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1.
Nat Commun ; 11(1): 615, 2020 01 30.
Artigo em Inglês | MEDLINE | ID: mdl-32001693

RESUMO

Angiogenesis induction into damaged sites has long been an unresolved issue. Local treatment with pro-angiogenic molecules has been the most common approach. However, this approach has critical side effects including inflammatory coupling, tumorous vascular activation, and off-target circulation. Here, the concept that a structure can guide desirable biological function is applied to physically engineer three-dimensional channel networks in implant sites, without any therapeutic treatment. Microchannel networks are generated in a gelatin hydrogel to overcome the diffusion limit of nutrients and oxygen three-dimensionally. Hydrogel implantation in mouse and porcine models of hindlimb ischemia rescues severely damaged tissues by the ingrowth of neighboring host vessels with microchannel perfusion. This effect is guided by microchannel size-specific regenerative macrophage polarization with the consequent functional recovery of endothelial cells. Multiple-site implantation reveals hypoxia and neighboring vessels as major causative factors of the beneficial function. This technique may contribute to the development of therapeutics for hypoxia/inflammatory-related diseases.


Assuntos
Indutores da Angiogênese/efeitos adversos , Gelatina/química , Gelatina/farmacologia , Hidrogéis/química , Hidrogéis/farmacologia , Isquemia/terapia , Animais , Modelos Animais de Doenças , Células Endoteliais/patologia , Desenho de Equipamento , Feminino , Membro Posterior/irrigação sanguínea , Membro Posterior/diagnóstico por imagem , Membro Posterior/patologia , Hidrogéis/uso terapêutico , Hipóxia , Isquemia/diagnóstico por imagem , Isquemia/patologia , Macrófagos , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Neovascularização Fisiológica/fisiologia , Doenças Vasculares Periféricas/patologia , Doenças Vasculares Periféricas/terapia , Próteses e Implantes , Suínos , Cicatrização
2.
Carbohydr Polym ; 231: 115770, 2020 Mar 01.
Artigo em Inglês | MEDLINE | ID: mdl-31888821

RESUMO

The development of long-lasting therapeutic drug delivery system is greatly desired for effective treatment of glaucoma, a chronic and multifactorial disease. Herein, the roles of aromatic ring number in phenolic compound-conjugated chitosan injectables are exploited for achieving an advanced drug carrier with potent anti-inflammatory and anti-oxidant properties. Low and high number of aromatic rings can induce deleterious impacts on the pharmaceutical applications of injectables, whereas the compound with a moderate ring number is proved as the most efficient agent for boosting drug delivery performances and endowing the chitosan injectables with therapeutic properties. Kaempferol-conjugated injectable formulation reveals a remarkable effectiveness for intracameral pilocarpine administration, which can alleviate progressive glaucoma via simultaneously exerting multiple pharmacological activities to suppress ocular hypertension, inflammation, and oxidative stress. These findings provide a significant advance in understanding structure-property relationship of the phenolic compound-conjugated chitosan injectables as long-lasting therapeutic drug delivery systems for medical management of glaucoma.


Assuntos
Quitosana/química , Sistemas de Liberação de Medicamentos , Quempferóis/farmacologia , Fenóis/química , Animais , Quitosana/farmacologia , Gelatina/farmacologia , Glaucoma/tratamento farmacológico , Humanos , Hipertensão/tratamento farmacológico , Inflamação/tratamento farmacológico , Quempferóis/química , Estresse Oxidativo/efeitos dos fármacos , Fenóis/farmacologia , Espectroscopia de Infravermelho com Transformada de Fourier , Relação Estrutura-Atividade
3.
Mater Sci Eng C Mater Biol Appl ; 108: 110432, 2020 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-31923974

RESUMO

Coaxial electrospinning with the ability to use simultaneously two separate solvents provides a promising strategy for drug delivery. Nevertheless, controlled release of hydrophilic and sensitive therapeutics from slow biodegradable polymers is still challenging. To address this gap, we fabricated core-sheath fibers for dual delivery of lysozyme, as a model protein, and phenytoin sodium as a small therapeutic molecule. The sheath was processed by a gelatin solution while the core fibers were fabricated from an aqueous gelatin/PVA solution. Microstructural studies by transmission and scanning electron microscopy reveal the formation of homogeneous core-sheath nanofibers with an outer and inner diameter of 180 ± 48 nm and 106 ± 30 nm, respectively. Thermal gravimetric analysis determines that the mass loss of the core-sheath fibers fall between the mass loss values of individual sheath and core fibers. Swelling studies indicate higher water absorption of the core-sheath mat compared to the separate sheath and core membranes. In vitro drug release studies in Phosphate Buffered Saline (PBS) determine sustained release of the therapeutics from the core-sheath structure. The release trails three stages including non-Fickian diffusion at the early stage followed by the Fickian diffusion mechanism. The present study shows a useful approach to design core-sheath nanofibrous membranes with controlled and programmable drug release profiles.


