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Sample records for chitosan composite scaffold

  1. Chitosan composite three dimensional macrospheric scaffolds for bone tissue engineering.

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    Vyas, Veena; Kaur, Tejinder; Thirugnanam, Arunachalam

    2017-11-01

    The present work deals with the fabrication of chitosan composite scaffolds with controllable and predictable internal architecture for bone tissue engineering. Chitosan (CS) based composites were developed by varying montmorillonite (MMT) and hydroxyapatite (HA) combinations to fabricate macrospheric three dimensional (3D) scaffolds by direct agglomeration of the sintered macrospheres. The fabricated CS, CS/MMT, CS/HA and CS/MMT/HA 3D scaffolds were characterized for their physicochemical, biological and mechanical properties. The XRD and ATR-FTIR studies confirmed the presence of the individual constituents and the molecular interaction between them, respectively. The reinforcement with HA and MMT showed reduced swelling and degradation rate. It was found that in comparison to pure CS, the CS/HA/MMT composites exhibited improved hemocompatibility and protein adsorption. The sintering of the macrospheres controlled the swelling ability of the scaffolds which played an important role in maintaining the mechanical strength of the 3D scaffolds. The CS/HA/MMT composite scaffold showed 14 folds increase in the compressive strength when compared to pure CS scaffolds. The fabricated scaffolds were also found to encourage the MG 63 cell proliferation. Hence, from the above studies it can be concluded that the CS/HA/MMT composite 3D macrospheric scaffolds have wider and more practical application in bone tissue regeneration applications. Copyright © 2017 Elsevier B.V. All rights reserved.

  2. Development of porous Ti6Al4V/chitosan sponge composite scaffold for orthopedic applications

    International Nuclear Information System (INIS)

    Guo, Miao; Li, Xiang

    2016-01-01

    A novel composite scaffold consisting of porous Ti6Al4V part filled with chitosan sponge was fabricated using a combination of electron beam melting and freeze-drying. The mechanical properties of porous Ti6Al4V part were examined via compressive test. The ultimate compressive strength was 85.35 ± 8.68 MPa and the compressive modulus was 2.26 ± 0.42 GPa. The microstructure of composite scaffold was characterized using scanning electron microscopy. The chitosan sponge filled in Ti6Al4V part exhibited highly porous and well-interconnected micro-pore architecture. The osteoblastic cells were seeded on scaffolds to test their seeding efficiency and biocompatibility. Significantly higher cell seeding efficiency was found on composite scaffold. The biological response of osteoblasts on composite scaffolds was superior in terms of improved cell attachment, higher proliferation, and well-spread morphology in relation to porous Ti6Al4V part. These results suggest that the Ti6Al4V/chitosan composite scaffold is potentially useful as a biomedical scaffold for orthopedic applications. - Highlights: • A novel composite scaffold with sufficient mechanical properties and favorable cell affinity environment was developed. • Significantly higher cell seeding efficiency was found on composite scaffold. • The osteoblasts on composite scaffolds showed well-spread morphology, improved cell attachment and higher proliferation.

  3. Fabrication of chitin-chitosan/nano TiO2-composite scaffolds for tissue engineering applications.

    Science.gov (United States)

    Jayakumar, R; Ramachandran, Roshni; Divyarani, V V; Chennazhi, K P; Tamura, H; Nair, S V

    2011-03-01

    In this study, we prepared chitin-chitosan/nano TiO(2) composite scaffolds using lyophilization technique for bone tissue engineering. The prepared composite scaffold was characterized using SEM, XRD, FTIR and TGA. In addition, swelling, degradation and biomineralization capability of the composite scaffolds were evaluated. The developed composite scaffold showed controlled swelling and degradation when compared to the control scaffold. Cytocompatibility of the scaffold was assessed by MTT assay and cell attachment studies using osteoblast-like cells (MG-63), fibroblast cells (L929) and human mesenchymal stem cells (hMSCs). Results indicated no sign of toxicity and cells were found attached to the pore walls within the scaffolds. These results suggested that the developed composite scaffold possess the prerequisites for tissue engineering scaffolds and it can be used for tissue engineering applications. Copyright © 2010 Elsevier B.V. All rights reserved.

  4. Fabrication and biocompatibility of poly(L-lactic acid) and chitosan composite scaffolds with hierarchical microstructures

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    Lou, Tao, E-mail: taolou72@aliyun.com [College of Chemistry and Chemical Engineering, Qingdao University, Qingdao 266071 (China); Wang, Xuejun [College of Chemistry and Chemical Engineering, Qingdao University, Qingdao 266071 (China); Yan, Xu [College of Physics & Collaborative Innovation Center for Low-Dimensional Nanomaterials and Optoelectronic Devices, Qingdao University, Qingdao 266071 (China); Miao, Yu [Department of Mechanical Engineering, Columbia University, New York, NY 10027 (United States); Long, Yun-Ze, E-mail: yunzelong@163.com [College of Physics & Collaborative Innovation Center for Low-Dimensional Nanomaterials and Optoelectronic Devices, Qingdao University, Qingdao 266071 (China); Yin, Hai-Lei [Department of Osteology, No. 401 Hospital of P. L. A., Qingdao 266071 (China); Sun, Bin [College of Physics & Collaborative Innovation Center for Low-Dimensional Nanomaterials and Optoelectronic Devices, Qingdao University, Qingdao 266071 (China); Song, Guojun [College of Chemistry and Chemical Engineering, Qingdao University, Qingdao 266071 (China)

    2016-07-01

    The scaffold microstructure is crucial to reconstruct tissue normal functions. In this article, poly(L-lactic acid) and chitosan fiber (PLLA/CTSF) composite scaffolds with hierarchical microstructures both in fiber and pore sizes were successfully fabricated by combining thermal induced phase separation and salt leaching techniques. The composite scaffolds consisted of a nanofibrous PLLA matrix with diameter of 50–500 nm, and chitosan fibers with diameter of about 20 μm were homogenously distributed in the PLLA matrix as a microsized reinforcer. The composite scaffolds also had high porosity (> 94%) and hierarchical pore size, which were consisted of both micropores (50 nm–10 μm) and macropores (50–300 μm). By tailoring the microstructure and chemical composition, the mechanical property, pH buffer and protein adsorption capacity of the composite scaffold were improved significantly compared with those of PLLA scaffold. Cell culture results also revealed that the PLLA/CTSF composite scaffolds supported MG-63 osteoblast proliferation and penetration. - Highlights: • Composite scaffolds fabricated by combining thermal induced phase separation and salt leaching techniques • Hierarchical microstructure both in fiber and pore sizes • The scaffold microenvironment facilitates the protein adsorption, cell proliferation and penetration.

  5. Radiation synthesis of gelatin/CM-chitosan/{beta}-tricalcium phosphate composite scaffold for bone tissue engineering

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    Zhou Ying [College of Engineering, Peking University, Beijing 100871 (China); Center for Biomedical Materials and Tissue Engineering, Academy for Advanced Interdisciplinary Studies, Peking University, Beijing 100871 (China); Xu Ling, E-mail: lingxu@pku.edu.cn [College of Engineering, Peking University, Beijing 100871 (China); Center for Biomedical Materials and Tissue Engineering, Academy for Advanced Interdisciplinary Studies, Peking University, Beijing 100871 (China); Zhang Xiangmei; Zhao Yinghui [Center for Biomedical Materials and Tissue Engineering, Academy for Advanced Interdisciplinary Studies, Peking University, Beijing 100871 (China); Wei Shicheng, E-mail: sc-wei@pku.edu.cn [Center for Biomedical Materials and Tissue Engineering, Academy for Advanced Interdisciplinary Studies, Peking University, Beijing 100871 (China); Department of Oral and Maxillofacial Surgery, School and Hospital of Stomatology, Peking University, Beijing 100081 (China); Zhai Maolin [Beijing National Laboratory for Molecular Sciences, Department of Applied Chemistry, College of Chemistry and Molecular Engineering, Peking University, Beijing 100871 (China)

    2012-05-01

    A series of biodegradable composite scaffolds was fabricated from an aqueous solution of gelatin, carboxymethyl chitosan (CM-chitosan) and {beta}-tricalcium phosphate ({beta}-TCP) by radiation-induced crosslinking at ambient temperature. Ultrasonic treatment on the polymer solutions significantly influenced the distribution of {beta}-TCP particles. An ultrasonic time of 20 min, followed by 30 kGy irradiation induced a crosslinked scaffold with homogeneous distribution of {beta}-TCP particles, interconnected porous structure, sound swelling capacity and mechanical strength. Fourier Transform Infrared Spectroscopy and X-ray Diffraction analysis indicated that {beta}-TCP successfully incorporated with the network of gelatin and CM-chitosan. In vivo implantation of the scaffold into the mandible of beagle dog revealed that the scaffolds had excellent biocompatibility and the presence of {beta}-TCP can accelerate bone regeneration. The comprehensive results of this study paved way for the application of gelatin/CM-chitosan/{beta}-TCP composite scaffolds as candidate of bone tissue engineering material. - Highlights: Black-Right-Pointing-Pointer Radiation induced a crosslinked scaffold with interconnected porous structure. Black-Right-Pointing-Pointer Ultrasonic time of 20 min led to homogenerously distribution of {beta}-TCP. Black-Right-Pointing-Pointer Increasing amount of {beta}-TCP would restrict the swelling properties. Black-Right-Pointing-Pointer Proper fraction of {beta}-TCP will promote the mechanical properties of the scaffolds. Black-Right-Pointing-Pointer Hybrid of {beta}-TCP promoted the bone regeneration of the mandibles of beagle dogs.

  6. Performance of PRP Associated with Porous Chitosan as a Composite Scaffold for Regenerative Medicine

    Directory of Open Access Journals (Sweden)

    Andréa Arruda Martins Shimojo

    2015-01-01

    Full Text Available This study aimed to evaluate the in vitro performance of activated platelet-rich plasma associated with porous sponges of chitosan as a composite scaffold for proliferation and osteogenic differentiation of human adipose tissue-derived mesenchymal stem cells. The sponges were prepared by controlled freezing (−20, −80, or −196°C and lyophilization of chitosan solutions (1, 2, or 3% w/v. The platelet-rich plasma was obtained from controlled centrifugation of whole blood and activated with calcium and autologous serum. The composite scaffolds were prepared by embedding the sponges with the activated platelet-rich plasma. The results showed the performance of the scaffolds was superior to that of activated platelet-rich plasma alone, in terms of delaying the release of growth factors and increased proliferation of the stem cells. The best preparation conditions of chitosan composite scaffolds that coordinated the physicochemical and mechanical properties and cell proliferation were 3% (w/v chitosan and a −20°C freezing temperature, while −196°C favored osteogenic differentiation. Although the composite scaffolds are promising for regenerative medicine, the structures require stabilization to prevent the collapse observed after five days.

  7. Diatomite reinforced chitosan composite membrane as potential scaffold for guided bone regeneration.

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    Tamburaci, Sedef; Tihminlioglu, Funda

    2017-11-01

    In this study, natural silica source, diatomite, incorporated novel chitosan based composite membranes were fabricated and characterized for bone tissue engineering applications as possible bone regeneration membrane. The effect of diatomite loading on the mechanical, morphological, chemical, thermal and surface properties, wettability and in vitro cytotoxicity and cell proliferation on of composite membranes were investigated and observed by tensile test, atomic force microscopy (AFM), Fourier transform infrared spectroscopy (FTIR), thermal gravimetric analysis (TGA), protein adsorption assay, air/water contact angle analysis and WST-1 respectively. Swelling studies were also performed by water absorption capacity determination. Results showed that incorporation of diatomite to the chitosan matrix increased the surface roughness, swelling capacity and tensile modulus of membranes. An increase of about 52% in Young's modulus was achieved for 10wt% diatomite composite membranes compared with chitosan membranes. High cell viability results were obtained with indirect extraction method. Besides, in vitro cell proliferation and ALP activity results showed that diatom incorporation significantly increased the ALP activity of Saos-2 cells cultured on chitosan membranes. The novel composite membranes prepared in the present study with tunable properties can be considered as a potential candidate as a scaffold in view of its enhanced physical & chemical properties as well as biological activities for bone tissue engineering applications. Copyright © 2017 Elsevier B.V. All rights reserved.

  8. A composite chitosan-gelatin bi-layered, biomimetic macroporous scaffold for blood vessel tissue engineering.

    Science.gov (United States)

    Badhe, Ravindra V; Bijukumar, Divya; Chejara, Dharmesh R; Mabrouk, Mostafa; Choonara, Yahya E; Kumar, Pradeep; du Toit, Lisa C; Kondiah, Pierre P D; Pillay, Viness

    2017-02-10

    A composite chitosan-gelatin macroporous hydrogel-based scaffold with bi-layered tubular architecture was engineered by solvent casting-co-particulate leaching. The scaffold constituted an inner macroporous layer concealed by a non-porous outer layer mimicking the 3D matrix of blood vessels with cellular adhesion and proliferation. The scaffold was evaluated for its morphological, physicochemical, physicomechanical and biodurability properties employing SEM, FTIR, DSC, XRD, porositometry, rheology and texture analysis. The fluid uptake and biodegradation in the presence of lysozymes was also investigated. Cellular attachment and proliferation was analysed using human dermal fibroblasts (HDF-a) seeded onto the scaffold and evaluated by MTT assay, SEM, and confocal microscopy. Results demonstrated that the scaffold had a desirable tensile strength=95.81±11kPa, elongation at break 112.5±13%, porosity 82% and pores between 100 and 230μm, 50% in vitro biodegradation at day 16 and proliferated fibroblasts over 20 days. These results demonstrate that scaffold may be an excellent tubular archetype for blood vessel tissue engineering. Copyright © 2016 Elsevier Ltd. All rights reserved.

  9. Preparation and comparative characterization of keratin–chitosan and keratin–gelatin composite scaffolds for tissue engineering applications

    International Nuclear Information System (INIS)

    Balaji, S.; Kumar, Ramadhar; Sripriya, R.; Kakkar, Prachi; Ramesh, D. Vijaya; Reddy, P. Neela Kanta; Sehgal, P.K.

    2012-01-01

    We report fabrication of three dimensional scaffolds with well interconnected matrix of high porosity using keratin, chitosan and gelatin for tissue engineering and other biomedical applications. Scaffolds were fabricated using porous Keratin–Gelatin (KG), Keratin–Chitosan (KC) composites. The morphology of both KG and KC was investigated using SEM. The scaffolds showed high porosity with interconnected pores in the range of 20–100 μm. They were further tested by FTIR, DSC, CD, tensile strength measurement, water uptake and swelling behavior. In vitro cell adhesion and cell proliferation tests were carried out to study the biocompatibility behavior and their application as an artificial skin substitute. Both KG and KC composite scaffolds showed similar properties and patterns for cell proliferation. Due to rapid degradation of gelatin in KG, we found that it has limited application as compared to KC scaffold. We conclude that KC scaffold owing to its slow degradation and antibacterial properties would be a better substrate for tissue engineering and other biomedical application. Highlights: ► Extraction of reduced keratin from horn meal. ► Preparation of keratin–gelatin and keratin–chitosan composite scaffolds. ► Characterizations of the composite scaffolds. ► Comparative cytotoxicity analysis on NIH3T3 fibroblasts.

  10. Development of core-shell coaxially electrospun composite PCL/chitosan scaffolds.

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    Surucu, Seda; Turkoglu Sasmazel, Hilal

    2016-11-01

    This study was related to combining of synthetic Poly (ε-caprolactone) (PCL) and natural chitosan polymers to develop three dimensional (3D) PCL/chitosan core-shell scaffolds for tissue engineering applications. The scaffolds were fabricated with coaxial electrospinning technique and the characterizations of the samples were done by thickness and contact angle (CA) measurements, scanning electron microscopy (SEM), transmission electron microscopy (TEM), X-Ray Photoelectron Spectroscopy (XPS) analyses, mechanical and PBS absorption and shrinkage tests. The average inter-fiber diameter values were calculated for PCL (0.717±0.001μm), chitosan (0.660±0.007μm) and PCL/chitosan core-shell scaffolds (0.412±0.003μm), also the average inter-fiber pore size values exhibited decreases of 66.91% and 61.90% for the PCL and chitosan scaffolds respectively, compared to PCL/chitosan core-shell ones. XPS analysis of the PCL/chitosan core-shell structures exhibited the characteristic peaks of PCL and chitosan polymers. The cell culture studies (MTT assay, Confocal Laser Scanning Microscope (CLSM) and SEM analyses) carried out with L929 ATCC CCL-1 mouse fibroblast cell line proved that the biocompatibility performance of the scaffolds. The obtained results showed that the created micro/nano fibrous structure of the PCL/chitosan core-shell scaffolds in this study increased the cell viability and proliferation on/within scaffolds. Copyright © 2016 Elsevier B.V. All rights reserved.

  11. Stabilization of porous chitosan improves the performance of its association with platelet-rich plasma as a composite scaffold

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    Shimojo, A.A.M., E-mail: lshimojo51@gmail.com; Perez, A.G.M.; Galdames, S.E.M.; Brissac, I.C.S.; Santana, M.H.A.

    2016-03-01

    This study offers innovative perspectives for optimizing of scaffolds based on correlation structure–function aimed the regenerative medicine. Thus, we evaluated in vitro performance of stabilized porous chitosan (SPCHTs) associated with activated platelet-rich plasma (aP-PRP) as a composite scaffold for the proliferation and osteogenic differentiation of human adipose-derived mesenchymal stem cells (h-AdMSCs). The porous structure of chitosan (PCHT) was prepared similarly to solid sponges by controlled freezing (− 20 °C) and lyophilization of a 3% (w/v) chitosan solution. Stabilization was performed by treating the PCHT with sodium hydroxide (TNaOH), an ethanol series (TEtOH) or by crosslinking with tripolyphosphate (CTPP). The aP-PRP was obtained from the controlled centrifugation of whole blood and activated with autologous serum and calcium. Imaging of the structures showed fibrin networks inside and on the surface of SPCHTs as a consequence of electrostatic interactions. SPCHTs were non-cytotoxic, and the porosity, pore size and Young's modulus were approximately 96%, 145 μm and 1.5 MPa for TNaOH and TEtOH and 94%, 110 μm and 1.8 MPa for CTPP, respectively. Stabilization maintained the integrity of the SPCHTs for at least 10 days of cultivation. SPCHTs showed controlled release of the growth factors TGF-β1 and PDGF-AB. Although generating different patterns, all of the stabilization treatments improved the proliferation of seeded h-AdMSCs on the composite scaffold compared to aP-PRP alone, and differentiation of the composite scaffold treated with TEtOH was significantly higher than for non-stabilized PCHT. We conclude that the composite scaffolds improved the in vitro performance of PRP and have potential in regenerative medicine. - Highlights: • Stabilization maintains the integrity of the chitosan scaffolds for at least 10 days. • Fibrin networks on the chitosan scaffolds were referred to electrostatic interactions. • Stabilized chitosan

  12. Stabilization of porous chitosan improves the performance of its association with platelet-rich plasma as a composite scaffold

    International Nuclear Information System (INIS)

    Shimojo, A.A.M.; Perez, A.G.M.; Galdames, S.E.M.; Brissac, I.C.S.; Santana, M.H.A.

    2016-01-01

    This study offers innovative perspectives for optimizing of scaffolds based on correlation structure–function aimed the regenerative medicine. Thus, we evaluated in vitro performance of stabilized porous chitosan (SPCHTs) associated with activated platelet-rich plasma (aP-PRP) as a composite scaffold for the proliferation and osteogenic differentiation of human adipose-derived mesenchymal stem cells (h-AdMSCs). The porous structure of chitosan (PCHT) was prepared similarly to solid sponges by controlled freezing (− 20 °C) and lyophilization of a 3% (w/v) chitosan solution. Stabilization was performed by treating the PCHT with sodium hydroxide (TNaOH), an ethanol series (TEtOH) or by crosslinking with tripolyphosphate (CTPP). The aP-PRP was obtained from the controlled centrifugation of whole blood and activated with autologous serum and calcium. Imaging of the structures showed fibrin networks inside and on the surface of SPCHTs as a consequence of electrostatic interactions. SPCHTs were non-cytotoxic, and the porosity, pore size and Young's modulus were approximately 96%, 145 μm and 1.5 MPa for TNaOH and TEtOH and 94%, 110 μm and 1.8 MPa for CTPP, respectively. Stabilization maintained the integrity of the SPCHTs for at least 10 days of cultivation. SPCHTs showed controlled release of the growth factors TGF-β1 and PDGF-AB. Although generating different patterns, all of the stabilization treatments improved the proliferation of seeded h-AdMSCs on the composite scaffold compared to aP-PRP alone, and differentiation of the composite scaffold treated with TEtOH was significantly higher than for non-stabilized PCHT. We conclude that the composite scaffolds improved the in vitro performance of PRP and have potential in regenerative medicine. - Highlights: • Stabilization maintains the integrity of the chitosan scaffolds for at least 10 days. • Fibrin networks on the chitosan scaffolds were referred to electrostatic interactions. • Stabilized chitosan

  13. Production of Composite Scaffold Containing Silk Fibroin, Chitosan, and Gelatin for 3D Cell Culture and Bone Tissue Regeneration.

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    Li, Jianqing; Wang, Qiuke; Gu, Yebo; Zhu, Yu; Chen, Liang; Chen, Yunfeng

    2017-11-08

    BACKGROUND Bone tissue engineering, a powerful tool to treat bone defects, is highly dependent on use of scaffolds. Both silk fibroin (SF) and chitosan (Cs) are biocompatible and actively studied for reconstruction of tissue engineering. Gelatin (Gel) is also widely applied in the biomedical field due to its low antigenicity and physicochemical stability. MATERIAL AND METHODS In this study, 4 different types of scaffolds were constructed - SF, SF/Cs, SF/Gel, and SF/Cs/Gel - and we compared their physical and chemical properties as well as biological characterization of these scaffolds to determine the most suitable scaffold for use in bone regeneration. First, these scaffolds were produced via chemical cross-linking method and freeze-drying technique. Next, the characterization of internal structure was studied using scanning electron microscopy and the porosity was evaluated by liquid displacement method. Then, we compared physicochemical properties such as water absorption rate and degradation property. Finally, MC3T3-E1 cells were inoculated on the scaffolds to study the biocompatibility and osteogenesis of the three-dimensional (3D) scaffolds in vitro. RESULTS The composite scaffold formed by all 3 components was the best for use in bone regeneration. CONCLUSIONS We conclude that the best scaffold among the 4 studied for MC3T3-E1 cells is our SF/Cs/Gel scaffold, suggesting a new choice for bone regeneration that can be used to treat bone defects or fractures in clinical practice.

  14. The influence of phosphorylation and freezing temperature on the mechanical properties of hydroxyapatite/chitosan composite as bone scaffold biomaterial

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    Albab, Muh Fadhil; Giovani, Nicholas; Yuwono, Akhmad Herman; Sofyan, Nofrijon; Ramahdita, Ghiska; Whulanza, Yudan

    2018-02-01

    Biomaterials composite of hydroxyapatite/chitosan is a preeminent material for medical applications including bone scaffold. To improve its mechanical properties, the chitosan as the matrix needs to be modified with particular chemical agents. One of the methods is phosphorylation of chitosan by using orthophosphoric acid prior to the biomaterials fabrication. In the current study, biomaterials with the weight composition of 70% hydroxyapatite (HA) and 30% phosphorylated chitosan have been fabricated using thermally induced phase separation (TIPS) method with freezing temperature variation of -20, -30, -40 and -80°C prior to three day-freeze drying. The results obtained by this work showed that the highest compression modulus of 376.9 kPa, highest compressive strength of 38.4 kPa and biggest pore size of 48.24 µm were achieved in the freezing temperature of -20°C. In comparison to non-phosphorylated chitosan/hydroxyapatite, the modification of chitosan using orthophosphoric acid in this work has been found to increase the compressive strength of composite up to 5.5 times.

