RESUMO
The ability of human tissues to self-repair is limited, which motivates the scientific community to explore new and better therapeutic approaches to tissue regeneration. The present manuscript provides a comparative study between a marine-based composite biomaterial, and another composed of well-established counterparts for bone tissue regeneration. Blue shark skin collagen was combined with bioapatite obtained from blue shark's teeth (mColl:BAp), while bovine collagen was combined with synthetic hydroxyapatite (bColl:Ap) to produce 3D composite scaffolds by freeze-drying. Collagens showed similar profiles, while apatite particles differed in their composition, being the marine bioapatite a fluoride-enriched ceramic. The marine-sourced biomaterials presented higher porosities, improved mechanical properties, and slower degradation rates when compared to synthetic apatite-reinforced bovine collagen. The in vivo performance regarding bone tissue regeneration was evaluated in defects created in femoral condyles in New Zealand rabbits twelve weeks post-surgery. Micro-CT results showed that mColl:BAp implanted condyles had a slower degradation and an higher tissue formation (17.9 ± 6.9 %) when compared with bColl:Ap implanted ones (12.9 ± 7.6 %). The histomorphometry analysis provided supporting evidence, confirming the observed trend by quantifying 13.1 ± 7.9 % of new tissue formation for mColl:BAp composites and 10.4 ± 3.2 % for bColl:Ap composites, suggesting the potential use of marine biomaterials for bone regeneration.
Assuntos
Materiais Biocompatíveis , Alicerces Teciduais , Humanos , Animais , Coelhos , Bovinos , Materiais Biocompatíveis/uso terapêutico , Apatitas , Regeneração Óssea , Colágeno/farmacologiaRESUMO
Collagen is an important material for biomedical research, but using mammalian tissue-derived collagen carries the risk of zoonotic disease transmission. Marine organisms, such as farmed tilapia, have emerged as a safe alternative source of collagen for biomedical research. However, the tilapia collagen products for biomedical research are rare, and their biological functions remain largely unexamined. In this study, we characterized a commercial tilapia skin collagen using SDS-PAGE and fibril formation assays and evaluated its effects on skin fibroblast adhesion, proliferation, and migration, comparing it with commercial collagen from rat tails, porcine skin, and bovine skin. The results showed that tilapia skin collagen is a type I collagen, similar to rat tail collagen, and has a faster fibril formation rate and better-promoting effects on cell migration than porcine and bovine skin collagen. We also confirmed its application in a 3D culture for kidney cells' spherical cyst formation, fibroblast-induced gel contraction, and tumor spheroid interfacial invasion. Furthermore, we demonstrated that the freeze-dried tilapia skin collagen scaffold improved wound closure in a mouse excisional wound model, similar to commercial porcine or bovine collagen wound dressings. In conclusion, tilapia skin collagen is an ideal biomaterial for biomedical research.
Assuntos
Pesquisa Biomédica , Tilápia , Camundongos , Ratos , Suínos , Animais , Bovinos , Mamíferos , Colágeno/farmacologia , Pele , Modelos Animais de DoençasRESUMO
Osteoporosis is a worldwide disease resulting in the increase of bone fragility and enhanced fracture risk in adults. In the context of osteoporotic fractures, bone tissue engineering (BTE), i.e., the use of bone substitutes combining biomaterials, cells, and other factors, is considered a potential alternative to conventional treatments. Innovative scaffolds need to be tested in in vitro systems where the simultaneous presence of osteoblasts (OBs) and osteoclasts (OCs), the two main players of bone remodeling, is required to mimic their crosstalk and molecular cooperation. To this aim, two composite materials were developed, based on type I collagen, and containing either strontium-enriched mesoporous bioactive glasses or rod-like hydroxyapatite nanoparticles. The developed nanostructured systems underwent genipin chemical crosslinking and were then tested with an indirect co-culture of human trabecular bone-derived OBs and buffy coat-derived OC precursors, for 2-3 weeks. The favorable structural and biological properties of the materials proved to successfully support the viability, adhesion, and differentiation of cells, encouraging a further investigation of the developed bioactive systems as biomaterial inks for the 3D printing of more complex scaffolds for BTE.
