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Pure magnesium (Mg) granules were implanted into the tibial medullary cavity of osteomyelitis modeled animals after debridement, and the animals without implant were taken as the control group. The antibacterial and osteogenic effects on bone tissue during Mg degradation were evaluated through detecting Mg ions, counting bacteria culture in peripheral blood, histology and iconography. The results showed that there was no significant difference for the concentration of serum Mg between the preoperative and postoperative animals within 5 weeks, maintaining in the normal range, and the number of bacteria in bone tissue of the Mg implant group was significantly lower than that of the control group. Mg implantation showed good biocompatibility no harmful to the liver, spleen, kidney and other organs in the modeled animals. In addition, the formation rate of new bone tissues around the implanted Mg was faster, indicating that the degradation of Mg could also promote the osteogenic process with good biocompatibility.
Assuntos
Antibacterianos/química , Materiais Biocompatíveis/química , Magnésio/química , Osteomielite/tratamento farmacológico , Staphylococcus aureus/efeitos dos fármacos , Animais , Substitutos Ósseos/química , Osso e Ossos/metabolismo , Modelos Animais de Doenças , Feminino , Masculino , Osteogênese , Próteses e Implantes , Coelhos , Infecções Estafilocócicas/microbiologia , Tíbia/patologia , Fatores de TempoRESUMO
OBJECTIVE: In this study, we aimed to analyze the risk factors of bone cement leakage in percutaneous kyphoplasty (PKP) treatment of osteoporotic vertebral compression fractures (OVCFs), and provide suggestions for reducing bone cement leakage. METHODS: A retrospective study of 517 cases of OVCFs treated with PKP were divided into 2 groups according to whether they had bone cement leakage or not, leakage group (n = 72) and non-leakage group (n = 445). The risk factors of leakage were systematically analyzed using clinical baseline data, imaging observation data, and surgery-related factors. To select the statistically significant results (P < 0.05) among the risk factors mentioned above, we used the binary logistic regression method to identify the main risk factors. RESULTS: The univariate analysis of clinical baseline dataï¼imaging observation data, and surgery-related factors showed that bone mineral density (BMD) (P < 0.001), hypertension (P < 0.05), injury factors (P < 0.01), cortical defect (P < 0.001), grade of vertebral compression (P < 0.001), surgical approach (P < 0.05), stage of bone cement injection (P < 0.01), and balloon pressure (P < 0.05) were the risk factors for bone cement leakage. The recovery rate of vertebral height, and the Cobb angle correction rate were lower in the bone cement leakage group (P < 0.001). The correction effect of kyphosis after operation was limited. Binary logistic analysis results showed that BMD (odds ratio [OR] 5.605), cortical defect (OR 3.115), and stage of bone cement injection (OR 2.385) were bone cement leakage-independent risk factors. CONCLUSIONS: Impairment of BMD value, defects of cortical bone, and inappropriate stage of bone cement injection will increase the risk of bone cement leakage in PKP treatment and limit PKP effects.
