Background Cu-Al-Ni shape memory alloys (SMAs) possess two-way shape memory effects, superelasticity, and damping capacity. Nonetheless, Cu-Al-Ni SMAs remain promising candidates for use in biomedical applications, as they are more economical and machinable than other SMAs. Ensuring the biocompatibility of Cu-Al-Ni SMAs is crucial to their development for biomedical applications. Therefore, this study aimed to assess the toxicity of Cu-Al-Ni SMAs using a Probit dose–response model and augmented simplex design. Methods In this study, the effects of Cu2+, Al3+ and Ni2+ metal ions on bacteria (Escherichia coli DH5α) using Probit dose–response analysis and augmented simplex design to assess the actual toxicity of the Cu-Al-Ni SMAs. Results Extraction and repetition of Escherichia coli DH5α solutions with high Cu2+ ion concentrations and 30-hour incubation demonstrated that Escherichia coli DH5α was able to alter its growth mechanisms in response to toxins. Metal ions leached from Cu-Al-Ni SMAs appeared in a multitude of compositions with varying degrees of toxicity, and those appearing close to a saddle region identified in the contour plot of the augmented simplex model were identified as candidates for elevated toxicity levels. When the Cu-13.5Al-4Ni SMA plate was immersed in Ringer's solution, the selective leaching rate of Ni2+ ions far exceeded that of Cu2+ and Al3+. The number of Cu2+, Al3+ and Ni2+ ions leached from Cu-Al-Ni SMAs increased with immersion time; however, at higher ratios, toxicity interactions among the metal ions had the effect of gradually reducing overall toxicity levels with regard to Escherichia coli DH5α. Conclusions The quantities of Cu2+, Al3+ and Ni2+ ions leached from the Cu-13.5Al-4Ni SMA plate increased with immersion time, the toxicity interactions associated with these compositions reduced the actual toxicity to Escherichia coli DH5α.
{"title":"Toxicity Assessment and Selective Leaching Characteristics of Cu-Al-Ni Shape Memory Alloys in Biomaterials Applications","authors":"Shih-Hang Chang, Bor-Yann Chen, Jin-Xiang Lin","doi":"10.5301/jabfm.5000245","DOIUrl":"https://doi.org/10.5301/jabfm.5000245","url":null,"abstract":"Background Cu-Al-Ni shape memory alloys (SMAs) possess two-way shape memory effects, superelasticity, and damping capacity. Nonetheless, Cu-Al-Ni SMAs remain promising candidates for use in biomedical applications, as they are more economical and machinable than other SMAs. Ensuring the biocompatibility of Cu-Al-Ni SMAs is crucial to their development for biomedical applications. Therefore, this study aimed to assess the toxicity of Cu-Al-Ni SMAs using a Probit dose–response model and augmented simplex design. Methods In this study, the effects of Cu2+, Al3+ and Ni2+ metal ions on bacteria (Escherichia coli DH5α) using Probit dose–response analysis and augmented simplex design to assess the actual toxicity of the Cu-Al-Ni SMAs. Results Extraction and repetition of Escherichia coli DH5α solutions with high Cu2+ ion concentrations and 30-hour incubation demonstrated that Escherichia coli DH5α was able to alter its growth mechanisms in response to toxins. Metal ions leached from Cu-Al-Ni SMAs appeared in a multitude of compositions with varying degrees of toxicity, and those appearing close to a saddle region identified in the contour plot of the augmented simplex model were identified as candidates for elevated toxicity levels. When the Cu-13.5Al-4Ni SMA plate was immersed in Ringer's solution, the selective leaching rate of Ni2+ ions far exceeded that of Cu2+ and Al3+. The number of Cu2+, Al3+ and Ni2+ ions leached from Cu-Al-Ni SMAs increased with immersion time; however, at higher ratios, toxicity interactions among the metal ions had the effect of gradually reducing overall toxicity levels with regard to Escherichia coli DH5α. Conclusions The quantities of Cu2+, Al3+ and Ni2+ ions leached from the Cu-13.5Al-4Ni SMA plate increased with immersion time, the toxicity interactions associated with these compositions reduced the actual toxicity to Escherichia coli DH5α.","PeriodicalId":51074,"journal":{"name":"Journal of Applied Biomaterials & Biomechanics","volume":"14 1","pages":"59 - 64"},"PeriodicalIF":0.0,"publicationDate":"2016-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.5301/jabfm.5000245","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"70591261","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
J. Franková, V. Pivodová, H. Vágnerová, J. Juráňová, J. Ulrichová