  15. Preparation and biological properties of a novel composite scaffold of nano-hydroxyapatite/chitosan/carboxymethyl cellulose for bone tissue engineering

    Directory of Open Access Journals (Sweden)

    Chengdong Xiong

    2009-07-01

    Full Text Available Abstract In this study, we report the physico-chemical and biological properties of a novel biodegradable composite scaffold made of nano-hydroxyapatite and natural derived polymers of chitosan and carboxymethyl cellulose, namely, n-HA/CS/CMC, which was prepared by freeze-drying method. The physico-chemical properties of n-HA/CS/CMC scaffold were tested by infrared absorption spectra (IR, transmission electron microscope(TEM, scanning electron microscope(SEM, universal material testing machine and phosphate buffer solution (PBS soaking experiment. Besides, the biological properties were evaluated by MG63 cells and Mesenchymal stem cells (MSCs culture experiment in vitro and a short period implantation study in vivo. The results show that the composite scaffold is mainly formed through the ionic crossing-linking of the two polyions between CS and CMC, and n-HA is incorporated into the polyelectrolyte matrix of CS-CMC without agglomeration, which endows the scaffold with good physico-chemical properties such as highly interconnected porous structure, high compressive strength and good structural stability and degradation. More important, the results of cells attached, proliferated on the scaffold indicate that the scaffold is non-toxic and has good cell biocompatibility, and the results of implantation experiment in vivo further confirm that the scaffold has good tissue biocompatibility. All the above results suggest that the novel degradable n-HA/CS/CMC composite scaffold has a great potential to be used as bone tissue engineering material.

  16. Preparation and biological properties of a novel composite scaffold of nano-hydroxyapatite/chitosan/carboxymethyl cellulose for bone tissue engineering

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    Liuyun, Jiang; Yubao, Li; Chengdong, Xiong

    2009-01-01

    In this study, we report the physico-chemical and biological properties of a novel biodegradable composite scaffold made of nano-hydroxyapatite and natural derived polymers of chitosan and carboxymethyl cellulose, namely, n-HA/CS/CMC, which was prepared by freeze-drying method. The physico-chemical properties of n-HA/CS/CMC scaffold were tested by infrared absorption spectra (IR), transmission electron microscope(TEM), scanning electron microscope(SEM), universal material testing machine and phosphate buffer solution (PBS) soaking experiment. Besides, the biological properties were evaluated by MG63 cells and Mesenchymal stem cells (MSCs) culture experiment in vitro and a short period implantation study in vivo. The results show that the composite scaffold is mainly formed through the ionic crossing-linking of the two polyions between CS and CMC, and n-HA is incorporated into the polyelectrolyte matrix of CS-CMC without agglomeration, which endows the scaffold with good physico-chemical properties such as highly interconnected porous structure, high compressive strength and good structural stability and degradation. More important, the results of cells attached, proliferated on the scaffold indicate that the scaffold is non-toxic and has good cell biocompatibility, and the results of implantation experiment in vivo further confirm that the scaffold has good tissue biocompatibility. All the above results suggest that the novel degradable n-HA/CS/CMC composite scaffold has a great potential to be used as bone tissue engineering material. PMID:19594953

  17. Anti-infective efficacy, cytocompatibility and biocompatibility of a 3D-printed osteoconductive composite scaffold functionalized with quaternized chitosan.

    Science.gov (United States)

    Yang, Ying; Yang, Shengbing; Wang, Yugang; Yu, Zhifeng; Ao, Haiyong; Zhang, Hongbo; Qin, Ling; Guillaume, Olivier; Eglin, David; Richards, R Geoff; Tang, Tingting

    2016-12-01

    Contaminated or infected bone defects remain serious challenges in clinical trauma and orthopaedics, and a bone substitute with both osteoconductivity and antibacterial properties represents an improvement for treatment strategy. In this study, quaternized chitosan (hydroxypropyltrimethyl ammonium chloride chitosan, HACC) was grafted to 3D-printed scaffolds composed of polylactide-co-glycolide (PLGA) and hydroxyapatite (HA), in order to design bone engineering scaffolds endowed with antibacterial and osteoconductive properties. We found that both the PLGA/HA/HACC and PLGA/HACC composite scaffolds decreased bacterial adhesion and biofilm formation under in vitro and in vivo conditions. Additionally, ATP leakage assay indicated that immobilizing HACC on the scaffolds could effectively disrupt microbial membranes. Using human bone marrow-derived mesenchymal stem cells (hBMSCs), we demonstrated that HA incorporated scaffolds, including PLGA/HA and PLGA/HA/HACC, favoured cell attachment, proliferation, spreading and osteogenic differentiation compared to HA-free PLGA or PLGA/HACC scaffolds. Finally, an in vivo biocompatibility assay conducted on rats, showed that HA incorporated scaffolds (including PLGA/HA and PLGA/HA/HACC scaffolds) exhibited good neovascularization and tissue integration. Taken together, our findings support the approach for developing porous PLGA/HA/HACC composite scaffold with potential clinical application in the treatment of infected bone. Although plenty of conductive scaffold biomaterials have been exploited to improve bone regeneration under infection, potential tissue toxicity under high concentration and antibiotic-resistance are their main deficiencies. This study indicated that HACC-grafted PLGA/HA composite scaffold prepared using an innovative 3D-printing technique and covalent grafting strategy showed significantly enhanced antibacterial activities, especially against the antibiotic-resistant strains, together with good osteogenic

  18. In vitro cytocompatibility evaluation of chitosan/graphene oxide 3D scaffold composites designed for bone tissue engineering.

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    Dinescu, Sorina; Ionita, Mariana; Pandele, Andreea Madalina; Galateanu, Bianca; Iovu, Horia; Ardelean, Aurel; Costache, Marieta; Hermenean, Anca

    2014-01-01

    Extensively studied nowadays, graphene oxide (GO) has a benefic effect on cell proliferation and differentiation, thus holding promise for bone tissue engineering (BTE) approaches. The aim of this study was not only to design a chitosan 3D scaffold improved with GO for optimal BTE, but also to analyze its physicochemical properties and to evaluate its cytocompatibility and ability to support cell metabolic activity and proliferation. Overall results show that the addition of GO in the scaffold's composition improved mechanical properties and pore formation and enhanced the bioactivity of the scaffold material for tissue engineering. The new developed CHT/GO 3 wt% scaffold could be a potential candidate for further in vitro and in vivo osteogenesis studies and BTE approaches.

  19. A composite scaffold of MSC affinity peptide-modified demineralized bone matrix particles and chitosan hydrogel for cartilage regeneration

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    Meng, Qingyang; Man, Zhentao; Dai, Linghui; Huang, Hongjie; Zhang, Xin; Hu, Xiaoqing; Shao, Zhenxing; Zhu, Jingxian; Zhang, Jiying; Fu, Xin; Duan, Xiaoning; Ao, Yingfang

    2015-12-01

    Articular cartilage injury is still a significant challenge because of the poor intrinsic healing potential of cartilage. Stem cell-based tissue engineering is a promising technique for cartilage repair. As cartilage defects are usually irregular in clinical settings, scaffolds with moldability that can fill any shape of cartilage defects and closely integrate with the host cartilage are desirable. In this study, we constructed a composite scaffold combining mesenchymal stem cells (MSCs) E7 affinity peptide-modified demineralized bone matrix (DBM) particles and chitosan (CS) hydrogel for cartilage engineering. This solid-supported composite scaffold exhibited appropriate porosity, which provided a 3D microenvironment that supports cell adhesion and proliferation. Cell proliferation and DNA content analysis indicated that the DBM-E7/CS scaffold promoted better rat bone marrow-derived MSCs (BMMSCs) survival than the CS or DBM/CS groups. Meanwhile, the DBM-E7/CS scaffold increased matrix production and improved chondrogenic differentiation ability of BMMSCs in vitro. Furthermore, after implantation in vivo for four weeks, compared to those in control groups, the regenerated issue in the DBM-E7/CS group exhibited translucent and superior cartilage-like structures, as indicated by gross observation, histological examination, and assessment of matrix staining. Overall, the functional composite scaffold of DBM-E7/CS is a promising option for repairing irregularly shaped cartilage defects.

  20. Manipulation of chemical composition and architecture of non-biodegradable poly(ethylene terephthalate)/chitosan fibrous scaffolds and their effects on L929 cell behavior

    International Nuclear Information System (INIS)

    Veleirinho, Beatriz; Berti, Fernanda V.; Dias, Paulo F.; Maraschin, Marcelo; Ribeiro-do-Valle, Rosa M.; Lopes-da-Silva, José A.

    2013-01-01

    Microporous, non-woven fibrous scaffolds made of poly(ethylene terephthalate) and chitosan were produced by electrospinning. Fiber morphology, diameter, pore size, and wettability were manipulated by varying the chemical composition of the electrospinning solution, i.e. chitosan concentration and molecular weight, and by post-electrospinning treatment with glutaraldehyde. In vitro studies were conducted using a fibroblast cell line toward a comprehensive understanding of how scaffolds characteristics can modulate the cell behavior, i.e. viability, adhesion, proliferation, extracellular matrix secretion, and three-dimensional colonization. Substantial differences were found as a result of scaffold morphological changes. Higher levels of adhesion, spreading, and superficial proliferation were achieved for scaffolds with smaller fiber and pore diameters while cell penetration and internal colonization were enhanced for scaffolds with larger pores. Additionally, the available area for cell adhesion, which is related to fiber and pore size, was a crucial factor for the viability of L929 cells. This paper provides significant insights for the development and optimization of electrospun scaffolds toward an improved biological performance. Highlights: ► Hybrid PET/chitosan mats were produced by electrospinning. ► Scaffold architecture was manipulated by changing composition of the spun solution. ► The scaffolds showed in vitro biocompatibility to L929 cells. ► Smaller fiber diameters and pore areas allowed for higher levels of cell adhesion and proliferation. ► A 3D cell colonization was achieved for scaffolds with higher fiber diameters.

  1. Advances in allogenic bone graft processing and usage: preparation and evaluation of chitosan-demineralized cancellous bone powder composite scaffolds as a bone graft substitute

    International Nuclear Information System (INIS)

    Yongyudh Vajaradul

    2008-01-01

    Full text: Demineralized bone matrix (DBM) is currently used by surgeons. It usually exists as a lyophilized powder which is difficult to handle and operated. In this study, we try to improve these disadvantages by combining DBM with a biomaterial. It focuses on a natural biodegradable polymer, chitosan, to act as a temporary matrix for bone growth that easily prepare in any size and shape by using tissue engineering knowledge to get a proper temporary matrix. Thus, the development of chitosan-demineralized bone powder composite scaffold is an alternative way. Polymeric scaffold has been demonstrated to have great potential for tissue engineering because the scaffold or three dimension (3D) construct provides the necessary support for cells to proliferate, extracellular matrix deposition and vascularization of neo-tissue. Moreover, chitosan, a natural cationic polymer which its structural is similar to extracellular matrix glycosaminoblycans, is biodegradable, biocompatible, non-antigenic and biofunctional. It can enhance osteoblast cells proliferation and mineral matrix deposition in culture. The first study was to fabricate and analyze composite scaffold composed of either chitosan-demineralized cancellous bone powders or chitosan-demineralized cancellous cartilage bone powders in a ratio 50:50 and 70:30 w/w (chitosan : bone powders) based on physical properties composing of average pore diameter, mechanical integrity and swelling property. Secondly, scaffolds were evaluated in term of biological properties composing of their ability to support neo osteogenesis, including assessments of cell attachment and viability, cell morphology, and the biosynthesis of extracellular matrix. Results indicated that chitosan-demineralized cancellous bone powder composite scaffolds possessing an interconnecting, porous structure could be easily created through a simple freezing and lyophilization process. (Author)

  2. Sustained Local Release of NGF from a Chitosan-Sericin Composite Scaffold for Treating Chronic Nerve Compression.

    Science.gov (United States)

    Zhang, Lei; Yang, Wen; Tao, Kaixiong; Song, Yu; Xie, Hongjian; Wang, Jian; Li, Xiaolin; Shuai, Xiaoming; Gao, Jinbo; Chang, Panpan; Wang, Guobin; Wang, Zheng; Wang, Lin

    2017-02-01

    Chronic nerve compression (CNC), a common form of peripheral nerve injury, always leads to chronic peripheral nerve pain and dysfunction. Current available treatments for CNC are ineffective as they usually aim to alleviate symptoms at the acute phase with limited capability toward restoring injured nerve function. New approaches for effective recovery of CNC injury are highly desired. Here we report for the first time a tissue-engineered approach for the repair of CNC. A genipin cross-linked chitosan-sericin 3D scaffold for delivering nerve growth factor (NGF) was designed and fabricated. This scaffold combines the advantages of both chitosan and sericin, such as high porosity, adjustable mechanical properties and swelling ratios, the ability of supporting Schwann cells growth, and improving nerve regeneration. The degradation products of the composite scaffold upregulate the mRNA levels of the genes important for facilitating nerve function recovery, including glial-derived neurotrophic factor (GDNF), early growth response 2 (EGR2), and neural cell adhesion molecule (NCAM) in Schwann cells, while down-regulating two inflammatory genes' mRNA levels in macrophages, tumor necrosis factor alpha (TNF-α), and interleukin-1 beta (IL-1β). Importantly, our tissue-engineered strategy achieves significant nerve functional recovery in a preclinical CNC animal model by decreasing neuralgia, improving nerve conduction velocity (NCV), accelerating microstructure restoration, and attenuating gastrocnemius muscles dystrophy. Together, this work suggests a promising clinical alternative for treating chronic peripheral nerve compression injury.

  3. Effects of flow configuration on bone tissue engineering using human mesenchymal stem cells in 3D chitosan composite scaffolds.

    Science.gov (United States)

    Sellgren, Katelyn L; Ma, Teng

    2015-08-01

    Perfusion bioreactor plays important role in supporting 3D bone construct development. Scaffolds of chitosan composites have been studied to support bone tissue regeneration from osteogenic progenitor cells including human mesenchymal stem cells (hMSC). In this study, porous scaffolds of hydroxyapatite (H), chitosan (C), and gelatin (G) were fabricated by phase-separation and press-fitted in the perfusion bioreactor system where media flow is configured either parallel or transverse with respect to the scaffolds to investigate the impact of flow configuration on hMSC proliferation and osteogenic differentiation. The in vitro results showed that the interstitial flow in the transverse flow (TF) constructs reduced cell growth during the first week of culture but improved spatial cell distribution and early onset of osteogenic differentiation measured by alkaline phosphatase and expression of osteogenic genes. After 14 days of bioreactor culture, the TF constructs have comparable cell number but higher expression of bone markers genes and proteins compared to the parallel flow constructs. To evaluate ectopic bone formation, the HCG constructs seeded with hMSCs pre-cultured under two flow configurations for 7 days were implanted in CD-1 nude mice. While Masson's Trichrom staining revealed bone formation in both constructs, the TF constructs have improved spatial cell and osteoid distribution throughout the 2.0 mm constructs. The results highlight the divergent effects of media flow over the course of construct development and suggest that the flow configuration is an important parameter regulating the cellular events leading to bone construct formation in the HCG scaffolds. © 2014 Wiley Periodicals, Inc.

  4. Ag-loaded MgSrFe-layered double hydroxide/chitosan composite scaffold with enhanced osteogenic and antibacterial property for bone engineering tissue.

    Science.gov (United States)

    Cao, Dandan; Xu, Zhengliang; Chen, Yixuan; Ke, Qinfei; Zhang, Changqing; Guo, Yaping

    2018-02-01

    Bone tissue engineering scaffolds for the reconstruction of large bone defects should simultaneously promote osteogenic differentiation and avoid postoperative infection. Herein, we develop, for the first time, Ag-loaded MgSrFe-layered double hydroxide/chitosan (Ag-MgSrFe/CS) composite scaffold. This scaffold exhibits three-dimensional interconnected macroporous structure with a pore size of 100-300 μm. The layered double hydroxide nanoplates in the Ag-MgSrFe/CS show lateral sizes of 200-400 nm and thicknesses of ∼50 nm, and the Ag nanoparticles with particle sizes of ∼20 nm are uniformly dispersed on the scaffold surfaces. Human bone marrow-derived mesenchymal stem cells (hBMSCs) present good adhesion, spreading, and proliferation on the Ag-MgSrFe/CS composite scaffold, suggesting that the Ag and Sr elements in the composite scaffold have no toxicity to hBMSCs. When compared with MgFe/CS composite scaffold, the Ag-MgSrFe/CS composite scaffold has better osteogenic property. The released Sr 2+ ions from the composite scaffold enhance the alkaline phosphatase activity of hBMSCs, promote the extracellular matrix mineralization, and increase the expression levels of osteogenic-related RUNX2 and BMP-2. Moreover, the Ag-MgSrFe/CS composite scaffold possesses good antibacterial property because the Ag nanoparticles in the composite scaffold effectively prevent biofilm formation against S. aureus. Hence, the Ag-MgSrFe/CS composite scaffold with excellent osteoinductivity and antibacterial property has a great potential for bone tissue engineering. © 2017 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 106B: 863-873, 2018. © 2017 Wiley Periodicals, Inc.

  5. Manipulation of chemical composition and architecture of non-biodegradable poly(ethylene terephthalate)/chitosan fibrous scaffolds and their effects on L929 cell behavior

    Energy Technology Data Exchange (ETDEWEB)

    Veleirinho, Beatriz [QOPNA Research Unit, Department of Chemistry, University of Aveiro, 3810-193 Aveiro (Portugal); Berti, Fernanda V. [Integrated Technologies Laboratory, Chemical and Food Engineering Department, Federal University of Santa Catarina, 88040-900 Florianopolis (Brazil); Dias, Paulo F. [Department of Cell Biology, Embryology and Genetics, Federal University of Santa Catarina, 88040-900 Florianopolis (Brazil); Maraschin, Marcelo [Department of Plant Science, Federal University of Santa Catarina, 88040-900 Florianopolis (Brazil); Ribeiro-do-Valle, Rosa M. [Department of Pharmacology, Federal University of Santa Catarina, 88040-900 Florianopolis (Brazil); Lopes-da-Silva, Jose A., E-mail: jals@ua.pt [QOPNA Research Unit, Department of Chemistry, University of Aveiro, 3810-193 Aveiro (Portugal)

    2013-01-01

    Microporous, non-woven fibrous scaffolds made of poly(ethylene terephthalate) and chitosan were produced by electrospinning. Fiber morphology, diameter, pore size, and wettability were manipulated by varying the chemical composition of the electrospinning solution, i.e. chitosan concentration and molecular weight, and by post-electrospinning treatment with glutaraldehyde. In vitro studies were conducted using a fibroblast cell line toward a comprehensive understanding of how scaffolds characteristics can modulate the cell behavior, i.e. viability, adhesion, proliferation, extracellular matrix secretion, and three-dimensional colonization. Substantial differences were found as a result of scaffold morphological changes. Higher levels of adhesion, spreading, and superficial proliferation were achieved for scaffolds with smaller fiber and pore diameters while cell penetration and internal colonization were enhanced for scaffolds with larger pores. Additionally, the available area for cell adhesion, which is related to fiber and pore size, was a crucial factor for the viability of L929 cells. This paper provides significant insights for the development and optimization of electrospun scaffolds toward an improved biological performance. Highlights: Black-Right-Pointing-Pointer Hybrid PET/chitosan mats were produced by electrospinning. Black-Right-Pointing-Pointer Scaffold architecture was manipulated by changing composition of the spun solution. Black-Right-Pointing-Pointer The scaffolds showed in vitro biocompatibility to L929 cells. Black-Right-Pointing-Pointer Smaller fiber diameters and pore areas allowed for higher levels of cell adhesion and proliferation. Black-Right-Pointing-Pointer A 3D cell colonization was achieved for scaffolds with higher fiber diameters.

  6. Development of various composition multicomponent chitosan/fish collagen/glycerin 3D porous scaffolds: Effect on morphology, mechanical strength, biostability and cytocompatibility.

    Science.gov (United States)

    Ullah, Saleem; Zainol, Ismail; Chowdhury, Shiplu Roy; Fauzi, M B

    2018-05-01

    The various composition multicomponent chitosan/fish collagen/glycerin 3D porous scaffolds were developed and investigated the effect of various composition chitosan/fish collagen/glycerin on scaffolds morphology, mechanical strength, biostability and cytocompatibility. The scaffolds were fabricated via freeze-drying technique. The effects of various compositions consisting in 3D scaffolds were investigated via FT-IR analysis, porosity, swelling and mechanical tests, and effect on the morphology of scaffolds investigated microscopically. The biostability and cytocompatibility tests were used to explore the ability of scaffolds to use for tissue engineering application. The average pore sizes of scaffolds were in range of 100.73±27.62-116.01±52.06, porosity 71.72±3.46-91.17±2.42%, tensile modulus in dry environment 1.47±0.08-0.17±0.03MPa, tensile modulus in wet environment 0.32±0.03-0.14±0.04MPa and biodegradation rate (at day 30) 60.38±0.70-83.48±0.28%. In vitro culture of human fibroblasts and keratinocytes showed that the various composition multicomponent 3D scaffolds were good cytocompatibility however, the scaffolds contained high amount of fish collagen excellently facilitated cell proliferation and adhesion. It was found that the high amount fish collagen and glycerin scaffolds have high porosity, enough mechanical strength and biostability, and excellent cytocompatibility. Copyright © 2018 Elsevier B.V. All rights reserved.

  7. A dual-application poly (dl-lactic-co-glycolic) acid (PLGA)-chitosan composite scaffold for potential use in bone tissue engineering.

    Science.gov (United States)

    Boukari, Yamina; Qutachi, Omar; Scurr, David J; Morris, Andrew P; Doughty, Stephen W; Billa, Nashiru

    2017-11-01

    The development of patient-friendly alternatives to bone-graft procedures is the driving force for new frontiers in bone tissue engineering. Poly (dl-lactic-co-glycolic acid) (PLGA) and chitosan are well-studied and easy-to-process polymers from which scaffolds can be fabricated. In this study, a novel dual-application scaffold system was formulated from porous PLGA and protein-loaded PLGA/chitosan microspheres. Physicochemical and in vitro protein release attributes were established. The therapeutic relevance, cytocompatibility with primary human mesenchymal stem cells (hMSCs) and osteogenic properties were tested. There was a significant reduction in burst release from the composite PLGA/chitosan microspheres compared with PLGA alone. Scaffolds sintered from porous microspheres at 37 °C were significantly stronger than the PLGA control, with compressive strengths of 0.846 ± 0.272 MPa and 0.406 ± 0.265 MPa, respectively (p < 0.05). The formulation also sintered at 37 °C following injection through a needle, demonstrating its injectable potential. The scaffolds demonstrated cytocompatibility, with increased cell numbers observed over an 8-day study period. Von Kossa and immunostaining of the hMSC-scaffolds confirmed their osteogenic potential with the ability to sinter at 37 °C in situ.

  8. Fabrication, characterization and in vitro drug release behavior of electrospun PLGA/chitosan nanofibrous scaffold

    Energy Technology Data Exchange (ETDEWEB)

    Meng, Z.X.; Zheng, W.; Li, L. [Center for Biomedical Materials and Engineering, Harbin Engineering University, Harbin 150001 (China); Zheng, Y.F., E-mail: yfzheng@pku.edu.cn [Center for Biomedical Materials and Engineering, Harbin Engineering University, Harbin 150001 (China); Department of Advanced Materials and Nanotechnology, College of Engineering, Peking University, Beijing 100871 (China)

    2011-02-15

    Graphical abstract: The fenbufen loaded PLGA/chitosan nanofibrous scaffolds were fabricated by electrospinning. The hydrophilicity of nanofibrous scaffold was enhanced with the increase of chitosan content. The drug release also is accelerated with chitosan increasing because the higher hydrophilicity makes drug diffusing from scaffold more easily. Research highlights: {yields} The average diameter increased with the increase of chitosan content and then decreased. {yields} The release rate of fenbufen increased with the increase of chitosan. {yields} The aligned nanofibrous scaffold exhibits lower drug release rate. {yields} The drug release could be controlled by crosslinking in glutaraldehyde vapor. - Abstract: In this study both aligned and randomly oriented poly(D,L-lactide-co-glycolide) (PLGA)/chitosan nanofibrous scaffold have been prepared by electrospinning. The ratio of PLGA to chitosan was adjusted to get smooth nanofiber surface. Morphological characterization using scanning electron microscopy showed that the aligned nanofiber diameter distribution obtained by electrospinning of polymer blend increased with the increase of chitosan content which was similar to that of randomly oriented nanofibers. The release characteristic of model drug fenbufen (FBF) from the FBF-loaded aligned and randomly oriented PLGA and PLGA/chitosan nanofibrous scaffolds was investigated. The drug release rate increased with the increase of chitosan content because the addition of chitosan enhanced the hydrophilicity of the PLGA/chitosan composite scaffold. Moreover, for the aligned PLGA/chitosan nanofibrous scaffold the release rate was lower than that of randomly oriented PLGA/chitosan nanofibrous scaffold, which indicated that the nanofiber arrangement would influence the release behavior. In addition, crosslinking in glutaraldehyde vapor would decrease the burst release of FBF from FBF-loaded PLGA/chitosan nanofibrous scaffold with a PLGA/chitosan ratio less than 9/1, which

  9. Fabrication, characterization and in vitro drug release behavior of electrospun PLGA/chitosan nanofibrous scaffold

    International Nuclear Information System (INIS)

    Meng, Z.X.; Zheng, W.; Li, L.; Zheng, Y.F.