Assuntos
Biomimética , Osso e Ossos/citologia , Colágeno/farmacologia , Nanoestruturas/química , Fosfatase Alcalina/metabolismo , Animais , Bovinos , Adesão Celular/efeitos dos fármacos , Morte Celular/efeitos dos fármacos , Diferenciação Celular/efeitos dos fármacos , Forma Celular/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Técnicas de Cocultura , Elasticidade , Humanos , Hidrólise , Osteoclastos/citologia , Osteoclastos/efeitos dos fármacos , Fosfatase Ácida Resistente a Tartarato/metabolismo , ViscosidadeRESUMO
Human induced pluripotent stem cells (hiPSCs) are considered to be one of the most promising cell resources for regenerative medicine. HiPSCs usually maintain their pluripotency when they are cultured on feeder cell layers or are attached to a cell-adhesive extracellular matrix. In this study, we developed a culture system based on UV/ozone modification for conventional cell culture plastics to generate a suitable surface condition for hiPSCs. Time of flight secondary ion mass spectrometry (ToF-SIMS) was carried out to elucidate the relationship between hiPSC adhesion and UV/ozone irradiation-induced changes to surface chemistry of cell culture plastics. Cell culture plastics with modified surfaces enabled growth of a feeder-free hiPSC culture with markedly reduced cell-adhesive matrix coating. Our cell culture system using UV/ozone-modified cell culture plastics may produce clinically relevant hiPSCs at low costs, and can be easily scaled up in culture systems to produce a large number of hiPSCs.
Assuntos
Técnicas de Cultura de Células/economia , Análise Custo-Benefício , Células-Tronco Pluripotentes Induzidas/citologia , Ozônio/farmacologia , Plásticos/farmacologia , Raios Ultravioleta , Fosfatase Alcalina/metabolismo , Adesão Celular/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Proliferação de Células/efeitos da radiação , Células Cultivadas , Colágeno/farmacologia , Combinação de Medicamentos , Humanos , Células-Tronco Pluripotentes Induzidas/efeitos dos fármacos , Células-Tronco Pluripotentes Induzidas/efeitos da radiação , Cariótipo , Laminina/farmacologia , Células-Tronco Pluripotentes/citologia , Células-Tronco Pluripotentes/efeitos dos fármacos , Células-Tronco Pluripotentes/metabolismo , Poliestirenos , Proteoglicanas/farmacologia , Propriedades de SuperfícieRESUMO
OBJECTIVE: Recombinant collagen (rCOL)-hyaluronic acid (HA) composite scaffolds were prepared to thoroughly investigate their biological properties. METHODS: The rCOL and HA composite scaffolds were formulated via lyophilization. The scaffolds were characterized for various materials properties, including porosity, surface modification, and degradation rates. Biological properties such as in vitro cytotoxicity, cell adhesion, proliferation and migration effects were also evaluated. RESULTS: The water absorption, mechanical strength, degradation resistance and thermal stability of the prepared rCOL-HA composites were improved over that of the control studied. Scanning electron microscopy (SEM) revealed that the composites formed a three-dimensional network structure with uniform pore distribution. The cytotoxicity of the composites was minimal (grade I) and the material showed strong adhesion and proliferation effects when grown with mouse fibroblasts, particularly the composite material of rCOL (5% HA) group (P < 0.05). CONCLUSION: The rCOL-HA composite prepared via lyophilization after cross-linking is characterized by high porosity, high water absorption, and good interaction between the material and cells, as well as good biodegradability. Compared with rCOL materials, rCOL-HA has increased mechanical strength, water absorption and thermal stability. The biocompatibility and fibroblast proliferation of rCOL-HA have excellent biological performance, providing a new material for wound healing applications.
Assuntos
Colágeno/farmacologia , Ácido Hialurônico/farmacologia , Proteínas Recombinantes/farmacologia , Alicerces Teciduais/química , Absorção Fisico-Química , Animais , Materiais Biocompatíveis/farmacologia , Varredura Diferencial de Calorimetria , Adesão Celular/efeitos dos fármacos , Linhagem Celular , Movimento Celular/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Colágeno/química , Ácido Hialurônico/química , Camundongos , Porosidade , Espectroscopia de Infravermelho com Transformada de Fourier , Resistência à Tração , Água/químicaRESUMO
During menstruation, endometrial hemostasis is achieved by platelet aggregation, fibrin deposition, and thrombus formation that interact with local endocrine and immunological factors which cause termination of menstrual bleeding. Interactions between steroidal sex hormones and platelet functions are not well understood. The aim of this study was to evaluate the effect of platelet function during the menstrual cycle and luteal phase in women of reproductive age. The cross-sectional study on women of reproductive age included 44 healthy women. Platelet function was assessed by PFA-100TM analyzer with collagen/epinephrine and collagen/ADP cartridges during the menstrual cycle and luteal phase. There were no significant differences in platelet function between menstruation and ovulatory phase. Platelet activity in Arab collagen/epinephrine cartridge increased during menstruation compared to non-Arab ethnic subjects and no significant differences in platelet function were found when using collagen/ADP cartridge. This study suggested modulation in platelet functions during menstruation and luteal phase in women of reproductive age. Further studies, including a large number of subjects, platelet genetic and progesterone factors change in platelet clotting associated to menstrual cycle should be conducted.