Assuntos
Gelatina , Muramidase , Nanofibras/química , Fenitoína , Álcool de Polivinil , Animais , Linhagem Celular , Preparações de Ação Retardada/química , Preparações de Ação Retardada/farmacocinética , Preparações de Ação Retardada/farmacologia , Gelatina/química , Gelatina/farmacologia , Camundongos , Muramidase/química , Muramidase/farmacocinética , Muramidase/farmacologia , Fenitoína/química , Fenitoína/farmacocinética , Fenitoína/farmacologia , Álcool de Polivinil/química , Álcool de Polivinil/farmacologia
4.
Mater Sci Eng C Mater Biol Appl ; 107: 110296, 2020 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-31761169

RESUMO

Nanofibrous scaffolds composed of polycaprolactone (PCL) and gelatin (Ge) were obtained through a hydrolytic assisted electrospinning process. The PCL-to-Ge proportion (100/0 to 20/80), as well as the dissolution time (24, 48, 72, 96, 120 h) into a 1:1 formic/acetic acid solvent before electrospinning were modified to obtain the different samples. A strong influence of these factors on the physicochemical properties of the scaffolds was observed. Higher Ge percentage reduced crystallinity, allowed a uniform morphology and increased water contact angle. The increase in the dissolution time considerably reduced the molar mass and, subsequently, fibre diameter and crystallinity were affected. During in vitro biocompatibility tests, higher cell adhesion and proliferation were found for the 60/40, 50/50 and 40/60 PCL/Ge compositions that was corroborated by MTT assay, fluorescence and microscopy. A weakened structure, more labile to the in vitro degradation in physiologic conditions was found for these compositions with higher dissolution times (72 and 96 h). Particularly, the 40/60 PCL/Ge scaffolds revealed an interesting progressive degradation behaviour as a function of the dissolution time. Moreover, these scaffolds were non-inflammatory, as revealed by the pyrogen test and after the 15-day subcutaneous in vivo implantation in mice. Finally, a reduction of the scar tissue area after infarction was found for the 40/60 PCL/Ge scaffolds electrospun after 72 h implanted in rat hearts. These results are especially interesting and represent a feasible way to avoid undesired inflammatory reactions during the scaffold assimilation.


Assuntos
Gelatina , Poliésteres , Tecidos Suporte/química , Animais , Adesão Celular/efeitos dos fármacos , Linhagem Celular , Proliferação de Células/efeitos dos fármacos , Células Cultivadas , Modelos Animais de Doenças , Gelatina/química , Gelatina/farmacologia , Humanos , Leucócitos Mononucleares/efeitos dos fármacos , Teste de Materiais , Camundongos , Camundongos Endogâmicos C57BL , Infarto do Miocárdio/patologia , Miocárdio/patologia , Nanofibras/química , Poliésteres/química , Poliésteres/farmacologia , Ratos , Ratos Wistar , Engenharia Tecidual/métodos
5.
Int J Food Microbiol ; 312: 108375, 2020 Jan 02.
Artigo em Inglês | MEDLINE | ID: mdl-31669767

RESUMO

Recently, oxo-biodegradable polymers have attracted much attention due to taking less time to break down after disposal in comparison to ordinary polymers. Polyvinyl alcohol/gelatin (PVA/G) nanocomposite films, containing ZnO, TiO2 or ZnO/TiO2 nanoparticles supported on 4A zeolite (4A z), are novel active packaging that can control the release of antimicrobial compounds. The present study assessed the efficacy of PVA/G nanocomposite films with 1.5% (w/w) ZnO/4A z (treatment 1), 1.5% (w/w) TiO2/4A z (treatment 2), or 1% (w/w) ZnO, TiO2/4A z (treatment 3) in controlling the microbial load and maintaining the sensory qualities of white shrimp during storage at 4 ±â€¯1 °C. Firstly, the optimum concentration of each material for addition to the film was determined by micro-dilution and disc diffusion. Secondly, the specimens were checked for total viable count (TVC), as well as the counts of each of Pseudomonas spp., Enterobacteriaceae, Shewanella putrefaciens, inoculated Staphylococcus aureus, Listeria monocytogenes, and Escherichia coli O157:H7. According to the results, the PVA/G nanocomposite films containing treatments 1-3 significantly decreased the number of bacteria in the treatment group in comparison to the control group (P < .05). The results of the antimicrobial activity of the three treatments by using the disc diffusion method revealed that the inhibition zone varied from 8.11 ±â€¯0.02 to 12.63 ±â€¯0.04 mm. Also it should be noted that, the finding of micro-dilution test varied from 1 ±â€¯0.01 to 3 ±â€¯0.01. The ZnO, TiO2/4A z nanocomposite had a significantly greater antimicrobial impact against Gram-negative bacteria compared to Gram-positive bacteria (P < .05). Finally, the microbiological and sensory investigation of the efficacy of the PVA/G nanocomposite films as active packaging materials revealed a considerable improvement in shrimp shelf life (12 days) in comparison to the control (6 days). Therefore, these nanocomposite films can be used as novel active packaging in the maintenance of the microbial load and sensory qualities of shrimp.