    2011-01-01

    Graphical abstract: The fenbufen loaded PLGA/chitosan nanofibrous scaffolds were fabricated by electrospinning. The hydrophilicity of nanofibrous scaffold was enhanced with the increase of chitosan content. The drug release also is accelerated with chitosan increasing because the higher hydrophilicity makes drug diffusing from scaffold more easily. Research highlights: → The average diameter increased with the increase of chitosan content and then decreased. → The release rate of fenbufen increased with the increase of chitosan. → The aligned nanofibrous scaffold exhibits lower drug release rate. → The drug release could be controlled by crosslinking in glutaraldehyde vapor. - Abstract: In this study both aligned and randomly oriented poly(D,L-lactide-co-glycolide) (PLGA)/chitosan nanofibrous scaffold have been prepared by electrospinning. The ratio of PLGA to chitosan was adjusted to get smooth nanofiber surface. Morphological characterization using scanning electron microscopy showed that the aligned nanofiber diameter distribution obtained by electrospinning of polymer blend increased with the increase of chitosan content which was similar to that of randomly oriented nanofibers. The release characteristic of model drug fenbufen (FBF) from the FBF-loaded aligned and randomly oriented PLGA and PLGA/chitosan nanofibrous scaffolds was investigated. The drug release rate increased with the increase of chitosan content because the addition of chitosan enhanced the hydrophilicity of the PLGA/chitosan composite scaffold. Moreover, for the aligned PLGA/chitosan nanofibrous scaffold the release rate was lower than that of randomly oriented PLGA/chitosan nanofibrous scaffold, which indicated that the nanofiber arrangement would influence the release behavior. In addition, crosslinking in glutaraldehyde vapor would decrease the burst release of FBF from FBF-loaded PLGA/chitosan nanofibrous scaffold with a PLGA/chitosan ratio less than 9/1, which would be beneficial

  10. Preparation of aminated chitosan/alginate scaffold containing halloysite nanotubes with improved cell attachment.

    Science.gov (United States)

    Amir Afshar, Hamideh; Ghaee, Azadeh

    2016-10-20

    The chemical nature of biomaterials play important role in cell attachment, proliferation and migration in tissue engineering. Chitosan and alginate are biodegradable and biocompatible polymers used as scaffolds for various medical and clinical applications. Amine groups of chitosan scaffolds play an important role in cell attachment and water adsorption but also associate with alginate carboxyl groups via electrostatic interactions and hydrogen bonding, consequently the activity of amine groups in the scaffold decreases. In this study, chitosan/alginate/halloysite nanotube (HNTs) composite scaffolds were prepared using a freeze-drying method. Amine treatment on the scaffold occurred through chemical methods, which in turn caused the hydroxyl groups to be replaced with carboxyl groups in chitosan and alginate, after which a reaction between ethylenediamine, 1-ethyl-3,(3-dimethylaminopropyl) carbodiimide (EDC) and scaffold triggered the amine groups to connect to the carboxyl groups of chitosan and alginate. The chemical structure, morphology and mechanical properties of the composite scaffolds were investigated by FTIR, CHNS, SEM/EDS and compression tests. The electrostatic attraction and hydrogen bonding between chitosan, alginate and halloysite was confirmed by FTIR spectroscopy. Chitosan/alginate/halloysite scaffolds exhibit significant enhancement in compressive strength compared with chitosan/alginate scaffolds. CHNS and EDS perfectly illustrate that amine groups were effectively introduced in the aminated scaffold. The growth and cell attachment of L929 cells as well as the cytotoxicity of the scaffolds were investigated by SEM and Alamar Blue (AB). The results indicated that the aminated chitosan/alginate/halloysite scaffold has better cell growth and cell adherence in comparison to that of chitosan/alginate/halloysite samples. Aminated chitosan/alginate/halloysite composite scaffolds exhibit great potential for applications in tissue engineering, ideally in

  11. Fabrication and In Vitro Evaluation of Nanosized Hydroxyapatite/Chitosan-Based Tissue Engineering Scaffolds

    Directory of Open Access Journals (Sweden)

    Tao Sun

    2014-01-01

    Full Text Available Composite scaffolds based on biodegradable natural polymer and osteoconductive hydroxyapatite (HA nanoparticles can be promising for a variety of tissue engineering (TE applications. This study addressed the fabrication of three-dimensional (3D porous composite scaffolds composed of HA and chitosan fabricated via thermally induced phase separation and freeze-drying technique. The scaffolds produced were subsequently characterized in terms of microstructure, porosity, and mechanical property. In vitro degradation and in vitro biological evaluation were also investigated. The scaffolds were highly porous and had interconnected pore structures. The pore sizes ranged from several microns to a few hundred microns. The incorporated HA nanoparticles were well mixed and physically coexisted with chitosan in composite scaffold structures. The addition of 10% (w/w HA nanoparticles to chitosan enhanced the compressive mechanical properties of composite scaffold compared to pure chitosan scaffold. In vitro degradation results in phosphate buffered saline (PBS showed slower uptake properties of composite scaffolds. Moreover, the scaffolds showed positive response to mouse fibroblast L929 cells attachment. Overall, the findings suggest that HA/chitosan composite scaffolds could be suitable for TE applications.

  12. A novel chitosan-tussah silk fibroin/nano-hydroxyapatite composite bone scaffold platform with tunable mechanical strength in a wide range.

    Science.gov (United States)

    Ran, Jiabing; Hu, Jingxiao; Sun, Guanglin; Chen, Si; Jiang, Pei; Shen, Xinyu; Tong, Hua

    2016-12-01

    Currently, great efforts have been made to enhance the mechanical strength of bone tissue engineering (BTE) scaffolds, which are composed of biopolymeric matrices and inorganic nano-fillers. But the tunability of mechanical strength in a wide range for BTE scaffolds has seldom been investigated in spite of the great importance of this performance. In this work, a chitosan-tussah silk fibroin/hydroxyapatite (CS-TSF/HAp) hydrogel was synthesized by using a novel in situ precipitation method. Through in situ inducing the conformation transition of TSF in the CS-TSF/HAp hydrogel, which could be monitored by XRD, FT-IR, TGA, and DTA, the elastic modulus and fracture strength of the final CS-TSF/HAp composite could be tailored in a wide range without changing its composition, morphology, roughness, and crystal structures. The elastic modulus of the CS-TSF/HAp composite ranged from ∼250 to ∼400MPa while its fracture strength ranged from ∼45 to ∼100MPa. In order to clarify the rationale behind this process, a speculative explanation was provided. In vitro cell culture indicated that MC3T3-E1 cells cultured on the CS-TSF/HAp composite had positive adhesion, proliferation, and differentiation potential. We believed that the CS-TSF/HAp composite could be used as an ideal scaffold platform for cell culture and implantation of bone reconstruction. Copyright © 2016 Elsevier B.V. All rights reserved.

  13. Biocompatibility of chitosan/Mimosa tenuiflora scaffolds for tissue engineering

    Energy Technology Data Exchange (ETDEWEB)

    Martel-Estrada, Santos Adriana [Instituto de arquitectura diseño y arte, Universidad Autónoma de Ciudad Juárez, Ave. Del Charro #610 norte, Col. Partido Romero, C.P. 32320 Cd. Juárez, Chihuahua (Mexico); Rodríguez-Espinoza, Brenda [Instituto de Ciencias Biomédicas, Universidad Autónoma de Ciudad Juárez, Anillo envolvente del PRONAF y Estocolmo, C.P. 32320 Cd. Juárez, Chihuahua (Mexico); Santos-Rodríguez, Elí [ICTP Meso-American Centre for Theoretical Physics (ICTP-MCTP)/Universidad Autónoma de Chiapas, Ciudad Universitaria, Carretera Zapata Km. 4, Real del Bosque (Terán), C.P. 29040 Tuxtla Gutiérrez, Chiapas (Mexico); Jiménez-Vega, Florinda [Instituto de Ciencias Biomédicas, Universidad Autónoma de Ciudad Juárez, Anillo envolvente del PRONAF y Estocolmo, C.P. 32320 Cd. Juárez, Chihuahua (Mexico); García-Casillas, Perla E.; Martínez-Pérez, Carlos A. [Instituto de Ingeniería y Tecnología, Universidad Autónoma de Ciudad Juárez, Ave. Del Charro #610 norte, Col. Partido Romero, C.P. 32320 Cd. Juárez, Chihuahua (Mexico); and others

    2015-09-15

    Highlights: • The porosity of the composites allow biological processes for the cell adaptation on the scaffolds. • The composites improve the viability and proliferation of cells. • Composition of the scaffold plays an important role in the biocompatibility. • The results indicate that Mimosa Tenuiflora can induce the differentiation of osteoblast cells. - Abstract: In search of a plant that exhibits osteogenic activity, Mimosa tenuiflora (M. tenuiflora) cortex represents the opportunity to create a biomaterial that, together with the chitosan, is osteoconductive and promote better and rapid regeneration of bone tissue. Thus, the composite of chitosan/M. tenuiflora cortex fabricated will have properties of biocompatibility and allow the osteoblast proliferation. Composites were developed with different concentrations of chitosan/M. tenuiflora cortex (w/w) using thermally induced phase separation technique (TIPS). To analyze the effects of composite on osteoblasts, primary cultures, each sample was collected on days 1, 3 and 7 after seeding. The evaluation of composites consisted of viability and proliferation tests in which we observed the metabolic activity of the cells using MTT reagent and determined the DNA concentration by means of fluorescence. The expression of the marker alkaline phosphatase (ALP) using p-nitrophenyl phosphate was examined, allowing the observation to the activity of proliferation and differentiation of osteoblastic cells. Moreover, an analysis of biomineralization was performed using scanning electron microscopy (SEM), energy dispersive spectroscopy, infrared spectroscopy and X-ray diffraction. The results showed that 80/20 chitosan/M. tenuiflora cortex biocomposite has the best performance with osteoblasts compared to biomaterials 100/0 and 70/30 chitosan/M. tenuiflora composites. Finally, it was determined that the composite of chitosan/M. tenuiflora cortex presents no cytotoxicity and increases the capacity of the osteoblasts

  14. In vitro osteoclastogenesis on textile chitosan scaffold

    Directory of Open Access Journals (Sweden)

    C Heinemann

    2010-02-01

    Full Text Available Textile chitosan fibre scaffolds were evaluated in terms of interaction with osteoclast-like cells, derived from human primary monocytes. Part of the scaffolds was further modified by coating with fibrillar collagen type I in order to make the surface biocompatible. Monocytes were cultured directly on the scaffolds in the presence of macrophage colony stimulating factor (M-CSF and receptor activator of nuclear factor kappaB ligand (RANKL for up to 18 days. Confocal laser scanning microscopy (CLSM as well as scanning electron microscopy (SEM revealed the formation of multinuclear osteoclast-like cells on both the raw chitosan fibres and the collagen-coated scaffolds. The modified surface supported the osteoclastogenesis. Differentiation towards the osteoclastic lineage was confirmed by the microscopic detection of cathepsin K, tartrate resistant acid phosphatase (TRAP, acidic compartments using 3-(2,4-dinitroanillino-3’-amino-N-methyldipropylamine (DAMP, immunological detection of TRAP isoform 5b, and analysis of gene expression of the osteoclastic markers TRAP, cathepsin K, vitronectin receptor, and calcitonin receptor using reverse transcription-polymerase chain reaction (RT-PCR. The feature of the collagen-coated but also of the raw chitosan fibre scaffolds to support attachment and differentiation of human monocytes facilitates cell-induced material resorption – one main requirement for successful bone tissue engineering.

  15. Nature derived scaffolds for tissue engineering applications: Design and fabrication of a composite scaffold incorporating chitosan-g-d,l-lactic acid and cellulose nanocrystals from Lactuca sativa L. cv green leaf.

    Science.gov (United States)

    Ko, Sung Won; Soriano, Juan Paolo E; Lee, Ji Yeon; Unnithan, Afeesh Rajan; Park, Chan Hee; Kim, Cheol Sang

    2018-04-15

    Through exhaustive extraction via successive alkali and bleaching treatments cellulose was isolated from lettuce. The isolated cellulose was hydrolyzed using 64wt% H 2 SO 4 at 55°C under constant stirring for 1h to obtain cellulose nanocrystals (CNCs). Characterizations such as SEM, TEM, FTIR, TGA and XRD were done in order to determine differences in the physico-chemical characteristics of cellulose after each treatment step. The isolated CNCs have mean dimensions of 237±26, 33±12 and 32±7nm in length, thickness and height, respectively. These nanocrystals were incorporated to the formulations that were used to fabricate different chitosan-g-d,l-lactic acid (CgLA) scaffolds. Amide linkage formation between chitosan and lactic acid and further removal of water was facilitated by oven-drying under vacuum at 80°C. Results show that an increase in the concentration of CNCs added, increase in porosity, degradability, drug release property and cell viability were observed from the fabricated composite scaffolds. These results can provide information on how nanofillers such as CNCs can alter the properties of tissue scaffolds through the chemical properties and interactions they provide. Moreover, these characteristics can give new properties that are necessary for certain tissue engineering applications. Copyright © 2017 Elsevier B.V. All rights reserved.

  16. Chitosan/bioactive glass nanoparticles scaffolds with shape memory properties.

    Science.gov (United States)

    Correia, Cristina O; Leite, Álvaro J; Mano, João F

    2015-06-05

    We propose a combination of chitosan (CHT) with bioactive glass nanoparticles (BG-NPs) in order to produce CHT/BG-NPs scaffolds that combine the shape memory properties of chitosan and the biomineralization ability of BG-NPs for applications in bone regeneration. The addition of BG-NPs prepared by a sol-gel route to the CHT polymeric matrix improved the bioactivity of the nanocomposite scaffold, as seen by the precipitation of bone-like apatite layer upon immersion in simulated body fluid (SBF). Shape memory tests were carried out while the samples were immersed in varying compositions of water/ethanol mixtures. Dehydration with ethanol enables to fix a temporary shape of a deformed scaffold that recovers the initial geometry upon water uptake. The scaffolds present good shape memory properties characterized by a recovery ratio of 87.5% for CHT and 89.9% for CHT/BG-NPs and a fixity ratio of 97.2% for CHT and 98.2% for CHT/BG-NPs (for 30% compressive deformation). The applicability of such structures was demonstrated by a good geometrical accommodation of a previously compressed scaffold in a bone defect. The results indicate that the developed CHT/BG-NPs nanocomposite scaffolds have potential for being applied in bone tissue engineering. Copyright © 2015 Elsevier Ltd. All rights reserved.

  17. Chitosan based metallic nanocomposite scaffolds as antimicrobial wound dressings.

    Science.gov (United States)

    Mohandas, Annapoorna; Deepthi, S; Biswas, Raja; Jayakumar, R

    2018-09-01

    Chitosan based nanocomposite scaffolds have attracted wider applications in medicine, in the area of drug delivery, tissue engineering and wound healing. Chitosan matrix incorporated with nanometallic components has immense potential in the area of wound dressings due to its antimicrobial properties. This review focuses on the different combinations of Chitosan metal nanocomposites such as Chitosan/nAg, Chitosan/nAu, Chitosan/nCu, Chitosan/nZnO and Chitosan/nTiO 2 towards enhancement of healing or infection control with special reference to the antimicrobial mechanism of action and toxicity.

  18. Mechanical properties, biological activity and protein controlled release by poly(vinyl alcohol)-bioglass/chitosan-collagen composite scaffolds: a bone tissue engineering applications.

    Science.gov (United States)

    Pon-On, Weeraphat; Charoenphandhu, Narattaphol; Teerapornpuntakit, Jarinthorn; Thongbunchoo, Jirawan; Krishnamra, Nateetip; Tang, I-Ming

    2014-05-01

    In the present study, composite scaffolds made with different weight ratios (0.5:1, 1:1 and 2:1) of bioactive glass (15Ca:80Si:5P) (BG)/polyvinyl alcohol (PVA) (PVABG) and chitosan (Chi)/collagen (Col) (ChiCol) were prepared by three mechanical freeze-thaw followed by freeze-drying to obtain the porous scaffolds. The mechanical properties and the in vitro biocompatibility of the composite scaffolds to simulated body fluid (SBF) and to rat osteoblast-like UMR-106 cells were investigated. The results from the studies indicated that the porosity and compressive strength were controlled by the weight ratio of PVABG:ChiCol. The highest compressive modulus of the composites made was 214.64 MPa which was for the 1:1 weight ratio PVABG:ChiCol. Mineralization study in SBF showed the formation of apatite crystals on the PVABG:ChiCol surface after 7 days of incubation. In vitro cell availability and proliferation tests confirmed the osteoblast attachment and growth on the PVABG:ChiCol surface. MTT and ALP tests on the 1:1 weight ratio PVABG:ChiCol composite indicated that the UMR-106 cells were viable. Alkaline phosphatase activity was found to increase with increasing culturing time. In addition, we showed the potential of PVABG:ChiCol drug delivery through PBS solution studies. 81.14% of BSA loading had been achieved and controlled release for over four weeks was observed. Our results indicated that the PVABG:ChiCol composites, especially the 1:1 weight ratio composite exhibited significantly improved mechanical, mineral deposition, biological properties and controlled release. This made them potential candidates for bone tissue engineering applications. Copyright © 2014 Elsevier B.V. All rights reserved.

  19. Nerve growth factor loaded heparin/chitosan scaffolds for accelerating peripheral nerve regeneration.

    Science.gov (United States)

    Li, Guicai; Xiao, Qinzhi; Zhang, Luzhong; Zhao, Yahong; Yang, Yumin

    2017-09-01

    Artificial chitosan scaffolds have been widely investigated for peripheral nerve regeneration. However, the effect was not as good as that of autologous grafts and therefore could not meet the clinical requirement. In the present study, the nerve growth factor (NGF) loaded heparin/chitosan scaffolds were fabricated via electrostatic interaction for further improving nerve regeneration. The physicochemical properties including morphology, wettability and composition were measured. The heparin immobilization, NGF loading and release were quantitatively and qualitatively characterized, respectively. The effect of NGF loaded heparin/chitosan scaffolds on nerve regeneration was evaluated by Schwann cells culture for different periods. The results showed that the heparin immobilization and NGF loading did not cause the change of bulk properties of chitosan scaffolds except for morphology and wettability. The pre-immobilization of heparin in chitosan scaffolds could enhance the stability of subsequently loaded NGF. The NGF loaded heparin/chitosan scaffolds could obviously improve the attachment and proliferation of Schwann cells in vitro. More importantly, the NGF loaded heparin/chitosan scaffolds could effectively promote the morphology development of Schwann cells. The study may provide a useful experimental basis to design and develop artificial implants for peripheral nerve regeneration and other tissue regeneration. Copyright © 2017 Elsevier Ltd. All rights reserved.

  20. Biochemical properties of Hemigraphis alternata incorporated chitosan hydrogel scaffold.

    Science.gov (United States)

    Annapoorna, M; Sudheesh Kumar, P T; Lakshman, Lakshmi R; Lakshmanan, Vinoth-Kumar; Nair, Shantikumar V; Jayakumar, R

    2013-02-15

    In this work, Hemigraphis alternata extract incorporated chitosan scaffold was synthesized and characterized for wound healing. The antibacterial activity of Hemigraphis incorporated chitosan scaffold (HIC) against Escherichia coli and Staphylococcus aureus was evaluated which showed a reduction in total colony forming units by 45-folds toward E. coli and 25-fold against S. aureus respectively. Cell viability studies using Human Dermal Fibroblast cells (HDF) showed 90% viability even at 48 h when compared to the chitosan control. The herbal scaffold made from chitosan was highly haemostatic and antibacterial. The obtained results were in support that the herbal scaffold can be effectively applied for infectious wounds. Copyright © 2012 Elsevier Ltd. All rights reserved.

  1. Biomimetic Composite Scaffold for Breast Reconstruction Following Tumor Resection

    National Research Council Canada - National Science Library

    Patrick, Jr, Charles W

    2005-01-01

    ... of life and outcomes are markedly improved. We hypothesized that a novel composite material consisting of silk fibroin and chitosan will act as a biomimetic scaffold amenable to in vivo adipogenesis. The specific aims (SAs...

  2. Patient-Derived Human Induced Pluripotent Stem Cells From Gingival Fibroblasts Composited With Defined Nanohydroxyapatite/Chitosan/Gelatin Porous Scaffolds as Potential Bone Graft Substitutes.

    Science.gov (United States)

    Ji, Jun; Tong, Xin; Huang, Xiaofeng; Zhang, Junfeng; Qin, Haiyan; Hu, Qingang

    2016-01-01

    (hiPSCs) were established from clinically easily derived human gingival fibroblasts (hGFs) and defined nanohydroxyapatite/chitosan/gelatin (HCG) scaffolds. hiPSCs derived from hGFs had better osteogenesis capability than that of hGFs. More interestingly, osteogenic differentiation of hiPSCs from hGFs was elevated significantly when composited with HCG-311 scaffolds in vitro and in vivo. The present study has uncovered the important role of different nHA ratios in HCG scaffolds in osteogenesis induction of hiPSCs derived from hGFs. This technique could serve as a potential innovative approach for bone tissue engineering, especially large bone regeneration clinically. ©AlphaMed Press.

  3. Cytocompatibility of chitosan and collagen-chitosan scaffolds for tissue engineering

    Directory of Open Access Journals (Sweden)

    Ligia L. Fernandes

    2011-01-01

    Full Text Available In this work, chitosan and collagen-chitosan porous scaffolds were produced by the freeze drying method and characterized as potential skin substitutes. Their beneficial effects on soft tissues justify the choice of both collagen and chitosan. Samples were characterized using scanning electron microscope, Fourier Transform InfraRed Spectroscopy (FTIR and thermogravimetry (TG. The in vitro cytocompatibility of chitosan and collagen-chitosan scaffolds was evaluated with three different assays. Phenol and titanium powder were used as positive and negative controls, respectively. Scanning electron microscopy revealed the highly interconnected porous structure of the scaffolds. The addition of collagen to chitosan increased both pore diameter and porosity of the scaffolds. Results of FTIR and TG analysis indicate that the two polymers interact yielding a miscible blend with intermediate thermal degradation properties. The reduction of XTT ((2,3-bis[2-methyloxy-4-nitro-5-sulfophenyl]-2H-tetrazolium-5-carboxanilide and the uptake of Neutral Red (NR were not affected by the blend or by the chitosan scaffold extracts, but the blend and the titanium powder presented greater incorporation of Crystal Violet (CV than phenol and chitosan alone. In conclusion, collagen-chitosan scaffolds produced by freeze-drying methods were cytocompatible and presented mixed properties of each component with intermediate thermal degradation properties.

  4. Oxygen Plasma Treatment on 3D-Printed Chitosan/Gelatin/Hydroxyapatite Scaffolds for Bone Tissue Engineering.

    Science.gov (United States)

    Lee, Chang-Min; Yang, Seong-Won; Jung, Sang-Chul; Kim, Byung-Hoon

    2017-04-01

    The 3D hydroxyapatite/gelatin/chitosan composite scaffolds were fabricated by 3D printing technique. The scaffolds were treated by oxygen plasma to improve the bioactivity and its surface characterization and in vitro cell culture were investigated. The scaffolds exhibited the good porosity and interconnectivity of pores. After oxygen plasma etching, roughness and wettability on the scaffolds surface are increased. Plasma treated scaffolds showed higher proliferation than that of untreated scaffolds. Oxygen plasma treatment could be used as potential tool to enhance the biocompatibility on the 3D composite scaffolds.