Background Nanoparticles are widely used in different technological fields, one of which is medicine. Because of their antibacterial properties, silver nanoparticles (AgNPs) are used in several types of wound dressings for the treatment of burns and nonhealing wounds, but their influence on each component of the wound-healing process remains unclear. In the present study, we evaluated the effects of AgNPs on normal human dermal fibroblasts (NHDFs) and normal human epidermal keratinocytes (NHEKs). Both cell types are important for wound healing, including with regard to inflammation, proliferation and tissue remodeling. Each phase of wound healing can be characterized by the secretion of cytokines, chemokines and growth factors. Methods The production of inflammatory parameters (tumor necrosis factor α [TNF-α], interleukin-6 [IL-6], IL-8 and IL-12 and cyclooxygenase-2 [COX-2]), angiogenesis parameters (vascular endothelial growth factor [VEGF], granulocyte macrophage colony-stimulating factor) and matrix metalloproteinases (MMP-1, MMP-2, MMP-3 and MMP-9) by NHDFs and NHEKs were examined by ELISA or Western blot after 24 and 48 hours of incubation with AgNPs. Results We found that AgNPs decreased some inflammatory cytokines (TNF-α and IL-12) and growth factors (VEGF) that were produced by NHDFs and NHEKs after 24 and 48 hours and decreased the expression of COX-2 after 24 hours but only at the highest concentration of AgNPs (25 parts per million). Conclusions The results indicate that NHEKs are more susceptible to the application of AgNPs than NHDFs, and AgNPs may be useful for medical applications for the treatment of wounds.
{"title":"Effects of Silver Nanoparticles on Primary Cell Cultures of Fibroblasts and Keratinocytes in a Wound-Healing Model","authors":"J. Franková, V. Pivodová, H. Vágnerová, J. Juráňová, J. Ulrichová","doi":"10.5301/jabfm.5000268","DOIUrl":"https://doi.org/10.5301/jabfm.5000268","url":null,"abstract":"Background Nanoparticles are widely used in different technological fields, one of which is medicine. Because of their antibacterial properties, silver nanoparticles (AgNPs) are used in several types of wound dressings for the treatment of burns and nonhealing wounds, but their influence on each component of the wound-healing process remains unclear. In the present study, we evaluated the effects of AgNPs on normal human dermal fibroblasts (NHDFs) and normal human epidermal keratinocytes (NHEKs). Both cell types are important for wound healing, including with regard to inflammation, proliferation and tissue remodeling. Each phase of wound healing can be characterized by the secretion of cytokines, chemokines and growth factors. Methods The production of inflammatory parameters (tumor necrosis factor α [TNF-α], interleukin-6 [IL-6], IL-8 and IL-12 and cyclooxygenase-2 [COX-2]), angiogenesis parameters (vascular endothelial growth factor [VEGF], granulocyte macrophage colony-stimulating factor) and matrix metalloproteinases (MMP-1, MMP-2, MMP-3 and MMP-9) by NHDFs and NHEKs were examined by ELISA or Western blot after 24 and 48 hours of incubation with AgNPs. Results We found that AgNPs decreased some inflammatory cytokines (TNF-α and IL-12) and growth factors (VEGF) that were produced by NHDFs and NHEKs after 24 and 48 hours and decreased the expression of COX-2 after 24 hours but only at the highest concentration of AgNPs (25 parts per million). Conclusions The results indicate that NHEKs are more susceptible to the application of AgNPs than NHDFs, and AgNPs may be useful for medical applications for the treatment of wounds.","PeriodicalId":51074,"journal":{"name":"Journal of Applied Biomaterials & Biomechanics","volume":"14 1","pages":"137 - 142"},"PeriodicalIF":0.0,"publicationDate":"2016-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.5301/jabfm.5000268","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"70592330","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Claudia Ottardi, L. La Barbera, L. Pietrogrande, T. Villa
Background Vertebral compression fractures occur in the thoracolumbar junction, causing the collapse of the vertebral body. For their treatment, vertebroplasty and kyphoplasty are used, but it is still unknown which technique is to be preferred. Methods Finite element models of the thoracic spine were developed to evaluate the outcomes of vertebroplasty and kyphoplasty. A mild and severe collapse of T10 treated with vertebroplasty or kyphoplasty was studied. Stresses on the endplates and intradiscal pressures were extrapolated to determine the stress distribution in the adjacent structures. Results The validation ensured a correct stiffness and a proper kinematic of each functional spinal unit. The results demonstrated that a consolidation following vertebroplasty caused slight variations of intradiscal pressures and stresses. If a kyphoplasty was performed after a mild collapse of the vertebral body, a 25% stress reduction on endplates was found. In cases of severe collapse, when a partial height restoration was achieved, a 15% stress reduction was obtained, while with a full recovery of the anterior wall of the collapsed vertebra, there was a further reduction of 40%. Conclusions To reduce the stresses on the adjacent endplates and the risk of fracture, the results suggest a kyphoplasty is to be preferred, trying to restore the initial vertebral body height.