Assuntos
Plaquetas/fisiologia , Menstruação/fisiologia , Ovulação/fisiologia , Adulto , Colágeno/farmacologia , Estudos Transversais , Epinefrina/farmacologia , Feminino , Humanos , Pré-Menopausa/fisiologiaRESUMO
Efforts on bioengineering are directed towards the construction of biocompatible scaffolds and the determination of the most favorable microenvironment, which will better support cell proliferation and differentiation. Perfusion bioreactors are attracting growing attention as an effective, modern tool in tissue engineering. A natural biomaterial extensively used in regenerative medicine with outstanding biocompatibility, biodegradability and non-toxic characteristics, is collagen, a structural protein with undisputed beneficial characteristics. This is a study designed according to the above considerations. 3D printed polycaprolactone (PCL) scaffolds with rectangular pores were coated with collagen either as a coating on the scaffold's trabeculae, or as a gel-cell solution penetrating scaffolds' pores. We employed histological, molecular and imaging techniques to analyze colonization, proliferation and chondrogenic differentiation of Adipose Derived Mesenchymal Stem Cells (ADMSCs). Two different differentiation culture media were employed to test chondrogenic differentiation on gelated and non gelated PCL scaffolds in static and in perfusion bioreactors dynamic culture conditions. In dynamic culture, non gelated scaffolds combined with our in house TGF-ß2 based medium, augmented chondrogenic differentiation performance, which overall was significantly less favorable compared to StemPro™ propriety medium. The beneficial mechanical stimulus of dynamic culture, appears to outgrow the disadvantage of the "weaker" TGF-ß2 medium used for chondrogenic differentiation. Even though cells in static culture grew well on the scaffold, there was limited penetration inside the construct, so the purpose of the 3D culture was not fully served. In contrast dynamic culture achieved better penetration and uniform distribution of the cells within the scaffold.
Assuntos
Cartilagem/efeitos dos fármacos , Condrogênese/efeitos dos fármacos , Colágeno/farmacologia , Poliésteres/farmacologia , Engenharia Tecidual/métodos , Alicerces Teciduais , Agrecanas/genética , Agrecanas/metabolismo , Materiais Biocompatíveis , Biomarcadores/metabolismo , Reatores Biológicos , Cartilagem/citologia , Cartilagem/metabolismo , Diferenciação Celular/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Condrócitos/citologia , Condrócitos/efeitos dos fármacos , Condrócitos/metabolismo , Condrogênese/genética , Colágeno/química , Meios de Cultura/química , Meios de Cultura/farmacologia , Expressão Gênica , Humanos , Células-Tronco Mesenquimais/citologia , Células-Tronco Mesenquimais/efeitos dos fármacos , Células-Tronco Mesenquimais/metabolismo , Poliésteres/química , Porosidade , Cultura Primária de Células , Impressão Tridimensional , Regeneração/genética , Fatores de Transcrição SOX9/genética , Fatores de Transcrição SOX9/metabolismo , Propriedades de SuperfícieRESUMO
Pancreatic ß-cells in the islets of Langerhans secrete insulin in response to the rise in glucose levels following food intake. The hypoglycemic action of insulin applies a strong evolutionary brake on ß-cell division. However, under some conditions ß-cells can be stimulated to enter cell cycle progression and divide, for example following exposure to increased glucose levels or during pregnancy. Here, a protocol is described for the isolation of rat adult islets of Langerhans, followed by culture of intact islets in Matrigel and measurement of ß-cell replication by the incorporation of ethynyldeoxyuridine (EdU). EdU positive cells are revealed by a click reaction, nuclei are visualized using a DNA-binding fluorophore (Hoechst 33342), and ß-cells are identified using immunofluorescence detection.