  5. Scanning electron microscopy and swelling test of shrimp shell chitosan and chitosan-RGD scaffolds

    Science.gov (United States)

    Mandacan, M. C.; Yuniastuti, M.; Amir, L. R.; Idrus, E.; Suniarti, D. F.

    2017-08-01

    Shrimp shell chitosan and chitosan-RGD scaffold membranes are produced to be biocompatible with tissue engineering. Nonetheless, their architectural properties have not yet been studied. Analyze the architectural properties of chitosan and chitosan-RGD scaffolds. Analyze pore count and size, interpore distance, and porosity (using SEM testing and ImageJ analysis) and water absorption (using a swelling test). The properties of the chitosan and chitosan-RGD scaffolds were as follows, respectively. The pore counts were 225 and 153; pore size, 171.4 μam and 180.2 μam interpore distance, 105.7 μam and 101.4 μam porosity, 22% and 10.2%; and water absorption, 9.1 mgH2O/mgScaffold and 19.3 mgH2O/mgScaffold. The shrimp shell chitosan-RGD membrane scaffold was found to have architectural properties that make it more conducive to use in tissue engineering.

  6. Enzymatically biomineralized chitosan scaffolds for tissue-engineering applications.

    NARCIS (Netherlands)

    Dash, M.; Samal, S.K.; Douglas, T.E.L.; Schaubroeck, D.; Leeuwenburgh, S.C.G.; Voort, P. van der; Declercq, H.A.; Dubruel, P.

    2017-01-01

    Porous biodegradable scaffolds represent promising candidates for tissue-engineering applications because of their capability to be preseeded with cells. We report an uncrosslinked chitosan scaffold designed with the aim of inducing and supporting enzyme-mediated formation of apatite minerals in the

  7. 3D printing biodegradable scaffolds with chitosan materials for tissue engineering

    Science.gov (United States)

    Bardakova, K. N.; Demina, T. S.; Grebenik, E. A.; Minaev, N. V.; Akopova, T. A.; Bagratashvili, V. N.; Timashev, P. S.

    2018-04-01

    Chitosan-g-oligo (L,L-lactide) copolymer was synthesized through a solvent-free reaction in an extruder. Three-dimensional scaffolds based on photosensitive composition contained the synthetized copolymer were formed by two-photon polymerization. The optimum ratio of components, methods of preparation of photopolymerizable mixtures, parameters of the laser structuring and procedure of washing from unbound crosslinkers have been optimized. Chitosan scaffolds were non-cytotoxic and might therefore be a suitable candidate for treating spinal cord injuries and other neuronal degenerative diseases.

  8. Mechanical enhancement and in vitro biocompatibility of nanofibrous collagen-chitosan scaffolds for tissue engineering.

    Science.gov (United States)

    Zou, Fengjuan; Li, Runrun; Jiang, Jianjun; Mo, Xiumei; Gu, Guofeng; Guo, Zhongwu; Chen, Zonggang

    2017-12-01

    The collagen-chitosan complex with a three-dimensional nanofiber structure was fabricated to mimic native ECM for tissue repair and biomedical applications. Though the three-dimensional hierarchical fibrous structures of collagen-chitosan composites could provide more adequate stimulus to facilitate cell adhesion, migrate and proliferation, and thus have the potential as tissue engineering scaffolding, there are still limitations in their applications due to the insufficient mechanical properties of natural materials. Because poly (vinyl alcohol) (PVA) and thermoplastic polyurethane (TPU) as biocompatible synthetic polymers can offer excellent mechanical properties, they were introduced into the collagen-chitosan composites to fabricate the mixed collagen/chitosan/PVA fibers and a sandwich structure (collagen/chitosan-TPU-collagen/chitosan) of nanofiber in order to enhance the mechanical properties of the nanofibrous collagen-chitosan scaffold. The results showed that the tensile behavior of materials was enhanced to different degrees with the difference of collagen content in the fibers. Besides the Young's modulus had no obvious changes, both the break strength and the break elongation of materials were heightened after reinforced by PVA. For the collagen-chitosan nanofiber reinforced by TPU, both the break strength and the Young's modulus of materials were heightened in different degrees with the variety of collagen content in the fibers despite the decrease of the break elongation of materials to some extent. In vitro cell test demonstrated that the materials could provide adequate environment for cell adhesion and proliferation. All these indicated that the reinforced collagen-chitosan nanofiber could be as potential scaffold for tissue engineering according to the different mechanical requirements in clinic.

  9. Extraction and Characterization of Chitin and Chitosan from Blue Crab and Synthesis of Chitosan Cryogel Scaffolds

    Directory of Open Access Journals (Sweden)

    Nimet Bölgen

    2016-08-01

    Full Text Available Polymeric scaffolds produced by cryogelation technique have attracted increasing attention for tissue engineering applications. Cryogelation is a technique which enables to produce interconnected porous matrices from the frozen reaction mixtures of polymers or monomeric precursors. Chitosan is a biocompatible, biodegradable, nontoxic, antibacterial, antioxidant and antifungal natural polymer that is obtained by deacetylation of chitin, which is mostly found in the exoskeleton of many crustacean. In this study, chitin was isolated from the exoskeleton of blue crap (Callinectes sapidus using a chemical method. Callinectes sapidus samples were collected from a market, as a waste material after it has been consumed as food. Demineralization, deproteinization and decolorization steps were applied to the samples to obtain chitin. Chitosan was prepared from isolated chitin by deacetylation at high temperatures. The chemical compositon of crab shell, extracted chitin and chitosan were characterized with FTIR analyses. And also to determine the physicochemical and functional properties of the produced chitosan; solubility, water binding and fat binding analysis were performed. Chitosan cryogel scaffolds were prepared by crosslinking reaction at cryogenic conditions at constant amount of chitosan (1%, w/v with different ratios of glutaraldehyde (1, 3, and 6%, v/v as crosslinker. The chemical structure of the scaffolds were examined by FTIR. Also, the water uptake capacity of scaffolds have been determined. Collectively, the results suggested that the characterized chitosan cryogels can be potential scaffolds to be used in tissue engineering applications.

  10. Degradation behavior and compatibility of micro, nanoHA/chitosan scaffolds with interconnected spherical macropores.

    Science.gov (United States)

    Ruixin, Li; Cheng, Xu; Yingjie, Liu; Hao, Li; Caihong, Shi; Weihua, Su; Weining, An; Yinghai, Yuan; Xiaoli, Qin; Yunqiang, Xu; Xizheng, Zhang; Hui, Li

    2017-10-01

    Hydroxyapatite/Chitosan (HA/CS) composite have significant application in biomedical especially for bone replacement. Inorganic particle shape and size of composite affect the scaffold mechanical property, biological property, and degradation. The aim of this study was to fabricate HA/CS scaffold with good pore connectivity and analyze their biological, degradation properties. Microhydroxyapatite/chitosan (mHA/CS) and nanohydroxyapatite/chitosan (nHA/CS) composite scaffolds with interconnected spherical pore architectures were fabricated. Composite scaffolds structure parameters were analyzed using micro CT. Cell proliferation and morphology were tested and compared between two scaffolds using mouse osteoblastic cell line MC3T3-E1. To research the composite degradation in lysozyme PBS solution, degradation rate and reducing sugar content were tested, and scaffolds morphology were observed by SEM. The results showed that microHA and nanoHA were fabricated by being calcined and synthesis methods, and their infrared spectra are very similar. EDAX composition analysis demonstrated that both of microHA and nanoHA were calcium deficiency HA. Micro-CT results demonstrated the scaffolds had interconnected spherical pores, and the structure parameters were similar. Cell viabilities were significant increased with cultured time, but there were no significant difference between microHA/CS and nanoHA/CS scaffolds. Scaffold structure was gradually destroyed and inorganic composition HA particles are more prominent with degradation time. (1) Inorganic particle shape and size of composite affect the scaffold mechanical property, biological property, and degradation. NanoHA/CS and microHA/CS scaffolds with good pore connectivity were fabricated and their biological, degradation properties were studied in this manuscript. (2) The scaffold with interconnected porosity construct provides the necessary support for cells to proliferate and maintain their differentiated function, and

  11. Bone repair by cell-seeded 3D-bioplotted composite scaffolds made of collagen treated tricalciumphosphate or tricalciumphosphate-chitosan-collagen hydrogel or PLGA in ovine critical-sized calvarial defects.

    Science.gov (United States)

    Haberstroh, Kathrin; Ritter, Kathrin; Kuschnierz, Jens; Bormann, Kai-Hendrik; Kaps, Christian; Carvalho, Carlos; Mülhaupt, Rolf; Sittinger, Michael; Gellrich, Nils-Claudius

    2010-05-01

    The aim of this study was to investigate the osteogenic effect of three different cell-seeded 3D-bioplotted scaffolds in a ovine calvarial critical-size defect model. The choice of scaffold-materials was based on their applicability for 3D-bioplotting and respective possibility to produce tailor-made scaffolds for the use in cranio-facial surgery for the replacement of complex shaped boneparts. Scaffold raw-materials are known to be osteoinductive when being cell-seeded [poly(L-lactide-co-glycolide) (PLGA)] or having components with osteoinductive properties as tricalciumphosphate (TCP) or collagen (Col) or chitosan. The scaffold-materials PLGA, TCP/Col, and HYDR (TCP/Col/chitosan) were cell-seeded with osteoblast-like cells whether gained from bone (OLB) or from periost (OLP). In a prospective and randomized design nine sheep underwent osteotomy to create four critical-sized calvarial defects. Three animals each were assigned to the HYDR-, the TCP/Col-, or the PLGA-group. In each animal, one defect was treated with a cell-free, an OLB- or OLP-seeded group-specific scaffold, respectively. The fourth defect remained untreated as control (UD). Fourteen weeks later, animals were euthanized for histo-morphometrical analysis of the defect healing. OLB- and OLP-seeded HYDR and OLB-seeded TCP/Col scaffolds significantly increased the amount of newly formed bone (NFB) at the defect bottom and OLP-seeded HYDR also within the scaffold area, whereas PLGA-scaffolds showed lower rates. The relative density of NFB was markedly higher in the HYDR/OLB group compared to the corresponding PLGA group. TCP/Col had good stiffness to prepare complex structures by bioplotting but HYDR and PLGA were very soft. HYDR showed appropriate biodegradation, TCP/Col and PLGA seemed to be nearly undegraded after 14 weeks. 3D-bioplotted, cell-seeded HYDR and TCP/Col scaffolds increased the amount of NFB within ovine critical-size calvarial defects, but stiffness, respectively, biodegradation of

  12. Collagen-chitosan scaffold modified with Au and Ag nanoparticles: Synthesis and structure

    Energy Technology Data Exchange (ETDEWEB)

    Rubina, M.S.; Kamitov, E.E. [A.N. Nesmeyanov Institute of Organoelement Compounds, Russian Academy of Sciences, Moscow, 119991 Russian Federation (Russian Federation); Zubavichus, Ya. V.; Peters, G.S. [National Research center «Kurchatov Institute», Moscow, 123182 Russian Federation (Russian Federation); Naumkin, A.V. [A.N. Nesmeyanov Institute of Organoelement Compounds, Russian Academy of Sciences, Moscow, 119991 Russian Federation (Russian Federation); Suzer, S. [Department of Chemistry, Bilkent University, Ankara, 06800 Turkey (Turkey); Vasil’kov, A.Yu., E-mail: alexandervasilkov@yandex.ru [A.N. Nesmeyanov Institute of Organoelement Compounds, Russian Academy of Sciences, Moscow, 119991 Russian Federation (Russian Federation)

    2016-03-15

    Graphical abstract: - Highlights: • Biocompatible collagen-chitosan scaffolds were modified by Au and Ag nanoparticles via the metal-vapor synthesis. • Structural and morphological parameters of the nanocomposites were assessed using a set of modern instrumental techniques, including electron microscopy, X-ray diffraction, small-angle X-ray scattering, EXAFS, XPS. • Potential application of the nanocomposites are envisaged. - Abstract: Nowadays, the dermal biomimetic scaffolds are widely used in regenerative medicine. Collagen-chitosan scaffold one of these materials possesses antibacterial activity, good compatibility with living tissues and has been already used as a wound-healing material. In this article, collagen-chitosan scaffolds modified with Ag and Au nanoparticles have been synthesized using novel method - the metal-vapor synthesis. The nanocomposite materials are characterized by XPS, TEM, SEM and synchrotron radiation-based X-ray techniques. According to XRD data, the mean size of the nanoparticles (NPs) is 10.5 nm and 20.2 nm in Au-Collagen-Chitosan (Au-CollCh) and Ag-Collagen-Chitosan (Ag-CollCh) scaffolds, respectively in fair agreement with the TEM data. SAXS analysis of the composites reveals an asymmetric size distribution peaked at 10 nm for Au-CollCh and 25 nm for Ag-CollCh indicative of particle's aggregation. According to SEM data, the metal-carrying scaffolds have layered structure and the nanoparticles are rather uniformly distributed on the surface material. XPS data indicate that the metallic nanoparticles are in their unoxidized/neutral states and dominantly stabilized within the chitosan-rich domains.

  13. Collagen-chitosan scaffold modified with Au and Ag nanoparticles: Synthesis and structure

    International Nuclear Information System (INIS)

    Rubina, M.S.; Kamitov, E.E.; Zubavichus, Ya. V.; Peters, G.S.; Naumkin, A.V.; Suzer, S.; Vasil’kov, A.Yu.

    2016-01-01

    Graphical abstract: - Highlights: • Biocompatible collagen-chitosan scaffolds were modified by Au and Ag nanoparticles via the metal-vapor synthesis. • Structural and morphological parameters of the nanocomposites were assessed using a set of modern instrumental techniques, including electron microscopy, X-ray diffraction, small-angle X-ray scattering, EXAFS, XPS. • Potential application of the nanocomposites are envisaged. - Abstract: Nowadays, the dermal biomimetic scaffolds are widely used in regenerative medicine. Collagen-chitosan scaffold one of these materials possesses antibacterial activity, good compatibility with living tissues and has been already used as a wound-healing material. In this article, collagen-chitosan scaffolds modified with Ag and Au nanoparticles have been synthesized using novel method - the metal-vapor synthesis. The nanocomposite materials are characterized by XPS, TEM, SEM and synchrotron radiation-based X-ray techniques. According to XRD data, the mean size of the nanoparticles (NPs) is 10.5 nm and 20.2 nm in Au-Collagen-Chitosan (Au-CollCh) and Ag-Collagen-Chitosan (Ag-CollCh) scaffolds, respectively in fair agreement with the TEM data. SAXS analysis of the composites reveals an asymmetric size distribution peaked at 10 nm for Au-CollCh and 25 nm for Ag-CollCh indicative of particle's aggregation. According to SEM data, the metal-carrying scaffolds have layered structure and the nanoparticles are rather uniformly distributed on the surface material. XPS data indicate that the metallic nanoparticles are in their unoxidized/neutral states and dominantly stabilized within the chitosan-rich domains.

  14. Development of keratin–chitosan–gelatin composite scaffold for soft tissue engineering

    Energy Technology Data Exchange (ETDEWEB)

    Kakkar, Prachi [Central Leather Research Institute (Council of Scientific and Industrial Research), Adyar, Chennai 600020 (India); Verma, Sudhanshu; Manjubala, I. [Biomedical Engineering Division, School of Bio Sciences and Technology, VIT University, Vellore 632014 (India); Madhan, B., E-mail: bmadhan76@yahoo.co.in [Central Leather Research Institute (Council of Scientific and Industrial Research), Adyar, Chennai 600020 (India)

    2014-12-01

    Keratin has gained much attention in the recent past as a biomaterial for wound healing owing to its biocompatibility, biodegradability, intrinsic biological activity and presence of cellular binding motifs. In this paper, a novel biomimetic scaffold containing keratin, chitosan and gelatin was prepared by freeze drying method. The prepared keratin composite scaffold had good structural integrity. Fourier Transform Infrared (FTIR) spectroscopy showed the retention of the native structure of individual biopolymers (keratin, chitosan, and gelatin) used in the scaffold. Thermogravimetric Analysis (TGA) and Differential Scanning Calorimetry (DSC) results revealed a high thermal denaturation temperature of the scaffold (200–250 °C). The keratin composite scaffold exhibited tensile strength (96 kPa), compression strength (8.5 kPa) and water uptake capacity (> 1700%) comparable to that of a collagen scaffold, which was used as control. The morphology of the keratin composite scaffold observed using a Scanning Electron Microscope (SEM) exhibited good porosity and interconnectivity of pores. MTT assay using NIH 3T3 fibroblast cells demonstrated that the cell viability of the keratin composite scaffold was good. These observations suggest that the keratin–chitosan–gelatin composite scaffold is a promising alternative biomaterial for tissue engineering applications. - Highlights: • Fabrication of novel Keratin-Chitosan-Gelatin composite scaffold • Keratin composite scaffold shows excellent water uptake capacity and porosity • Keratin composite scaffold shows good thermal and physical stability • Biocompatibility of the developed scaffold is comparable to collagen scaffolds • Developed scaffold is a promising material for soft tissue engineering applications.

  15. Effect of enzymatic degradation of chitosan in polyhydroxybutyrate/chitosan/calcium phosphate composites on in vitro osteoblast response.

    Science.gov (United States)

    Giretova, Maria; Medvecky, Lubomir; Stulajterova, Radoslava; Sopcak, Tibor; Briancin, Jaroslav; Tatarkova, Monika

    2016-12-01

    Polyhydroxybutyrate/chitosan/calcium phosphate composites are interesting biomaterials for utilization in regenerative medicine and they may by applied in reconstruction of deeper subchondral defects. Insufficient informations were found in recent papers about the influence of lysozyme degradation of chitosan in calcium phosphate/chitosan based composites on in vitro cytotoxicity and proliferation activity of osteoblasts. The effect of enzymatic chitosan degradation on osteoblasts proliferation was studied on composite films in which the porosity of origin 3D scaffolds was eliminated and the surface texture was modified. The significantly enhanced proliferation activity with faster population growth of osteoblasts were found on enzymatically degraded biopolymer composite films with α-tricalcium phosphate and nanohydroxyapatite. No cytotoxicity of composite films prepared from lysozyme degraded scaffolds containing a large fraction of low molecular weight chitosans (LMWC), was revealed after 10 days of cultivation. Contrary to above in the higher cytotoxicity origin untreated nanohydroxyapatite films and porous composite scaffolds. The results showed that the synergistic effect of surface distribution, morphology of nanohydroxyapatite particles, microtopography and the presence of LMWC due to chitosan degradation in composite films were responsible for compensation of the cytotoxicity of nanohydroxyapatite composite films or porous composite scaffolds.

  16. PEDOT:PSS-Containing Nanohydroxyapatite/Chitosan Conductive Bionanocomposite Scaffold: Fabrication and Evaluation

    Directory of Open Access Journals (Sweden)

    Alireza Lari

    2016-01-01

    Full Text Available Conductive poly(3,4-ethylenedioxythiophene-poly(4-styrene sulfonate (PEDOT:PSS was incorporated into nanohydroxyapatite/chitosan (nHA/CS composite scaffolds through a freezing and lyophilization technique. The bionanocomposite conductive scaffold was then characterized using several techniques. A scanning electron microscope image showed that the nHA and PEDOT:PSS were dispersed homogeneously in the chitosan matrix, which was also confirmed by energy-dispersive X-ray (EDX analysis. The conductive properties were measured using a digital multimeter. The weight loss and water-uptake properties of the bionanocomposite scaffolds were studied in vitro. An in vitro cell cytotoxicity test was carried out using mouse fibroblast (L929 cells cultured onto the scaffolds. Using a freezing and lyophilization technique, it was possible to fabricate three-dimensional, highly porous, and interconnected PEDOT:PSS/nHA/CS scaffolds with good handling properties. The porosity was 74% and the scaffold’s conductivity was 9.72±0.78 μS. The surface roughness was increased with the incorporation of nHA and PEDOT:PSS into the CS scaffold. The compressive mechanical properties increased significantly with the incorporation of nHA but did not change significantly with the incorporation of PEDOT:PSS. The PEDOT:PSS-containing nHA/CS scaffold exhibited significantly higher cell attachment. The PEDOT:PSS/nHA/CS scaffold could be a potential bionanocomposite conductive scaffold for tissue engineering.

  17. Preparation and Evaluation of Gelatin-Chitosan-Nanobioglass 3D Porous Scaffold for Bone Tissue Engineering

    Directory of Open Access Journals (Sweden)

    Kanchan Maji

    2016-01-01

    Full Text Available The aim of the present study was to prepare and characterize bioglass-natural biopolymer based composite scaffold and evaluate its bone regeneration ability. Bioactive glass nanoparticles (58S in the size range of 20–30 nm were synthesized using sol-gel method. Porous scaffolds with varying bioglass composition from 10 to 30 wt% in chitosan, gelatin matrix were fabricated using the method of freeze drying of its slurry at 40 wt% solids loading. Samples were cross-linked with glutaraldehyde to obtain interconnected porous 3D microstructure with improved mechanical strength. The prepared scaffolds exhibited >80% porosity with a mean pore size range between 100 and 300 microns. Scaffold containing 30 wt% bioglass (GCB 30 showed a maximum compressive strength of 2.2±0.1 MPa. Swelling and degradation studies showed that the scaffold had excellent properties of hydrophilicity and biodegradability. GCB 30 scaffold was shown to be noncytotoxic and supported mesenchymal stem cell attachment, proliferation, and differentiation as indicated by MTT assay and RUNX-2 expression. Higher cellular activity was observed in GCB 30 scaffold as compared to GCB 0 scaffold suggesting the fact that 58S bioglass nanoparticles addition into the scaffold promoted better cell adhesion, proliferation, and differentiation. Thus, the study showed that the developed composite scaffolds are potential candidates for regenerating damaged bone tissue.

  18. In vitro evaluation of chitosan/poly(lactic acid-glycolic acid) sintered microsphere scaffolds for bone tissue engineering.

    Science.gov (United States)

    Jiang, Tao; Abdel-Fattah, Wafa I; Laurencin, Cato T

    2006-10-01

    A three-dimensional (3-D) scaffold is one of the major components in many tissue engineering approaches. We developed novel 3-D chitosan/poly(lactic acid-glycolic acid) (PLAGA) composite porous scaffolds by sintering together composite chitosan/PLAGA microspheres for bone tissue engineering applications. Pore sizes, pore volume, and mechanical properties of the scaffolds can be manipulated by controlling fabrication parameters, including sintering temperature and sintering time. The sintered microsphere scaffolds had a total pore volume between 28% and 37% with median pore size in the range 170-200microm. The compressive modulus and compressive strength of the scaffolds are in the range of trabecular bone making them suitable as scaffolds for load-bearing bone tissue engineering. In addition, MC3T3-E1 osteoblast-like cells proliferated well on the composite scaffolds as compared to PLAGA scaffolds. It was also shown that the presence of chitosan on microsphere surfaces increased the alkaline phosphatase activity of the cells cultured on the composite scaffolds and up-regulated gene expression of alkaline phosphatase, osteopontin, and bone sialoprotein.