{"title":"Vertebroplasty and Kyphoplasty for the Treatment of Thoracic Fractures in Osteoporotic Patients: A Finite Element Comparative Analysis","authors":"Claudia Ottardi, L. La Barbera, L. Pietrogrande, T. Villa","doi":"10.5301/jabfm.5000287","DOIUrl":"https://doi.org/10.5301/jabfm.5000287","url":null,"abstract":"Background Vertebral compression fractures occur in the thoracolumbar junction, causing the collapse of the vertebral body. For their treatment, vertebroplasty and kyphoplasty are used, but it is still unknown which technique is to be preferred. Methods Finite element models of the thoracic spine were developed to evaluate the outcomes of vertebroplasty and kyphoplasty. A mild and severe collapse of T10 treated with vertebroplasty or kyphoplasty was studied. Stresses on the endplates and intradiscal pressures were extrapolated to determine the stress distribution in the adjacent structures. Results The validation ensured a correct stiffness and a proper kinematic of each functional spinal unit. The results demonstrated that a consolidation following vertebroplasty caused slight variations of intradiscal pressures and stresses. If a kyphoplasty was performed after a mild collapse of the vertebral body, a 25% stress reduction on endplates was found. In cases of severe collapse, when a partial height restoration was achieved, a 15% stress reduction was obtained, while with a full recovery of the anterior wall of the collapsed vertebra, there was a further reduction of 40%. Conclusions To reduce the stresses on the adjacent endplates and the risk of fracture, the results suggest a kyphoplasty is to be preferred, trying to restore the initial vertebral body height.","PeriodicalId":51074,"journal":{"name":"Journal of Applied Biomaterials & Biomechanics","volume":"14 1","pages":"197 - 204"},"PeriodicalIF":0.0,"publicationDate":"2016-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.5301/jabfm.5000287","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"70592600","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Background Conductive biomaterials are an ideal biosubstrate for modifying cellular behaviors by conducting either internal or external electrical signals. In this study, based on a simple-preparation graphite exfoliation method in organic reagent, a nonfunctionalized graphene nanosheet film (NGNF) with high conductivity and large size was simply fabricated through spraying coating. The biocompatibility of the NGNF was carefully tested with primary cortical neuron cells, and its biocompatibility properties were compared with a chemical vapor deposition (CVD) graphene film. Methods Nonfunctionalized graphene nanosheet (NGN) was first exfoliated from graphite with a flat-tip ultrasonicator probe, and then spray-coated onto glass slide substrate to form the film. The morphology of NGNF was observed with light microscopy and SEM. The morphology and neuronal network formation of primary cortical neuron cells onto NGNF, as shown by DAPI and Alexa Fluor® 488 staining, were observed with fluorescent microscopy. Cell viability and proliferation were measured with MTT. Results NGNF had better cell biocompatibility than CVD graphene film. MTT test showed that NGNF exhibited no cytotoxicity. According to neuronal network formation at 7 days of cell culture, primary neuron cells aggregated into 50-μm “nuclei”; the average neurite number and length were 3 and 100 μm, respectively. However, these values were almost doubled after 14 days of cell culture. Conclusions These results may improve the use of NGNF as a conductive scaffold for nerve regeneration.
