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1.
J Chem Phys ; 158(24)2023 Jun 28.
Artigo em Inglês | MEDLINE | ID: mdl-37347137

RESUMO

This work follows a companion article, which will be referred to as Paper I [Campeggio et al., J. Chem. Phys. 158, 244104 (2023)] in which a quantum-stochastic Liouville equation for the description of the quantum-classical dynamics of a molecule in a dissipative bath has been formulated in curvilinear internal coordinates. In such an approach, the coordinates of the system are separated into three subsets: the quantum coordinates, the classical relevant nuclear degrees of freedom, and the classical irrelevant (bath) coordinates. The equation has been derived in natural internal coordinates, which are bond lengths, bond angles, and dihedral angles. The resulting equation needs to be parameterized. In particular, one needs to compute the potential energy surfaces, the friction tensor, and the rate constants for the nonradiative jumps among the quantum states. While standard methods exist for the calculation of energy and dissipative properties, an efficient evaluation of the transition rates needs to be developed. In this paper, an approximated treatment is introduced, which leads to a simple explicit formula with a single adjustable parameter. Such an approximated expression is compared with the exact calculation of transition rates obtained via molecular dynamics simulations. To make such a comparison possible, a simple sandbox system has been used, with two quantum states and a single internal coordinate (together with its conjugate momentum). Results show that the adjustable parameter, which is an effective decoherence time, can be parameterized from the effective relaxation times of the autocorrelation functions of the conjugated momenta of the relevant nuclear coordinates.


Assuntos
Simulação de Dinâmica Molecular , Teoria Quântica , Eletrônica , Movimento (Física)
2.
Pharmacogenomics J ; 10(5): 408-17, 2010 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-20029385

RESUMO

Atypical antipsychotics (APDs) are currently used in clinical practice for a variety of mental disorders such as schizophrenia, bipolar disorder and severe behavioral disturbances. A well-known disadvantage of using these compounds is a propensity for weight gain, resulting frequently in obesity. The mechanisms underlying pharmacologically induced weight gain are still controversial. The objective of this study was to evaluate in vitro the effects of different APDs on adipogenic events in cultured human pre-adipocytes and in rat muscle-derived stem cells (MDSCs), aiming to identify a common intracellular event contributable to these drugs. Culture behavior was evaluated in terms of cell proliferation, lipid accumulation, gene expression and morphological features. Results indicate that APDs influence adipogenic events through changes in the differentiation and proliferation of pre-adipocytes and MDSCs that are brought on by protein kinase C-ß (PKC-ß) activation. These data identify a signaling route that could be a potential target of pharmacological approaches for preventing the weight gain associated with APD treatment.


Assuntos
Adipócitos/efeitos dos fármacos , Adipogenia/efeitos dos fármacos , Antipsicóticos/efeitos adversos , Proteína Quinase C/metabolismo , Células-Tronco/efeitos dos fármacos , Aumento de Peso/efeitos dos fármacos , Adipócitos/citologia , Adipócitos/enzimologia , Adipogenia/genética , Animais , Animais Recém-Nascidos , Técnicas de Cultura de Células , Diferenciação Celular/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Células Cultivadas , Ativação Enzimática , Expressão Gênica/efeitos dos fármacos , Inativação Gênica/efeitos dos fármacos , Proteína Quinase C/antagonistas & inibidores , Proteína Quinase C/genética , Proteína Quinase C beta , RNA Interferente Pequeno/genética , RNA Interferente Pequeno/farmacologia , Ratos , Células-Tronco/citologia , Células-Tronco/enzimologia , Aumento de Peso/genética
3.
J Mater Sci Mater Med ; 20(1): 235-47, 2009 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-18758917

RESUMO

The present study describes the production of hyaluronan based porous microparticles by a semi-continuous gas anti-solvent (GAS) precipitation process to be used as a growth factor delivery system for in vivo treatment of ulcers. Operative process conditions, such as pressure, nozzle diameter and HYAFF11 solution concentrations, were adjusted to optimize particle production in terms of morphology and size. Scanning electron microscopy (SEM) and light scattering demonstrated that porous nano-structured particles with a size of 300 and 900 nm had a high specific surface suitable for absorption of growth factors from the aqueous environment within the polymeric matrix. Water acted as a plasticizer, enhancing growth factor absorption. Water contents within the HYAFF11 matrix were analyzed by differential scanning calorimetry (DSC). The absorption process was developed using fluorescence dyes and growth factors. Immunohistochemical analysis confirmed the high efficiency of absorption of growth factor and a mathematical model was generated to quantify and qualify the in vitro kinetics of growth factor release within the polymeric matrix. In vivo experiments were performed with the aim to optimize timed and focal release of PDGF to promote optimal tissue repair and regeneration of full-thickness wounds.