  19. Bone repair by periodontal ligament stem cell-seeded nanohydroxyapatite-chitosan scaffold

    Directory of Open Access Journals (Sweden)

    Ge S

    2012-10-01

    Full Text Available Shaohua Ge,1 Ning Zhao,1 Lu Wang,1 Meijiao Yu,1 Hong Liu,2 Aimei Song,1 Jing Huang,1 Guancong Wang,2 Pishan Yang11Key Laboratory of Oral Biomedicine of Shandong Province, Department of Periodontology, School of Stomatology, 2Center of Bio and Micro/Nano Functional Materials, State Key Laboratory of Crystal Materials, Shandong University, Jinan, ChinaBackground: A nanohydroxyapatite-coated chitosan scaffold has been developed in recent years, but the effect of this composite scaffold on the viability and differentiation of periodontal ligament stem cells (PDLSCs and bone repair is still unknown. This study explored the behavior of PDLSCs on a new nanohydroxyapatite-coated genipin-chitosan conjunction scaffold (HGCCS in vitro as compared with an uncoated genipin-chitosan framework, and evaluated the effect of PDLSC-seeded HGCCS on bone repair in vivo.Methods: Human PDLSCs were cultured and identified, seeded on a HGCCS and on a genipin-chitosan framework, and assessed by scanning electron microscopy, confocal laser scanning microscopy, MTT, alkaline phosphatase activity, and quantitative real-time polymerase chain reaction at different time intervals. Moreover, PDLSC-seeded scaffolds were used in a rat calvarial defect model, and new bone formation was assessed by hematoxylin and eosin staining at 12 weeks postoperatively.Results: PDLSCs were clonogenic and positive for STRO-1. They had the capacity to undergo osteogenic and adipogenic differentiation in vitro. When seeded on HGCCS, PDLSCs exhibited significantly greater viability, alkaline phosphatase activity, and upregulated the bone-related markers, bone sialoprotein, osteopontin, and osteocalcin to a greater extent compared with PDLSCs seeded on the genipin-chitosan framework. The use of PDLSC-seeded HGCCS promoted calvarial bone repair.Conclusion: This study demonstrates the potential of HGCCS combined with PDLSCs as a promising tool for bone regeneration.Keywords: periodontal ligament, stem

  20. Chitosan/poly(epsilon-caprolactone) blend scaffolds for cartilage repair

    NARCIS (Netherlands)

    Neves, Sara C.; Moreira Teixeira, Liliana; Moroni, Lorenzo; Reis, Rui L.; van Blitterswijk, Clemens; Alves, Natália M.; Karperien, Hermanus Bernardus Johannes; Mano, João F.

    2011-01-01

    Chitosan (CHT)/poly(ɛ-caprolactone) (PCL) blend 3D fiber-mesh scaffolds were studied as possible support structures for articular cartilage tissue (ACT) repair. Micro-fibers were obtained by wet-spinning of three different polymeric solutions: 100:0 (100CHT), 75:25 (75CHT) and 50:50 (50CHT) wt.%

  1. Incorporation of chitosan microspheres into collagen-chitosan scaffolds for the controlled release of nerve growth factor.

    Directory of Open Access Journals (Sweden)

    Wen Zeng

    Full Text Available Artifical nerve scaffold can be used as a promising alternative to autologous nerve grafts to enhance the repair of peripheral nerve defects. However, current nerve scaffolds lack efficient microstructure and neurotrophic support.Microsphere-Scaffold composite was developed by incorporating chitosan microspheres loaded with nerve growth factor (NGF-CMSs into collagen-chitosan scaffolds (CCH with longitudinally oriented microchannels (NGF-CMSs/CCH. The morphological characterizations, in vitro release kinetics study, neurite outgrowth assay, and bioactivity assay were evaluated. After that, a 15-mm-long sciatic nerve gap in rats was bridged by the NGF-CMSs/CCH, CCH physically absorbed NGF (NGF/CCH, CCH or nerve autograft. 16 weeks after implantation, electrophysiology, fluoro-gold retrograde tracing, and nerve morphometry were performed.The NGF-CMSs were evenly distributed throughout the longitudinally oriented microchannels of the scaffold. The NGF-CMSs/CCH was capable of sustained release of bioactive NGF within 28 days as compared with others in vitro. In vivo animal study demonstrated that the outcomes of NGF-CMSs/CCH were better than those of NGF/CCH or CCH.Our findings suggest that incorporation of NGF-CMSs into the CCH may be a promising tool in the repair of peripheral nerve defects.

  2. Collagen-chitosan scaffold - Lauric acid plasticizer for skin tissue engineering on burn cases

    Science.gov (United States)

    Widiyanti, Prihartini; Setyadi, Ewing Dian; Rudyardjo, Djony Izak

    2017-02-01

    The prevalence of burns in the world is more than 800 cases per one million people each year and this is the second highest cause of death due to trauma after traffic accident. Many studies are turning to skin substitute methods of tissue engineering. The purpose of this study is to determine the composition of the collagen, chitosan, and lauric acid scaffold, as well as knowing the results of the characterization of the scaffold. The synthesis of chitosan collagen lauric acid scaffold as a skin tissue was engineered using freeze dried method. Results from making of collagen chitosan lauric acid scaffold was characterized physically, biologically and mechanically by SEM, cytotoxicity, biodegradation, and tensile strength. From the morphology test, the result obtained is that pore diameter size ranges from 94.11 to 140.1 µm for samples A,B,C,D, which are in the range of normal pore size 63-150 µm, while sample E has value below the standard which is about 37.87 to 47.36 µm. From cytotoxicity assay, the result obtained is the percentage value of living cells between 20.11 to 21.51%. This value is below 50% the standard value of living cells. Incompatibility is made possible because of human error mainly the replication of washing process over the standard. Degradation testing obtained values of 19.44% - 40% by weight which are degraded during the 7 days of observation. Tensile test results obtained a range of values of 0.192 - 3.53 MPa. Only sample A (3.53 MPa) and B (1.935 MPa) meet the standard values of skin tissue scaffold that is 1-24 MPa. Based on the results of the characteristics of this study, composite chitosan collagen scaffold with lauric acid plasticizer has a potential candidate for skin tissue engineering for skin burns cases.

  3. Preparation and Evaluation of Gelatin-Chitosan-Nanobioglass 3D Porous Scaffold for Bone Tissue Engineering

    OpenAIRE

    Maji, Kanchan; Dasgupta, Sudip; Pramanik, Krishna; Bissoyi, Akalabya

    2016-01-01

    The aim of the present study was to prepare and characterize bioglass-natural biopolymer based composite scaffold and evaluate its bone regeneration ability. Bioactive glass nanoparticles (58S) in the size range of 20?30?nm were synthesized using sol-gel method. Porous scaffolds with varying bioglass composition from 10 to 30?wt% in chitosan, gelatin matrix were fabricated using the method of freeze drying of its slurry at 40?wt% solids loading. Samples were cross-linked with glutaraldehyde t...

  4. Injectable porous nano-hydroxyapatite/chitosan/tripolyphosphate scaffolds with improved compressive strength for bone regeneration

    Energy Technology Data Exchange (ETDEWEB)

    Uswatta, Suren P.; Okeke, Israel U. [Department of Bioengineering, The University of Toledo, Toledo, OH 43614 (United States); Jayasuriya, Ambalangodage C., E-mail: a.jayasuriya@utoledo.edu [Department of Bioengineering, The University of Toledo, Toledo, OH 43614 (United States); Department of Orthopaedic Surgery, The University of Toledo, Toledo, OH 43614 (United States)

    2016-12-01

    In this study we have fabricated porous injectable spherical scaffolds using chitosan biopolymer, sodium tripolyphosphate (TPP) and nano-hydroxyapatite (nHA). TPP was primarily used as an ionic crosslinker to crosslink nHA/chitosan droplets. We hypothesized that incorporating nHA into chitosan could support osteoconduction by emulating the mineralized cortical bone structure, and improve the Ultimate Compressive Strength (UCS) of the scaffolds. We prepared chitosan solutions with 0.5%, 1% and 2% (w/v) nHA concentration and used simple coacervation and lyophilization techniques to obtain spherical scaffolds. Lyophilized spherical scaffolds had a mean diameter of 1.33 mm (n = 25). Further, portion from each group lyophilized scaffolds were soaked and dried to obtain Lyophilized Soaked and Dried (LSD) scaffolds. LSD scaffolds had a mean diameter of 0.93 mm (n = 25) which is promising property for the injectability. Scanning Electron Microscopy images showed porous surface morphology and interconnected pore structures inside the scaffolds. Lyophilized and LSD scaffolds had surface pores < 10 and 2 μm, respectively. 2% nHA/chitosan LSD scaffolds exhibited UCS of 8.59 MPa compared to UCS of 2% nHA/chitosan lyophilized scaffolds at 3.93 MPa. Standardize UCS values were 79.98 MPa and 357 MPa for 2% nHA/chitosan lyophilized and LSD particles respectively. One-way ANOVA results showed a significant increase (p < 0.001) in UCS of 1% and 2% nHA/chitosan lyophilized scaffolds compared to 0% and 0.5% nHA/chitosan lyophilized scaffolds. Moreover, 2% nHA LSD scaffolds had significantly increased (p < 0.005) their mean UCS by 120% compared to 2% nHA lyophilized scaffolds. In a drawback, all scaffolds have lost their mechanical properties by 95% on the 2nd day when fully immersed in phosphate buffered saline. Additionally live and dead cell assay showed no cytotoxicity and excellent osteoblast attachment to both lyophilized and LSD scaffolds at the end of 14th day of in vitro

  5. Injectable porous nano-hydroxyapatite/chitosan/tripolyphosphate scaffolds with improved compressive strength for bone regeneration

    International Nuclear Information System (INIS)

    Uswatta, Suren P.; Okeke, Israel U.; Jayasuriya, Ambalangodage C.

    2016-01-01

    In this study we have fabricated porous injectable spherical scaffolds using chitosan biopolymer, sodium tripolyphosphate (TPP) and nano-hydroxyapatite (nHA). TPP was primarily used as an ionic crosslinker to crosslink nHA/chitosan droplets. We hypothesized that incorporating nHA into chitosan could support osteoconduction by emulating the mineralized cortical bone structure, and improve the Ultimate Compressive Strength (UCS) of the scaffolds. We prepared chitosan solutions with 0.5%, 1% and 2% (w/v) nHA concentration and used simple coacervation and lyophilization techniques to obtain spherical scaffolds. Lyophilized spherical scaffolds had a mean diameter of 1.33 mm (n = 25). Further, portion from each group lyophilized scaffolds were soaked and dried to obtain Lyophilized Soaked and Dried (LSD) scaffolds. LSD scaffolds had a mean diameter of 0.93 mm (n = 25) which is promising property for the injectability. Scanning Electron Microscopy images showed porous surface morphology and interconnected pore structures inside the scaffolds. Lyophilized and LSD scaffolds had surface pores < 10 and 2 μm, respectively. 2% nHA/chitosan LSD scaffolds exhibited UCS of 8.59 MPa compared to UCS of 2% nHA/chitosan lyophilized scaffolds at 3.93 MPa. Standardize UCS values were 79.98 MPa and 357 MPa for 2% nHA/chitosan lyophilized and LSD particles respectively. One-way ANOVA results showed a significant increase (p < 0.001) in UCS of 1% and 2% nHA/chitosan lyophilized scaffolds compared to 0% and 0.5% nHA/chitosan lyophilized scaffolds. Moreover, 2% nHA LSD scaffolds had significantly increased (p < 0.005) their mean UCS by 120% compared to 2% nHA lyophilized scaffolds. In a drawback, all scaffolds have lost their mechanical properties by 95% on the 2nd day when fully immersed in phosphate buffered saline. Additionally live and dead cell assay showed no cytotoxicity and excellent osteoblast attachment to both lyophilized and LSD scaffolds at the end of 14th day of in vitro

  6. Electrospun biomimetic scaffold of hydroxyapatite/chitosan supports enhanced osteogenic differentiation of mMSCs

    International Nuclear Information System (INIS)

    Peng Hongju; Feng Bei; Yuan Huihua; Zhang Yanzhong; Yin Zi; Liu Huanhuan; Chen Xiao; Ouyang Hongwei; Su Bo

    2012-01-01

    Engaging functional biomaterial scaffolds to regulate stem cell differentiation has drawn a great deal of attention in the tissue engineering and regenerative medicine community. In this study, biomimetic composite nanofibrous scaffolds of hydroxyapatite/chitosan (HAp/CTS) were prepared to investigate their capacity for inducing murine mesenchymal stem cells (mMSCs) to differentiate into the osteogenic lineage, in the absence and presence of an osteogenic supplementation (i.e., ascorbic acid, β-glycerol phosphate, and dexamethasone), respectively. Using electrospun chitosan (CTS) nanofibrous scaffolds as the control, cell morphology, growth, specific osteogenic genes expression, and quantified proteins secretion on the HAp/CTS scaffolds were sequentially examined and assessed. It appeared that the HAp/CTS scaffolds supported better attachment and proliferation of the mMSCs. Most noteworthy was that in the absence of the osteogenic supplementation, expression of osteogenic genes including collagen I (Col I), runt-related transcription factor 2 (Runx2), alkaline phosphatase (ALP), and osteocalcin (OCN) were significantly upregulated in mMSCs cultured on the HAp/CTS nanofibrous scaffolds. Also increased secretion of the osteogenesis protein markers of alkaline phosphatase and collagen confirmed that the HAp/CTS nanofibrous scaffold markedly promoted the osteogenic commitment in the mMSCs. Moreover, the presence of osteogenic supplementation proved an enhanced efficacy of mMSC osteogenesis on the HAp/CTS nanofibrous scaffolds. Collectively, this study demonstrated that the biomimetic nanofibrous HAp/CTS scaffolds could support and enhance the adhesion, proliferation, and particularly osteogenic differentiation of the mMSCs. It also substantiated the potential of using biomimetic nanofibrous scaffolds of HAp/CTS for functional bone repair and regeneration applications. (paper)

  7. Processing and characterization of chitosan/PVA and methylcellulose porous scaffolds for tissue engineering

    International Nuclear Information System (INIS)

    Kanimozhi, K.; Khaleel Basha, S.; Sugantha Kumari, V.

    2016-01-01

    Biomimetic porous scaffold chitosan/poly(vinyl alcohol) CS/PVA containing various amounts of methylcellulose (MC) (25%, 50% and 75%) incorporated in CS/PVA blend was successfully produced by a freeze drying method in the present study. The composite porous scaffold membranes were characterized by infrared spectroscopy (FTIR), X-ray diffraction (XRD), thermogravimetric analysis (TGA), differential scanning calorimetry (DSC), scanning electron microscopy (SEM), swelling degree, porosity, degradation of films in Hank's solution and the mechanical properties. Besides these characterizations, the antibacterial activity of the prepared scaffolds was tested, toward the bacterial species Staphylococcus aureus (S. aureus) and Escherichia coli (E. coli). FTIR, XRD and DSC demonstrated that there was strong intermolecular hydrogen bonding between the molecules of CS/PVA and MC. The crystalline microstructure of the scaffold membranes was not well developed. SEM images showed that the morphology and diameter of the scaffolds were mainly affected by the weight ratio of MC. By increasing the MC content in the hybrid scaffolds, their swelling capacity and porosity increased. The mechanical properties of these scaffolds in dry and swollen state were greatly improved with high swelling ratio. The elasticity of films was also significantly improved by the incorporation of MC, and the scaffolds could also bear a relative high tensile strength. These findings suggested that the developed scaffold possess the prerequisites and can be used as a scaffold for tissue engineering. - Highlights: • The porous scaffolds of CS/PVA containing different MC contents were fabricated. • Addition of MC improved the compatibility between CS and PVA. • The mechanical properties of these scaffolds were greatly improved with high swelling ratio. • Biocompatibility test showed that the different MC content scaffolds had no cytotoxicity.

  8. Processing and characterization of chitosan/PVA and methylcellulose porous scaffolds for tissue engineering

    Energy Technology Data Exchange (ETDEWEB)

    Kanimozhi, K. [Department of Chemistry, Auxilium College, Vellore 632 006 (India); Khaleel Basha, S. [Department of Biochemistry, C. Abdul Hakeem College, Melvisharam 632 509 (India); Sugantha Kumari, V., E-mail: sheenasahana04@gmail.com [Department of Chemistry, Auxilium College, Vellore 632 006 (India)

    2016-04-01

    Biomimetic porous scaffold chitosan/poly(vinyl alcohol) CS/PVA containing various amounts of methylcellulose (MC) (25%, 50% and 75%) incorporated in CS/PVA blend was successfully produced by a freeze drying method in the present study. The composite porous scaffold membranes were characterized by infrared spectroscopy (FTIR), X-ray diffraction (XRD), thermogravimetric analysis (TGA), differential scanning calorimetry (DSC), scanning electron microscopy (SEM), swelling degree, porosity, degradation of films in Hank's solution and the mechanical properties. Besides these characterizations, the antibacterial activity of the prepared scaffolds was tested, toward the bacterial species Staphylococcus aureus (S. aureus) and Escherichia coli (E. coli). FTIR, XRD and DSC demonstrated that there was strong intermolecular hydrogen bonding between the molecules of CS/PVA and MC. The crystalline microstructure of the scaffold membranes was not well developed. SEM images showed that the morphology and diameter of the scaffolds were mainly affected by the weight ratio of MC. By increasing the MC content in the hybrid scaffolds, their swelling capacity and porosity increased. The mechanical properties of these scaffolds in dry and swollen state were greatly improved with high swelling ratio. The elasticity of films was also significantly improved by the incorporation of MC, and the scaffolds could also bear a relative high tensile strength. These findings suggested that the developed scaffold possess the prerequisites and can be used as a scaffold for tissue engineering. - Highlights: • The porous scaffolds of CS/PVA containing different MC contents were fabricated. • Addition of MC improved the compatibility between CS and PVA. • The mechanical properties of these scaffolds were greatly improved with high swelling ratio. • Biocompatibility test showed that the different MC content scaffolds had no cytotoxicity.

  9. Improving effects of chitosan nanofiber scaffolds on osteoblast proliferation and maturation

    Science.gov (United States)

    Ho, Ming-Hua; Liao, Mei-Hsiu; Lin, Yi-Ling; Lai, Chien-Hao; Lin, Pei-I; Chen, Ruei-Ming

    2014-01-01

    Osteoblast maturation plays a key role in regulating osteogenesis. Electrospun nanofibrous products were reported to possess a high surface area and porosity. In this study, we developed chitosan nanofibers and examined the effects of nanofibrous scaffolds on osteoblast maturation and the possible mechanisms. Macro- and micro observations of the chitosan nanofibers revealed that these nanoproducts had a flat surface and well-distributed fibers with nanoscale diameters. Mouse osteoblasts were able to attach onto the chitosan nanofiber scaffolds, and the scaffolds degraded in a time-dependent manner. Analysis by scanning electron microscopy further showed mouse osteoblasts adhered onto the scaffolds along the nanofibers, and cell–cell communication was also detected. Mouse osteoblasts grew much better on chitosan nanofiber scaffolds than on chitosan films. In addition, human osteoblasts were able to adhere and grow on the chitosan nanofiber scaffolds. Interestingly, culturing human osteoblasts on chitosan nanofiber scaffolds time-dependently increased DNA replication and cell proliferation. In parallel, administration of human osteoblasts onto chitosan nanofibers significantly induced osteopontin, osteocalcin, and alkaline phosphatase (ALP) messenger (m)RNA expression. As to the mechanism, chitosan nanofibers triggered runt-related transcription factor 2 mRNA and protein syntheses. Consequently, results of ALP-, alizarin red-, and von Kossa-staining analyses showed that chitosan nanofibers improved osteoblast mineralization. Taken together, results of this study demonstrate that chitosan nanofibers can stimulate osteoblast proliferation and maturation via runt-related transcription factor 2-mediated regulation of osteoblast-associated osteopontin, osteocalcin, and ALP gene expression. PMID:25246786

  10. Characterization and Cell Culture of a Grafted Chitosan Scaffold for Tissue Engineering

    Directory of Open Access Journals (Sweden)

    Wen-Chuan Hsieh

    2015-01-01

    Full Text Available Poly(vinyl alcohol (PVA was grafted to chitosan to form a porous scaffold. The PVA-g-chitosan 3D scaffold was then observed by Fourier transform infrared spectroscopy (FT-IR. The water absorbency of PVA-g-chitosan was increased 370% by grafting. Scanning electron microscope (SEM observations of the material revealed that the 3D scaffold is highly porous when formed using a homogenizer at 300 rpm. Compression testing demonstrated that as the amount of chitosan increases, the strength of the 3D scaffold strength reached showed that, by increasing the amount of chitosan, the strength of the 3D scaffold could be increased to 16 × 10−1 MPa. Over 35 days of enzymatic degradation, the 3D scaffold was degraded by various enzymes at rates of up to 10%. In vitro tests showed good cell proliferation and growth in the 3D scaffold.

  11. A healing method of tympanic membrane perforations using three-dimensional porous chitosan scaffolds.

    Science.gov (United States)

    Kim, Jangho; Kim, Seung Won; Choi, Seong Jun; Lim, Ki Taek; Lee, Jong Bin; Seonwoo, Hoon; Choung, Pill-Hoon; Park, Keehyun; Cho, Chong-Su; Choung, Yun-Hoon; Chung, Jong Hoon

    2011-11-01

    Both surgical tympanoplasty and paper patch grafts are frequently procedured to heal tympanic membrane (TM) perforation or chronic otitis media, despite their many disadvantages. In this study, we report a new healing method of TM perforation by using three-dimensional (3D) porous chitosan scaffolds (3D chitosan scaffolds) as an alternative method to surgical treatment or paper patch graft. Various 3D chitosan scaffolds were prepared; and the structural characteristics, mechanical property, in vitro biocompatibility, and healing effects of the 3D chitosan scaffolds as an artificial TM in in vivo animal studies were investigated. A 3D chitosan scaffold of 5 wt.% chitosan concentration showed good proliferation of TM cells in an in vitro study, as well as suitable structural characteristics and mechanical property, as compared with either 1% or 3% chitosan. In in vivo animal studies, 3D chitosan scaffold were able to migrate through the pores and surfaces of TM cells, thus leading to more effective TM regeneration than paper patch technique. Histological observations demonstrated that the regenerated TM with the 3D chitosan scaffold consisted of three (epidermal, connective tissue, and mucosal) layers and were thicker than normal TMs. The 3D chitosan scaffold technique may be an optimal healing method used in lieu of surgical tympanoplasty in certain cases to heal perforated TMs.

  12. Characterization of collagen / chitosan films for skin regenerating scaffold

    International Nuclear Information System (INIS)

    Ismarul, I.N.; Ishak, Y.; Ismail, Z.; Mohd Shalihuddin, W.M.

    2004-01-01

    Various Proportions of chitosan/collagen films (70/30% to 95/05%) w/w were prepared and evaluated for its suitability as skin regenerating scaffold. Interactions between chitosan and collagen were studied using Fourier Transform Infrared spectroscopy (FTIR) and Differential Scanning Colorimetry (DSC). Scanning Electron Microscope (SEM) was used to investigate the morphology of the blend. Mechanical properties were evaluated using a Universal Testing Machine (UTM). The chitosan/collagen films were found to swell proportionally with time until it reaches equilibrium, FTIR spectroscopy indicated no chemical interaction between the components of the blends, DSC data indicated only one peak proving that these two materials are compatible at all proportions investigated. SEM micrographs also indicated good homogeneity between these two materials. (Author)

  13. Soft chitosan microbeads scaffold for 3D functional neuronal networks.

    Science.gov (United States)

    Tedesco, Maria Teresa; Di Lisa, Donatella; Massobrio, Paolo; Colistra, Nicolò; Pesce, Mattia; Catelani, Tiziano; Dellacasa, Elena; Raiteri, Roberto; Martinoia, Sergio; Pastorino, Laura

    2018-02-01

    The availability of 3D biomimetic in vitro neuronal networks of mammalian neurons represents a pivotal step for the development of brain-on-a-chip experimental models to study neuronal (dys)functions and particularly neuronal connectivity. The use of hydrogel-based scaffolds for 3D cell cultures has been extensively studied in the last years. However, limited work on biomimetic 3D neuronal cultures has been carried out to date. In this respect, here we investigated the use of a widely popular polysaccharide, chitosan (CHI), for the fabrication of a microbead based 3D scaffold to be coupled to primary neuronal cells. CHI microbeads were characterized by optical and atomic force microscopies. The cell/scaffold interaction was deeply characterized by transmission electron microscopy and by immunocytochemistry using confocal microscopy. Finally, a preliminary electrophysiological characterization by micro-electrode arrays was carried out. Copyright © 2017 Elsevier Ltd. All rights reserved.