{"title":"Growth and Follow-Up of Primary Cortical Neuron Cells on Nonfunctionalized Graphene Nanosheet Film","authors":"Shiyun Meng, Rong Peng","doi":"10.5301/jabfm.5000263","DOIUrl":"https://doi.org/10.5301/jabfm.5000263","url":null,"abstract":"Background Conductive biomaterials are an ideal biosubstrate for modifying cellular behaviors by conducting either internal or external electrical signals. In this study, based on a simple-preparation graphite exfoliation method in organic reagent, a nonfunctionalized graphene nanosheet film (NGNF) with high conductivity and large size was simply fabricated through spraying coating. The biocompatibility of the NGNF was carefully tested with primary cortical neuron cells, and its biocompatibility properties were compared with a chemical vapor deposition (CVD) graphene film. Methods Nonfunctionalized graphene nanosheet (NGN) was first exfoliated from graphite with a flat-tip ultrasonicator probe, and then spray-coated onto glass slide substrate to form the film. The morphology of NGNF was observed with light microscopy and SEM. The morphology and neuronal network formation of primary cortical neuron cells onto NGNF, as shown by DAPI and Alexa Fluor® 488 staining, were observed with fluorescent microscopy. Cell viability and proliferation were measured with MTT. Results NGNF had better cell biocompatibility than CVD graphene film. MTT test showed that NGNF exhibited no cytotoxicity. According to neuronal network formation at 7 days of cell culture, primary neuron cells aggregated into 50-μm “nuclei”; the average neurite number and length were 3 and 100 μm, respectively. However, these values were almost doubled after 14 days of cell culture. Conclusions These results may improve the use of NGNF as a conductive scaffold for nerve regeneration.","PeriodicalId":51074,"journal":{"name":"Journal of Applied Biomaterials & Biomechanics","volume":"42 1","pages":"26 - 34"},"PeriodicalIF":0.0,"publicationDate":"2016-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.5301/jabfm.5000263","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"70592013","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
C. Cristallini, E. Bellotti, F. Spezia, E. Rosellini, M. Cascone, N. Barbani
Background The accumulation of amyloid beta protein in the brain causes the cognitive impairment observed in neurodegenerative pathologies such as Alzheimer's disease. The present study aimed to test the hypothesis that a rapid removal of amyloid beta protein peptides from the blood by an extracorporeal purification system could represent an alternative solution for the treatment of patients suffering from this neurodegenerative disease. Methods In this regard, we investigated the specific recognition properties of a molecularly imprinted membrane based on poly(ethylene-co-vinyl alcohol) toward the amyloid beta protein fragment 25-35 (AbP), the more neurotoxic domain of amyloid beta protein. A chemical modification of the copolymer backbone using succinic anhydride was also performed to favor the formation of carboxylic groups and thus improve imprinting performance. Results The physico-chemical, morphological, mechanical and functional characterisations gave interesting results confirming the ability of imprinted membranes to in vitro rebind AbP. Conclusions This work represents a proof of concept regarding the development of a biocompatible polymer membrane capable of selectively removing amyloid beta peptide from the blood and consequently from the cerebrospinal fluid.