Assuntos
Antibacterianos/farmacologia , Embalagem de Alimentos/métodos , Armazenamento de Alimentos/métodos , Penaeidae/microbiologia , Titânio/farmacologia , Zeolitas/farmacologia , Óxido de Zinco/farmacologia , Animais , Carga Bacteriana/efeitos dos fármacos , Escherichia coli/efeitos dos fármacos , Gelatina/farmacologia , Listeria monocytogenes/efeitos dos fármacos , Nanocompostos , Nanopartículas , Álcool de Polivinil/farmacologia , Refrigeração , Staphylococcus aureus/efeitos dos fármacos
6.
Mater Sci Eng C Mater Biol Appl ; 106: 110116, 2020 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-31753329

RESUMO

Microcarrier cell scaffolds have potential as injectable cell delivery vehicles or as building blocks for tissue engineering. The use of small cell carriers allows for a 'bottom up' approach to tissue assembly when moulding microparticles into larger structures, which can facilitate the introduction of hierarchy by layering different matrices and cell types, while evenly distributing cells through the structure. In this work, silk fibroin (SF), purified from Bombyx mori cocoons, was blended with gelatin (G) to produce materials composed of varying ratios of the two components (SF: G 25:75, 50:50, and 75:25). Cell compatibility to these materials was first confirmed in two-dimensional culture and found to be equivalent to standard tissue culture plastic, and better than SF or G alone. The mechanical properties of the blends were investigated and the blended materials were found to have increased Young's moduli over SF alone. Microcarriers of SF/G blends with defined diameters were generated in a reproducible manner through the use of an axisymmetric flow focussing device, constructed from off-the-shelf parts and fittings. These SF/G microcarriers supported adhesion of rat mesenchymal stem cells with high degrees of efficiency under dynamic culture conditions and, after culturing in osteogenic differentiation medium, cells were shown to have characteristics typical of osteoblasts. This work illustrates that microcarriers composed of SF/G blends are promising building blocks for osteogenic tissue engineering.


Assuntos
Fibroínas/química , Gelatina/química , Engenharia Tecidual , Tecidos Suporte/química , Animais , Células da Medula Óssea/citologia , Adesão Celular/efeitos dos fármacos , Diferenciação Celular/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Células Cultivadas , Fibroínas/farmacologia , Gelatina/farmacologia , Células-Tronco Mesenquimais/citologia , Células-Tronco Mesenquimais/metabolismo , Osteogênese/efeitos dos fármacos , Ratos , Ratos Wistar
7.
Int J Nanomedicine ; 14: 9603-9617, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31824154

RESUMO

Purpose: Non-healing or slow healing chronic wounds are among serious complications of diabetes that eventually result in amputation of limbs and increased morbidities and mortalities. Chronic diabetic wounds show reduced blood vessel formation (lack of angiogenesis), inadequate cell proliferation and poor cell migration near wounds. In this paper, we report the development of a hydrogel-based novel wound dressing material loaded with reduced graphene oxide (rGO) to promote cell proliferation, cell migration and angiogenesis for wound healing applications. Methods: Gelatin-methacryloyl (GelMA) based hydrogels loaded with different concentrations of rGO were fabricated by UV crosslinking. Morphological and physical characterizations (porosity, degradation, and swelling) of rGO incorporated GelMA hydrogel was performed. In vitro cell proliferation, cell viability and cell migration potential of the hydrogels were analyzed by MTT assay, live/dead staining, and wound healing scratch assay respectively. Finally, in vivo chicken embryo angiogenesis (CEO) testing was performed to evaluate the angiogenic potential of the prepared hydrogel. Results: The experimental results showed that the developed hydrogel possessed enough porosity and exudate-absorbing capacity. The biocompatibility of prepared hydrogel on three different cell lines (3T3 fibroblasts, EA.hy926 endothelial cells, and HaCaT keratinocytes) was confirmed by in vitro cell culture studies (live/dead assay). The GelMA hydrogel containing 0.002% w/w rGO considerably increased the proliferation and migration of cells as evident from MTT assay and wound healing scratch assay. Furthermore, rGO impregnated GelMA hydrogel significantly enhanced the angiogenesis in the chick embryo model. Conclusion: The positive effect of 0.002% w/w rGO impregnated GelMA hydrogels on angiogenesis, cell migration and cell proliferation suggests that these formulations could be used as a functional wound healing material for the healing of chronic wounds.


Assuntos
Gelatina/farmacologia , Grafite/farmacologia , Hidrogéis/farmacologia , Metacrilatos/farmacologia , Neovascularização Fisiológica/efeitos dos fármacos , Células 3T3 , Animais , Sobrevivência Celular/efeitos dos fármacos , Embrião de Galinha , Células Endoteliais/efeitos dos fármacos , Fibroblastos/efeitos dos fármacos , Humanos , Camundongos , Oxirredução , Suínos , Cicatrização/efeitos dos fármacos , Difração de Raios X
8.
Bull Exp Biol Med ; 168(1): 95-98, 2019 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-31758379

RESUMO

The effects of composite fibroin-gelatin microparticles (100-250 µ) on the rate of wound healing and regeneration under conditions of contraction prevention were studied on the model of splinted full-thickness skin wound in a mouse. Subcutaneous injection of these particles into the defect area accelerated wound healing and promoted re-epithelialization and recovery of normal structure of the epidermis. In addition, the composite microparticles promoted the formation of connective tissue of characteristic structure, replacing the derma over the entire defect, and stimulated regeneration of subcutaneous muscle (panniculus carnosus) and skin appendages (sebaceous glands and hair follicles).