Assuntos
Materiais Biocompatíveis/química , Substâncias de Crescimento/administração & dosagem , Ácido Hialurônico/química , Nanopartículas/química , Nanopartículas/uso terapêutico , Úlcera Cutânea/terapia , Absorção , Animais , Masculino , Teste de Materiais , Microscopia Eletrônica de Varredura , Nanopartículas/ultraestrutura , Nanotecnologia/instrumentação , Fator de Crescimento Derivado de Plaquetas/administração & dosagem , Ratos , Ratos Wistar , Úlcera Cutânea/tratamento farmacológico , Úlcera Cutânea/patologia , Alicerces Teciduais , Fator de Crescimento Transformador beta/administração & dosagem , Água/química , Cicatrização/efeitos dos fármacos
4.
Eur J Histochem ; 51 Suppl 1: 1-8, 2007.
Artigo em Inglês | MEDLINE | ID: mdl-17703587

RESUMO

Hyaluronan-based scaffold were used for in vitro commitment of human and rat bone marrow mesenchymal stem cells (MSC). Cells were cultured either in monolayer and in 3D conditions up to 35 days. In order to monitor the differentiating processes molecular biology and morphological studies were performed at different time points. All the reported data supported the evidence that both human and rat MSC grown onto hyaluronan-derived three-dimensional scaffold were able to acquire a unique phenotype of chondrocytes and osteocytes depending on the presence of specific differentiation inducing factors added into the culture medium without significative differences in term of time expression of extracellular matrix proteins.


Assuntos
Células-Tronco Adultas/citologia , Condrócitos/citologia , Células-Tronco Mesenquimais/citologia , Células-Tronco Adultas/metabolismo , Células-Tronco Adultas/ultraestrutura , Animais , Células da Medula Óssea , Diferenciação Celular , Proliferação de Células , Células Cultivadas , Condrócitos/metabolismo , Condrogênese/fisiologia , Colágeno Tipo I/genética , Colágeno Tipo I/metabolismo , Colágeno Tipo II/genética , Colágeno Tipo II/metabolismo , Citometria de Fluxo , Humanos , Células-Tronco Mesenquimais/metabolismo , Células-Tronco Mesenquimais/ultraestrutura , Osteocalcina/genética , Osteocalcina/metabolismo , Osteogênese , Osteonectina/genética , Osteonectina/metabolismo , Osteopontina/genética , Osteopontina/metabolismo , Ratos , Ratos Endogâmicos F344 , Reação em Cadeia da Polimerase Via Transcriptase Reversa
5.
J Biomed Mater Res A ; 82(1): 213-21, 2007 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-17266017

RESUMO

In this study we coated a new biocompatible, nanostructured titanium alloy, Ti13Nb13Zr, with a thin layer of hydroxyapatite nanocrystals and we investigated the response of human bone-marrow-derived mesenchymal cells. The coating was realized using a slightly supersaturated CaP solution, which provokes a fast deposition of nanocrystalline hydroxyapatite. A thin layer of deposition is appreciable on the etched Ti13Nb13Zr substrates after just 1.5 h soaking in the CaP solution, and it reaches a thickness of 1-2 mum after 3 h soaking. The coating seems thinner than that deposited on Ti6Al4V, which was examined for comparison, likely because of the different roughness profiles of the two etched alloys, and it is constituted of elongated HA nanocrystals, with a mean length of about 100 nm. Mesenchymal stem cells were seeded onto coated and uncoated Ti alloys and cultured for up to 35 days. Cell morphology, proliferation and differentiation were evaluated. The cells display good adhesion and proliferation on the uncoated substrates, whereas the presence of hydroxyapatite coating slightly reduces cell proliferation and induces differentiation of MSCs towards a phenotypic osteoblastic lineage, in agreement with the increase of the expression of osteopontin, osteonectin and collagen type I, evaluated by means of rt-PCR. Type I collagen expression is higher in Ti13Nb13Zr MSC culture compared to Ti6Al4V, standing for a more efficient extracellular matrix deposition.