{"title":"A New Strategy to Reduce Amyloid Deposition using Peptide-Imprinted Membranes","authors":"C. Cristallini, E. Bellotti, F. Spezia, E. Rosellini, M. Cascone, N. Barbani","doi":"10.5301/jabfm.5000288","DOIUrl":"https://doi.org/10.5301/jabfm.5000288","url":null,"abstract":"Background The accumulation of amyloid beta protein in the brain causes the cognitive impairment observed in neurodegenerative pathologies such as Alzheimer's disease. The present study aimed to test the hypothesis that a rapid removal of amyloid beta protein peptides from the blood by an extracorporeal purification system could represent an alternative solution for the treatment of patients suffering from this neurodegenerative disease. Methods In this regard, we investigated the specific recognition properties of a molecularly imprinted membrane based on poly(ethylene-co-vinyl alcohol) toward the amyloid beta protein fragment 25-35 (AbP), the more neurotoxic domain of amyloid beta protein. A chemical modification of the copolymer backbone using succinic anhydride was also performed to favor the formation of carboxylic groups and thus improve imprinting performance. Results The physico-chemical, morphological, mechanical and functional characterisations gave interesting results confirming the ability of imprinted membranes to in vitro rebind AbP. Conclusions This work represents a proof of concept regarding the development of a biocompatible polymer membrane capable of selectively removing amyloid beta peptide from the blood and consequently from the cerebrospinal fluid.","PeriodicalId":51074,"journal":{"name":"Journal of Applied Biomaterials & Biomechanics","volume":"14 1","pages":"129 - 136"},"PeriodicalIF":0.0,"publicationDate":"2016-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.5301/jabfm.5000288","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"70592677","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Luca Coluccino, P. Stagnaro, M. Vassalli, S. Scaglione
Background The design of an appropriate microenvironment for stem cell differentiation constitutes a multitask mission and a critical step toward the clinical application of tissue substitutes. With the aim of producing a bioactive material for orthopedic applications, a transforming growth factor-β (TGF- β1)/hydroxyapatite (HA) association within an alginate-based scaffold was investigated. The bioactive scaffold was carefully designed to offer specific biochemical cues for an efficient and selective cell differentiation toward the bony and chondral lineages. Methods Highly porous alginate scaffolds were fabricated from a mixture of calcium cross-linked alginates by means of a freeze-drying technique. In the chondral layer, the TGF in citric acid was mixed with an alginate/alginate-sulfate solution. In the bony layer, HA granules were added as bioactive signal, to offer an osteoinductive surface to the cells. Optical and scanning electron microscopy analyses were performed to assess the macro-micro architecture of the biphasic scaffold. Different mechanical tests were conducted to evaluate the elastic modulus of the grafts. For the biological validation of the developed prototype, mesenchymal stem cells were loaded onto the samples; cellular adhesion, proliferation and in vivo biocompatibility were evaluated. Results and conclusions The results successfully demonstrated the efficacy of the designed osteochondral graft, which combined interesting functional properties and biomechanical performances, thus becoming a promising candidate for osteochondral tissue-engineering applications.
{"title":"Bioactive TGF-β1/HA Alginate-Based Scaffolds for Osteochondral Tissue Repair: Design, Realization and Multilevel Characterization","authors":"Luca Coluccino, P. Stagnaro, M. Vassalli, S. Scaglione","doi":"10.5301/jabfm.5000249","DOIUrl":"https://doi.org/10.5301/jabfm.5000249","url":null,"abstract":"Background The design of an appropriate microenvironment for stem cell differentiation constitutes a multitask mission and a critical step toward the clinical application of tissue substitutes. With the aim of producing a bioactive material for orthopedic applications, a transforming growth factor-β (TGF- β1)/hydroxyapatite (HA) association within an alginate-based scaffold was investigated. The bioactive scaffold was carefully designed to offer specific biochemical cues for an efficient and selective cell differentiation toward the bony and chondral lineages. Methods Highly porous alginate scaffolds were fabricated from a mixture of calcium cross-linked alginates by means of a freeze-drying technique. In the chondral layer, the TGF in citric acid was mixed with an alginate/alginate-sulfate solution. In the bony layer, HA granules were added as bioactive signal, to offer an osteoinductive surface to the cells. Optical and scanning electron microscopy analyses were performed to assess the macro-micro architecture of the biphasic scaffold. Different mechanical tests were conducted to evaluate the elastic modulus of the grafts. For the biological validation of the developed prototype, mesenchymal stem cells were loaded onto the samples; cellular adhesion, proliferation and in vivo biocompatibility were evaluated. Results and conclusions The results successfully demonstrated the efficacy of the designed osteochondral graft, which combined interesting functional properties and biomechanical performances, thus becoming a promising candidate for osteochondral tissue-engineering applications.","PeriodicalId":51074,"journal":{"name":"Journal of Applied Biomaterials & Biomechanics","volume":"14 1","pages":"42 - 52"},"PeriodicalIF":0.0,"publicationDate":"2015-12-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.5301/jabfm.5000249","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"70591417","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Titanium and its alloys are conventionally used to produce medical devices, but their biocompatibility has not yet been optimized. Surface modification, especially control of the surface roughness of titanium, is one strategy for improving biocompatibility and providing effective binding to hard tissue. However, the soft tissue compatibility of metallic materials is currently poorly understood, and effective techniques for tight binding between metal surfaces and soft tissue are still under development. Therefore, we here investigated whether the surface roughness of titanium affects fibroblast adhesion and proliferation. Our results showed that a surface roughness of ∼100 nm reduces fibroblast function. On such surfaces, distinct focal adhesion was not observed. These findings improve the general understanding of the binding compatibility between soft tissues and metallic materials.