Assuntos
Fibroínas/química , Fibroínas/farmacologia , Gelatina/química , Gelatina/farmacologia , Dermatopatias/tratamento farmacológico , Pele/metabolismo , Animais , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Regeneração/efeitos dos fármacos , Pele/efeitos dos fármacos , Cicatrização/efeitos dos fármacos
9.
Biomed Res Int ; 2019: 6749326, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31781634

RESUMO

Background: The treatment of chronic kidney diseases (CKDs) by different approaches using mesenchymal stem cells (MSCs) has made great strides. In this study, we aimed to explore the potential mechanism of gelatin microcryogels (GMs) as a cell therapeutic vector to block the progression of CKD. Methods: In vivo, the pedicled omentum valve with MSC-loaded GMs was packed onto 5/6 nephrectomized kidneys derived from rats. The therapeutic effects were evaluated. In vitro, TNF-α, TGF-ß, and MSCs were added to the medium of the HK-2 cell culture system, and key genes involved in anti-inflammatory and antifibrosis effects were evaluated by qPCR. Results: After 12 weeks of MSC transplantation, kidney functions, such as serum creatinine, urea nitrogen, and 24-hour urine protein, were significantly improved. The pedicled omentum valve was packed with MSC-loaded GMs onto the 5/6 nephrectomized kidney, and the expressions of collagen IV, α-SMA, and TGF-ß were all evaluated by immunohistochemical staining and western blot analysis. MSC-loaded-GMs also showed antifibrotic effects by inducing the upregulation of HO-1, BMP-7, and HGF and the downregulation of MCP-1 at the mRNA level. Four weeks after MSC-loaded GM treatment, we found that the mRNA levels of TNF-α and IL-6 were clearly reduced. MSC-conditional medium (MSC-CM) showed that the TNF-α-induced expression of IL-8 and IL-6 mRNA was reversed; E-cadherin mRNA was upregulated; and the TGF-ß-induced expression of collagen IV, α-SMA, and fibronectin (FN) mRNA in HK-2 cells was reduced. Conclusions: We demonstrated that the pedicled omentum valve packed with MSC-loaded GMs had a renal protective effect on the 5/6 nephrectomized kidney by observing the anti-inflammatory and antifibrosis effects.


Assuntos
Criogéis/farmacologia , Gelatina/farmacologia , Transplante de Células-Tronco Mesenquimais , Células-Tronco Mesenquimais/metabolismo , Insuficiência Renal Crônica , Animais , Fibrose , Masculino , Células-Tronco Mesenquimais/patologia , Ratos , Ratos Sprague-Dawley , Insuficiência Renal Crônica/metabolismo , Insuficiência Renal Crônica/patologia , Insuficiência Renal Crônica/terapia
10.
Mar Drugs ; 17(10)2019 Oct 03.
Artigo em Inglês | MEDLINE | ID: mdl-31623339

RESUMO

For full use of fish by-products, scale gelatin (TG) and antioxidant peptides (APs) of skipjack tuna (Katsuwonus pelamis) were prepared, and their properties were characterized using an amino acid analyzer, sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), Fourier transform infrared spectroscopy (FTIR), electrospray ionization mass spectrometers (ESI-MS), and radical scavenging assays. The results indicate that TG with a yield of 3.46 ± 0.27% contained Gly (327.9 ± 5.2 residues/1000 residues) as the major amino acid and its imino acid content was 196.1 residues/1000 residues. The structure of TG was more unstable than that of type I collagen from scales of skipjack tuna (TC) and TG was more suitable for preparation of hydrolysate by protease than mammalian gelatins. Therefore, TG was separately hydrolyzed under five proteases (pepsin, papain, trypsin, neutrase, and alcalase) and ten APs (TGP1-TGP10) were isolated from the alcalase-hydrolysate. Among them, TGP5, TGP7, and TGP9 with high antioxidant activity were identified as His-Gly-Pro-Hyp-Gly-Glu (TGP5), Asp-Gly-Pro-Lys-Gly-His (TGP7) and Met-Leu-Gly-Pro-Phe-Gly-Pro-Ser (TGP9), respectively. Furthermore, TGP5, TGP7, and TGP9 exhibited a high radical scavenging capability on 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical (EC50 values of 1.34, 0.54, and 0.67 mg/mL, respectively), hydroxyl radical (EC50 values of 1.03, 0.41, and 0.74 mg/mL, respectively), and superoxide anion radical (EC50 values of 1.19, 0.71, and 1.59 mg/mL, respectively). These results suggest that three APs (TGP5, TGP7, and TGP9), especially TGP7, have a strong antioxidant activity and could act as potential antioxidant ingredients applied in functional products.