Assuntos
Fraturas por Compressão , Cifoplastia , Fraturas por Osteoporose , Fraturas da Coluna Vertebral , Vertebroplastia , Humanos , Estudos Retrospectivos , Cifoplastia/métodos , Cimentos Ósseos/uso terapêutico , Fraturas por Osteoporose/cirurgia , Fraturas por Compressão/cirurgia , Fraturas da Coluna Vertebral/cirurgia , Fatores de Risco , Resultado do Tratamento , Vertebroplastia/efeitos adversosRESUMO
BACKGROUND: Titanium (Ti) is now widely used as implant material due to its excellent mechanical properties and superior biocompatibilities, while its inert bioactivities might lead to insufficient osseointegration, and limit its performance in dental applications. METHODS: We introduced a robust and simple approach of modifying titanium surfaces with polysaccharide complexes. Titanium samples were subjected to hydrothermal treatment to create a uniform porous structure on the surface, followed by coating with a bioinspired and self-assembly polydopamine layer. Strontium Eucommia Ulmoides Polysaccharide (EUP-Sr) complexes are then introduced to the polydopamine-coated porous titanium. Multiple morphological and physiochemical characterizations are employed for material evaluation, while cell proliferation and gene expression tests using macrophages, primary alveolar bone osteoblasts, and vascular endothelial cells are used to provide an overall insight into the functions of the product. The significances of statistical differences were analyzed using student's t-test. RESULTS: Microscopic and spectrometric characterizations confirmed that the Ti surface formed a porous structure with an adequate amount of EUP-Sr loading. The attachment was attributed to hydrogen bonding between the ubiquitous glycosidic linkage of the polysaccharide complex and the ring structure of polydopamine, yet the loaded EUP-Sr complex can be gradually released, consequently benefiting the neighboring microenvironment. Cell experiments showed no cytotoxicity of the material, and the product showed promising anti-inflammation, osseointegration, and angiogenesis properties, which were further confirmed by in vivo evaluations. CONCLUSION: We believe the EUP-Sr modified titanium implant is a promising candidate to be used in dental applications with notable osteoimmunomodulation and angiogenesis functions. And the novel technique proposed in this study would benefit the modification of metal/inorganic surfaces with polysaccharides for future research.
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Rapid and convenient wound healing is crucial for reducing potential post-traumatic wound complications. In this study, a temperature-sensitive polymer of poly-(HEMA-co-NIPAM) (PHN) was synthesized by free-radical polymerization, in which the solution quickly underwent a sol-gel transition above 29°C, thus responding to a typical body temperature and facilitating wound sealing. PHN solution incorporated with copper ions (PHN-Cu) not only exhibited excellent antibacterial properties, but also expedited wound closure and facilitated tissue angiogenesis. The in vivo and in vitro experiments showed that the PHN-Cu had a higher wound closure rate and demonstrated an ability to promote skin tissue angiogenesis. Such a versatile, convenient aqueous solution could enable nonprofessionals to promptly treat wounds in a short time after injury, thus providing suitable conditions for later treatment, and can be used as a convenient method to clean wounds.
Assuntos
Antibacterianos/química , Cobre/química , Poliaminas/química , Poli-Hidroxietil Metacrilato/análogos & derivados , Cicatrização/efeitos dos fármacos , Animais , Antibacterianos/farmacologia , Movimento Celular/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Cobre/farmacologia , Composição de Medicamentos , Escherichia coli/efeitos dos fármacos , Humanos , Masculino , Transição de Fase , Poli-Hidroxietil Metacrilato/química , Ratos Sprague-Dawley , Staphylococcus aureus/efeitos dos fármacos , Temperatura de Transição , Resultado do TratamentoRESUMO
To delay the degradation of magnesium alloys, silk fibroin as a natural organic polymer coating was fabricated on a 3-amino-propyltriethoxysilane (APTES) pretreated Mg-Zn-Ca alloy. APTES pretreatment coated the surface of magnesium alloys with amino groups, which can bond with functional groups in silk fibroin to form a compact coating/substrate interface. The influences of the APTES concentration and drying temperature on the coating adhesion and interface were investigated to explore the optimal parameters in the fabrication process. The nanoporous silk fibroin films completely covered the APTES pretreated Mg-Zn-Ca surface, which reached a thickness of ~7 µm. The chemical states for the coated Mg-Zn-Ca alloy were compared to those of the bare Mg-Zn-Ca alloy and the APTES pretreated Mg-Zn-Ca alloy to illustrate the coating mechanism. During in vitro degradation and electrochemical measurements in simulated body fluid (SBF), the samples with the silk fibroin coating showed remarkably improved corrosion resistance and a slower degradation rate compared to those of the bare samples, suggesting that the silk fibroin coating was an effective protection coating for the substrates and can delay the degradation of magnesium alloys. Moreover, a model for the in vitro degradation was proposed. In vitro cell experiments confirmed the excellent biocompatibility of silk fibroin coated Mg-Zn-Ca structure.