{"title":"Sub-Micrometer Scale Surface Roughness of Titanium Reduces Fibroblasts Function","authors":"S. Migita, So Okuyama, Kunitaka Araki","doi":"10.5301/jabfm.5000260","DOIUrl":"https://doi.org/10.5301/jabfm.5000260","url":null,"abstract":"Titanium and its alloys are conventionally used to produce medical devices, but their biocompatibility has not yet been optimized. Surface modification, especially control of the surface roughness of titanium, is one strategy for improving biocompatibility and providing effective binding to hard tissue. However, the soft tissue compatibility of metallic materials is currently poorly understood, and effective techniques for tight binding between metal surfaces and soft tissue are still under development. Therefore, we here investigated whether the surface roughness of titanium affects fibroblast adhesion and proliferation. Our results showed that a surface roughness of ∼100 nm reduces fibroblast function. On such surfaces, distinct focal adhesion was not observed. These findings improve the general understanding of the binding compatibility between soft tissues and metallic materials.","PeriodicalId":51074,"journal":{"name":"Journal of Applied Biomaterials & Biomechanics","volume":"14 1","pages":"65 - 69"},"PeriodicalIF":0.0,"publicationDate":"2015-12-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.5301/jabfm.5000260","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"70592165","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Purpose Widespread application of collagen-glycosaminoglycan dermal templates in the treatment of cutaneous defects has identified the interval between initial engraftment and skin graft application as important for improvement. The aim of this study was to evaluate the effect of hyaluronan supplementation of Integra® dermal template on human dermal fibroblasts and keratinocytes in both in vitro and ex vivo models. Methods This study utilized in vitro and ex vivo cell culture techniques to investigate supplementing Integra® Regeneration Template with hyaluronan (HA), as a strategy to decrease this interval. In vitro, Integra® was HA supplemented at 0.15, 1, 1.5 and 2 mg/mL−1. Primary human dermal fibroblast (PHDF) and keratinocyte proliferation, PHDF viability, migration and HA-induced signal transduction (phosphor-MAPK Array) were assessed. Ex vivo, wound models (wound diameter 4 mm) were created within 8 mm skin biopsies. Wounds were filled with Integra® or HA supplemented Integra®. Re-epithelialization was compared through hematoxylin and eosin-stained cross-sections at 7, 14 and 21 days in culture. Model viability was assessed through lactate dehydrogenase (LDH) assays. Results In vitro, PHDF and keratinocyte proliferation were enhanced significantly (p<0.001) when supplemented with HA. S-Phase and G2/M PHDFs in HA supplemented scaffolds increased. PHDF viability was enhanced to 72 hours culture with 1.5 mg/mL−1 HA (p = 0.016). PHDF migration was maximally enhanced at 1 mg/mL−1 and 1.5 mg/mL−1, whilst increased levels of phosphorylated Erk/MAPK proteins indicated increased metabolic activity. In ex vivo models, HA supplementation accelerated re-epithelialization at all concentrations. This ex vivo model provides a robust model for preclinical assessment of skin substitutes. Conclusions HA supplementation to Integra® demonstrates increased in vitro growth, viability and migration. Whilst ex vivo data suggest HA supplementation of Integra® may increase rapidity of wound closure.