Assuntos
Antioxidantes/farmacologia , Gelatina/farmacologia , Peptídeos/farmacologia , Atum/metabolismo , Aminoácidos/metabolismo , Animais , Colágeno Tipo I/metabolismo , Depuradores de Radicais Livres/farmacologia , Hidrólise/efeitos dos fármacos , Radical Hidroxila/metabolismo , Superóxidos/metabolismo
11.
Mar Drugs ; 17(10)2019 Oct 17.
Artigo em Inglês | MEDLINE | ID: mdl-31627413

RESUMO

In this study, we aimed to obtain gelatin from the marine snail Rapana venosa using acidic and enzymatic extraction methods and to characterize these natural products for cosmetic and pharmaceutical applications. Marine gelatins presented protein values and hydroxyproline content similar to those of commercial mammalian gelatin, but with higher melting temperatures. Their electrophoretic profile and Fourier transform infrared (FTIR) spectra revealed protein and absorption bands situated in the amide region, specific for gelatin molecule. Scanning electron microscopy (SEM) analysis showed significant differences in the structure of the lyophilized samples, depending on the type of gelatin. In vitro studies performed on human keratinocytes showed no cytotoxic effect of acid-extracted gelatin at all tested concentrations and moderate cytotoxicity of enzymatic extracted gelatin at concentrations higher than 0.5 mg/mL. Also, both marine gelatins favored keratinocyte cell adhesion. No irritant potential was recorded as the level of IL-1α and IL-6 proinflammatory cytokines released by HaCaT cells cultivated in the presence of marine gelatins was significantly reduced. Together, these data suggest that marine snails are an alternative source of gelatins with potential use in pharmaceutical and skincare products.


Assuntos
Organismos Aquáticos/química , Produtos Biológicos/química , Gelatina/química , Caramujos/química , Animais , Produtos Biológicos/farmacologia , Adesão Celular/efeitos dos fármacos , Linhagem Celular , Citocinas/metabolismo , Gelatina/farmacologia , Humanos , Inflamação/tratamento farmacológico , Inflamação/metabolismo , Queratinócitos/efeitos dos fármacos , Queratinócitos/metabolismo
12.
Mater Sci Eng C Mater Biol Appl ; 104: 109898, 2019 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-31499960

RESUMO

Surface of the implantable devices is the root cause of several complications such as infections, implant loosening and chronic inflammation. There is an urgent need for multifunctional coatings that can address these shortcomings simultaneously in a manner similar to the structures of extracellular matrix. Herein, we developed a coating system composed of ECM components and a naturally derived polypeptide. The interactions between the coating components create an environment that enables incorporation of an antimicrobial/angiogenic polypeptide. The film composition is based gelatin and hyaluronic acid modified with aldehyde groups (HA-Ald) that can react with poly (arginine) (PAR) through transient interactions. Nanoplasmon measurements demonstrated a significantly higher loading of PAR in films containing HA-Ald with longer retention of PAR in the structure. The presence of PAR not only provides to the film surface antimicrobial (contact-killing) properties but also increased endothelial cell-cell contacts (PECAM) and VEGFA gene expression and secretion by human vascular endothelial cells. This multifunctional coating can be easily applied to surface of implants where it can enact on several problems simultaneously.


Assuntos
Materiais Revestidos Biocompatíveis/farmacologia , Gelatina/farmacologia , Ácido Hialurônico/farmacologia , Peptídeos/farmacologia , Polímeros/farmacologia , Próteses e Implantes , Animais , Antibacterianos/farmacologia , Bovinos , Matriz Extracelular/metabolismo , Células Endoteliais da Veia Umbilical Humana/efeitos dos fármacos , Humanos , Fator A de Crescimento do Endotélio Vascular/metabolismo
13.
Mater Sci Eng C Mater Biol Appl ; 104: 109892, 2019 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-31499962

RESUMO

The bone-derived-osteoblast seeded biomaterials scaffold in tissue engineering, have displayed prominence in the treatment of the osseous medical condition. In vitro osteogenesis of rabbit osteoblast cell (rOb) from bone tissue (rT) and MSC-derived rOb from bone marrow (rM) on Gelatin-Hydroxyapatite (HG) based biomaterials was investigated. In this work, lyophilized biomaterial was prepared by the addition of amorphous chitosan ('C') to 'H' dispersed in 'G' matrix, to find its role in biomaterials biocompatibility. Isolated rOb seeded biomaterials were studied using CLSM and flow cytometry for proliferation potential. The biomaterial's core and surface morphology was studied from SEM-EDX and AFM respectively. Upon co-culture with HCG, rT over rM showed rabbit bone extracellular matrix (ECM) mimicking properties both in in vitro studies and biomaterials micro architecture. The in vitro metabolic behaviour was studied by Alamar Blue (AB) assay, DNA content using Hoechst 33258, potency via the activity of Alkaline Phosphatase (ALP), Calcium's relative content by Alizarin Red S (ARS) assay. A novel combination of biomaterials-cell interaction was observed when rT was co-cultured with HCG and proved effective in osteogenesis with regard to Bone Bioengineering.