Assuntos
Materiais Revestidos Biocompatíveis/química , Durapatita/química , Células-Tronco Mesenquimais/citologia , Nanopartículas/química , Titânio/química , Ligas/química , Sequência de Bases , Células da Medula Óssea/citologia , Células da Medula Óssea/metabolismo , Adesão Celular , Técnicas de Cultura de Células/métodos , Diferenciação Celular , Proliferação de Células , Colágeno Tipo I/genética , Primers do DNA/genética , Expressão Gênica , Humanos , Teste de Materiais , Células-Tronco Mesenquimais/metabolismo , Microscopia Eletrônica de Varredura , Osteoblastos/citologia , Osteoblastos/metabolismo , Osteonectina/genética , Osteopontina/genética
6.
Eur J Vasc Endovasc Surg ; 32(4): 411-7, 2006 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-16678455

RESUMO

OBJECTIVE: To evaluate a new Hyaluronan-based graft. MATERIAL AND METHODS: Hyaluronan-based grafts (HYAFF 11trade mark tube, diameter 2 mm, length 1.5 cm) were implanted in an end-to-end fashion in the abdominal aorta of 15 rats. Histology, immunohistochemistry and electron microscopy were used to evaluate the results at 7, 21, and 90 days. RESULTS: At day 7, new tissue was observed in the graft coming from both the proximal and distal ends of the aorta. The luminal surface of the regenerating tissue was covered by endothelial cells (CD34(+), VEGFR-2(+), vWF(+)). At day 21, regenerating tissue joined at the centre of the tube. The neo-vessel was formed by smooth muscle cells (Myosin Light Chain Kinase) as well as elastic, and collagen fibres. At day 90 a stable artery segment was formed and the biomaterial was almost completely degraded. Infiltration of neutrophils and lymphocytes was not observed. All animals survived the observation period and there were no signs of stenoses or aneurysms. CONCLUSION: The hyaluronan-based graft allowed complete regeneration of a newly formed vascular tube in which all the cellular and extracellular components are present and organized in a well defined architecture similar to native artery.


Assuntos
Implantes Absorvíveis , Prótese Vascular , Ácido Hialurônico , Engenharia Tecidual , Animais , Aorta Abdominal/cirurgia , Artérias/cirurgia , Materiais Biocompatíveis , Implante de Prótese Vascular , Células Cultivadas , Endotélio Vascular/citologia , Masculino , Ratos , Ratos Sprague-Dawley
7.
Virus Res ; 114(1-2): 1-5, 2005 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-15990190

RESUMO

With the aim to detect what kind of cells, in addition to erythroid progenitors, could be involved in the pathogenesis of B19 infection in some connective tissue diseases, primary cultures of human fibroblasts (HF) and endothelial cells (HUVEC) were exposed to a B19 positive serum (350 genome copies/cell). The presence of NS1 and VP1 mRNA, in both HF and HUVEC cultures 1, 2 and 6 days after the exposure, indicated infection by B19 virus. However, no significant increase of B19 DNA level in the infected HF and HUVEC cultures was detectable through the entire incubation period of 6 days. It is possible that HF and HUVEC are not permissive for B19 virus replication or, alternatively, that few cells only get infected by B19 virus. HF and HUVEC stimulation with different growth factors or cytokines could be required for a B19 productive infection to occur.


Assuntos
Células Endoteliais/virologia , Fibroblastos/virologia , Parvovirus B19 Humano/patogenicidade , Células Cultivadas , DNA Viral/análise , Humanos , Infecções por Parvoviridae/microbiologia , Parvovirus B19 Humano/genética , Parvovirus B19 Humano/isolamento & purificação , RNA Mensageiro/metabolismo , RNA Viral/metabolismo , Pele/citologia , Veias Umbilicais/citologia , Proteínas Virais/genética , Proteínas Virais/metabolismo
8.
Biomaterials ; 26(34): 7038-45, 2005 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-15993941