Assuntos
Ligas , Materiais Revestidos Biocompatíveis , Fibroínas , Teste de Materiais , Propilaminas , Silanos , Ligas/química , Ligas/farmacologia , Animais , Cálcio/química , Cálcio/farmacologia , Materiais Revestidos Biocompatíveis/química , Materiais Revestidos Biocompatíveis/farmacologia , Fibroínas/química , Fibroínas/farmacologia , Masculino , Manganês/química , Manganês/farmacologia , Camundongos , Porosidade , Propilaminas/química , Propilaminas/farmacologia , Silanos/química , Silanos/farmacologia , Zinco/química , Zinco/farmacologiaRESUMO
Traditional haemostatic materials generally have slow hemostasis rate and poor biocompatibility. This paper reports on the haemostatic properties and mechanism of silk fibroin (SF). SF-PEG sponge that could be solubilised and changed to gel form by blood was fabricated through mixing SF and polyethylene glycol (PEG, 1500 Da) followed by lyophilisation of the mixed solution. SF-PEG sponge, together with control samples of SF sponge (no PEG) and a commercially available haemostatic material, gelatine sponge, were subjected to the hemostasis tests using a liver trauma model of rabbit. The results showed that SF was superior to gelatine sponge in hemostasis time (136.17 ± 62.27 s and 249.83 ± 29.18 s) and blood loss (2.16 ± 1.27 g vs. 4.97 ± 1.44 g). Furthermore, in vitro experiments indicated SF-PEG sol-gel transition promoted platelet adhesion and aggregation, as well as platelet-fibrinogen interaction. Therefore, except for the physical blocking of bleeding port due to PEG-induced SF fast gelation, SF might also have an impact on blood coagulation process, a phenomenon that has not been reported before. In conclusion, SF is a new type of haemostatic material that might be able to meet the requirements of speed, efficiency and biosafety in a variety of clinical applications.
Assuntos
Fibroínas/química , Fibroínas/farmacologia , Hemostasia/efeitos dos fármacos , Polietileno/química , Animais , Géis , Humanos , Fígado/efeitos dos fármacos , Fígado/fisiologia , Estrutura Secundária de Proteína , Coelhos , Ratos , Ratos Wistar , Cicatrização/efeitos dos fármacosRESUMO
Magnesium and its alloys have generated considerable interest as one of the most promising biodegradable metals for biomedical bone implants. However, the enormous challenges are to improve their rapid corrosion excessively as well as to endow them with biocompatibility and biosafety. Herein, we introduce a natural silk fibroin protein coating to control the corrosion resistance and enhance the biocompatibility of MgZnCa alloy. To obtain a robust and reliable coated structure, different surface-activation processes are employed to increase the available functional groups on MgZnCa surfaces before coating. Compared to oxygen plasma activation, our unique vacuum ultraviolet-ozone (VUV/O3) activation method is effective in realizing uniform silk fibroin films as a protective barrier on MgZnCa alloy surfaces, and the nanoscratch test verified the superior adhesion strength of the silk fibroin-coated magnesium alloy structure. Long-term immersion results combined with electrochemical tests showed the preferable in vitro anticorrosion behavior and a low degradation rate of coated Mg alloy (1/8 times that of uncoated Mg alloy). Cell adhesion and cytotoxicity tests demonstrated that silk fibroin-coated MgZnCa presented improved biocompatibility with bone marrow mesenchymal stem cells. An animal study involving silk fibroin-coated MgZnCa implanted on one side of a rabbit spine for 180â¯days showed remarkably improved in vivo corrosion resistance, with 1/18 times the degradation rate of uncoated MgZnCa. These results not only comprehensively confirmed the validity of the VUV/O3-activation method as a coating strategy but also implied the tremendous potential of the modified Mg alloy for application as a degradable biomedical implant material. STATEMENT OF SIGNIFICANCE: MgZnCa alloy is a promising material in clinical implantation. Silk fibroin (SF) is a natural organic material with biocompatibility and biodegradability. To date, the combination of SF and MgZnCa alloy has exhibited considerable prospects for orthopedic applications. The realization of a direct coating is an enormous challenge because strong chemical bonds cannot be easily formed between organic and inorganic materials. To solve this bottleneck, we proposed a unique vacuum ultraviolet-ozone (VUV/O3) surface-activation method for the first time to modify the Mg alloy surface before SF coating, which significantly enhanced both in vitro and in vivo performance, such as superior biocompatibility and remarkably improved corrosion resistance of magnesium alloys (â¼1/18 the in vivo degradation rate of uncoated MgZnCa).
