoatrial/química , Tecido Conjuntivo/química , Fibroblastos/citologia , Junções Comunicantes/química , Junções Comunicantes/fisiologia , Humanos , Potenciais da Membrana , Modelos Cardiovasculares , Miócitos Cardíacos/química , Miócitos Cardíacos/citologia , Nó Sinoatrial/citologiaRESUMO
One of the most active inhibitors angiotensin-converting enzyme is quinapril that has a high affinity for tissue ACE, improves endothelial vasodilation, has a wide therapeutic range and beneficient influence on heart rate. A new biological active compound with antioxidant action that has endothelioprotective, cardioprotective, antiischemic action is angiolin. In experimental arterial hypertension in the animals blood serum the activity of collagenase, the content of free and protein connecting fractions of hydroxyproline and indicators that reflect the metabolism of glycosaminoglycans have been increased. Angiolin increases the activity of collagenase free and protein connecting fractions of hydroxyproline comparing to control. Concentration glycosoaminoglycan (GAG) also exceeds the standard data. Quinapril has similar to angiolin action directed effect to the connective tissue components, though losing as proteinconecting of hydroxiproline action. Cooperative application quinapril with angioline most effectively influence the metabolic processes stabilization in experimental animals.
Assuntos
Inibidores da Enzima Conversora de Angiotensina/farmacologia , Anti-Hipertensivos/farmacologia , Antioxidantes/farmacologia , Tecido Conjuntivo/efeitos dos fármacos , Hipertensão/tratamento farmacológico , Tetra-Hidroisoquinolinas/farmacologia , Animais , Colagenases/sangue , Tecido Conjuntivo/química , Tecido Conjuntivo/metabolismo , Combinação de Medicamentos , Sinergismo Farmacológico , Glicosaminoglicanos/sangue , Frequência Cardíaca/efeitos dos fármacos , Hidroxiprolina/sangue , Hipertensão/sangue , Hipertensão/fisiopatologia , Peptidil Dipeptidase A/sangue , Quinapril , Ratos , Ratos Endogâmicos SHR , Ratos Endogâmicos WKYRESUMO
The four types of platelet-derived growth factors (PDGFs) and the two types of PDGF receptors (PDGFRs, which belong to class III receptor tyrosine kinases) have important functions in the development of connective tissue cells. Recent structural studies have revealed novel mechanisms of PDGFs in propeptide loading and receptor recognition/activation. The detailed structural understanding of PDGF-PDGFR signaling has provided a template that can aid therapeutic intervention to counteract the aberrant signaling of this normally silent pathway, especially in proliferative diseases such as cancer. This review summarizes the advances in the PDGF system with a focus on relating the structural and functional understandings, and discusses the basic aspects of PDGFs and PDGFRs, the mechanisms of activation, and the insights into the therapeutic antagonism of PDGFRs. This article is part of a Special Issue entitled: Emerging recognition and activation mechanisms of receptor tyrosine kinases.
Assuntos
Fator de Crescimento Derivado de Plaquetas/química , Isoformas de Proteínas/química , Precursores de Proteínas/química , Receptores do Fator de Crescimento Derivado de Plaquetas/química , Plaquetas/química , Plaquetas/citologia , Plaquetas/fisiologia , Tecido Conjuntivo/química , Tecido Conjuntivo/fisiologia , Células do Tecido Conjuntivo/química , Células do Tecido Conjuntivo/citologia , Células do Tecido Conjuntivo/fisiologia , Regulação da Expressão Gênica , Humanos , Modelos Moleculares , Fator de Crescimento Derivado de Plaquetas/classificação , Fator de Crescimento Derivado de Plaquetas/fisiologia , Ligação Proteica , Isoformas de Proteínas/classificação , Isoformas de Proteínas/fisiologia , Precursores de Proteínas/metabolismo , Receptores do Fator de Crescimento Derivado de Plaquetas/classificação , Receptores do Fator de Crescimento Derivado de Plaquetas/fisiologia , Transdução de SinaisRESUMO
The functional properties of human pediatric detrusor smooth muscle are poorly described, in contrast to those of adult tissue. Characterization is necessary for more informed management options of bladder dysfunction in children. We therefore compared the histological, contractile, intracellular Ca2+ concentration responses and biomechanical properties of detrusor biopsy samples from pediatric (3-48 mo) and adults (40-60 yr) patients who had functionally normal bladders and were undergoing open surgery. The smooth muscle fraction of biopsies was isolated to measure proportions of smooth muscle and connective tissue (van Gieson stain); in muscle strips, isometric tension to contractile agonists or electrical field stimulation and their passive biomechanical properties; in isolated myocytes, intracellular Ca2+ concentration responses to agonists. Pediatric detrusor tissue compared with adult tissue