RESUMO

Tissue engineering is a promising approach to developing hepatic tissue suitable for the functional replacement of a failing liver. The aim of the present study was to investigate whether an extracellular cell matrix obtained from fibroblasts-cultured within scaffolds of hyaluronic acid (HYAFF) could influence the proliferation rate and survival of rat hepatocytes both during long-term culture and after in vivo transplantation. Cultures were evaluated by histological and morphological analysis, a proliferation assay and metabolic activity (albumin secretion). Hepatocytes cultured in extracellular matrix-enriched scaffolds exhibited a round cellular morphology and re-established cell-cell contacts, growing into aggregates of several cells along and/or among fibers in the fabric. Hepatocytes were able to secrete albumin up to 14 days in culture. In vivo results demonstrated the biocompatibility of HYAFF-11 implanted in nude mice, in which hepatocytes maintained small well-organised aggregates until the 35th day. In conclusion, the presence of a fibroblast-secreted extracellular matrix improved the biological properties of the hyaluronan scaffold, favoring the survival and morphological integrity of hepatocytes in vitro and in vivo.


Assuntos
Matriz Extracelular/metabolismo , Fibroblastos/metabolismo , Hepatócitos/citologia , Hepatócitos/transplante , Ácido Hialurônico/química , Fígado Artificial , Engenharia Tecidual/métodos , Animais , Materiais Biocompatíveis/química , Técnicas de Cultura de Células/métodos , Proliferação de Células , Sobrevivência Celular/fisiologia , Células Cultivadas , Hepatócitos/fisiologia , Humanos , Teste de Materiais , Camundongos , Camundongos SCID , Ratos , Ratos Wistar
9.
Osteoarthritis Cartilage ; 11(3): 208-16, 2003 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-12623292

RESUMO

OBJECTIVE: To identify the CD44-receptor-mediated effects of 5-7 x 10(5)MW hyaluronan (HA, Hyalgan) on cell viability in normal and damaged human chondrocyte primary cultures isolated from articular cartilage. DESIGN: Primary cultures of human chondrocytes were established from normal articular biopsies and expanded to the second culture passage. The dose-response effects of HA on the viability of normal cultures were identified. Chondrocytes were then treated with either hypoxanthine (2 mM) and xanthine oxidase (20-60 mU), or with activated polymorphonuclear leukocytes (PMNs) to induce injury. Damaged and control cells were then treated with 5-7 x 10(5)HA in the previously identified optimal dose of 0.05 mg/ml. Viability was assessed at specific time periods for the chemically and PMN-damaged cells. To identify if HA effects were mediated by the CD44 receptor, chondrocytes were incubated with anti-CD44 antibody at saturating concentrations (5 microg/ml for 100,000 cells) to produce a maximum inhibition of HA binding. Cells were evaluated using the MTT viability assay, histology, electron microscopy and immunohistochemistry. RESULTS: Direct addition of HA (optimal dose, 0.5 mg/ml) significantly increased cell survival in normal chondrocyte primary cultures (P<0.05). Similarly, addition of this same dose of HA to cultures of free radical-damaged chondrocytes, restored the viability to baseline conditions. Cell viability rates dropped significantly (P<0.05) when CD44 receptor binding was inhibited, indicating that cell growth was mediated by the CD44 receptor. CONCLUSIONS: HA (0.5 mg/ml of 5-7 x 10(5)) significantly increased the viability of normal human chondrocytes in primary culture and restored cell viability to near normal levels after oxidative cell injury.


Assuntos
Condrócitos/efeitos dos fármacos , Receptores de Hialuronatos/metabolismo , Ácido Hialurônico/farmacologia , Cartilagem Articular/metabolismo , Sobrevivência Celular/efeitos dos fármacos , Células Cultivadas , Relação Dose-Resposta a Droga , Humanos , Hipoxantina/farmacologia , Imuno-Histoquímica/métodos , Microscopia Eletrônica , Neutrófilos/fisiologia , Xantina Oxidase/farmacologia
10.
Biomaterials ; 24(7): 1205-11, 2003 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-12527261