Assuntos
Ligas , Materiais Revestidos Biocompatíveis , Teste de Materiais , Ligas/química , Ligas/farmacologia , Animais , Cálcio/química , Cálcio/farmacologia , Materiais Revestidos Biocompatíveis/química , Materiais Revestidos Biocompatíveis/farmacologia , Corrosão , Fibroínas/química , Fibroínas/farmacologia , Magnésio/química , Magnésio/farmacologia , Masculino , Coelhos , Zinco/química , Zinco/farmacologiaRESUMO
Tissue engineering (TE) is an emerging and promising strategy to heal tissue failure by integrating science and technology of materials, cells and growth factors. With the increasing of aging population, restoring musculoskeletal tissue has become the focus of TE. Among various materials tested in TE, silk fibroin (SF) is increasingly being recognized as a promising material. SF, a natural protein polymer with excellent physiochemical characteristics, has established a good reputation in terms of musculoskeletal tissue engineering (MTE). The present article provides an overview of SF and introduces various approaches of fabricating SF-based biomaterial followed by their applications in MTE.
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Materiais Biocompatíveis/química , Fibroínas/química , Engenharia Tecidual , Animais , Materiais Biocompatíveis/farmacologia , Cartilagem/fisiologia , Tecido Conjuntivo/fisiologia , Humanos , Hidrogéis/química , Músculo Esquelético/fisiologia , Regeneração/efeitos dos fármacosRESUMO
The effects of changing solution pH in the range of 3.6-10.0 during a one-step silk microsphere preparation process, by mixing silk and polyethylene glycol (PEG), was assessed. The microspheres prepared at low pH (3.6) showed a more homogeneous size (1-3µm) and less porous texture than those prepared at neutral pH. High pH (10.0) inhibited microsphere formation, yielding small and inhomogeneous microspheres. Compared to neutral pH, low pH also increased the content of silk crystalline ß-sheet structure from approx. 30% to above 40%. As a result, the microspheres produced at low pH were more thermally stable as well as resistant to chemical (8M urea) and enzymatic (protease XIV) degradation when compared to microspheres prepared at neutral pH. Doxorubicin hydrochloride (DOX) and curcumin (CUR) were successfully loaded in silk microspheres via control of solution pH. The loading efficiency of DOX was approx. 95% at pH7.0 and approx. 60% for CUR at pH3.6, attributed to charge-charge interactions and hydrophobic interactions between the silk and drug molecules, respectively. When PBS, pH7.4, was used as a medium for release studies, the pH3.6 microspheres released both drugs more slowly than the pH7.0 microspheres, likely due to the high content of crystalline ß-sheet structure that enhanced drug-silk interactions as well as restricted drug molecule diffusion.