Assuntos
Células Secretoras de Insulina/citologia , Ilhotas Pancreáticas/citologia , Animais , Divisão Celular/efeitos dos fármacos , Divisão Celular/fisiologia , Células Cultivadas , Colágeno/farmacologia , Combinação de Medicamentos , Ingestão de Alimentos/efeitos dos fármacos , Ingestão de Alimentos/fisiologia , Feminino , Glucose/metabolismo , Humanos , Insulina/metabolismo , Células Secretoras de Insulina/efeitos dos fármacos , Células Secretoras de Insulina/metabolismo , Ilhotas Pancreáticas/efeitos dos fármacos , Ilhotas Pancreáticas/metabolismo , Laminina/farmacologia , Gravidez , Proteoglicanas/farmacologia , Ratos , Ratos WistarRESUMO
3D-printing and additive manufacturing can be powerful techniques to design customized structures and produce synthetic bone grafts with multifunctional effects suitable for bone repair. In our work we aimed the development of novel multifunctionalized 3D printed poly(lactic acid) (PLA) scaffolds with bioinspired surface coatings able to reduce bacterial biofilm formation while favoring human bone marrow-derived mesenchymal stem cells (hMSCs) activity. For that purpose, 3D printing was used to prepare PLA scaffolds that were further multifunctionalized with collagen (Col), minocycline (MH) and bioinspired citrate- hydroxyapatite nanoparticles (cHA). PLA-Col-MH-cHA scaffolds provide a closer structural support approximation to native bone architecture with uniform macroporous, adequate wettability and an excellent compressive strength. The addition of MH resulted in an adequate antibiotic release profile that by being compatible with local drug delivery therapy was translated into antibacterial activities against Staphylococcus aureus, a main pathogen associated to bone-related infections. Subsequently, the hMSCs response to these scaffolds revealed that the incorporation of cHA significantly stimulated the adhesion, proliferation and osteogenesis-related gene expression (RUNX2, OCN and OPN) of hMSCs. Furthermore, the association of a bioinspired material (cHA) with the antibiotic MH resulted in a combined effect of an enhanced osteogenic activity. These findings, together with the antibiofilm activity depicted strengthen the appropriateness of this 3D-printed PLA-Col-MH-cHA scaffold for future use in bone repair. By targeting bone repair while mitigating the typical infections associated to bone implants, our 3D scaffolds deliver an integrated strategy with the combined effects further envisaging an increase in the success rate of bone-implanted devices.
Assuntos
Regeneração Óssea/efeitos dos fármacos , Colágeno/farmacologia , Durapatita/farmacologia , Minociclina/farmacologia , Nanopartículas/química , Poliésteres/farmacologia , Impressão Tridimensional , Engenharia Tecidual/métodos , Adsorção , Antibacterianos/farmacologia , Biofilmes/efeitos dos fármacos , Liberação Controlada de Fármacos , Regulação da Expressão Gênica/efeitos dos fármacos , Humanos , Células-Tronco Mesenquimais/citologia , Células-Tronco Mesenquimais/efeitos dos fármacos , Células-Tronco Mesenquimais/metabolismo , Células-Tronco Mesenquimais/ultraestrutura , Testes de Sensibilidade Microbiana , Osseointegração/efeitos dos fármacos , Osteogênese/efeitos dos fármacos , Osteogênese/genética , Staphylococcus aureus/efeitos dos fármacos , Alicerces Teciduais/químicaRESUMO
Chronic wounds (ie, wounds that fail to progress through a normal, orderly, timely sequence of repair) continue to pose significant clinical and economic burdens. A prospective, descriptive, 3-week post-marketing surveillance study was conducted across 3 wound care centers in the United States to evaluate the effectiveness of a collagen calcium alginate dressing on chronic wounds in conjunction with standard care (SC) practices (eg, offloading, debridement, compression) to support healing. Eligible participants had to be >18 years of age, have at least 1 chronic wound, and no known sensitivity to collagen. Demographic characteristics were recorded at the screening visit on case report forms. At each visit, wound-related pain was assessed using the Visual Analog Scale along with wound characteristics including size (using digital planimetry), wound exudate (minimal, moderate, heavy), and odor (none, mild). Participants were monitored for adverse events as well as infection based on signs and symptoms in and around the local wound bed, the deeper structures, and the surrounding skin. An intention-to-treat approach was used for all analyses. If an observation was missing, the last observation carried forward principle was used. For wounds that healed, pain and exudate were set to 0 (no pain/exudate) at visit 4. Descriptive, paired t tests and the Wilcoxon signed rank test were used to analyze the data. Of the 31 participants (15 men, 16 women, mean age 66.6 years), most (13, 42%) had a diabetic foot ulcer or venous leg ulcer (10, 32%); median duration of all wounds was 148 days. Thirty (30) patients completed the study. The mean number of comorbidities was 10.6 ± 6.3, and patients used a mean of 9.3 ± 5.64 prescription or over-the-counter medications. For all wounds combined, mean wound area was 4.8 ± 8.38 cm2 at baseline. At week 3, a decrease in wound area of 38.1% was noted (median: 45% ± 42.54; P = .006); 3 wounds healed completely. The change in wound exudate level from visit 1 to visit 4 was statistically significant (P = .006). No adverse events or infections occurred. In this population, the use of etiology-appropriate SC and a collagen calcium alginate dressing resulted in a decrease in wound area after 3 weeks of care. Longer-term studies to confirm these observations and controlled clinical studies to compare the effects of this dressing to other nongauze dressing treatments are needed.