RESUMO

Experiences coming from many cell-culture studies has brought about the concept that tissue and organ reconstruction should be performed in a three-dimensional environment as it normally occurs in vivo. As far as endothelial cell culture is concerned, it has been shown that angiogenesis can be successfully achieved only when cells are cultured in the presence of collagen-based matrices or basal membrane substrates. The aim of the present investigation is to demonstrate that human umbilical vein endothelial cells (HUVEC) can be grown and differentiated on an artificial dermis obtained by fibroblasts cultured on hyaluronic acid-based scaffolds. For this purpose, we have cultured HUVEC, retrieved by collagenase digestion of perfused human umbilical vein either alone and with fibroblast at 1/1 ratio into HYAFF-11 non-woven mesh. Cultures were maintained for up to 3 weeks. Samples were taken at different time points within this period for the MTT proliferation test and for immunohistochemical analysis. Our results demonstrate that hyaluronan-based biomaterials (HYAFF-11 NW mesh) represent a suitable substrate for HUVEC adhesion, proliferation and reorganization in microcapillary network.


Assuntos
Materiais Biocompatíveis , Endotélio Vascular/fisiologia , Fibronectinas/fisiologia , Pele Artificial , Veias Umbilicais/citologia , Adesão Celular/efeitos dos fármacos , Técnicas de Cultura de Células , Colágeno/análogos & derivados , Endotélio Vascular/citologia , Fibronectinas/biossíntese , Fibronectinas/farmacologia , Humanos , Queratinócitos/citologia
11.
J Mater Sci Mater Med ; 14(8): 727-9, 2003 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-15348416

RESUMO

In this study we report a preliminary investigation of the feasibility of non-woven/sponge fabrics of a hyaluronan derived biomaterials (benzyl ester of HA (HYAFF-11 FAB, Abano Terme, Italy) for the in vitro culture of rat hepatocytes and rat beta cells. Cell growth on hyaluronan derived biomaterials were tested in the presence of complete medium and in the presence of ECM (extracellular matrix) secreted by fibroblasts previously cultured into the scaffold. Hepatocytes and beta cells were extracted from rat liver/pancreas and seeded either on the HYAFF-11 scaffold alone, or on HYAFF-11 scaffold containing ECM. Direct assay of cell proliferation was performed with MTT test. For morphological observations samples were stained with hematoxylin and eosin. The results obtained by MTT test showed that hepatocytes cultivated in both the above described conditions were able to proliferate up to 14 days and Langerhans islet up to 21 days. After this time, cells started to undergo apoptosis. The morphological analyses showed cell aggregation in three-dimensional structures promoted by the fibers of the biomaterial. Our results confirmed that HYAFF-11 meshes represent a suitable scaffold for hepatocyte adhesion/Langerhans islet organization and proliferation. In particular, the presence of a fibroblast secreted extracellular matrix improves the biological property of the scaffold.

12.
Clin Exp Rheumatol ; 20(4): 445-54, 2002.
Artigo em Inglês | MEDLINE | ID: mdl-12175098

RESUMO

OBJECTIVE: Intraarticular injection of native hyaluronan (HA) or a cross-linked derivative are commonly utilized in the treatment of osteoarthritis. Unlike from native hyaluronan, the crosslinked HA derivative is a gel containing also other chemical entities. This study compares the local tolerability of these different preparations in normal rabbit knees, in order to provide further information on their biological effects. METHODS: Synovial fluids were aspirated after single or repeated weekly injections (up to three) of the therapeutic agents and cell count was determined in a Burker chamber and in an automatic cell counter. The percentage of the different cell types was determined by light microscopy in semithin sections of fixed synovial fluid cytocentrifugate. Fragments of synovial membrane were also morphologically analyzed. RESULTS: In the synovial membrane no signs of inflammation were evident either after a single or repeated injections of native Hyaluronan (Hyalgan or Artz). In addition, the cell recruitment and the percentage of cell types in the synovial fluid was not statistically different from saline treated joints. After 3 weekly injections of the crosslinked HA derivative (Hylan G-F20, Synvisc) about 50% of the treated joints appeared slightly inflamed and in these joints a statistically significantly higher cell content was determined in the synovial fluid compared to placebo and native Hyaluronan treatment. In addition an unexpectedly high percentage of eosinophils was found in the synovial fluid and in the synovial membrane of slightly inflamed joints treated with crosslinked HA. CONCLUSION: The data obtained after repeated intra-articular injections in normal rabbit knee joints confirm the safety profile of native Hyaluronan.