Assuntos
Polietilenoglicóis/química , Concentração de Íons de Hidrogênio , Microesferas , SedaRESUMO
Ultrasound (US) is a useful technique to monitor morphological and functional changes of biomaterial implants without sacrificing the animal. Contrast-enhanced ultrasound (CEUS) along with two-dimensional (2D) US were used to characterize the biodegradation and neovascularization of silk protein (8 wt%) hydrogel implants in rats. Cylinder-shaped silk hydrogel plugs were implanted into the space between the hind limb thigh muscles in Wistar rats (n = 6). The increase of echogenicity in 2D US revealed tissue-ingrowth-accompanied gel degradation over 18 weeks. The shape and size of the implanted gels remained qualitatively unchanged until week 15, as confirmed by Bland and Altman analysis and visualization of retrieved samples. Using CEUS, neovascularization was monitored by the presence of microbubbles in the gel area, and the dynamic vascularization process was indicated by the contrast enhancement values, which showed a relatively low level (< 5 dB) during weeks 1-8 and significantly increased levels (around 20 dB at week 15 and > 35 dB at week 18), suggesting that major vascularization had occurred in the gel implants by this time point. Histological and scanning electron microscopic analysis of explants revealed time-dependent increases in the pore size of the gel matrix, the presence of endothelial and red blood cells and the number of blood vessels in the gel implants, indicating that degradation and vascularization did occur in silk gel implants during the time period. The present study demonstrates the use of US imaging for monitoring of in vivo degradation and vascularization of silk implants in a non-destructive way. Copyright © 2015 John Wiley & Sons, Ltd.
Assuntos
Hidrogel de Polietilenoglicol-Dimetacrilato/farmacologia , Neovascularização Fisiológica/efeitos dos fármacos , Seda/farmacologia , Ultrassom/métodos , Animais , Meios de Contraste/química , Implantes Experimentais , Ratos Wistar , Seda/ultraestruturaRESUMO
Given the fabulous potential of promoting bone regeneration, BMP-2 has been investigated widely in the bone tissue engineering field. A sophisticated biomaterial loaded with BMP-2, which could avoid the required supraphysiological dose leading to high medical costs and risks of complications, has been considered as a promising strategy to treat non-healing bone defects. In this study, we developed a simple approach to engineer a composited hydrogel consisting polymeric particles (PLA/PLGA) used as a BMP-2 delivery vehicle. Compared with other groups, the introduction of PLA into PEG-silk gels endowed the hydrogel new physicochemical characteristics especially hydrophobicity which inhibited the burst release of BMP-2 and enhanced gel's structural stability. Moreover, such composited gels could stabilize entrapped proteins and maintain their bioactivity fully in vitro. In vivo, the bio-degradability experiment demonstrated this system was biocompatible and the reinforced hydrophobicity significantly decreased degradation rate, and in rat critical-sized cranial defects model, the gel containing PLA promoted the most bone formation. These findings demonstrated the introduction of PLA changed physicochemical features of gels more suitable as a BMP-2 carrier indicated by inducing bone regeneration efficiently in large bone defects at low delivered dose and this system may own translational potential.
Assuntos
Proteína Morfogenética Óssea 2/farmacologia , Regeneração Óssea/efeitos dos fármacos , Portadores de Fármacos/química , Hidrogel de Polietilenoglicol-Dimetacrilato/química , Seda/química , Animais , Proteína Morfogenética Óssea 2/química , Proteína Morfogenética Óssea 2/genética , Proteína Morfogenética Óssea 2/metabolismo , Osso e Ossos/patologia , Microscopia Eletrônica de Varredura , Ratos , Ratos Wistar , Proteínas Recombinantes/biossíntese , Proteínas Recombinantes/química , Proteínas Recombinantes/farmacologia , Espectroscopia de Infravermelho com Transformada de FourierRESUMO