showed several differences: a smaller smooth muscle-to-connective tissue ratio, similar contractures to carbachol or α,ß-methylene ATP when corrected for smooth muscle content, and similar intracellular Ca2+ transients to carbachol, α,ß-methylene ATP, raised K+ concentration or caffeine, but smaller nerve-mediated contractions and greater passive stiffness with slower stress relaxation. In particular, there were significant atropine-resistant nerve-mediated contractions in pediatric samples. Detrusor smooth muscle from functionally normal pediatric human bladders is less contractile than that from adult bladders and exhibits greater passive stiffness. Reduced bladder contractile function is not due to reduced smooth muscle contractility but to greater connective tissue deposition and to functional denervation. Significant atropine resistance in pediatric detrusor, unlike in adult tissue, demonstrates a different profile of functional neurotransmitter activation. These data have implications for the management of pediatric bladder function by therapeutic approaches.
Assuntos
Tecido Conjuntivo/fisiologia , Músculo Liso/fisiologia , Bexiga Urinária/fisiologia , Trifosfato de Adenosina/análogos & derivados , Trifosfato de Adenosina/farmacologia , Adulto , Fenômenos Biomecânicos , Cálcio/análise , Carbacol/farmacologia , Pré-Escolar , Tecido Conjuntivo/química , Estimulação Elétrica , Feminino , Humanos , Lactente , Masculino , Pessoa de Meia-Idade , Contração Muscular/fisiologia , Músculo Liso/química , Músculo Liso/efeitos dos fármacos , Bexiga Urinária/química , Bexiga Urinária/efeitos dos fármacosRESUMO
Muscle protein synthesis (MPS) rate is determined conventionally by obtaining two or more tissue biopsies during a primed, continuous infusion of a stable isotopically labeled amino acid. The purpose of the present study was to test whether tracer priming given as a flooding dose, thereby securing an instantaneous labeling of the tissue pools of free tracee amino acids, followed by a continuous infusion of the same tracer to maintain tracer isotopic steady state, could be used to determine the MPS rate over a prolonged period of time by obtaining only a single tissue biopsy. We showed that the tracer from the flood prime appeared immediately in the muscle free pool of amino acids and that this abundance could be kept constant by a subsequent continuous infusion of the tracer. When using phenylalanine as tracer, the flood-primed, continuous infusion protocol does not stimulate the MPS rate per se. In conclusion, the flood-primed, continuous infusion protocol using phenylalanine as tracer can validly be used to measure the protein synthesis rate in human in vivo experiments by obtaining only a single tissue biopsy after a prolonged infusion period.
Assuntos
Aminoácidos/química , Biópsia/métodos , Proteínas Musculares/biossíntese , Biossíntese de Proteínas/fisiologia , Traçadores Radioativos , Algoritmos , Colágeno/biossíntese , Colágeno/genética , Tecido Conjuntivo/química , Tecido Conjuntivo/metabolismo , Interpretação Estatística de Dados , Humanos , Infusões Intravenosas , Cetoácidos/sangue , Leucina/análise , Leucina/metabolismo , Masculino , Músculo Esquelético/química , Músculo Esquelético/metabolismo , Miofibrilas/metabolismo , Fenilalanina/sangue , Adulto JovemRESUMO
Previously, we found that oral autologous artificial connective tissue (AACT) had a different protein secretion profile to that of clot-embedded AACT. Other oral mucosa substitutes, having different cell types and scaffolds, had dissimilar secretion profiles of proteins (including that for AACT) that influence healing outcome; thus, to ascertain the profiles of factors secreted by artificial tissue and whether they are influenced by their microstructure might help in understanding their bioactivity. An important component of tissue microstructure is the fiber orientation of the scaffold used for manufacturing it. This work developed a surface plasmon resonance (SPR) methodology to quantify factors secreted by oral artificial connective tissue (ACT) in culture medium, and a method to manufacture unidirectional laminar collagen I scaffolds. The SPR methodology was used for assessing differences in the protein secretion profile of ACT made with collagen scaffolds having different fiber orientation (unidirectional vs multidirectional). Oral fibroblasts seeded onto unidirectional scaffolds increased the secretion of six factors involved in modulating healing compared to those seeded onto multidirectional scaffolds. Histological analysis of uni- and multidirectional ACT showed that cells differ in their alignment and morphology. This SPR-methodology led to nanoscale detection of paracrine factors and might be useful to study biomarkers of three-dimensional cell growth, cell differentiation, and wound-healing progression.