Assuntos
Adjuvantes Imunológicos/administração & dosagem , Ácido Hialurônico/administração & dosagem , Articulação do Joelho/efeitos dos fármacos , Animais , Contagem de Células , Eosinófilos/efeitos dos fármacos , Eosinófilos/patologia , Ácido Hialurônico/análogos & derivados , Injeções Intra-Articulares , Articulação do Joelho/patologia , Coelhos , Líquido Sinovial/citologia , Membrana Sinovial/efeitos dos fármacos , Membrana Sinovial/patologia
13.
Biomaterials ; 21(21): 2183-91, 2000 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-10985492

RESUMO

The objective of the present study was to demonstrate the safety and efficacy of a dermal replacement for cutaneous wounds of diverse origin. Autologous fibroblasts were cultured in fleece scaffolds made from benzyl esters of hyaluronic acid and applied onto cutaneous lesions. The cases presented are (1) skin removal for multiple epithelioma and (2) chronic deep decubitus ulcer. Dermal-like tissue applied by the surgeon elicited no adverse reactions, and was fully integrated and well-vascularized by 1-3 weeks. In Case 1, the material was fully integrated after 1 week, and after 3 weeks an epidermal autograft was overlaid which showed good take with excellent integration observed after 4 weeks. At 12 months, skin demonstrated visual normo-elastic properties and no signs of excessive scarring. In Case 2, 2-3 weeks after the dermal implant was applied, the wound was invaded with granulation tissue and healing occurred by secondary intention. The ulcer was healed by 8 weeks, with the biomaterial completely resorbed and a complete re-epithelialization over the dermal-like tissue. These results suggest that autologous fibroblast culture in hyaluronan-derived scaffolds may be successfully grafted in diverse cutaneous pathologies and constitute a suitable bed for further epidermal implantation.


Assuntos
Implantes Absorvíveis , Materiais Biocompatíveis , Técnicas de Cultura de Células/instrumentação , Ácido Hialurônico/uso terapêutico , Pele Artificial , Biodegradação Ambiental , Carcinoma/cirurgia , Células Cultivadas , Cicatriz/prevenção & controle , Elasticidade , Matriz Extracelular , Neoplasias Faciais/cirurgia , Fibroblastos/transplante , Humanos , Ácido Hialurônico/análogos & derivados , Masculino , Teste de Materiais , Pessoa de Meia-Idade , Projetos Piloto , Complicações Pós-Operatórias/cirurgia , Úlcera por Pressão/cirurgia , Segurança , Transplante de Pele , Transplante Autólogo , Cicatrização
14.
J Biomed Mater Res ; 50(2): 101-9, 2000 May.
Artigo em Inglês | MEDLINE | ID: mdl-10679672

RESUMO

The tolerability and safety of hyaluronan-based three-dimensional scaffolds as a culture vehicle for mesenchymal progenitor cells was investigated in this pilot study. The proliferation patterns and extracellular matrix production of rabbit and human mesenchymal, bone-marrow-derived progenitors first were characterized in vitro. Subsequently rabbit autologous cells were cultured in this hyaluronan-based scaffold and implanted in a full-thickness osteochondral lesion. In vitro histologic findings showed that mesenchymal progenitor cells adhered and proliferated onto the hyaluronan-derived scaffold. Human stem cells were shown to produce the main extracellular matrix molecules, accompanied by an occasional synthesis of mature type II collagen. In vivo data demonstrated that the biomaterial, with or without mesenchymal progenitors, did not elicit any inflammatory response and was completely degraded within 4 months after implantation. With regard to the efficacy of this cell therapy, even among the small number of animals tested there was histologic evidence that lesions filled with the biomaterial, either seeded or unseeded with cells, achieved a faster and better healing compared to empty controls. The present data suggest that the hyaluronan-based scaffolds are well tolerated and safe and may be a valuable delivery vehicle for tissue engineering in the repair of articular cartilage defects.