Silk hydrogels for tissue repair are usually pre-formed via chemical or physical treatments from silk solutions. For many medical applications, it is desirable to utilize injectable silk hydrogels at high concentrations (>8%) to avoid surgical implantation and to achieve slow in vivo degradation of the gel. In the present study, injectable silk solutions that formed hydrogels in situ were generated by mixing silk with low-molecular-weight polyethylene glycol (PEG), especially PEG300 and 400 (molecular weight 300 and 400g mol(-1)). Gelation time was dependent on the concentration and molecular weight of PEG. When the concentration of PEG in the gel reached 40-45%, gelation time was less than 30min, as revealed by measurements of optical density and rheological studies, with kinetics of PEG400 faster than PEG300. Gelation was accompanied by structural changes in silk, leading to the conversion from random coil in solution to crystalline ß-sheets in the gels, based on circular dichroism, attenuated total reflection Fourier transform infrared spectroscopy and X-ray diffraction. The modulus (127.5kPa) and yield strength (11.5kPa) determined were comparable to those of sonication-induced hydrogels at the same concentrations of silk. The time-dependent injectability of 15% PEG-silk hydrogel through 27G needles showed a gradual increase of compression forces from â¼10 to 50N within 60min. The growth of human mesenchymal stem cells on the PEG-silk hydrogels was hindered, likely due to the presence of PEG, which grew after a 5 day delay, presumably while the PEG solubilized away from the gel. When 5% PEG-silk hydrogel was subcutaneously injected in rats, significant degradation and tissue in-growth took place after 20 days, as revealed by ultrasound imaging and histological analysis. No significant inflammation around the gel was observed. The features of injectability, slow degradation and low initial cell attachment suggests that these PEG-silk hydrogels are of interest for many biomedical applications, such as anti-fouling and anti-adhesion.
Assuntos
Hidrogéis , Polietilenoglicóis , Seda , Animais , Cinética , Microscopia Eletrônica de Varredura , Ratos , Ratos Wistar , Espectroscopia de Infravermelho com Transformada de Fourier , Difração de Raios XRESUMO
BACKGROUND: Peripheral nerve injury is a common trauma, but presents a significant challenge to the clinic. Silk-based materials have recently become an important biomaterial for tissue engineering applications due to silk's biocompatibility and impressive mechanical and degradative properties. In the present study, a silk fibroin peptide (SF16) was designed and used as a component of the hydrogel scaffold for the repair of peripheral nerve injury. METHODS: The SF16 peptide's structure was characterized using spectrophotometry and atomic force microscopy, and the SF16 hydrogel was analyzed using scanning electron microscopy. The effects of the SF16 hydrogel on the viability and growth of live cells was first assessed in vitro, on PC12 cells. The in vivo test model involved the repair of a nerve gap with tubular nerve guides, through which it was possible to identify if the SF16 hydrogel would have the potential to enhance nerve regeneration. In this model physiological saline was set as the negative control, and collagen as the positive control. Walking track analysis and electrophysiological methods were used to evaluate the functional recovery of the nerve at 4 and 8 weeks after surgery. RESULTS: Analysis of the SF16 peptide's characteristics indicated that it consisted of a well-defined secondary structure and exhibited self-assembly. Results of scanning electron microscopy showed that the peptide based hydrogel may represent a porous scaffold that is viable for repair of peripheral nerve injury. Analysis of cell culture also supported that the hydrogel was an effective matrix to maintain the viability, morphology and proliferation of PC12 cells. Electrophysiology demonstrated that the use of the hydrogel scaffold (SF16 or collagen) resulted in a significant improvement in amplitude recovery in the in vivo model compared to physiological saline. Moreover, nerve cells in the SF16 hydrogel group displayed greater axon density, larger average axon diameter and thicker myelin compared to those of the group that received physiological saline. CONCLUSION: The SF16 hydrogel scaffold may promote excellent axonal regeneration and functional recovery after peripheral nerve injury, and the SF16 peptide may be a candidate for nerve tissue engineering applications.