Assuntos
Alginatos/farmacologia , Bandagens/normas , Doença Crônica/terapia , Cicatrização/efeitos dos fármacos , Idoso , Idoso de 80 Anos ou mais , Alginatos/uso terapêutico , Bandagens/estatística & dados numéricos , Doença Crônica/enfermagem , Colágeno/farmacologia , Colágeno/uso terapêutico , Angiopatias Diabéticas/complicações , Angiopatias Diabéticas/fisiopatologia , Feminino , Humanos , Masculino , Marketing de Serviços de Saúde/métodos , Pessoa de Meia-Idade , Vigilância da População/métodos , Estudos Prospectivos , Estados Unidos , Úlcera Varicosa/complicações , Úlcera Varicosa/fisiopatologia , Escala Visual AnalógicaRESUMO
In tissue engineering applications, vascularization can be accomplished by coimplantation of tissue forming cells and endothelial cells (ECs), whereby the latter are able to form functional blood vessels. The use of three-dimensional (3D) bioprinting technologies has the potential to improve the classical tissue engineering approach because these will allow the generation of scaffolds with high spatial control of endothelial cell allocation. This study focuses on a side by side comparison of popular commercially available bioprinting hydrogels (Matrigel, fibrin, collagen, gelatin, agarose, Pluronic F-127, alginate, and alginate/gelatin) in the context of their physicochemical parameters, their swelling/degradation characteristics, their biological effects on vasculogenesis-related EC parameters and their printability. The aim of this study was to identify the most suitable hydrogel or hydrogel combination for inkjet printing of ECs to build prevascularized tissue constructs. Most tested hydrogels displayed physicochemical characteristics suitable for inkjet printing. However, Pluronic F-127 and the alginate/gelatin blend were rapidly degraded when incubated in cell culture medium. Agarose, Pluronic F-127, alginate and alginate/gelatin hydrogels turned out to be unsuitable for bioprinting of ECs because of their non-adherent properties and/or their incapability to support EC proliferation. Gelatin was able to support EC proliferation and viability but was unable to support endothelial cell sprouting. Our experiments revealed fibrin and collagen to be most suitable for bioprinting of ECs, because these hydrogels showed acceptable swelling/degradation characteristics, supported vasculogenesis-related EC parameters and showed good printability. Moreover, ECs in constructs of preformed spheroids survived the printing process and formed capillary-like cords. © 2017 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 106A: 935-947, 2018.
Assuntos
Bioimpressão/métodos , Células Endoteliais da Veia Umbilical Humana/citologia , Hidrogéis/farmacologia , Animais , Morte Celular/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Colágeno/farmacologia , Elasticidade , Fibrina/farmacologia , Células Endoteliais da Veia Umbilical Humana/efeitos dos fármacos , Células Endoteliais da Veia Umbilical Humana/metabolismo , Humanos , Camundongos , Neovascularização Fisiológica/efeitos dos fármacos , Ratos , Reprodutibilidade dos Testes , Esferoides Celulares/citologia , Esferoides Celulares/efeitos dos fármacos , Tensão Superficial , ViscosidadeRESUMO
The growing interest in the use of recyclable and biodegradable natural materials has become a relevant topic in pharmaceutics. In this work, we suggest the use and valorization of natural horny skeleton of marine sponges (Porifera, Dictyoceratida) as bio-based dressing for topical drug delivery. Biomaterial characterization focusing on morpho-functional traits, swelling behavior, fluid uptake performances, glycosaminoglycans content and composition and microbiological quality assessment was carried out to investigate the collagenic skeleton properties. After grinding and sieving processes, l-cysteine hydrochloride-loaded formulations were designed in form of powder or polymeric film by testing various drug concentrations and different drying parameters. Drug content, SEM analyses and in vitro permeation studies were performed to test the suitability of skeleton-based formulations. To this respect, drying time and temperature are key parameters for skeleton-mediated drug crystallization. Consequently, this behavior seems to influence drug loading and permeation profiles of formulations. The high percentages of drug are found after absorption into sponge powder and in vitro permeation studies demonstrate that cysteine is released more slowly than the pure drug within 1h. Such a system is attractive because it combines the known healing properties of cysteine with the advantageous potentials of the collagen/proteoglycan network, which can act as biocompatible carrier able to absorb the excess of the wound exudate while releasing the drug. Furthermore, due to its glycosaminoglycans content, natural sponge skeletal scaffold might act as bioactive-biomimetic carrier regulating the wound healing processes.