Assuntos
Materiais Biocompatíveis , Biopolímeros , Células da Medula Óssea/efeitos dos fármacos , Portadores de Fármacos , Sistemas de Liberação de Medicamentos , Ácido Hialurônico , Animais , Humanos , Mesoderma/efeitos dos fármacos , Microesferas , Coelhos
15.
Ostomy Wound Manage ; 46(9): 44-8, 2000 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-11189540

RESUMO

This paper presents a dermal replacement technique for cutaneous wounds of diverse origin. An autologous tissue-engineered dermal-like structure was implanted in one patient with an acute surgical excision of a nevus and in another patient with a chronic post-traumatic ulcer. The dermal constructs were well integrated with the wound area, and, at 3 weeks, an epidermal covering was implanted. In both cases, 3 to 8 weeks after autografting, good integration of the transplanted epidermis with the surrounding native skin and a good take over the dermal-like tissue occurred. The use of this technology, despite its high cost, is increasing. Clinicians choose it because it helps heal critical wounds for which current therapies have failed.


Assuntos
Nevo Pigmentado/cirurgia , Transplante de Pele/métodos , Ferimentos e Lesões/cirurgia , Doença Aguda , Adolescente , Doença Crônica , Fibroblastos/citologia , Fibroblastos/transplante , Humanos , Masculino , Pessoa de Meia-Idade
16.
J Burn Care Rehabil ; 20(4): 282-91, 1999.
Artigo em Inglês | MEDLINE | ID: mdl-10425590

RESUMO

The current study was designed to investigate the biological role of small extracellular matrix fragments in wound healing. Human burn eschar tissue was digested with bacterial collagenase, and small aminoacidic fragments were inoculated in both human dermal fibroblast cultures and polyvinyl alcohol sponges implanted subcutaneously in the rat. Proliferation assays on cell cultures and biochemical and histologic analyses of the animal model were then performed. Results showed that fibroblasts treated with low concentrations of eschar fragments duplicated significantly faster than controls. Biochemical and histologic data from sponge implants showed that the inflammatory response was augmented by eschar-derived fragments at postoperative day 2, whereas protein and hydroxyproline synthesis were decreased at day 14. In conclusion, these data substantiate that the application of bacterial collagenase to débride necrotic tissue may have an indirect healing effect resulting from the local release of bioactive matrix-derived fragments.


Assuntos
Queimaduras/terapia , Proteínas da Matriz Extracelular/farmacologia , Colagenase Microbiana/metabolismo , Cicatrização/fisiologia , Animais , Queimaduras/microbiologia , Divisão Celular , Proteínas da Matriz Extracelular/metabolismo , Fibroblastos/fisiologia , Humanos , Inflamação , Masculino , Colagenase Microbiana/uso terapêutico , Ratos , Ratos Sprague-Dawley
17.
J Biomed Mater Res ; 46(3): 337-46, 1999 Sep 05.
Artigo em Inglês | MEDLINE | ID: mdl-10397990

RESUMO

Articular cartilage has a very limited self-repairing capacity; thus, chondral lesions normally result in chronic degeneration and, eventually, osteoarthritis development. Currently, tissue engineering offers a new tool for the clinical treatment of osteochondral defects. The present investigation aimed to develop an in vitro engineered cartilage using a new class of semisynthetic scaffolds. Two nonwoven meshes of hyaluronan esters (Hyaff(R) derivatives) were seeded with sternal chick embryo chondrocytes cultured for up to 21 days, after which time they were assessed for both the cellular growth profile and histological features. Avian chondrocytes easily adhered and proliferated onto hyaluronan-based scaffolds, demonstrating a significant preference for the fully esterified benzylic form. Histochemical staining revealed the presence of a neosynthesized glycosaminoglycan-rich extracellular matrix, and immunohistochemistry confirmed the deposition of collagen type II. Moreover, ultrastructural observations supported evidence that chondrocytes grown onto a hyaluronan-derived three-dimensional scaffold maintained their unique phenotype and organization in a cartilage-like extracellular matrix. These findings support the further pursuit of a transplantable engineered cartilage using human chondrocytes for the regeneration of chondral lesions.