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Hidrogel de Polietilenoglicol-Dimetacrilato/química , Regeneração Nervosa/efeitos dos fármacos , Peptídeos/química , Alicerces Teciduais/química , Animais , Técnicas de Cultura de Células , Linhagem Celular Tumoral , Feminino , Fibroínas/química , Fibroínas/farmacologia , Hidrogel de Polietilenoglicol-Dimetacrilato/farmacologia , Tamanho da Partícula , Peptídeos/farmacologia , Ratos , Ratos Sprague-Dawley , Nervo Isquiático/efeitos dos fármacos , Nervo Isquiático/lesões , Nervo Isquiático/ultraestruturaRESUMO
In regenerative medicine, accumulating evidence demonstrates that the property of substrates monitors neural stem cells behavior. However, how stem cells sense and interpret biochemical and topographical cues remains elusive. This study aimed to explore the mechanism how nanofibrous scaffold modulated stem cells behavior. Spinal cord derived neural progenitor cells (NPCs) were cultured on electrospun aligned and randomly oriented collagen nanofibrous scaffolds. A 30% increase in proliferation and an elevation of BrdU incorporation were observed in NPCs on collagen nanofibers, compared to that on collagen-coated surface. In particular, NPCs expanded faster on aligned nanofibers in comparison with that on randomly oriented nanofibers. Moreover, an alteration in cell cycle progression with a reduced percentage of cells in G0/G1 phase and increased cell proliferation index (S phase plus G2/M phase) was also detected in NPCs cultured on collagen nanofibers. Incubating NPCs with anti-ß1 integrin antibody or U1026 (an inhibitor of mitogen-activated protein kinase kinase, MEK) eliminated the altered cell cycle dynamics and BrdU incorporation induced by collagen nanofibers. In addition, cyclin D1 and cyclin dependent kinase 2 (CDK2), downstream genes of ß1 integrin/mitogen-activated protein kinase (MAPK) pathway that control G1/S phase transition, were correspondingly regulated by nanofibers. Collectively, these data suggested that the property of substrate modulated NPCs proliferation by promoting cell cycle through ß1 integrin/MAPK pathway. Our findings provide a better understanding of the interaction between NPCs and the substrate and therefore will pave way for regenerative medicine.
Assuntos
Colágeno/metabolismo , Colágeno/ultraestrutura , Integrina beta1/metabolismo , Sistema de Sinalização das MAP Quinases/fisiologia , Proteínas Quinases Ativadas por Mitógeno/metabolismo , Nanofibras/ultraestrutura , Células-Tronco Neurais/fisiologia , Animais , Materiais Biocompatíveis/química , Materiais Biocompatíveis/metabolismo , Ciclo Celular/fisiologia , Proliferação de Células , Células Cultivadas , Nanofibras/química , Regeneração Nervosa , Células-Tronco Neurais/citologia , Ratos , Ratos Sprague-Dawley , Medula Espinal/citologiaRESUMO
PAA/gelatin nanoparticles, with interpolymer complexes of gelatin and polyacrylic acid (PAA) as the cores and gelatin as the shells, were prepared via facile polymerization of AA on gelatin template. The morphology change of the nanoparticles during the reaction was traced by a combined use of dynamic light scattering (DLS) and atomic force microscopy (AFM) techniques, which revealed a discrepancy among the structure of the nanoparticles formed at different stages of the reaction: as the reaction proceeds, nanoparticles with larger compact cores and thinner shells are produced. The resultant nanoparticles are multi-responsive. Especially, they exhibit a significant temperature-dependent size change: upon raising the temperature from 25 degrees C, the nanoparticle size decreases monotonically until reaching equilibrium at about 40 degrees C. This temperature-dependence of the nanoparticle size was found to be reversible provided the nanoparticle solution was cooled at a low temperature (4 degrees C). The thermo-sensitivity of the nanoparticles is attributed to the thermo-induced sol-gel transition of the gelatin shells. In addition, the nanoparticles were further converted to hollow spheres via successive locking the shell structure by the reaction of gelatin with cross-linker glutaraldehyde, and cavitation of the cross-linked nanoparticles by switching the medium from acidic to neutral. The cavitation process was monitored by DLS, which indicated a mass decrease and size shrinkage. AFM and transmission electron microscopy (TEM) were used to trace the morphology change of the nanoparticles during the cavitation. The hollow structure was confirmed by TEM observation.