  14. Preparation, characterization and biological test of 3D-scaffolds based on chitosan, fibroin and hydroxyapatite for bone tissue engineering

    International Nuclear Information System (INIS)

    Lima, Paulo Autran Leite; Resende, Cristiane Xavier; Dulce de Almeida Soares, Glória; Anselme, Karine; Almeida, Luís Eduardo

    2013-01-01

    This work describes the preparation and characterization of porous 3D-scaffolds based on chitosan (CHI), chitosan/silk fibroin (CHI/SF) and chitosan/silk fibroin/hydroxyapatite (CHI/SF/HA) by freeze drying. The biomaterials were characterized by X-ray diffraction, attenuated total reflection Fourier transform infrared spectroscopy, thermogravimetric analysis, differential scanning calorimetry, scanning electron microscopy and energy dispersive spectroscopy. In addition, studies of porosity, pore size, contact angle and biological response of SaOs-2osteoblastic cells were performed. The CHI scaffolds have a porosity of 94.2 ± 0.9%, which is statistically higher than the one presented by CHI/SF/HA scaffolds, 89.7 ± 2.6%. Although all scaffolds were able to promote adhesion, growth and maintenance of osteogenic differentiation of SaOs-2 cells, the new 3D-scaffold based on CHI/SF/HA showed a significantly higher cell growth at 7 days and 21 days and the level of alkaline phosphatase at 14 and 21 days was statistically superior compared to other tested materials. - Highlights: • Preparation of 3D-scaffolds based on CHI, with or without addition of SF and HA. • Scaffolds exhibited interconnected porous structure (pore size superior to 50 μm). • The tripolyphosphate did not induce any significant cytotoxic response. • The CHI/SF/HA composite showed a higher cell growth and ALP activity

  15. Heavy Metal Removal by Chitosan and Chitosan Composite

    International Nuclear Information System (INIS)

    Abdel-Mohdy, F.A.; El-Sawy, S.; Ibrahim, M.S.

    2005-01-01

    Radiation grafting of diethyl aminoethyl methacrylate (DEAEMA) on chitosan to impart ion exchange properties and to be used for the separation of metal ions from waste water, was carried out. The effect of experimental conditions such as monomer concentration and the radiation dose on grafting were studied. On using chitosan, grafted chitosan and some chitosan composites in metal ion removal they show high up-take capacity for Cu 2+ and lower uptake capacities for the other divalent metal ions used (Zn and Co). Competitive study, performed with solutions containing mixture of metal salts, showed high selectivity for Cu 2+ than the other metal ion. Limited grafting of DEAEMA polymer -containing specific functional groups-onto the chitosan backbone improves the sorption performance

  16. Effects of Chitosan Alkali Pretreatment on the Preparation of Electrospun PCL/Chitosan Blend Nanofibrous Scaffolds for Tissue Engineering Application

    Directory of Open Access Journals (Sweden)

    Fatemeh Roozbahani

    2013-01-01

    Full Text Available Recently, nanofibrous scaffolds have been used in the field of biomedical engineering as wound dressings, tissue engineering scaffolds, and drug delivery applications. The electrospun nanofibrous scaffolds can be used as carriers for several types of drugs, genes, and growth factors. PCL is one of the most commonly applied synthetic polymers for medical use because of its biocompatibility and slow biodegradability. PCL is hydrophobic and has no cell recognition sites on its structure. Electrospinning of chitosan and PCL blend was investigated in formic acid/acetic acid as the solvent with different PCL/chitosan ratios. High viscosity of chitosan solutions makes difficulties in the electrospinning process. Strong hydrogen bonds in a 3D network in acidic condition prevent the movement of polymeric chains exposed to the electrical field. Consequently, the amount of chitosan in PCL/chitosan blend was limited and more challenging when the concentration of PCL increases. The treatment of chitosan in alkali condition under high temperature reduced its molecular weight. Longer treatment time further decreased the molecular weight of chitosan and hence its viscosity. Electrospinning of PCL/chitosan blend was possible at higher chitosan ratio, and SEM images showed a decrease in fiber diameter and narrower distribution with increase in the chitosan ratio.

  17. Characterization and Cell Culture of a Grafted Chitosan Scaffold for Tissue Engineering

    OpenAIRE

    Hsieh, Wen-Chuan; Liau, Jiun-Jia; Li, Yi-Jhong

    2015-01-01

    Poly(vinyl alcohol) (PVA) was grafted to chitosan to form a porous scaffold. The PVA-g-chitosan 3D scaffold was then observed by Fourier transform infrared spectroscopy (FT-IR). The water absorbency of PVA-g-chitosan was increased 370% by grafting. Scanning electron microscope (SEM) observations of the material revealed that the 3D scaffold is highly porous when formed using a homogenizer at 300 rpm. Compression testing demonstrated that as the amount of chitosan increases, the strength of t...

  18. Anti-biofouling 3D porous systems: the blend effect of oxazoline-based oligomers on chitosan scaffolds.

    Science.gov (United States)

    Correia, Vanessa G; Coelho, Margarida; Barroso, Telma; Raje, Vivek P; Bonifácio, Vasco D B; Casimiro, Teresa; Pinho, Mariana G; Aguiar-Ricardo, Ana

    2013-01-01

    The production, characterization and anti-biofouling activity of 3D porous scaffolds combining different blends of chitosan and oxazoline-based antimicrobial oligomers is reported. The incorporation of ammonium quaternized oligo(2-oxazoline)s into the composition of the scaffold enhances the stability of the chitosan scaffold under physiological conditions as well as its ability to repel protein adsorption. The blended scaffolds showed mean pore sizes in the range of 18-32 μm, a good pore interconnectivity and high porosity, as well as a large surface area, ultimate key features for anti-biofouling applications. Bovine serum albumin (BSA) adhesion profiles showed that the composition of the scaffolds plays a critical role in the chitosan-oligooxazoline system. Oligobisoxazoline-enriched scaffolds (20% w/w, CB8020) decreased protein adsorption (BSA) by up to 70%. Moreover, 1 mg of CB8020 was able to kill 99.9% of Escherichia coli cells upon contact, demonstrating its potential as promising material for production of tailored non-fouling 3D structures to be used in the construction of novel devices with applications in the biomedical field and water treatment processes.

  19. Effect of strontium addition and chitosan concentration variation on cytotoxicity of chitosan-alginate-carbonate apatite based bone scaffold

    Science.gov (United States)

    Perkasa, Rilis Eka; Umniati, B. Sri; Sunendar, Bambang

    2017-09-01

    Bone scaffold is one of the most important component in bone tissue engineering. Basically, bone scaffold is a biocompatible structure designed to replace broken bone tissue temporarily. Unlike conventional bone replacements, an advanced bone scaffold should be bioactive (e.g: supporting bone growth) and biodegradable as new bone tissue grow, while retain its mechanical properties similarity with bone. It is also possible to add more bioactive substrates to bone scaffold to further support its performance. One of the substrate is strontium, an element that could improve the ability of the bone to repair itself. However, it must be noted that excessive consumption of strontium could lead to toxicity and diseases, such as osteomalacia and hypocalcemia. This research aimed to investigate the effect of strontium addition to the cytotoxic property of chitosan-alginate-carbonate apatite bone scaffold. The amount of strontium added to the bone scaffold was 5% molar of the carbonate apatite content. As a control, bone scaffold without stronsium (0% molar) were also made. The effect of chitosan concentration variation on the cytotoxicity were also observed, where the concentration varies on 1% and 3% w/v of chitosan solution. The results showed an optimum result on bone scaffold sample with 5% molar of strontium and 3% chitosan, where 87.67% cells in the performed MTS-Assay cytotoxicity testing survived. This showed that the use of up to 5% molar addition of strontium and 3% chitosan could enhance the survivability of the cell.

  20. Chitosan scaffold as an alternative adsorbent for the removal of hazardous food dyes from aqueous solutions.

    Science.gov (United States)

    Esquerdo, V M; Cadaval, T R S; Dotto, G L; Pinto, L A A

    2014-06-15

    The dye adsorption with chitosan is considered an eco-friendly alternative technology in relation to the existing water treatment technologies. However, the application of chitosan for dyes removal is limited, due to its low surface area and porosity. Then we prepared a chitosan scaffold with a megaporous structure as an alternative adsorbent to remove food dyes from solutions. The chitosan scaffold was characterized by infrared spectroscopy, scanning electron microscopy and structural characteristics. The potential of chitosan scaffold to remove five food dyes from solutions was investigated by equilibrium isotherms and thermodynamic study. The scaffold-dyes interactions were elucidated, and desorption studies were carried out. The chitosan scaffold presented pore sizes from 50 to 200 μm, porosity of 92.2±1.2% and specific surface area of 1135±2 m(2) g(-1). The two-step Langmuir model was suitable to represent the equilibrium data. The adsorption was spontaneous, favorable, exothermic and enthalpy-controlled process. Electrostatic interactions occurred between chitosan scaffold and dyes. Desorption was possible with NaOH solution (0.10 mol L(-1)). The chitosan megaporous scaffold showed good structural characteristics and high adsorption capacities (788-3316 mg g(-1)). Copyright © 2014 Elsevier Inc. All rights reserved.

  1. Development of Chitosan Scaffolds with Enhanced Mechanical Properties for Intestinal Tissue Engineering Applications.

    Science.gov (United States)

    Zakhem, Elie; Bitar, Khalil N

    2015-10-13

    Massive resections of segments of the gastrointestinal (GI) tract lead to intestinal discontinuity. Functional tubular replacements are needed. Different scaffolds were designed for intestinal tissue engineering application. However, none of the studies have evaluated the mechanical properties of the scaffolds. We have previously shown the biocompatibility of chitosan as a natural material in intestinal tissue engineering. Our scaffolds demonstrated weak mechanical properties. In this study, we enhanced the mechanical strength of the scaffolds with the use of chitosan fibers. Chitosan fibers were circumferentially-aligned around the tubular chitosan scaffolds either from the luminal side or from the outer side or both. Tensile strength, tensile strain, and Young's modulus were significantly increased in the scaffolds with fibers when compared with scaffolds without fibers. Burst pressure was also increased. The biocompatibility of the scaffolds was maintained as demonstrated by the adhesion of smooth muscle cells around the different kinds of scaffolds. The chitosan scaffolds with fibers provided a better candidate for intestinal tissue engineering. The novelty of this study was in the design of the fibers in a specific alignment and their incorporation within the scaffolds.

  2. Characterization of gelatin/chitosan scaffold blended with aloe vera and snail mucus for biomedical purpose.

    Science.gov (United States)

    López Angulo, Daniel Enrique; do Amaral Sobral, Paulo José

    2016-11-01

    Biologically active scaffolds used in tissue engineering and regenerative medicine have been generating promising results in skin replacement. The present study aims to test the hypothesis that the incorporation of Aloe vera and snail mucus into scaffolds based on gelatin and chitosan could improve their structure, composition and biodegradability, with a potential effect on bioactivity. Homogeneous pore diameter as well as pore walls in the composite scaffold could be seen in the SEM image. The pores in the scaffolds were interconnected and their sizes ranged from 93 to 296μm. The addition of Aloe vera and snail mucus enlarged the mean pore size with increased porosity and caused changes in the pore architecture. The FTIR analysis has shown good affinity and interaction between the matrix and the Aloe, which may decrease water-binding sites, so this fact hindered the water absorption capacity of the material. The mechanical properties could explain the highest swelling capacity of the snail scaffold, because the high percentage of elongation could facilitate the entry of liquid in it, generating a matrix with plenty of fluid retention. The real innovation in the present work could be the use of these substances (Aloe and snail mucus) for tissue engineering. Copyright © 2016 Elsevier B.V. All rights reserved.

  3. Fabrication and Characteristics of Chitosan Sponge as a Tissue Engineering Scaffold

    Directory of Open Access Journals (Sweden)

    Takeshi Ikeda

    2014-01-01

    Full Text Available Cells, growth factors, and scaffolds are the three main factors required to create a tissue-engineered construct. After the appearance of bovine spongiform encephalopathy (BSE, considerable attention has therefore been focused on nonbovine materials. In this study, we examined the properties of a chitosan porous scaffold. A porous chitosan sponge was prepared by the controlled freezing and lyophilization of different concentrations of chitosan solutions. The materials were examined by scanning electron microscopy, and the porosity, tensile strength, and basic fibroblast growth factor (bFGF release profiles from chitosan sponge were examined in vitro. The morphology of the chitosan scaffolds presented a typical microporous structure, with the pore size ranging from 50 to 200 μm. The porosity of chitosan scaffolds with different concentrations was approximately 75–85%. A decreasing tendency for porosity was observed as the concentration of the chitosan increased. The relationship between the tensile properties and chitosan concentration indicated that the ultimate tensile strength for the sponge increased with a higher concentration. The in vitro bFGF release study showed that the higher the concentration of chitosan solution became, the longer the releasing time of the bFGF from the chitosan sponge was.

  4. Biomimetic mineral-organic composite scaffolds with controlled internal architecture.

    Science.gov (United States)

    Manjubala, I; Woesz, Alexander; Pilz, Christine; Rumpler, Monika; Fratzl-Zelman, Nadja; Roschger, Paul; Stampfl, Juergen; Fratzl, Peter

    2005-12-01

    Bone and cartilage generation by three-dimensional scaffolds is one of the promising techniques in tissue engineering. One approach is to generate histologically and functionally normal tissue by delivering healthy cells in biocompatible scaffolds. These scaffolds provide the necessary support for cells to proliferate and maintain their differentiated function, and their architecture defines the ultimate shape. Rapid prototyping (RP) is a technology by which a complex 3-dimensional (3D) structure can be produced indirectly from computer aided design (CAD). The present study aims at developing a 3D organic-inorganic composite scaffold with defined internal architecture by a RP method utilizing a 3D printer to produce wax molds. The composite scaffolds consisting of chitosan and hydroxyapatite were prepared using soluble wax molds. The behaviour and response of MC3T3-E1 pre-osteoblast cells on the scaffolds was studied. During a culture period of two and three weeks, cell proliferation and in-growth were observed by phase contrast light microscopy, histological staining and electron microscopy. The Giemsa and Gömöri staining of the cells cultured on scaffolds showed that the cells proliferated not only on the surface, but also filled the micro pores of the scaffolds and produced extracellular matrix within the pores. The electron micrographs showed that the cells covering the surface of the struts were flattened and grew from the periphery into the middle region of the pores.

  5. Fabrication of chitosan/gallic acid 3D microporous scaffold for tissue engineering applications.

    Science.gov (United States)

    Thangavel, Ponrasu; Ramachandran, Balaji; Muthuvijayan, Vignesh

    2016-05-01

    This study explores the potential of gallic acid incorporated chitosan (CS/GA) 3D scaffolds for tissue engineering applications. Scaffolds were prepared by freezing and lyophilization technique and characterized. FTIR spectra confirmed the presence of GA in chitosan (CS) gel. DSC and TGA analysis revealed that the structure of chitosan was not altered due to the incorporation of GA, but thermal stability was significantly increased compared to the CS scaffold. SEM micrographs showed smooth, homogeneous, and microporous architecture of the scaffolds with good interconnectivity. CS/GA scaffolds exhibited approximately 90% porosity on average, increased swelling (600-900%) and controlled biodegradation (15-40%) in PBS (pH 7.4 at 37°C) with 1 mg/mL of lysozyme. CS/GA scaffolds showed 2-4 fold decrease in CFUs (p < 0.05) for both gram positive and gram negative bacteria compared to the CS scaffold. Cytotoxicity of these scaffolds was evaluated using NIH 3T3 L1 fibroblast cells. CS/GA 0.25% scaffold showed similar viability with CS scaffold at 24 and 48 h. CS/GA scaffolds (0.5-1.0%) showed 60-75% viability at 24 h and 90% at 48 h. SEM images showed that an increased cell attachment was observed for CS/GA scaffolds compared to CS scaffolds. These findings authenticate that CS/GA scaffolds were cytocompatible and would be useful for tissue engineering applications. © 2015 Wiley Periodicals, Inc.

  6. Characterization of Silk Fibroin/Chitosan 3D Porous Scaffold and In Vitro Cytology.

    Directory of Open Access Journals (Sweden)

    Shuguang Zeng

    Full Text Available Bone tissue engineering is a powerful tool to treat bone defects caused by trauma, infection, tumors and other factors. Both silk fibroin (SF and chitosan (CS are non-toxic and have good biocompatibility, but are poor biological scaffolds when used alone. In this study, the microscopic structure and related properties of SF/CS composite scaffolds with different component ratios were examined. The scaffold material most suitable for osteoblast growth was determined, and these results offer an experimental basis for the future reconstruction of bone defects. First, via freeze-drying and chemical crosslinking methods, SF/CS composites with different component ratios were prepared and their structure was characterized. Changes in the internal structure of the SF and CS mixture were observed, confirming that the mutual modification between the two components was complete and stable. The internal structure of the composite material was porous and three-dimensional with a porosity above 90%. We next studied the pore size, swelling ratio, water absorption ratio, degradation and in vitro cell proliferation. For the 40% SF-60% CS group, the pore size of the scaffold was suitable for the growth of osteoblasts, and the rate of degradation was steady. This favors the early adhesion, growth and proliferation of MG-63 cells. In addition to good biocompatibility and satisfactory cell affinity, this material promotes the secretion of extracellular matrix materials by osteoblasts. Thus, 40% SF-60% CS is a good material for bone tissue engineering.

  7. Characterization of Silk Fibroin/Chitosan 3D Porous Scaffold and In Vitro Cytology.

    Science.gov (United States)

    Zeng, Shuguang; Liu, Lei; Shi, Yong; Qiu, Junqi; Fang, Wei; Rong, Mingdeng; Guo, Zehong; Gao, Wenfeng

    2015-01-01

    Bone tissue engineering is a powerful tool to treat bone defects caused by trauma, infection, tumors and other factors. Both silk fibroin (SF) and chitosan (CS) are non-toxic and have good biocompatibility, but are poor biological scaffolds when used alone. In this study, the microscopic structure and related properties of SF/CS composite scaffolds with different component ratios were examined. The scaffold material most suitable for osteoblast growth was determined, and these results offer an experimental basis for the future reconstruction of bone defects. First, via freeze-drying and chemical crosslinking methods, SF/CS composites with different component ratios were prepared and their structure was characterized. Changes in the internal structure of the SF and CS mixture were observed, confirming that the mutual modification between the two components was complete and stable. The internal structure of the composite material was porous and three-dimensional with a porosity above 90%. We next studied the pore size, swelling ratio, water absorption ratio, degradation and in vitro cell proliferation. For the 40% SF-60% CS group, the pore size of the scaffold was suitable for the growth of osteoblasts, and the rate of degradation was steady. This favors the early adhesion, growth and proliferation of MG-63 cells. In addition to good biocompatibility and satisfactory cell affinity, this material promotes the secretion of extracellular matrix materials by osteoblasts. Thus, 40% SF-60% CS is a good material for bone tissue engineering.

  8. Morphology and characterization of 3D micro-porous structured chitosan scaffolds for tissue engineering.

    Science.gov (United States)

    Hsieh, Wen-Chuan; Chang, Chih-Pong; Lin, Shang-Ming

    2007-06-15

    This research studies the morphology and characterization of three-dimensional (3D) micro-porous structures produced from biodegradable chitosan for use as scaffolds for cells culture. The chitosan 3D micro-porous structures were produced by a simple liquid hardening method, which includes the processes of foaming by mechanical stirring without any chemical foaming agent added, and hardening by NaOH cross linking. The pore size and porosity were controlled with mechanical stirring strength. This study includes the morphology of chitosan scaffolds, the characterization of mechanical properties, water absorption properties and in vitro enzymatic degradation of the 3D micro-porous structures. The results show that chitosan 3D micro-porous structures were successfully produced. Better formation samples were obtained when chitosan concentration is at 1-3%, and concentration of NaOH is at 5%. Faster stirring rate would produce samples of smaller pore diameter, but when rotation speed reaches 4000 rpm and higher the changes in pore size is minimal. Water absorption would reduce along with the decrease of chitosan scaffolds' pore diameter. From stress-strain analysis, chitosan scaffolds' mechanical properties are improved when it has smaller pore diameter. From in vitro enzymatic degradation results, it shows that the disintegration rate of chitosan scaffolds would increase along with the processing time increase, but approaching equilibrium when the disintegration rate reaches about 20%.

  9. Microwave-induced biomimetic approach for hydroxyapatite coatings of chitosan scaffolds.

    Science.gov (United States)

    Kaynak Bayrak, Gökçe; Demirtaş, T Tolga; Gümüşderelioğlu, Menemşe

    2017-02-10

    Simulated body fluid (SBF) can form calcium phosphates on osteoinductive materials, so it is widely used for coating of bone scaffolds to mimic natural extracellular matrix (ECM). However, difficulties of bulk coating in 3D scaffolds and the necessity of long process times are the common problems for coating with SBF. In the present study, a microwave-assisted process was developed for rapid and internal coating of chitosan scaffolds. The scaffolds were fabricated as superporous hydrogel (SPH) by combining microwave irradiation and gas foaming methods. Then, they were immersed into 10x  SBF-like solution and homogenous bone-like hydroxyapatite (HA) coating was achieved by microwave treatment at 600W without the need of any nucleating agent. Cell culture studies with MC3T3-E1 preosteoblasts showed that microwave-assisted biomimetic HA coating process could be evaluated as an efficient and rapid method to obtain composite scaffolds for bone tissue engineering. Copyright © 2016 Elsevier Ltd. All rights reserved.

  10. Design and Fabrication of Biodegradable Porous Chitosan/Gelatin/Tricalcium Phosphate Hybrid Scaffolds for Tissue Engineering

    Directory of Open Access Journals (Sweden)

    Y. Mohammadi

    2007-08-01

    Full Text Available In this study, based on a biomimetic approach, novel 3D biodegradable porous hybrid scaffolds consisting of chitosan, gelatin, and tricalcium phosphate were developed for bone and cartilage tissue engineering. Macroporous chitosan/ gelatin/β-TCP scaffolds were prepared through the process of freeze-gelation/solid-liquid phase separation. The results showed that the prepared scaffolds are highly porous, with porosities larger than 80%, and have interconnected pores. Biocompatibility studies were successfully performed by in vitro and in vivo assays. Moreover, the attachment, migration, and proliferation of chondrocytes on these unique temporary scaffolds were examined to determine their potentials in tissue engineering applications.

  11. A review on chitosan centred scaffolds and their applications in tissue engineering.

    Science.gov (United States)

    Ahmed, Shakeel; Annu; Sheikh, Javed; Ali, Akbar

    2018-05-03

    The diversity and availability of biopolymer and increased clinical demand for safe scaffolds lead to an increased interest in fabricating scaffolds in order to achieve fruitful progress in tissue engineering. Due to biocompatibility, biodegradability, inherent antimicrobial character, chitosan has drawn ample consideration in recent years. Chitosan is a biopolymer obtained by de-acetylation of chitin extracted from shells of crustaceans and fungi. Due to the presence of reactive functionality in the molecular chain chitosan can be modified either chemically or physically to fabricate the tailor-made scaffolds having desired properties for tissue engineering centered applications. In this review chitosan, its properties and role either virgin, chemically or physically modified, 2D or 3D scaffolds for tissue engineering application have been highlighted. Copyright © 2017. Published by Elsevier B.V.

  12. Hybrid chitosan-ß-glycerol phosphate-gelatin nano-/micro fibrous scaffolds with suitable mechanical and biological properties for tissue engineering.

    Science.gov (United States)

    Lotfi, Marzieh; Bagherzadeh, Roohollah; Naderi-Meshkin, Hojjat; Mahdipour, Elahe; Mafinezhad, Asghar; Sadeghnia, Hamid Reza; Esmaily, Habibollah; Maleki, Masoud; Hasssanzadeh, Halimeh; Ghayaour-Mobarhan, Majid; Bidkhori, Hamid Reza; Bahrami, Ahmad Reza

    2016-03-01

    Scaffold-based tissue engineering is considered as a promising approach in the regenerative medicine. Graft instability of collagen, by causing poor mechanical properties and rapid degradation, and their hard handling remains major challenges to be addressed. In this research, a composite structured nano-/microfibrous scaffold, made from a mixture of chitosan-ß-glycerol phosphate-gelatin (chitosan-GP-gelatin) using a standard electrospinning set-up was developed. Gelatin-acid acetic and chitosan ß-glycerol phosphate-HCL solutions were prepared at ratios of 30/70, 50/50, 70/30 (w/w) and their mechanical and biological properties were engineered. Furthermore, the pore structure of the fabricated nanofibrous scaffolds was investigated and predicted using a theoretical model. Higher gelatin concentrations in the polymer blend resulted in significant increase in mean pore size and its distribution. Interaction between the scaffold and the contained cells was also monitored and compared in the test and control groups. Scaffolds with higher chitosan concentrations showed higher rate of cell attachment with better proliferation property, compared with gelatin-only scaffolds. The fabricated scaffolds, unlike many other natural polymers, also exhibit non-toxic and biodegradable properties in the grafted tissues. In conclusion, the data clearly showed that the fabricated biomaterial is a biologically compatible scaffold with potential to serve as a proper platform for retaining the cultured cells for further application in cell-based tissue engineering, especially in wound healing practices. These results suggested the potential of using mesoporous composite chitosan-GP-gelatin fibrous scaffolds for engineering three-dimensional tissues with different inherent cell characteristics. © 2015 Wiley Periodicals, Inc.