Assuntos
Materiais Biocompatíveis/química , Materiais Biocompatíveis/farmacologia , Gelatina/química , Gelatina/farmacologia , Osteoblastos/efeitos dos fármacos , Fosfatase Alcalina/química , Animais , Bioengenharia/métodos , Osso e Ossos/efeitos dos fármacos , Diferenciação Celular/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Células Cultivadas , Quitosana/química , Técnicas de Cocultura/métodos , Durapatita/química , Células-Tronco Mesenquimais/efeitos dos fármacos , Osteogênese/efeitos dos fármacos , Coelhos , Engenharia Tecidual/métodos , Tecidos Suporte/química
14.
Mater Sci Eng C Mater Biol Appl ; 104: 109933, 2019 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-31499966

RESUMO

Osteoporotic bone represents - particularly in case of fractures - difficult conditions for its regeneration. In the present study, the focus was put on a degradable bone substitute material of gelatin-modified calcium and strontium phosphates facing the special demands of osteoporotic bone. The release of strontium ions from the material ought to stimulate osteoblastogenesis either direct by ion release or indirect after material resorption by increased presence and activity of osteoclasts, which subsequently stimulate osteoblasts. A new porous material was produced from calcium phosphate, strontium phosphate and a mixed phase of calcium/strontium phosphate precipitated in presence of gelatin. Initially, ion release was analyzed in standard­calcium containing (2.0 mM) and low-calcium (0.4 mM) minimum essential medium. The cultivation of human peripheral blood mononuclear cells next to the material led to formation of osteoclast-like cells, able to migrate, fuse, and differentiate. Especially, the mixed gelatin-modified calcium/strontium phosphate allowed osteoclastogenesis as proven morphologically and by real-time quantitative polymerase chain reaction (RT-qPCR). It was precisely this material that led to the best osteoblastic reaction of human bone marrow stromal cells cultured on the material. The investigations of the bone substitute material indicate active involvement in the balance of cells of the bone morphogenetic unit.


Assuntos
Materiais Biocompatíveis/farmacologia , Fosfatos de Cálcio/farmacologia , Gelatina/farmacologia , Osteoblastos/efeitos dos fármacos , Osteoclastos/efeitos dos fármacos , Fosfatos/farmacologia , Estrôncio/farmacologia , Animais , Contagem de Células , Diferenciação Celular/efeitos dos fármacos , Células Cultivadas , Regulação da Expressão Gênica/efeitos dos fármacos , Humanos , Células-Tronco Mesenquimais/citologia , Células-Tronco Mesenquimais/efeitos dos fármacos , Minerais/metabolismo , Monócitos/citologia , Monócitos/efeitos dos fármacos , Osteoblastos/citologia , Osteoclastos/citologia , Osteogênese/efeitos dos fármacos , Suínos
15.
Arch Environ Contam Toxicol ; 77(3): 452-460, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31451889

RESUMO

The adoption of genetically modified (GM) crops has occurred rapidly in the United States. The transfer of GM corn byproducts from agricultural fields to nearby streams after harvest is significant and occurs well into the post-harvest year. These corn leaves, stems, and cobs then become a detrital food source for organisms, such as shredders in the stream ecosystem. Considering that the nontarget effects of Bt corn have been observed in some terrestrial organisms, we assessed whether Bt toxins affect an important aquatic organism, juvenile F. rusticus crayfish. Juvenile crayfish were fed six distinct diet treatments: two varieties of Bt corn, two non-Bt controls of herbicide tolerant corn, and two controls: fish gelatin and river detritus. Juveniles were fed these diets while housed in flow-through artificial streams that received natural stream water from a local source. Specific growth rate and survivorship of the crayfish were measured throughout the study. Juveniles fed corn diets grew significantly less and had reduced survival compared with juveniles fed fish gelatin or river detritus diets. Furthermore, juveniles fed one Bt variety of corn (VT Triple Pro®) exhibited significantly less growth than those fed one of the herbicide tolerant varieties (Roundup Ready 2®). Our study shows that corn inputs to streams may be detrimental to the growth and survivorship of juvenile crayfish and that certain Bt varieties may exacerbate these negative effects. These effects on crayfish will have repercussions for the entire ecosystem, because crayfish are conduits of energy between many trophic levels.