Assuntos
Colágeno Tipo I/química , Tecido Conjuntivo/química , Mucosa Bucal/química , Proteínas/análise , Meios de Cultivo Condicionados , Ensaio de Imunoadsorção Enzimática , Fibroblastos/química , Mucosa Bucal/citologia , Ressonância de Plasmônio de SuperfícieRESUMO
BACKGROUND: Keratocystic odontogenic tumor (KCOT) is a benign tumor that arises sporadically or associated with nevoid basal cell carcinoma syndrome (NBCCS). Its locally aggressive behavior contrasts with its cystic histological appearance. To better understand the interaction between tumor cells and the stroma, the present study aimed to evaluate and compare the immunohistochemical expression of matrix metalloproteinases (MMP-1, -2, and -9), the cellular proliferation index (Ki-67), and the presence of myofibroblasts (MFs) in KCOTs. METHODS: Eleven cases of isolated KCOT (G1) and 12 cases of KCOT associated with NBCCS (G2) were selected for an immunohistochemical investigation of the proteins MMP-1, MMP-2, MMP-9, Ki-67, and α-smooth muscle actin (α-SMA) in MFs. A group of 6 pericoronal follicles (G3) was included as a normal odontogenic tissue control. RESULTS: Significant differences between the G3 and G1/G2 groups regarding the expression of MMP-1, MMP-9 (in connective tissue), and Ki-67 were observed. In KCOT, there was a positive correlation between the Ki-67 antigen and MMP-1 and between MFs and MMP-1 in the parenchyma. No statistical differences were found between G1 and G2 groups. CONCLUSIONS: MMP-1, MMP-9, and proliferative activity appear to play important roles in KCOT pathogenesis. The increased proliferative activity with KCOT was associated with elevated MMP-1 production in the parenchyma, which influenced the growth of the lesion in association with an increased number of MFs.
Assuntos
Saco Dentário/química , Antígeno Ki-67/análise , Metaloproteinase 1 da Matriz/análise , Metaloproteinase 2 da Matriz/análise , Metaloproteinase 9 da Matriz/análise , Miofibroblastos/patologia , Tumores Odontogênicos/química , Actinas/análise , Síndrome do Nevo Basocelular/metabolismo , Síndrome do Nevo Basocelular/patologia , Proliferação de Células , Tecido Conjuntivo/química , Tecido Conjuntivo/patologia , Saco Dentário/patologia , Epitélio/química , Epitélio/patologia , Humanos , Imuno-Histoquímica , Queratinas/análise , Tumores Odontogênicos/patologia , Células Estromais/química , Células Estromais/patologiaRESUMO
BACKGROUND: We have demonstrated the induction of perlecan-rich stroma of oral squamous cell carcinoma (SCC) on and after its start of invasion. However, it remains unknown how such a neoplastic stroma is actually arranged in tumor tissues. METHODS: To this end, tissue microarray samples, in which keratin and perlecan were contrastively labeled by immunohistochemistry, were three-dimensionally analyzed using digital images and image analysis software to demonstrate the relationship between SCC foci and the perlecan-positive stromal space or that between carcinoma in situ (CIS) and invasive SCC foci. RESULTS: The three-dimensional (3D) reconstruction demonstrated three kinds of perlecan profiles for inside (I) and outside (O) areas of the carcinoma cell focus: mode 1, I(+)/O(-) ; mode 2, I(+)/O(+) ; and mode 3, I(-)/O(+). Mode 1 was seen in CIS as well as SCC tumor massifs in the surface part. Mode 2 was seen in small SCC foci, which seemed isolated in 2D sections but were mostly continuous with the tumor massif in 3D reconstructions. Mode 3 was limited to small SCC foci, which were truly segregated from the tumor massif. CONCLUSIONS: The results indicated that the 2D SCC focus isolation could not be regarded as invasion but that the SCC foci surrounded by perlecan-positive stroma (modes 2 and 3) could be regarded as a more objective measure for invasion of SCC. This is the first 3D tissue-level demonstration of the neoplastic stroma space induced with oral SCC invasion, the presence of which we have predicted based on our previous 2D and tissue culture evidence.