  13. Modified n-HA/PA66 scaffolds with chitosan coating for bone tissue engineering: cell stimulation and drug release.

    Science.gov (United States)

    Zou, Qin; Li, Junfeng; Niu, Lulu; Zuo, Yi; Li, Jidong; Li, Yubao

    2017-09-01

    The dipping-drying procedure and cross-linking method were used to make drug-loaded chitosan (CS) coating on nano-hydroxyapatite/polyamide66 (nHA/PA66) composite porous scaffold, endowing the scaffold controlled drug release functionality. The prefabricated scaffold was immersed into an aqueous drug/CS solution in a vacuum condition and then crosslinked by vanillin. The structure, porosity, composition, compressive strength, swelling ratio, drug release and cytocompatibility of the pristine and coating scaffolds were investigated. After coating, the scaffold porosity and pore interconnection were slightly decreased. Cytocompatibility performance was observed through an in vitro experiment based on cell attachment and the MTT assay by MG63 cells which revealed positive cell viability and increasing proliferation over the 11-day period in vitro. The drug could effectively release from the coated scaffold in a controlled fashion and the release rate was sustained for a long period and highly dependent on coating swelling, suggesting the possibility of a controlled drug release. Our results demonstrate that the scaffold with drug-loaded crosslinked CS coating can be used as a simple technique to render the surfaces of synthetic scaffolds active, thus enabling them to be a promising high performance biomaterial in bone tissue engineering.

  14. Self-assembly model, hepatocytes attachment and inflammatory response for silk fibroin/chitosan scaffolds

    International Nuclear Information System (INIS)

    She Zhending; Feng Qingling; Liu Weiqiang

    2009-01-01

    Silk fibroin is an attractive natural fibrous protein for biomedical application due to its good biocompatibility and high tensile strength. Silk fibroin is apt to form a sheet-like structure during the freeze-drying process, which is not suitable for the scaffold of tissue engineering. In our former study, the adding of chitosan promoted the self-assembly of silk fibroin/chitosan (SFCS) into a three-dimensional (3D) homogeneous porous structure. In this study, a model of the self-assembly is proposed; furthermore, hepatocytes attachment and inflammatory response for the SFCS scaffold were examined. The rigid chain of chitosan may be used as a template for β-sheet formation of silk fibroin, and this may break the sheet structure of the silk fibroin scaffold and promote the formation of a 3D porous structure of the SFCS scaffold. Compared with the polylactic glycolic acid scaffold, the SFCS scaffold further facilitates the attachment of hepatocytes. To investigate the inflammatory response, SFCS scaffolds were implanted into the greater omentum of rats. From the results of implantation, we could demonstrate in vivo that the implantation of SFCS scaffolds resulted in only slight inflammation. Keeping the good histocompatibility and combining the advantages of both fibroin and chitosan, the SFCS scaffold could be a prominent candidate for soft tissue engineering, for example, in the liver.

  15. Self-assembly model, hepatocytes attachment and inflammatory response for silk fibroin/chitosan scaffolds

    Energy Technology Data Exchange (ETDEWEB)

    She Zhending; Feng Qingling [State Key Laboratory of New Ceramics and Fine Processing, Department of Materials Science and Engineering, Tsinghua University, Beijing 100084 (China); Liu Weiqiang, E-mail: biomater@mail.tsinghua.edu.c [Center for Advanced Materials and Biotechnology, Research Institute of Tsinghua University in Shenzhen, Shenzhen 518057 (China)

    2009-08-15

    Silk fibroin is an attractive natural fibrous protein for biomedical application due to its good biocompatibility and high tensile strength. Silk fibroin is apt to form a sheet-like structure during the freeze-drying process, which is not suitable for the scaffold of tissue engineering. In our former study, the adding of chitosan promoted the self-assembly of silk fibroin/chitosan (SFCS) into a three-dimensional (3D) homogeneous porous structure. In this study, a model of the self-assembly is proposed; furthermore, hepatocytes attachment and inflammatory response for the SFCS scaffold were examined. The rigid chain of chitosan may be used as a template for beta-sheet formation of silk fibroin, and this may break the sheet structure of the silk fibroin scaffold and promote the formation of a 3D porous structure of the SFCS scaffold. Compared with the polylactic glycolic acid scaffold, the SFCS scaffold further facilitates the attachment of hepatocytes. To investigate the inflammatory response, SFCS scaffolds were implanted into the greater omentum of rats. From the results of implantation, we could demonstrate in vivo that the implantation of SFCS scaffolds resulted in only slight inflammation. Keeping the good histocompatibility and combining the advantages of both fibroin and chitosan, the SFCS scaffold could be a prominent candidate for soft tissue engineering, for example, in the liver.

  16. Preparation and characterization of collagen/PLA, chitosan/PLA, and collagen/chitosan/PLA hybrid scaffolds for cartilage tissue engineering.

    Science.gov (United States)

    Haaparanta, Anne-Marie; Järvinen, Elina; Cengiz, Ibrahim Fatih; Ellä, Ville; Kokkonen, Harri T; Kiviranta, Ilkka; Kellomäki, Minna

    2014-04-01

    In this study, three-dimensional (3D) porous scaffolds were developed for the repair of articular cartilage defects. Novel collagen/polylactide (PLA), chitosan/PLA, and collagen/chitosan/PLA hybrid scaffolds were fabricated by combining freeze-dried natural components and synthetic PLA mesh, where the 3D PLA mesh gives mechanical strength, and the natural polymers, collagen and/or chitosan, mimic the natural cartilage tissue environment of chondrocytes. In total, eight scaffold types were studied: four hybrid structures containing collagen and/or chitosan with PLA, and four parallel plain scaffolds with only collagen and/or chitosan. The potential of these types of scaffolds for cartilage tissue engineering applications were determined by the analysis of the microstructure, water uptake, mechanical strength, and the viability and attachment of adult bovine chondrocytes to the scaffolds. The manufacturing method used was found to be applicable for the manufacturing of hybrid scaffolds with highly porous 3D structures. All the hybrid scaffolds showed a highly porous structure with open pores throughout the scaffold. Collagen was found to bind water inside the structure in all collagen-containing scaffolds better than the chitosan-containing scaffolds, and the plain collagen scaffolds had the highest water absorption. The stiffness of the scaffold was improved by the hybrid structure compared to plain scaffolds. The cell viability and attachment was good in all scaffolds, however, the collagen hybrid scaffolds showed the best penetration of cells into the scaffold. Our results show that from the studied scaffolds the collagen/PLA hybrids are the most promising scaffolds from this group for cartilage tissue engineering.

  17. Physical characterization and modeling of chitosan/peg blends for injectable scaffolds.

    Science.gov (United States)

    Lima, Daniel B; Almeida, Renata D; Pasquali, Matheus; Borges, Sílvia P; Fook, Marcus L; Lisboa, Hugo M

    2018-06-01

    Injectable scaffolds find many applications on the biomedical field due to several advantages on preformed scaffolds such as being able to fill any defect can be used in minimal invasion surgeries and are ready to use products. The most critical parameter for an injectable scaffold usage is its injectability, which can be related with rheological properties. Therefore, the objective of the present work was to increase knowledge about the critical parameters influencing injectability of biopolymers used for injectable scaffolds. Rheological and mechanical properties of a biopolymer blend in combination with injectability tests for a given design space controlled by the concentrations of both polymers and temperatures was made. Then those results were modeled to better understand the impact of parameters on injectability. The biopolymer blend chosen was Chitosan physically blended with Poly(ethylene glycol) where variations of both polymer concentrations and molecular weights were tested. Rheological and mechanical properties of all samples were determined, together with the injection force using a compression test at different injection conditions. All solutions were clear and transparent suggesting perfect miscibility. Rheological results were modeled using Ostwald-Waelle law and revealed a shear thinning pseudo-plastic solution at any composition and temperature, being chitosan concentration the most influencing variable. Compression tests results revealed mean injection forces ranging from 9.9 ± 0.06N to 29.9 ± 0.65N and it was possible to accurately estimate those results. Simulations revealed draw speed as the most influencing parameter. Cell viability tests revealed a non-cytotoxic biopolymer blend. Copyright © 2018 Elsevier Ltd. All rights reserved.

  18. Fabrication of Chitosan Silk-based Tracheal Scaffold Using Freeze-Casting Method

    Science.gov (United States)

    Nematollahi, Zeinab; Tafazzoli-Shadpour, Mohammad; Zamanian, Ali; Seyedsalehi, Amir; Mohammad-Behgam, Shadmehr; Ghorbani, Fariba; Mirahmadi, Fereshte

    2017-01-01

    Background: Since the treatments of long tracheal lesions are associated with some limitations, tissue engineered trachea is considered as an alternative option. This study aimed at preparing a composite scaffold, based on natural and synthetic materials for tracheal tissue engineering. Methods: Nine chitosan silk-based scaffolds were fabricated using three freezing rates (0.5, 1, and 2°C/min) and glutaraldehyde (GA) concentrations (0, 0.4, and 0.8 wt%). Samples were characterized, and scaffolds having mechanical properties compatible with those of human trachea and proper biodegradability were selected for chondrocyte cell seeding and subsequent biological assessments. Results: The pore sizes were highly influenced by the freezing rate and varied from 135.3×372.1 to 37.8×83.4 µm. Swelling and biodegradability behaviors were more affected by GA rather than freezing rate. Tensile strength raised from 120 kPa to 350 kPa by an increment of freezing rate and GA concentration. In addition, marked stiffening was demonstrated by increasing elastic modulus from 1.5 MPa to 12.2 MPa. Samples having 1 and 2°C/min of freezing rate and 0.8 wt% GA concentration made a non-toxic, porous structure with tensile strength and elastic modulus in the range of human trachea, facilitating the chondrocyte proliferation. The results of 21-day cell culture indicated that glycosaminoglycans content was significantly higher for the rate of 2°C/min (12.04 µg/min) rather than the other (9.6 µg/min). Conclusion: A homogenous porous structure was created by freeze drying. This allows the fabrication of a chitosan silk scaffold cross-linked by GA for cartilage tissue regeneration with application in tracheal regeneration. PMID:28131109

  19. Functionalization of chitosan/poly(lactic acid-glycolic acid) sintered microsphere scaffolds via surface heparinization for bone tissue engineering.

    Science.gov (United States)

    Jiang, Tao; Khan, Yusuf; Nair, Lakshmi S; Abdel-Fattah, Wafa I; Laurencin, Cato T

    2010-06-01

    Scaffolds exhibiting biological recognition and specificity play an important role in tissue engineering and regenerative medicine. The bioactivity of scaffolds in turn influences, directs, or manipulates cellular responses. In this study, chitosan/poly(lactic acid-co-glycolic acid) (chitosan/PLAGA) sintered microsphere scaffolds were functionalized via heparin immobilization. Heparin was successfully immobilized on chitosan/PLAGA scaffolds with controllable loading efficiency. Mechanical testing showed that heparinization of chitosan/PLAGA scaffolds did not significantly alter the mechanical properties and porous structures. In addition, the heparinized chitosan/PLAGA scaffolds possessed a compressive modulus of 403.98 +/- 19.53 MPa and a compressive strength of 9.83 +/- 0.94 MPa, which are in the range of human trabecular bone. Furthermore, the heparinized chitosan/PLAGA scaffolds had an interconnected porous structure with a total pore volume of 30.93 +/- 0.90% and a median pore size of 172.33 +/- 5.89 mum. The effect of immobilized heparin on osteoblast-like MC3T3-E1 cell growth was investigated. MC3T3-E1 cells proliferated three dimensionally throughout the porous structure of the scaffolds. Heparinized chitosan/PLAGA scaffolds with low heparin loading (1.7 microg/scaffold) were shown to be capable of stimulating MC3T3-E1 cell proliferation by MTS assay and cell differentiation as evidenced by elevated osteocalcin expression when compared with nonheparinized chitosan/PLAGA scaffold and chitosan/PLAGA scaffold with high heparin loading (14.1 microg/scaffold). This study demonstrated the potential of functionalizing chitosan/PLAGA scaffolds via heparinization with improved cell functions for bone tissue engineering applications.

  20. The potential of chitosan combined with chicken shank collagen as scaffold on bone defect regeneration process in Rattus norvegicus

    Directory of Open Access Journals (Sweden)

    Fitria Rahmitasari

    2016-12-01

    Full Text Available Background: In the field of dentistry, alveolar bone damage can be caused by periodontal disease, traumatic injury due to tooth extraction, cyst enucleation, and tumor surgery. One of the ways to regenerate the bone defect is using graft scaffold. Thus, combination of chitosan and collagen can stimulate osteogenesis. Purpose: The aim of this study was to examine the potential of chitosan combined with chicken shank collagen on bone defect regeneration process. Method: Twelve Rattus norvegicus were prepared as animal models in this research. A bone defect was intentionally created at both of the right and left femoral bones of the models. Next, 24 samples were divided into four groups, namely Group 1 using chitosan – collagen scaffold (50:50, Group 2 using chitosan collagen-scaffold (80:20, Group 3 using chitosan scaffold only, and Control Group using 3% CMC-Na. On 14th day, those animals were sacrificed, and histopathological anatomy examination was conducted to observe osteoclast cells. In addition, immunohistochemistry examination was also performed to observe RANKL expressions. Result: There was a significant difference in RANKL expressions among the groups, except between Group 3 using chitosan scaffold only and control group (p value > 0.05. The highest expression of RANKL was found in Group 1 with chitosan – collagen scaffold (50:50, followed by Group 2 with chitosan-collagen scaffold (80:20. Moreover, there was also a significant difference in osteoclast generation, except between Group 1 using chitosan – collagen scaffold (50:50 and Group 2 using chitosan-collagen scaffold (80:20, p value 0.05. Less osteoclast was found in the groups using chitosan – collagen scaffold (Group 1 and Group 2. Conclusion: Combination of chitosan and chicken shank collagen scaffold can improve regeneration process of bone defect in Rattus novergicus animals through increasing of RANKL expressions, and decreasing of osteoclast.

  1. Chitosan-g-lactide copolymers for fabrication of 3D scaffolds for tissue engineering

    Science.gov (United States)

    Demina, T. S.; Zaytseva-Zotova, D. S.; Timashev, P. S.; Bagratashvili, V. N.; Bardakova, K. N.; Sevrin, Ch; Svidchenko, E. A.; Surin, N. M.; Markvicheva, E. A.; Grandfils, Ch; Akopova, T. A.

    2015-07-01

    Chitosan-g-oligo (L, D-lactide) copolymers were synthesized and assessed to fabricate a number of 3D scaffolds using a variety of technologies such as oil/water emulsion evaporation technique, freeze-drying and two-photon photopolymerization. Solid-state copolymerization method allowed us to graft up to 160 wt-% of oligolactide onto chitosan backbone via chitosan amino group acetylation with substitution degree reaching up to 0.41. Grafting of hydrophobic oligolactide side chains with polymerization degree up to 10 results in chitosan amphiphilic properties. The synthesized chitosan-g-lactide copolymers were used to design 3D scaffolds for tissue engineering such as spherical microparticles and macroporous hydrogels.

  2. Improving effects of chitosan nanofiber scaffolds on osteoblast proliferation and maturation

    Directory of Open Access Journals (Sweden)

    Ho MH

    2014-09-01

    Full Text Available Ming-Hua Ho,1,2 Mei-Hsiu Liao,3 Yi-Ling Lin,2 Chien-Hao Lai,3 Pei-I Lin,3 Ruei-Ming Chen2–4 1Department of Chemical Engineering, National Taiwan University of Science and Technology, Taipei, Taiwan; 2Cell Physiology and Molecular Image Research Center and Department of Anesthesiology, Wan Fang Hospital, 3Graduate Institute of Medical Sciences, Taipei Medical University, Taipei, Taiwan; 4Anesthetics and Toxicology Research Center, Taipei Medical University Hospital, Taipei, Taiwan Abstract: Osteoblast maturation plays a key role in regulating osteogenesis. Electrospun nanofibrous products were reported to possess a high surface area and porosity. In this study, we developed chitosan nanofibers and examined the effects of nanofibrous scaffolds on osteoblast maturation and the possible mechanisms. Macro- and micro observations of the chitosan nanofibers revealed that these nanoproducts had a flat surface and well-distributed fibers with nanoscale diameters. Mouse osteoblasts were able to attach onto the chitosan nanofiber scaffolds, and the scaffolds degraded in a time-dependent manner. Analysis by scanning electron microscopy further showed mouse osteoblasts adhered onto the scaffolds along the nanofibers, and cell–cell communication was also detected. Mouse osteoblasts grew much better on chitosan nanofiber scaffolds than on chitosan films. In addition, human osteoblasts were able to adhere and grow on the chitosan nanofiber scaffolds. Interestingly, culturing human osteoblasts on chitosan nanofiber scaffolds time-dependently increased DNA replication and cell proliferation. In parallel, administration of human osteoblasts onto chitosan nanofibers significantly induced osteopontin, osteocalcin, and alkaline phosphatase (ALP messenger (mRNA expression. As to the mechanism, chitosan nanofibers triggered runt-related transcription factor 2 mRNA and protein syntheses. Consequently, results of ALP-, alizarin red-, and von Kossa-staining analyses

  3. Bioactive Sr(II/Chitosan/Poly(ε-caprolactone Scaffolds for Craniofacial Tissue Regeneration. In Vitro and In Vivo Behavior

    Directory of Open Access Journals (Sweden)

    Itzia Rodríguez-Méndez

    2018-03-01

    Full Text Available In craniofacial tissue regeneration, the current gold standard treatment is autologous bone grafting, however, it presents some disadvantages. Although new alternatives have emerged there is still an urgent demand of biodegradable scaffolds to act as extracellular matrix in the regeneration process. A potentially useful element in bone regeneration is strontium. It is known to promote stimulation of osteoblasts while inhibiting osteoclasts resorption, leading to neoformed bone. The present paper reports the preparation and characterization of strontium (Sr containing hybrid scaffolds formed by a matrix of ionically cross-linked chitosan and microparticles of poly(ε-caprolactone (PCL. These scaffolds of relatively facile fabrication were seeded with osteoblast-like cells (MG-63 and human bone marrow mesenchymal stem cells (hBMSCs for application in craniofacial tissue regeneration. Membrane scaffolds were prepared using chitosan:PCL ratios of 1:2 and 1:1 and 5 wt % Sr salts. Characterization was performed addressing physico-chemical properties, swelling behavior, in vitro biological performance and in vivo biocompatibility. Overall, the composition, microstructure and swelling degree (≈245% of scaffolds combine with the adequate dimensional stability, lack of toxicity, osteogenic activity in MG-63 cells and hBMSCs, along with the in vivo biocompatibility in rats allow considering this system as a promising biomaterial for the treatment of craniofacial tissue regeneration.

  4. Comparison of cadmium adsorption onto chitosan and epichlorohydrin crosslinked chitosan/eggshell composite

    Science.gov (United States)

    Rahmi; Marlina; Nisfayati

    2018-05-01

    The use of chitosan and epichlorohydrin crosslinked chitosan/eggshell composite for cadmium adsorption from water were investigated. The factors affecting adsorption such as pH and contact time were considered. The results showed that the optimum pH of adsorption was pH = 6.0 and the equilibrium time of adsorption was 40 min. The adsorption isotherm of Cd ions onto chitosan and composite were well fitted to Langmuir equation. The maximum adsorption capacity (fitting by Langmuir model) of chitosan and composite were 1.008 and 11.7647 mg/g, respectively. Adsorption performance of composite after regeneration was better than chitosan.

  5. Microwave-induced porosity and bioactivation of chitosan-PEGDA scaffolds: morphology, mechanical properties and osteogenic differentiation.

    Science.gov (United States)

    Demitri, Christian; Giuri, Antonella; De Benedictis, Vincenzo Maria; Raucci, Maria Grazia; Giugliano, Daniela; Sannino, Alessandro; Ambrosio, Luigi

    2017-01-01

    In this study, a new foaming method, based on physical foaming combined with microwave-induced curing, is proposed in combination with a surface bioactivation to develop scaffold for bone tissue regeneration. In the first step of the process, a stable physical foaming was induced using a surfactant (Pluronic) as blowing agent of a homogeneous blend of Chitosan and polyethylene glycol diacrylate (PEGDA700) solutions. In the second step, the porous structure of the foaming was chemically stabilized by radical polymerization induced by homogeneous heating of the sample in a microwave reactor. In this step, 2,2-azobis[2-(2-imidazolin-2yl)propane]dihydrochloride was used as thermoinitiator (TI). Chitosan and PEGDA were mixed in different blends to investigate the influence of the composition on the final properties of the material. The chemical properties of each sample were evaluated by infrared attenuated total reflectance analysis, before and after curing in order to maximize reaction yield and optimize kinetic parameters (i.e. time curing, microwave power). Absorption capacity, elastic modulus, porosity and morphology of the porous structure were measured for each sample. The stability of materials was evaluated in vitro by degradation test in phosphate-buffered saline. To improve the bioactivity and biological properties of chitosan scaffold, a biomineralization process was used. Biological characterization was carried out with the aim to prove the effect of biomineralization scaffold on human mesenchymal stem cells behaviour. Copyright © 2016 John Wiley & Sons, Ltd. Copyright © 2016 John Wiley & Sons, Ltd.

  6. Preparation and characterization of chitosan-heparin composite matrices for blood contacting tissue engineering

    International Nuclear Information System (INIS)

    He Qing; Gong Kai; Gong Yandao; Zhang Xiufang; Ao Qiang; Zhang Lihai; Hu Min

    2010-01-01

    Chitosan has been widely used for biomaterial scaffolds in tissue engineering because of its good mechanical properties and cytocompatibility. However, the poor blood compatibility of chitosan has greatly limited its biomedical utilization, especially for blood contacting tissue engineering. In this study, we exploited a polymer blending procedure to heparinize the chitosan material under simple and mild conditions to improve its antithrombogenic property. By an optimized procedure, a macroscopically homogeneous chitosan-heparin (Chi-Hep) blended suspension was obtained, with which Chi-Hep composite films and porous scaffolds were fabricated. X-ray photoelectron spectroscopy and sulfur elemental analysis confirmed the successful immobilization of heparin in the composite matrices (i.e. films and porous scaffolds). Toluidine blue staining indicated that heparin was distributed homogeneously in the composite matrices. Only a small amount of heparin was released from the matrices during incubation in normal saline for 10 days. The composite matrices showed improved blood compatibility, as well as good mechanical properties and endothelial cell compatibility. These results suggest that the Chi-Hep composite matrices are promising candidates for blood contacting tissue engineering.

  7. Preparation and characterization of three-dimensional scaffolds based on hydroxypropyl chitosan-graft-graphene oxide.

    Science.gov (United States)

    Sivashankari, P R; Moorthi, A; Abudhahir, K Mohamed; Prabaharan, M

    2018-04-15

    Hydroxypropyl chitosan (HPCH), a water soluble derivative of chitosan, is widely considered for tissue engineering and wound healing applications due to its biocompatibility and biodegradability. Graphene oxide (GO) is a carbon-based nanomaterial which is capable of imparting desired properties to the scaffolds. Hence, the integration of GO into HPCH could allow for the production of HPCH-based scaffolds with improved swelling character, mechanical strength, and stability aimed at being used in tissue engineering. In this study, hydroxypropyl chitosan-graft-graphene oxide (HPCH-g-GO) with varying GO content (0.5, 1, 3 and 4wt.%) was prepared using HPCH and GO as a tissue engineering scaffold material. The formation of HPCH-g-GO was confirmed by FTIR and XRD analysis. Using the HPCH-g-GO as a matrix material and glutaraldehyde as a crosslinking agent, the three dimensional (3D) porous scaffolds were fabricated by the freeze-drying method. The HPCH-g-GO scaffolds exhibited uniform porosity as observed in SEM analysis. The pore size and porosity reduced as the content of GO was increased. These scaffolds presented good swelling capacity, water retention ability, mechanical strength and in vitro degradation properties. The HPCH-g-GO scaffolds irrespective of their GO content demonstrated good cell viability when compared to control. Altogether, these results suggest that HPCH-g-GO scaffolds can be used as potential tissue engineering material. Copyright © 2017 Elsevier B.V. All rights reserved.