Assuntos
Ração Animal/efeitos adversos , Astacoidea/crescimento & desenvolvimento , Plantas Geneticamente Modificadas/toxicidade , Zea mays/genética , Animais , Astacoidea/efeitos dos fármacos , Proteínas de Bactérias/genética , Proteínas de Bactérias/toxicidade , Produtos Agrícolas , Ecossistema , Endotoxinas/genética , Endotoxinas/toxicidade , Gelatina/farmacologia , Proteínas Hemolisinas/genética , Proteínas Hemolisinas/toxicidade , Folhas de Planta/genética , Rios
16.
Vet Anaesth Analg ; 46(5): 579-586, 2019 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-31401048

RESUMO

OBJECTIVE: To determine if low molecular weight synthetic colloid fluids administered to dogs interfere with refractometric estimates of total plasma protein (TPPr) and urine osmolality (UOsm). STUDY DESIGN: Experimental study. ANIMALS: Eighteen healthy Greyhound dogs. METHODS: Anaesthetized Greyhounds subjected to haemorrhage for 60 minutes were given 80 mL kg-1 of Plasma-Lyte 148 (CRYST), or 20 mL kg-1 of hydroxyethyl starch 130/0.4 (HES) or succinylated gelatine (GELO) (n = 6 per group) intravenously over 20 minutes. Refractometric (TPPr) and biuret total plasma protein (TPPb) were measured before haemorrhage (Baseline), at end of shock (Shock), immediately (T20), then 40 minutes (T60), 100 minutes (T120) and 160 minutes (T180) after fluid administration. Urine specific gravity (USG) and UOsm were measured at all time points except T20. Estimated UOsm (eUOsm) was calculated from USG. Bias and limits of agreement (LOA) for TPPr versus TPPb, and eUOsm versus UOsm were calculated at each time point. RESULTS: For dogs given CRYST and GELO, median TPPr and TPPb decreased in parallel, with a small consistent TPP bias (CRYST range of bias, 0.38-0.67 g dL-1; GELO range of bias, 0.42-0.58 g dL-1). Dogs given HES showed divergence between median TPPr and TPPb after T20, with a peak bias at T20 of 1.62 g dL-1 (LOA 1.29-1.95). Dogs given HES and GELO had markedly increased USG [HES peak median USG at T180 of 1.119 (Q1-Q3 1.103-1.122); GELO peak median USG at T120 of 1.114 (Q1-Q3 1.082-1.119)], with large increases in bias between eUOsm and UOsm [HES peak bias at T60 of 2995 mOsm kg-1 (LOA 2032-3958 mOsm kg-1); GELO peak bias at T120 of 2465 mOsm kg-1 (LOA 940-3990 mOsm kg-1)]. CONCLUSIONS AND CLINICAL RELEVANCE: Administration of HES and GELO to dogs with haemorrhagic shock interferes with refractometric measurements for at least 3 hours after administration.


Assuntos
Doenças do Cão/tratamento farmacológico , Gelatina/uso terapêutico , Derivados de Hidroxietil Amido/uso terapêutico , Choque Hemorrágico/veterinária , Succinatos/uso terapêutico , Animais , Cães , Feminino , Hidratação/veterinária , Gelatina/administração & dosagem , Gelatina/farmacologia , Derivados de Hidroxietil Amido/administração & dosagem , Derivados de Hidroxietil Amido/farmacologia , Masculino , Refratometria/veterinária , Choque Hemorrágico/tratamento farmacológico , Succinatos/administração & dosagem , Succinatos/farmacologia , Urina/química
17.
Biomed Pharmacother ; 118: 109291, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31401395

RESUMO

As a health-care food and traditional Chinese medicine, E'jiao, from the skin of Equus animus L, has been used to nourish blood in China for more than 2000 years. In modern medicine, there are also evidences indicate it has a beneficial effect on chemotherapy-caused blood deficiency. However, its mechanism of action for blood invigoration remains unclear. In the present study, we investigated the hematopoietic effect of E'jiao in 5-Fluorouracil-treated mice. In addition to the counting of bone marrow nucleated cells (BMNCs), flow cytometry was used to detect the population of hematopoietic stem cells (HSCs), and colony-forming unit (CFU) was used to assay the differentiation ability of hematopoietic progenitor cells (HPCs). Gene expression profiles of bone marrow cells were obtained from RNA sequencing (RNA-seq) and differentially expressed genes (DEGs) were analyzed with an emphasis on hematopoiesis-related pathways. The results show that E'jiao promotes the proliferation of both BMNCs and HSCs, as well as the differentiation of HPCs. By providing a hematopoiesis-related molecular regulatory network of E'jiao, we point out that the mechanism of E'jiao is associated with pathways including ECM-receptor interaction, Wnt signaling pathway, PI3K-Akt signaling pathway, TGF-beta signaling pathway, Hematopoietic cell lineage and Osteoclast differentiation, in which Ibsp, Col1a1, Col1a2, Notum, Sost, Dkk1, Irx5, Irx3 and Dcn are the key regulatory molecules. These findings provide valuable molecular basis for the mechanism of action of E'jiao.