Assuntos
Organismos Aquáticos/química , Materiais Biocompatíveis/farmacologia , Colágeno/farmacologia , Sistemas de Liberação de Medicamentos , Invenções , Poríferos/química , Esqueleto/química , Administração Tópica , Alginatos/farmacologia , Animais , Colágeno/ultraestrutura , Cisteína/análise , Ácido Glucurônico/farmacologia , Glicosaminoglicanos/análise , Ácidos Hexurônicos/farmacologia , Microscopia Eletrônica de Varredura , Permeabilidade , Pós , Esterilização , Água/químicaRESUMO
AIM: To develop a biological scaffold that could be moulded to reproduce the geometry of a gutta-percha point with precision and allow the differentiation of mesenchymal stem cells into osteoblasts to be used as a regenerative endodontic material. METHODOLOGY: A collagen/alginate composite scaffold was cast into a sodium alginate mould to produce a gutta-percha point-like cone. Prior to gelation, the cone was seeded with human stem cells from the apical papilla (SCAPs) to evaluate cell/scaffold interactions. The reconstructed tissue was characterized after 8 days in culture. Elastic modulus, tissue compaction and cell differentiation were assessed. Student t-tests and the Mann-Whitney U test were performed. RESULTS: The fabrication method developed enabled the shape of a gutta-percha point to be mimicked with great accuracy and reproducibility (P = 0.31). Stem cells seeded into this composite scaffold were able to spread, survive and proliferate (P < 0.001). Moreover, they were able to differentiate into osteoblasts and produce calcified osseous extracellular matrix (P < 0.001). The construct showed no significant contraction after 8 days, preserving its shape and tip diameter (P = 0.58). CONCLUSIONS: The composite scaffold could present substantial benefits compared to synthetic materials. It could provide a favourable healing environment in the root canal conducive for regenerative endodontics and is therefore appropriate to be evaluated in vivo in further studies.
Assuntos
Alginatos/farmacologia , Diferenciação Celular/efeitos dos fármacos , Colágeno/farmacologia , Células-Tronco Mesenquimais/citologia , Osteoblastos/citologia , Alicerces Teciduais/química , Ácido Glucurônico/farmacologia , Guta-Percha , Ácidos Hexurônicos/farmacologia , Humanos , RegeneraçãoRESUMO
OBJECTIVE: To histomorphometrically compare the use of collagen-stabilized anorganic bovine bone (ABBM-C) (test) to anorganic bovine bone + autogenous bone (ABBM + AB) (control) in maxillary sinus augmentation. MATERIALS AND METHODS: Nine sheep underwent bilateral sinus augmentation. Each sinus was randomized to receive either control or test bone graft. Three animals were sacrificed at 8 weeks, and six animals were sacrificed at 16 weeks post-grafting. The 18 sinuses were processed for histomorphometry, which assessed the area fraction of new bone (%NB), residual graft (%RG) and soft tissue components (% STM), as well as graft particle osseointegration (% OI), within three zones equally distributed from the augmented sinus floor. RESULTS: At week 16, a significant increase in %NB was evident across all three zones in the control group when compared to week 8. A significantly greater %NB was evident in the control group when compared to the test group in zones 2 (P < 0.001) and 3 (P < 0.001). There was a significant increase in %OI in week 16 when compared to week 8 across all three zones in the control group (P < 0.001). %OI in the control group was significantly greater across all three zones when compared to the test group at week 16 (P < 0.001). Zone was found to be a significant main effect (P < 0.001) that was independent of time and treatment with decreasing %OI in distant zones. %RG did not significantly change with time for both groups. There was a significant reduction in %ST in week 16 when compared to week 8 across all three zones in the control group (P < 0.001). %ST in the test group was significantly greater across all zones when compared to the control group at week 16 (P < 0.001). CONCLUSION: Both groups exhibited very similar histomorphometric measurements in the zones proximal to the resident sinus wall. The % NB and % OI were greatest in the zones proximal to resident bony walls and gradually decreased as the distance from the proximal walls increased. There was greater % NB and % OI in the control group when compared to the test group in the distant zone.