{"title":"In Vitro and Ex vivo Analysis of Hyaluronan Supplementation of Integra® Dermal Template on Human Dermal Fibroblasts and Keratinocytes","authors":"T. Hodgkinson, A. Bayat","doi":"10.5301/jabfm.5000259","DOIUrl":"https://doi.org/10.5301/jabfm.5000259","url":null,"abstract":"Purpose Widespread application of collagen-glycosaminoglycan dermal templates in the treatment of cutaneous defects has identified the interval between initial engraftment and skin graft application as important for improvement. The aim of this study was to evaluate the effect of hyaluronan supplementation of Integra® dermal template on human dermal fibroblasts and keratinocytes in both in vitro and ex vivo models. Methods This study utilized in vitro and ex vivo cell culture techniques to investigate supplementing Integra® Regeneration Template with hyaluronan (HA), as a strategy to decrease this interval. In vitro, Integra® was HA supplemented at 0.15, 1, 1.5 and 2 mg/mL−1. Primary human dermal fibroblast (PHDF) and keratinocyte proliferation, PHDF viability, migration and HA-induced signal transduction (phosphor-MAPK Array) were assessed. Ex vivo, wound models (wound diameter 4 mm) were created within 8 mm skin biopsies. Wounds were filled with Integra® or HA supplemented Integra®. Re-epithelialization was compared through hematoxylin and eosin-stained cross-sections at 7, 14 and 21 days in culture. Model viability was assessed through lactate dehydrogenase (LDH) assays. Results In vitro, PHDF and keratinocyte proliferation were enhanced significantly (p<0.001) when supplemented with HA. S-Phase and G2/M PHDFs in HA supplemented scaffolds increased. PHDF viability was enhanced to 72 hours culture with 1.5 mg/mL−1 HA (p = 0.016). PHDF migration was maximally enhanced at 1 mg/mL−1 and 1.5 mg/mL−1, whilst increased levels of phosphorylated Erk/MAPK proteins indicated increased metabolic activity. In ex vivo models, HA supplementation accelerated re-epithelialization at all concentrations. This ex vivo model provides a robust model for preclinical assessment of skin substitutes. Conclusions HA supplementation to Integra® demonstrates increased in vitro growth, viability and migration. Whilst ex vivo data suggest HA supplementation of Integra® may increase rapidity of wound closure.","PeriodicalId":51074,"journal":{"name":"Journal of Applied Biomaterials & Biomechanics","volume":"14 1","pages":"18 - 9"},"PeriodicalIF":0.0,"publicationDate":"2015-12-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.5301/jabfm.5000259","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"70591402","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Purpose Bacterial cellulose (BC) is an interesting biomaterial found application in various fields due to its novel characteristics like purity, water holding capacity, degree of polymerization and mechanical strength. BC as wound dressing material has limitation because it has no antimicrobial activity. To circumvent this problem, the present study was carried out by impregnation of silver on bacterial cellulose surface. Methods Bacterial cellulose was produced by Gluconoacetobacter hansenii (strain NCIM 2529) by shaking culture method. The sodium borohydride and classical Tollens reaction was used for silver nanoparticle synthesis. Results The effectiveness of sodium borohydride method compared with Tollens reaction was evaluated on the basis of silver nanoparticle formation and its impregnation on BC as evidenced by UV-Vis spectrum analysis, FE-SEM-EDS analysis and FT-IR spectrum. The potential of nano silver impregnated BC was determined for sustained release antimicrobial wound dressing material by swelling ratio, mechanical properties and antimicrobial activity against Staphylococcus aureus. Conclusions Thus the nanosilver impregnated bacterial cellulose as promising antimicrobial wound dressing material was evidenced.