Assuntos
Fluoruracila/farmacologia , Gelatina/farmacologia , Perfilação da Expressão Gênica , Hematopoese/efeitos dos fármacos , Análise de Sequência de RNA , Animais , Feminino , Redes Reguladoras de Genes/efeitos dos fármacos , Hematopoese/genética , Camundongos , Camundongos Endogâmicos BALB C
18.
Biomed Pharmacother ; 117: 109183, 2019 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-31261029

RESUMO

This study aimed to fabricate the potential therapeutic scaffold to efficiently and safely fastening skin wound healing. A biocompatible grafting polymer-based thermal sensitive hybrid hydrogel (Chitosan-P123, CP) containing gelatin and curcumin was designed to be suitable stiffness for tissue regeneration. A detailed in the rheological study found that the encapsulated agents induced the change in the stiffness of the hydrogel from the hard to the soft. Especial, the thermally induced phase transition of CP hydrogel was governed by the participant of gelatin rather than curcumin. For example, at 25 wt% gelatin, CP hydrogel exhibited a unique gel-sol-gel transition following the function of temperature. Moreover, in vitro investigation revealed that the hybrid hydrogel provides the capacity of especially induced curcumin release with a sustainable rate as well as the excellent biocompatibility scaffold. Altogether with in vivo study, the hybrid hydrogel highlighted the advance of the dual synergistic of curcumin and gelatin in development of smart scaffold system, which promoted the efficacy in the regeneration of the structure and the barrier's function of damaged skin such as wound or skin cancer.


Assuntos
Quitosana/química , Curcumina/farmacologia , Gelatina/farmacologia , Hidrogéis/farmacologia , Temperatura , Cicatrização/efeitos dos fármacos , Animais , Células Cultivadas , Liberação Controlada de Fármacos , Sinergismo Farmacológico , Humanos , Masculino , Camundongos , Nanopartículas/química , Transição de Fase , Polímeros/síntese química , Polímeros/química , Espectroscopia de Prótons por Ressonância Magnética , Termogravimetria
19.
Mater Sci Eng C Mater Biol Appl ; 103: 109768, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31349413

RESUMO

Currently graphene-doped electrospun scaffolds have been a matter of great interest to be exploited in biomedical fields such as tissue engineering and drug delivery applications. The main objective of this paper is to evaluate the effect of graphene on biological properties of PCL/gelatin nanofibrous mats. SEM analysis was conducted to investigate the morphology of the electrospun nanofibers. The in-vitro cellular proliferation of PC12 cells on nanofibrous web was also investigated. Electrospun PCL/gelatin/graphene nanofibrous mats exhibited 99% antibacterial properties against gram-positive and gram-negative bacteria. Drug release studies indicated that the π-π stacking interaction between TCH and graphene has led to the far better controlled release of TCH from electrospun PCL/gelatin/graphene compared to PCL/gelatin nanofibrous scaffolds. These superior properties along with an improvement in hydrophilicity and biodegradation features has made the nanofibers a promising candidate to be used as electrically conductive scaffolds in neural tissue engineering as well as controlled drug delivery.


Assuntos
Antibacterianos , Proliferação de Células/efeitos dos fármacos , Bactérias Gram-Negativas/crescimento & desenvolvimento , Bactérias Gram-Positivas/crescimento & desenvolvimento , Nanofibras/química , Tecido Nervoso , Engenharia Tecidual , Animais , Antibacterianos/química , Antibacterianos/farmacologia , Preparações de Ação Retardada/química , Preparações de Ação Retardada/farmacologia , Gelatina/química , Gelatina/farmacologia , Grafite/química , Grafite/farmacologia , Tecido Nervoso/citologia , Tecido Nervoso/metabolismo , Células PC12 , Poliésteres/química , Poliésteres/farmacologia , Ratos
20.
Int J Biol Macromol ; 137: 392-404, 2019 Sep 15.
Artigo em Inglês | MEDLINE | ID: mdl-31233795

RESUMO

Gelatin is a biocompatible and biodegradable natural polymer obtained by collagen. Gelatin nanofibers meet all the necessary requirements when used as wound dressing material. However, their lack of antimicrobial properties limits their use. The purpose of this study is to expand the field of use of gelatin by providing it with antimicrobial properties. For this purpose, poly([2-(methacryloyloxy)ethyl] trimethylammonium chloride) (PMETAC), was used. In this study, the polymers were dissolved in formic acid-acetic acid and nanofibers were synthesized by electrospinning. The obtained nanofibers were characterized with SEM, FTIR, and TGA. The antibacterial effect, degradation tests, and cell viability, adhesion and proliferation were investigated. The SEM studies show that the nanofibers are homogeneous and smooth. At the end of 14 days, all nanofibers lost >90% of their mass. The nanofibers containing PMETAC showed good bactericidal activity against Staphylococcus aureus, Escherichia coli, methicillin-resistant Staphylococcus aureus and Acinetobacter baumannii. MTT test demonstrated that low doses of the nanofibers were biocompatible. The cell adhesion study has been shown that many cells attachment and proliferate on the surface of nanofibers. It has been found that the obtained nanofibers can be used safely and effectively as antimicrobial wound dressing material.


Assuntos
Antibacterianos/química , Antibacterianos/farmacologia , Bandagens/microbiologia , Gelatina/química , Gelatina/farmacologia , Nanofibras/química , Cicatrização/efeitos dos fármacos , Animais , Antibacterianos/toxicidade , Adesão Celular/efeitos dos fármacos , Linhagem Celular , Proliferação de Células/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Gelatina/toxicidade , Camundongos
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