Assuntos
Carcinoma de Células Escamosas/química , Proteoglicanas de Heparan Sulfato/química , Processamento de Imagem Assistida por Computador/métodos , Imageamento Tridimensional/métodos , Neoplasias da Língua/química , Idoso , Idoso de 80 Anos ou mais , Carcinoma in Situ/química , Carcinoma in Situ/patologia , Carcinoma de Células Escamosas/patologia , Tecido Conjuntivo/química , Tecido Conjuntivo/patologia , Epitélio/química , Epitélio/patologia , Feminino , Humanos , Imuno-Histoquímica , Queratina-13/análise , Queratina-17/análise , Queratina-19/análise , Antígeno Ki-67/análise , Masculino , Pessoa de Meia-Idade , Invasividade Neoplásica , Lesões Pré-Cancerosas/química , Lesões Pré-Cancerosas/patologia , Análise Serial de Tecidos , Neoplasias da Língua/patologiaRESUMO
In the 40 years since Elsdale and Bard's analysis of fibroblast culture in collagen gels we have moved far beyond the concept that such 3D fibril network systems are better models than monolayer cultures. This review analyses key aspects of that progression of models, against a background of what exactly each model system tries to mimic. This story tracks our increasing understanding of fibroblast responses to soft collagen gels, in particularly their cytoskeletal contraction, migration and integrin attachment. The focus on fibroblast mechano-function has generated models designed to directly measure the overall force generated by fibroblast populations, their reaction to external loads and the role of the matrix structure. Key steps along this evolution of 3D collagen models have been designed to mimic normal skin, wound repair, tissue morphogenesis and remodelling, growth and contracture during scarring/fibrosis. As new models are developed to understand cell-mechanical function in connective tissues the collagen material has become progressively more important, now being engineered to mimic more complex aspects of native extracellular matrix structure. These have included collagen fibril density, alignment and hierarchical structure, controlling material stiffness and anisotropy. But of these, tissue-like collagen density is key in that it contributes to control of the others. It is concluded that across this 40 year window major progress has been made towards establishing a family of 3D experimental collagen tissue-models, suitable to investigate normal and pathological fibroblast mechano-functions.
Assuntos
Colágeno/química , Células do Tecido Conjuntivo/química , Tecido Conjuntivo/química , Animais , Anisotropia , Células Cultivadas , Células do Tecido Conjuntivo/citologia , Matriz Extracelular/química , Fibroblastos/química , Fibroblastos/citologia , Géis , Humanos , Integrinas/química , Integrinas/metabolismo , Estresse MecânicoRESUMO
Collagens are the most abundant components of the extracellular matrix and many types of soft tissues. Elastin is another major component of certain soft tissues, such as arterial walls and ligaments. Many other molecules, though lower in quantity, function as essential components of the extracellular matrix in soft tissues. Some of these are reviewed in this chapter. Besides their basic structure, biochemistry and physiology, their roles in disorders of soft tissues are discussed only briefly as most chapters in this volume deal with relevant individual compounds. Fibronectin with its muldomain structure plays a role of "master organizer" in matrix assembly as it forms a bridge between cell surface receptors, e.g., integrins, and compounds such collagen, proteoglycans and other focal adhesion molecules. It also plays an essential role in the assembly of fibrillin-1 into a structured network. Laminins contribute to the structure of the extracellular matrix (ECM) and modulate cellular functions such as adhesion, differentiation, migration, stability of phenotype, and resistance towards apoptosis. Though the primary role of fibrinogen is in clot formation, after conversion to fibrin by thrombin, it also binds to a variety of compounds, particularly to various growth factors, and as such fibrinogen is a player in cardiovascular and extracellular matrix physiology. Elastin, an insoluble polymer of the monomeric soluble precursor tropoelastin, is the main component of elastic fibers in