Assuntos
Transplante Ósseo/métodos , Colágeno/farmacologia , Seio Maxilar/patologia , Minerais/farmacologia , Levantamento do Assoalho do Seio Maxilar/métodos , Animais , Bovinos , Seio Maxilar/cirurgia , Modelos Animais , OvinosRESUMO
OBJECTIVE: In the present study we evaluated the use of four commercially available fluorescent probes to monitor disease activity in murine CIA and its suppression during glucocorticoid therapy. METHODS: Arthritis was induced in male DBA/1 mice by immunization with type II collagen in Complete Freund's Adjuvant, followed by a boost of collagen in PBS. Four fluorescent probes from PerkinElmer in combination [ProSense 750 fluorescent activatable sensor technology (FAST) with Neutrophil Elastase 680 FAST and MMPSense 750 FAST with CatK 680 FAST] were used to monitor disease development from day 5 through to day 40 post-immunization. Fluorescence generated in vivo by the probes was correlated with clinical and histological score and paw measurements. RESULTS: The fluorescence intensity emitted by each probe was shown to correlate with the conventional measurements of disease. The highest degree of correlation was observed with ProSense 750 FAST in combination with Neutrophil Elastase 680 FAST; these probes were then used to successfully assess CIA suppression during dexamethasone treatment. CONCLUSION: We have demonstrated that longitudinal non-invasive duplexed optical fluorescence imaging provides a simple assessment of arthritic disease activity within the joints of mice following the induction of CIA and may represent a powerful tool to monitor the efficacy of drug treatments in preclinical studies.
Assuntos
Artrite Experimental/diagnóstico , Artrite Experimental/tratamento farmacológico , Dexametasona/farmacologia , Imagem Óptica/métodos , Animais , Antirreumáticos/farmacologia , Colágeno/farmacologia , Modelos Animais de Doenças , Masculino , Camundongos , Camundongos Endogâmicos DBA , Imagem Óptica/instrumentação , Distribuição Aleatória , Sensibilidade e Especificidade , Resultado do TratamentoRESUMO
Electrical impedance aggregometry (EIA) has gained popularity in clinical and research applications. Nonhuman primates are used to study disease and drug-related mechanisms that affect hemostasis, therefore establishing normal EIA parameters are necessary. The anticoagulants sodium heparin, hirudin and sodium citrate and three agonists, ADP, ASPI, and collagen were evaluated. Whole blood from 12 adult male rhesus macaques was collected to evaluate anticoagulants, sodium heparin, hirudin and sodium citrate using three agonists (ADP, ASPI and collagen), on the Multiplate® 5.0 Analyzer. Platelet function was reported for three parameters: Area under the curve (AUC), aggregation, and aggregation velocity. There was a significant difference in mean AUC between citrate and heparin samples, and citrate and hirudin samples regardless of the agonist used. There was no difference in AUC between heparin and hirudin. ADP-activated samples showed an increase in impedance with hirudin samples compared to citrate. Furthermore, heparin and hirudin out-perform citrate as the anticoagulant for EIA in the macaque. Finally, this study demonstrates the utility of the Multiplate® system in this model and provides important insight into anticoagulant choice when using EIA.
Assuntos
Plaquetas/efeitos dos fármacos , Plaquetas/metabolismo , Ácido Cítrico/farmacologia , Fibrinolíticos/farmacologia , Heparina/farmacologia , Hirudinas/farmacologia , Agregação Plaquetária/efeitos dos fármacos , Animais , Anticoagulantes/farmacologia , Área Sob a Curva , Colágeno/farmacologia , Macaca mulatta , Masculino , Ativação Plaquetária , Testes de Função Plaquetária , Sensibilidade e EspecificidadeRESUMO
Collagen hydrogels have been widely investigated as scaffolds for vascular tissue engineering due in part to the capacity of collagen to promote robust cell adhesion and elongation. However, collagen hydrogels display relatively low stiffness and strength, are thrombogenic, and are highly susceptible to cell-mediated contraction. In the current work, we develop and characterize a sequentially-formed interpenetrating network (IPN) that retains the benefits of collagen, but which displays enhanced mechanical stiffness and strength, improved thromboresistance, high physical stability and resistance to contraction. In this strategy, we first form a collagen hydrogel, infuse this hydrogel with poly(ethylene glycol) diacrylate (PEGDA), and subsequently crosslink the PEGDA by exposure to longwave UV light. These collagen-PEGDA IPNs allow for cell encapsulation during the fabrication process with greater than 90% cell viability via inclusion of cells within the collagen hydrogel precursor solution. Furthermore, the degree of cell spreading within the IPNs can be tuned from rounded to fully elongated by varying the time delay between the formation of the cell-laden collagen hydrogel and the formation of the PEGDA network. We also demonstrate that these collagen-PEGDA IPNs are able to support the initial stages of smooth muscle cell lineage progression by elongated human mesenchymal stems cells.