{"title":"In Situ Development of Nanosilver-Impregnated Bacterial Cellulose for Sustainable Released Antimicrobial Wound Dressing","authors":"B. Mohite, S. Patil","doi":"10.5301/jabfm.5000257","DOIUrl":"https://doi.org/10.5301/jabfm.5000257","url":null,"abstract":"Purpose Bacterial cellulose (BC) is an interesting biomaterial found application in various fields due to its novel characteristics like purity, water holding capacity, degree of polymerization and mechanical strength. BC as wound dressing material has limitation because it has no antimicrobial activity. To circumvent this problem, the present study was carried out by impregnation of silver on bacterial cellulose surface. Methods Bacterial cellulose was produced by Gluconoacetobacter hansenii (strain NCIM 2529) by shaking culture method. The sodium borohydride and classical Tollens reaction was used for silver nanoparticle synthesis. Results The effectiveness of sodium borohydride method compared with Tollens reaction was evaluated on the basis of silver nanoparticle formation and its impregnation on BC as evidenced by UV-Vis spectrum analysis, FE-SEM-EDS analysis and FT-IR spectrum. The potential of nano silver impregnated BC was determined for sustained release antimicrobial wound dressing material by swelling ratio, mechanical properties and antimicrobial activity against Staphylococcus aureus. Conclusions Thus the nanosilver impregnated bacterial cellulose as promising antimicrobial wound dressing material was evidenced.","PeriodicalId":51074,"journal":{"name":"Journal of Applied Biomaterials & Biomechanics","volume":"14 1","pages":"53 - 58"},"PeriodicalIF":0.0,"publicationDate":"2015-12-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.5301/jabfm.5000257","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"70591638","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
J. Nowotny, Jana Farack, C. Vater, M. Johnsen, M. Gelinsky, T. Tonn, P. Kasten
Purpose Tissue regeneration can be improved by local application of autologous bone marrow derived progenitor cells (BMSC) and platelet rich plasma (PRP). However, there is a lack of standardized application procedures for clinical use. Therefore, a technique in accordance with the guidelines for advanced therapies medical products of the European Medicine Agency was developed and established. Methods In detail, a process for the isolation and formulation of autologous bone marrow cells (BMC) and PRP in a clinical setting was validated. To investigate the influence of storage time and temperature on gel formation and gel stability, different concentrations of BMC were stored with and without additional platelets, thrombin and fibrinogen and analyzed over a period of 28 days. In addition, cell vitality using a live-dead staining and migration ability of human mesenchymal stem cells (hMSC) in the gel clot was investigated. Results For an optimized stable gel clot, human BMC and PRP should be combined with 10% to 20% fibrinogen (9 mg/mL to 18 mg/mL) and 5% to 20% thrombin (25 I.E. to 100 I.E.). Both freshly prepared and stored cells for 1 to 7 days had a stable consistence over 28 days at 37°C. Different platelet concentrations did not influence gel clot formation. The ratio of living cells did not decrease significantly over the observation period of 5 days in the live-dead staining. Conclusions The study identified an optimal gel texture for local application of BMC and PRP. Seeded hMSC could migrate therein and were able to survive to initiate a healing cascade.
{"title":"Translation of Cell Therapy into Clinical Practice: Validation of an Application Procedure for Bone Marrow Progenitor Cells and Platelet Rich Plasma","authors":"J. Nowotny, Jana Farack, C. Vater, M. Johnsen, M. Gelinsky, T. Tonn, P. Kasten","doi":"10.5301/jabfm.5000255","DOIUrl":"https://doi.org/10.5301/jabfm.5000255","url":null,"abstract":"Purpose Tissue regeneration can be improved by local application of autologous bone marrow derived progenitor cells (BMSC) and platelet rich plasma (PRP). However, there is a lack of standardized application procedures for clinical use. Therefore, a technique in accordance with the guidelines for advanced therapies medical products of the European Medicine Agency was developed and established. Methods In detail, a process for the isolation and formulation of autologous bone marrow cells (BMC) and PRP in a clinical setting was validated. To investigate the influence of storage time and temperature on gel formation and gel stability, different concentrations of BMC were stored with and without additional platelets, thrombin and fibrinogen and analyzed over a period of 28 days. In addition, cell vitality using a live-dead staining and migration ability of human mesenchymal stem cells (hMSC) in the gel clot was investigated. Results For an optimized stable gel clot, human BMC and PRP should be combined with 10% to 20% fibrinogen (9 mg/mL to 18 mg/mL) and 5% to 20% thrombin (25 I.E. to 100 I.E.). Both freshly prepared and stored cells for 1 to 7 days had a stable consistence over 28 days at 37°C. Different platelet concentrations did not influence gel clot formation. The ratio of living cells did not decrease significantly over the observation period of 5 days in the live-dead staining. Conclusions The study identified an optimal gel texture for local application of BMC and PRP. Seeded hMSC could migrate therein and were able to survive to initiate a healing cascade.","PeriodicalId":51074,"journal":{"name":"Journal of Applied Biomaterials & Biomechanics","volume":"33 1","pages":"1 - 8"},"PeriodicalIF":0.0,"publicationDate":"2015-12-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.5301/jabfm.5000255","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"70591587","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
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