matrix tissue where it provides elastic recoil and resilience to a variety of connective tissues, e.g., aorta and ligaments. Elastic fibers regulate activity of TGFßs through their association with fibrillin microfibrils. Elastin also plays a role in cell adhesion, cell migration, and has the ability to participate in cell signaling. Mutations in the elastin gene lead to cutis laxa. Fibrillins represent the predominant core of the microfibrils in elastic as well as non-elastic extracellular matrixes, and interact closely with tropoelastin and integrins. Not only do microfibrils provide structural integrity of specific organ systems, but they also provide a scaffold for elastogenesis in elastic tissues. Fibrillin is important for the assembly of elastin into elastic fibers. Mutations in the fibrillin-1 gene are closely associated with Marfan syndrome. Fibulins are tightly connected with basement membranes, elastic fibers and other components of extracellular matrix and participate in formation of elastic fibers. Tenascins are ECM polymorphic glycoproteins found in many connective tissues in the body. Their expression is regulated by mechanical stress both during development and in adulthood. Tenascins mediate both inflammatory and fibrotic processes to enable effective tissue repair and play roles in pathogenesis of Ehlers-Danlos, heart disease, and regeneration and recovery of musculo-tendinous tissue. One of the roles of thrombospondin 1 is activation of TGFß. Increased expression of thrombospondin and TGFß activity was observed in fibrotic skin disorders such as keloids and scleroderma. Cartilage oligomeric matrix protein (COMP) or thrombospondin-5 is primarily present in the cartilage. High levels of COMP are present in fibrotic scars and systemic sclerosis of the skin, and in tendon, especially with physical activity, loading and post-injury. It plays a role in vascular wall remodeling and has been found in atherosclerotic plaques as well.
Assuntos
Tecido Conjuntivo/química , Tecido Conjuntivo/ultraestrutura , Matriz Extracelular/química , Matriz Extracelular/ultraestrutura , Proteínas de Ligação ao Cálcio/fisiologia , Proteínas de Ligação ao Cálcio/ultraestrutura , Tecido Conjuntivo/metabolismo , Tecido Conjuntivo/fisiopatologia , Elastina/fisiologia , Elastina/ultraestrutura , Matriz Extracelular/metabolismo , Fibrilina-1 , Fibrilinas , Fibrinogênio/fisiologia , Fibrinogênio/ultraestrutura , Fibronectinas/fisiologia , Fibronectinas/ultraestrutura , Humanos , Laminina/fisiologia , Laminina/ultraestrutura , Proteínas dos Microfilamentos/fisiologia , Proteínas dos Microfilamentos/ultraestrutura , Ligação Proteica , Estrutura Secundária de Proteína , Estrutura Terciária de Proteína , Tenascina/fisiologia , Tenascina/ultraestrutura , Trombospondinas/fisiologia , Trombospondinas/ultraestruturaRESUMO
Connective tissue is an important component of meat products that provides support to animal muscles. Hydrogels are considered a promising alternative to connective tissues and simulate actual products by adjusting the gel texture and mouthfeel. This study used soybean protein isolate (SPI), corn starch (CS), konjac glucomannan (KGM), and seaweed powder (SP) as raw materials to examine the effect of different added SP and KGM concentrations on the gel texture. The G' of the gel increased five-fold when the SP and KGM concentration was increased from 1 % to 3 %. The results of mechanical property tests showed that with the addition of SP, the gel hardness increased from 316.00 g to 1827.23 g and the tensile strength increased from 0.027 MPa to 0.089 MPa. Sensory evaluation showed that the samples with 2 % SP and KGM presented the highest overall acceptability score and the most significant similarity to real connective tissue. The connective tissue simulants exhibited excellent water-holding capacity (>90 %), significantly increasing their juiciness. SEM indicated that 2 % KGM addition improved gel network structure stability. The results demonstrate the potential of seaweed polysaccharide-derived hydrogels as connective tissue mimics. This provides a new strategy for the preparation of high mechanical strength hydrogels and lays the foundation for structural diversification of plant-based meat.