Assuntos
Prótese Vascular , Colágeno/farmacologia , Teste de Materiais/métodos , Polietilenoglicóis/farmacologia , Engenharia Tecidual , Animais , Linhagem da Célula/efeitos dos fármacos , Forma Celular/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Regulação da Expressão Gênica/efeitos dos fármacos , Humanos , Células-Tronco Mesenquimais/citologia , Células-Tronco Mesenquimais/efeitos dos fármacos , Peso Molecular , Miócitos de Músculo Liso/citologia , Miócitos de Músculo Liso/efeitos dos fármacos , Fenótipo , Ratos , Reologia/efeitos dos fármacos , Sus scrofa , Resistência à Tração/efeitos dos fármacos , Trombose/patologiaRESUMO
BACKGROUND: Dietary mineral deficiency, hypertension and diabetes have become serious human health problems. Dietary approaches are increasingly being investigated to address these issues. Identification of food-derived biological peptides has become an important approach to control such diseases. Peptides generated from aquatic byproducts have been shown to possess biological activities. RESULTS: Significantly higher copper-chelating activity was observed on simulated hydrolysis of intact collagen. The collagen hydrolysate generated in the gastric stage exhibited moderate angiotensin-converting enzyme (ACE)-inhibitory activity with an IC50 value of 2.92 ± 0.22 mg mL(-1), which significantly decreased to 0.49 ± 0.02 mg mL(-1) after intestinal digestion. The dipeptidyl peptidase (DPP) IV-inhibitory potency of the collagen hydrolysate generated directly following simulated gastrointestinal digestion (SGID) (IC50 2.59 ± 0.04 mg mL(-1)) was significantly lower than that of the collagen tryptic hydrolysate (CTH) (IC50 1.53 ± 0.01 mg mL(-1)). The antioxidant activities of collagen and CTH using the ferric-reducing antioxidant power (FRAP) assay were 0.87 ± 0.10 and 1.27 ± 0.03 µmol Trolox equivalent (TE) g(-1) respectively after SGID. CONCLUSION: This study identifies collagen as a good and inexpensive substrate for the generation of biologically active peptides with potential applications as functional ingredients in the management of chronic illness and mineral deficiency problems.
Assuntos
Colágeno/farmacologia , Proteínas Alimentares/farmacologia , Gadiformes , Peptídeos/farmacologia , Hidrolisados de Proteína/farmacologia , Alimentos Marinhos , Pele/química , Inibidores da Enzima Conversora de Angiotensina/metabolismo , Inibidores da Enzima Conversora de Angiotensina/farmacologia , Animais , Antioxidantes/metabolismo , Antioxidantes/farmacologia , Quelantes/metabolismo , Quelantes/farmacologia , Colágeno/metabolismo , Proteínas Alimentares/metabolismo , Digestão , Inibidores da Dipeptidil Peptidase IV/metabolismo , Inibidores da Dipeptidil Peptidase IV/farmacologia , Proteínas de Peixes/metabolismo , Proteínas de Peixes/farmacologia , Humanos , Mucosa Intestinal/metabolismo , Peptídeos/metabolismo , Hidrolisados de Proteína/metabolismoRESUMO
Collagens are the most abundant high molecular weight proteins in both invertebrate and vertebrate organisms, including mammals, and possess mainly a structural role, existing different types according with their specific organization in distinct tissues. From this, they have been elected as one of the key biological materials in tissue regeneration approaches. Also, industry is constantly searching for new natural sources of collagen and upgraded methodologies for their production. The most common sources are from bovine and porcine origin, but other ways are making their route, such as recombinant production, but also extraction from marine organisms like fish. Different organisms have been proposed and explored for collagen extraction, allowing the sustainable production of different types of collagens, with properties depending on the kind of organism (and their natural environment) and extraction methodology. Such variety of collagen properties has been further investigated in different ways to render a wide range of applications. The present review aims to shed some light on the contribution of marine collagens for the scientific and technological development of this sector, stressing the opportunities and challenges that they are and most probably will be facing to assume a role as an alternative source for industrial exploitation.