Assuntos
Engenharia Biomédica , Agregação Celular/fisiologia , Condrócitos/citologia , Ácido Hialurônico , Teste de Materiais , Animais , Biodegradação Ambiental , Células Cultivadas , Embrião de Galinha , Matriz Extracelular , Histocitoquímica , Humanos , Imuno-Histoquímica , Microscopia Eletrônica , Próteses e Implantes
18.
J Mater Sci Mater Med ; 10(10/11): 683-8, 1999.
Artigo em Inglês | MEDLINE | ID: mdl-15347986

RESUMO

Tissue engineering offers the possibility to reconstruct tissue substitutes in order to replace lost or damaged tissues. The availability of appropriate biomaterial devices is essential to allow in vitro cultured cells to behave as in the original tissues in vivo. In our studies we utilized a seminatural biomaterial made up by the benzyl ester of hyaluronan to grow keratinocytes, fibroblasts and chondrocytes. Keratinocytes and fibroblasts were isolated from human foreskin. Cells were separately cultured on two different hyaluronan based biomaterial devices for the first 15 days and then co-cultured for an additional period of 2 weeks. Keratinocytes gave rise to a well-differentiated epithelial layer, while fibroblasts were able to synthesize all the main extracellular molecules inside the biomaterial spaces, forming dermal-like tissues. When these two tissues were co-cultured, a skin equivalent was formed with a dermal-epidermal junction. Chondrocytes were obtained from chick-embryo sterna and cultured for 21 days inside a non-woven scaffolding made up of a hyaluronan-based biomaterial. Cells were able to organize themselves into nodules embedded in a dense metachromatic substance in which type II collagen was present. Data from this study suggest that this novel class of hyaluronan derived biomaterials is suitable for different cell culture and in vitro tissue reconstruction.

19.
J Biomed Mater Res ; 40(2): 187-94, 1998 May.
Artigo em Inglês | MEDLINE | ID: mdl-9549613

RESUMO

Coverage of large, full-thickness burns presents a challenge for the surgeon due to the lack of availability of the patient's own skin. Currently, tissue engineering offers the possibility of performing a suitable therapeutic wound coverage after early burn excision by using cultured keratinocyte sheets supported by a dermal layer. The aim of this study was to develop and characterize a skin substitute composed of both epidermal and dermal elements. For this purpose we grew keratinocytes and fibroblasts separately for 15 days within two different types of biomaterials. Cells then were co-cultured for an additional period of 15 days, after which samples were taken and processed with either classic or immunohistochemical stainings. Results showed that (1) human fibroblasts and keratinocytes can be cultured on hyaluronic acid-derived biomaterials and that (2) the pattern of expression of particular dermal-epidermal molecules is similar to that found in normal skin. The data from this study suggest that our skin equivalent might be useful in the treatment of both burns and chronic wounds.


Assuntos
Pele Artificial , Materiais Biocompatíveis , Queimaduras/terapia , Divisão Celular , Técnicas de Cocultura , Proteínas da Matriz Extracelular/metabolismo , Fibroblastos/citologia , Fibroblastos/metabolismo , Humanos , Imuno-Histoquímica , Queratinócitos/citologia , Queratinócitos/metabolismo
20.
Semin Arthritis Rheum ; 26(1): 492-501, 1996 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-8870117

RESUMO

OBJECTIVE: To compare hyaluronan and its alpha-methyl prednisolone derivative (HYC-141) with respect to their potential to directly cause tissue damage and to protect tissues from attack by reactive oxygen species. METHODS: Cartilage samples and chondrocytes were isolated from 15-day chick embryos and were exposed to enzyme-based and activated inflammatory cell-free radical generating systems in the presence of varying concentrations of alpha-methyl prednisolone, hyaluronan, and HYC-141. Tissue and cell vitality was measured by determining the incorporation of radioactive sulphate into newly synthesized glycosaminoglycans. RESULTS: Only alpha-methyl prednisolone caused a significant decrease in biosynthetic activity. All the tested substances were capable, to some extent, of protecting tissues and cells from damage by reactive oxygen species; HYC-141 demonstrated the greatest protective effect. CONCLUSION: These data suggest that HYC-141 may possess certain advantages over the individual component molecules in the local treatment of arthropathies.


Assuntos
Anti-Inflamatórios/farmacologia , Antioxidantes/farmacologia , Cartilagem Articular/citologia , Ácido Hialurônico/farmacologia , Prednisolona/farmacologia , Animais , Células Cultivadas/efeitos dos fármacos , Embrião de Galinha , Relação Dose-Resposta a Droga , Radicais Livres/metabolismo , Glicoproteínas/biossíntese , Ácido Hialurônico/análogos & derivados , Esterno/citologia , Radioisótopos de Enxofre
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