  8. ALP gene expression in cDNA samples from bone tissue engineering using a HA/TCP/Chitosan scaffold

    Science.gov (United States)

    Stephanie, N.; Katarina, H.; Amir, L. R.; Gunawan, H. A.

    2017-08-01

    This study examined the potential use of hydroxyapatite (HA)/tricalcium phosphate (TCP)/Chitosan as a bone tissue engineering scaffold. The potential for using HA/TCP/chitosan as a scaffold was analyzed by measuring expression of the ALP osteogenic gene in cDNA from bone biopsies from four Macaque nemestrina. Experimental conditions included control (untreated), treatment with HA/TCP 70:30, HA/TCP 50:50, and HA/TCP/chitosan. cDNA samples were measured quantitively with Real-Time PCR (qPCR) and semi-quantitively by gel electrophoresis. There were no significant differences in ALP gene expression between treatment subjects after two weeks, but the HA/TCP/chitosan treatment gave the highest level of expression after four weeks. The scaffold using the HA/TCP/chitosan combination induced a higher level of expression of the osteogenic gene ALP than did scaffold without chitosan.

  9. Rapid adhesion and proliferation of keratinocytes on the gold colloid/chitosan film scaffold

    International Nuclear Information System (INIS)

    Zhang Yi; He Hong; Gao Wenjuan; Lu Shuangyun; Liu Yang; Gu Haiying

    2009-01-01

    The gold colloid/chitosan film scaffold, which could enhance the attached ratio and accelerate proliferation of newborn mice keratinocytes, was fabricated by nanotechnology and self-assembly technology. This nanometer scaffold was characterized by atomic force microscopy (AFM) and scanning electron microscopy (SEM). The keratinocytes were cultured and observed on three different extracellular matrices (ECM): gold colloid/chitosan film scaffold, chitosan film and cell culture plastic (control groups). 6 h, 12 h, 24 h after inoculation, the cell attached ratios were calculated respectively. In comparison to control groups, this scaffold could significantly (P < 0.01) increase the attached ratio of keratinocytes and promote their growth. Meanwhile, there were not any fusiform fibroblasts growing on this scaffold. The rapidly proliferating keratinocytes were indentified and characterized by immunohistochemistry and transmissive electron microscope (TEM), which showed the cells maintain their biological activity well. The results indicated that gold colloid/chitosan film scaffold was nontoxic to keratinocytes, and was a good candidate for wound dressing in skin tissue engineering.

  10. Chitosan-based hydrogel tissue scaffolds made by 3D plotting promotes osteoblast proliferation and mineralization.

    Science.gov (United States)

    Liu, I-Hsin; Chang, Shih-Hsin; Lin, Hsin-Yi

    2015-05-13

    A 3D plotting system was used to make chitosan-based tissue scaffolds with interconnected pores using pure chitosan (C) and chitosan cross-linked with pectin (CP) and genipin (CG). A freeze-dried chitosan scaffold (CF/D) was made to compare with C, to observe the effects of structural differences. The fiber size, pore size, porosity, compression strength, swelling ratio, drug release efficacy, and cumulative weight loss of the scaffolds were measured. Osteoblasts were cultured on the scaffolds and their proliferation, type I collagen production, alkaline phosphatase activity, calcium deposition, and morphology were observed. C had a lower swelling ratio, degradation, porosity and drug release efficacy and a higher compressional stiffness and cell proliferation compared to CF/D (p 3D-plotted samples, cells on CP exhibited the highest degree of mineralization after 21 d (p 3D-plotted scaffolds were stronger, less likely to degrade and better promoted osteoblast cell proliferation in vitro compared to the freeze-dried scaffolds. C, CP and CG were structurally similar, and the different crosslinking caused significant changes in their physical and biological performances.

  11. Collagen/chitosan based two-compartment and bi-functional dermal scaffolds for skin regeneration

    Energy Technology Data Exchange (ETDEWEB)

    Wang, Feng [Department of Plastic Surgery and Burns, Shenzhen Second People' s Hospital, Shenzhen 518035 (China); Wang, Mingbo [Key Laboratory of Biomedical Materials and Implants, Research Institute of Tsinghua University in Shenzhen, Shenzhen 518057 (China); She, Zhending [Key Laboratory of Biomedical Materials and Implants, Research Institute of Tsinghua University in Shenzhen, Shenzhen 518057 (China); Shenzhen Lando Biomaterials Co., Ltd., Shenzhen 518057 (China); Fan, Kunwu; Xu, Cheng [Department of Plastic Surgery and Burns, Shenzhen Second People' s Hospital, Shenzhen 518035 (China); Chu, Bin; Chen, Changsheng [Key Laboratory of Biomedical Materials and Implants, Research Institute of Tsinghua University in Shenzhen, Shenzhen 518057 (China); Shi, Shengjun, E-mail: shengjunshi@yahoo.com [The Burns Department of Zhujiang Hospital, Southern Medical University, Guangzhou 510280 (China); Tan, Rongwei, E-mail: tanrw@landobiom.com [Key Laboratory of Biomedical Materials and Implants, Research Institute of Tsinghua University in Shenzhen, Shenzhen 518057 (China); Shenzhen Lando Biomaterials Co., Ltd., Shenzhen 518057 (China)

    2015-07-01

    Inspired from the sophisticated bilayer structures of natural dermis, here, we reported collagen/chitosan based two-compartment and bi-functional dermal scaffolds. Two functions refer to mediating rapid angiogenesis based on recombinant human vascular endothelial growth factor (rhVEGF) and antibacterial from gentamicin, which were encapsulated in PLGA microspheres. The gentamicin and rhVEGF encapsulated PLGA microspheres were further combined with collagen/chitosan mixtures in low (lower layer) and high (upper layer) concentrations, and molded to generate the two-compartment and bi-functional scaffolds. Based on morphology and pore structure analyses, it was found that the scaffold has a distinct double layered porous and connective structure with PLGA microspheres encapsulated. Statistical analysis indicated that the pores in the upper layer and in the lower layer have great variations in diameter, indicative of a two-compartment structure. The release profiles of gentamicin and rhVEGF exceeded 28 and 49 days, respectively. In vitro culture of mouse fibroblasts showed that the scaffold can facilitate cell adhesion and proliferation. Moreover, the scaffold can obviously inhibit proliferation of Staphylococcus aureus and Serratia marcescens, exhibiting its unique antibacterial effect. The two-compartment and bi-functional dermal scaffolds can be a promising candidate for skin regeneration. - Highlights: • The dermal scaffold is inspired from the bilayer structures of natural dermis. • The dermal scaffold has two-compartment structures. • The dermal scaffold containing VEGF and gentamicin encapsulated PLGA microspheres • The dermal scaffold can facilitate cell adhesion and proliferation.

  12. Development of novel hybrid poly(l-lactide)/chitosan scaffolds using the rapid freeze prototyping technique

    Energy Technology Data Exchange (ETDEWEB)

    Zhu, N; Chen, X B [Division of Biomedical Engineering, University of Saskatchewan, Saskatoon, Saskatchewan (Canada); Li, M G [Department of Mechanical Engineering, University of Saskatchewan, Saskatoon, Saskatchewan (Canada); Cooper, D, E-mail: xbc719@mail.usask.ca [Department of Anatomy and Cell Biology, University of Saskatchewan, Saskatoon, Saskatchewan (Canada)

    2011-09-15

    Engineered scaffolds have been shown to be critical to various tissue engineering applications. This paper presents the development of a novel three-dimensional scaffold made from a mixture of chitosan microspheres (CMs) and poly(l-lactide) by means of the rapid freeze prototyping (RFP) technique. The CMs were used to encapsulate bovine serum albumin (BSA) and improve the scaffold mechanical properties. Experiments to examine the BSA release were carried out; the BSA release could be controlled by adjusting the crosslink degree of the CMs and prolonged after the CMs were embedded into the PLLA scaffolds, while the examination of the mechanical properties of the scaffolds illustrates that they depend on the ratio of CMs to PLLA in the scaffolds as well as the cryogenic temperature used in the RFP fabrication process. The chemical characteristics of the PLLA/chitosan scaffolds were evaluated by Fourier transform infrared (FTIR) spectroscopy. The morphological and pore structure of the scaffolds were also examined by scanning electron microscopy and micro-tomography. The results obtained show that the scaffolds have higher porosity and enhanced pore size distribution compared to those fabricated by the dispensing-based rapid prototyping technique. This study demonstrates that the novel scaffolds have not only enhanced porous structure and mechanical properties but also showed the potential to preserve the bioactivities of the biomolecules and to control the biomolecule distribution and release rate.

  13. Collagen/chitosan based two-compartment and bi-functional dermal scaffolds for skin regeneration

    International Nuclear Information System (INIS)

    Wang, Feng; Wang, Mingbo; She, Zhending; Fan, Kunwu; Xu, Cheng; Chu, Bin; Chen, Changsheng; Shi, Shengjun; Tan, Rongwei

    2015-01-01

    Inspired from the sophisticated bilayer structures of natural dermis, here, we reported collagen/chitosan based two-compartment and bi-functional dermal scaffolds. Two functions refer to mediating rapid angiogenesis based on recombinant human vascular endothelial growth factor (rhVEGF) and antibacterial from gentamicin, which were encapsulated in PLGA microspheres. The gentamicin and rhVEGF encapsulated PLGA microspheres were further combined with collagen/chitosan mixtures in low (lower layer) and high (upper layer) concentrations, and molded to generate the two-compartment and bi-functional scaffolds. Based on morphology and pore structure analyses, it was found that the scaffold has a distinct double layered porous and connective structure with PLGA microspheres encapsulated. Statistical analysis indicated that the pores in the upper layer and in the lower layer have great variations in diameter, indicative of a two-compartment structure. The release profiles of gentamicin and rhVEGF exceeded 28 and 49 days, respectively. In vitro culture of mouse fibroblasts showed that the scaffold can facilitate cell adhesion and proliferation. Moreover, the scaffold can obviously inhibit proliferation of Staphylococcus aureus and Serratia marcescens, exhibiting its unique antibacterial effect. The two-compartment and bi-functional dermal scaffolds can be a promising candidate for skin regeneration. - Highlights: • The dermal scaffold is inspired from the bilayer structures of natural dermis. • The dermal scaffold has two-compartment structures. • The dermal scaffold containing VEGF and gentamicin encapsulated PLGA microspheres • The dermal scaffold can facilitate cell adhesion and proliferation

  14. Novel fiber-based pure chitosan scaffold for tendon augmentation: biomechanical and cell biological evaluation.

    Science.gov (United States)

    Nowotny, J; Aibibu, D; Farack, J; Nimtschke, U; Hild, M; Gelinsky, M; Kasten, P; Cherif, Ch

    2016-07-01

    One possibility to improve the mechanical properties after tendon ruptures is augmentation with a scaffold. Based on wet spinning technology, chitosan fibres were processed to a novel pure high-grade multifilament yarn with reproducible quality. The fibres were braided to obtain a 3D tendon scaffold. The CS fibres and scaffolds were evaluated biomechanically and compared to human supraspinatus (SSP) tendons. For the cytobiological characterization, in vitro cell culture experiments with human mesenchymal stem cells (hMSC) were performed. Three types of 3D circular braided scaffolds were fabricated. Significantly, higher ultimate stress values were measured for scaffold with larger filament yarn, compared to scaffold with smaller filament yarn. During cultivation over 28 days, the cells showed in dependence of isolation method and/or donor a doubling or tripling of the cell number or even a six-fold increase on the CS scaffold, which was comparable to the control (polystyrene) or in the case of cells obtained from human biceps tendon even higher proliferation rates. After 14 days, the scaffold surface was covered homogeneously with a cell layer. In summary, the present work demonstrates that braided chitosan scaffolds constitute a straightforward approach for designing tendon analogues, maintaining important flexibility in scaffold design and providing favourable mechanical properties of the resulting construct.

  15. Fabrication and evaluation of a sustained-release chitosan-based scaffold embedded with PLGA microspheres

    International Nuclear Information System (INIS)

    Song, Kedong; Liu, Yingchao; Macedo, Hugo M.; Jiang, Lili; Li, Chao; Mei, Guanyu; Liu, Tianqing

    2013-01-01

    Nutrient depletion within three-dimensional (3D) scaffolds is one of the major hurdles in the use of this technology to grow cells for applications in tissue engineering. In order to help in addressing it, we herein propose to use the controlled release of encapsulated nutrients within polymer microspheres into chitosan-based 3D scaffolds, wherein the microspheres are embedded. This method has allowed maintaining a stable concentration of nutrients within the scaffolds over the long term. The polymer microspheres were prepared using multiple emulsions (w/o/w), in which bovine serum albumin (BSA) and poly (lactic-co-glycolic) acid (PLGA) were regarded as the protein pattern and the exoperidium material, respectively. These were then mixed with a chitosan solution in order to form the scaffolds by cryo-desiccation. The release of BSA, entrapped within the embedded microspheres, was monitored with time using a BCA kit. The morphology and structure of the PLGA microspheres containing BSA before and after embedding within the scaffold were observed under a scanning electron microscope (SEM). These had a round shape with diameters in the range of 27–55 μm, whereas the chitosan-based scaffolds had a uniform porous structure with the microspheres uniformly dispersed within their 3D structure and without any morphological change. In addition, the porosity, water absorption and degradation rate at 37 °C in an aqueous environment of 1% chitosan-based scaffolds were (92.99 ± 2.51) %, (89.66 ± 0.66) % and (73.77 ± 3.21) %, respectively. The studies of BSA release from the embedded microspheres have shown a sustained and cumulative tendency with little initial burst, with (20.24 ± 0.83) % of the initial amount released after 168 h (an average rate of 0.12%/h). The protein concentration within the chitosan-based scaffolds after 168 h was found to be (11.44 ± 1.81) × 10 −2 mg/mL. This novel chitosan-based scaffold embedded with PLGA microspheres has proven to be a

  16. Fabrication and evaluation of a sustained-release chitosan-based scaffold embedded with PLGA microspheres

    Energy Technology Data Exchange (ETDEWEB)

    Song, Kedong, E-mail: kedongsong@dlut.edu.cn [Dalian R and D Center for Stem Cell and Tissue Engineering, State Key Laboratory of Fine Chemicals, Dalian University of Technology, Dalian 116024 (China); Liu, Yingchao [Dalian R and D Center for Stem Cell and Tissue Engineering, State Key Laboratory of Fine Chemicals, Dalian University of Technology, Dalian 116024 (China); Macedo, Hugo M. [Biological Systems Engineering Laboratory, Department of Chemical Engineering, Department of Chemical Engineering, South Kensington Campus, London SW7 2AZ (United Kingdom); Jiang, Lili; Li, Chao; Mei, Guanyu [Dalian R and D Center for Stem Cell and Tissue Engineering, State Key Laboratory of Fine Chemicals, Dalian University of Technology, Dalian 116024 (China); Liu, Tianqing, E-mail: liutq@dlut.edu.cn [Dalian R and D Center for Stem Cell and Tissue Engineering, State Key Laboratory of Fine Chemicals, Dalian University of Technology, Dalian 116024 (China)

    2013-04-01

    Nutrient depletion within three-dimensional (3D) scaffolds is one of the major hurdles in the use of this technology to grow cells for applications in tissue engineering. In order to help in addressing it, we herein propose to use the controlled release of encapsulated nutrients within polymer microspheres into chitosan-based 3D scaffolds, wherein the microspheres are embedded. This method has allowed maintaining a stable concentration of nutrients within the scaffolds over the long term. The polymer microspheres were prepared using multiple emulsions (w/o/w), in which bovine serum albumin (BSA) and poly (lactic-co-glycolic) acid (PLGA) were regarded as the protein pattern and the exoperidium material, respectively. These were then mixed with a chitosan solution in order to form the scaffolds by cryo-desiccation. The release of BSA, entrapped within the embedded microspheres, was monitored with time using a BCA kit. The morphology and structure of the PLGA microspheres containing BSA before and after embedding within the scaffold were observed under a scanning electron microscope (SEM). These had a round shape with diameters in the range of 27–55 μm, whereas the chitosan-based scaffolds had a uniform porous structure with the microspheres uniformly dispersed within their 3D structure and without any morphological change. In addition, the porosity, water absorption and degradation rate at 37 °C in an aqueous environment of 1% chitosan-based scaffolds were (92.99 ± 2.51) %, (89.66 ± 0.66) % and (73.77 ± 3.21) %, respectively. The studies of BSA release from the embedded microspheres have shown a sustained and cumulative tendency with little initial burst, with (20.24 ± 0.83) % of the initial amount released after 168 h (an average rate of 0.12%/h). The protein concentration within the chitosan-based scaffolds after 168 h was found to be (11.44 ± 1.81) × 10{sup −2} mg/mL. This novel chitosan-based scaffold embedded with PLGA microspheres has proven to be a

  17. Chitosan scaffolds induce human dental pulp stem cells to neural differentiation: potential roles for spinal cord injury therapy.

    Science.gov (United States)

    Zhang, Jinlong; Lu, Xiaohui; Feng, Guijuan; Gu, Zhifeng; Sun, Yuyu; Bao, Guofeng; Xu, Guanhua; Lu, Yuanzhou; Chen, Jiajia; Xu, Lingfeng; Feng, Xingmei; Cui, Zhiming

    2016-10-01

    Cell-based transplantation strategies hold great potential for spinal cord injury (SCI) repair. Chitosan scaffolds have therapeutic benefits for spinal cord regeneration. Human dental pulp stem cells (DPSCs) are abundant available stem cells with low immunological incompatibility and can be considered for cell replacement therapy. The purpose of this study is to investigate the role of chitosan scaffolds in the neural differentiation of DPSCs in vitro and to assess the supportive effects of chitosan scaffolds in an animal model of SCI. DPSCs were incubated with chitosan scaffolds. Cell viability and the secretion of neurotrophic factors were analyzed. DPSCs incubated with chitosan scaffolds were treated with neural differentiation medium for 14 days and then neural genes and protein markers were analyzed by Western blot and reverse transcription plus the polymerase chain reaction. Our study revealed a higher cell viability and neural differentiation in the DPSC/chitosan-scaffold group. Compared with the control group, the levels of BDNF, GDNF, b-NGF, and NT-3 were significantly increased in the DPSC/chitosan-scaffold group. The Wnt/β-catenin signaling pathway played a key role in the neural differentiation of DPSCs combined with chitosan scaffolds. Transplantation of DPSCs together with chitosan scaffolds into an SCI rat model resulted in the marked recovery of hind limb locomotor functions. Thus, chitosan scaffolds were non-cytotoxic and provided a conducive and favorable microenvironment for the survival and neural differentiation of DPSCs. Transplantation of DPSCs might therefore be a suitable candidate for treating SCI and other neuronal degenerative diseases.

  18. Reinforced nanohydroxyapatite/polyamide66 scaffolds by chitosan coating for bone tissue engineering.

    Science.gov (United States)

    Huang, Di; Zuo, Yi; Zou, Qin; Wang, Yanying; Gao, Shibo; Wang, Xiaoyan; Liu, Haohuai; Li, Yubao

    2012-01-01

    High porosity of scaffold is always accompanied by poor mechanical property; the aim of this study was to enhance the strength and modulus of the highly porous scaffold of nanohydroxyapatite/polyamide66 (n-HA/PA66) by coating chitosan (CS) and to investigate the effect of CS content on the scaffold physical properties and cytological properties. The results show that CS coating can reinforce the scaffold effectively. The compress modulus and strength of the CS coated n-HA/PA66 scaffolds are improved to 32.71 and 2.38 MPa, respectively, being about six times and five times of those of the uncoated scaffolds. Meanwhile, the scaffolds still exhibit a highly interconnected porous structure and the porosity is approximate about 78%, slightly lower than the value (84%) of uncoated scaffold. The cytological properties of scaffolds were also studied in vitro by cocultured with osteoblast-like MG63 cells. The cytological experiments demonstrate that the reinforced scaffolds display favorable cytocompatibility and have no significant difference with the uncoated n-HA/PA66 scaffolds. The CS reinforced n-HA/PA66 scaffolds can meet the basic mechanical requirement of bone tissue engineering scaffold, presenting a potential for biomedical application in bone reconstruction and repair. Copyright © 2011 Wiley Periodicals, Inc.

  19. Injectable and microporous scaffold of densely-packed, growth factor-encapsulating chitosan microgels.

    Science.gov (United States)

    Riederer, Michael S; Requist, Brennan D; Payne, Karin A; Way, J Douglas; Krebs, Melissa D

    2016-11-05

    In this work, an emulsion crosslinking method was developed to produce chitosan-genipin microgels which acted as an injectable and microporous scaffold. Chitosan was characterized with respect to pH by light scattering and aqueous titration. Microgels were characterized with swelling, light scattering, and rheometry of densely-packed microgel solutions. The results suggest that as chitosan becomes increasingly deprotonated above the pKa, repulsive forces diminish and intermolecular attractions cause pH-responsive chain aggregation; leading to microgel-microgel aggregation as well. The microgels with the most chitosan and least cross-linker showed the highest yield stress and a storage modulus of 16kPa when condensed as a microgel paste at pH 7.4. Two oppositely-charged growth factors could be encapsulated into the microgels and endothelial cells were able to proliferate into the 3D microgel scaffold. This work motivates further research on the applications of the chitosan microgel scaffold as an injectable and microporous scaffold in regenerative medicine. Copyright © 2016 Elsevier Ltd. All rights reserved.

  20. Physico-functional and mechanical properties of chitosan and calcium salts incorporated fish gelatin scaffolds.

    Science.gov (United States)

    Jeevithan, E; Jeya Shakila, R; Varatharajakumar, A; Jeyasekaran, G; Sukumar, D

    2013-09-01

    Four types of fish gelatin scaffolds viz. gelatin (G), gelatin-chitosan (GC), gelatin-calcium acetate (GCA) and gelatin-chitosan-calcium acetate (GCCA) prepared were investigated for their functional properties, biomechanical strength, microstructural changes in relation to biodegradation. GC scaffold was superior with pH 3.15 and viscosity 9.40 cP. Chitosan and calcium acetate improved tensile strength (TS) and Young's modulus (YM), but lowered elongation at break (EAB). GCCA scaffold possessed moderate TS of 19.6 MPa, EAB of 4.76% and YM of 185 MPa. Foaming ability ratio of GC scaffold was high (3.41). GCA and GCCA scaffolds remained for 4 days before complete in vitro biodegradation. GC scaffold had larger cavities (180-300 μm) that were responsible for low swelling ratios and shrinkage factor. GCCA scaffold with moderate swelling rates, mechanical, functional properties and lowered biodegradation rate were found more suitable for biomedical applications. Copyright © 2013 Elsevier B.V. All rights reserved.

  1. 3D chitosan-gelatin-chondroitin porous scaffold improves osteogenic differentiation of mesenchymal stem cells

    Energy Technology Data Exchange (ETDEWEB)

    Machado, C B [Department of Biochemistry and Immunology, Institute of Biological Sciences, Federal University of Minas Gerais (Brazil); Ventura, J M G [Department of Ceramics and Glass Engineering, University of Aveiro (Portugal); Lemos, A F [Department of Ceramics and Glass Engineering, University of Aveiro (Portugal); Ferreira, J M F [Department of Ceramics and Glass Engineering, University of Aveiro (Portugal); Leite, M F [Department of Physiology and Biophysics, Institute of Biological Sciences, Federal University of Minas Gerais (Brazil); Goes, A M [Department of Biochemistry and Immunology, Institute of Biological Sciences, Federal University of Minas Gerais (Brazil)

    2007-06-01

    A porous 3D scaffold was developed to support and enhance the differentiation process of mesenchymal stem cells (MSC) into osteoblasts in vitro. The 3D scaffold was made with chitosan, gelatin and chondroitin and it was crosslinked by EDAC. The scaffold physicochemical properties were evaluated. SEM revealed the high porosity and interconnection of pores in the scaffold; rheological measurements show that the scaffold exhibits a characteristic behavior of strong gels. The elastic modulus found in compressive tests of the crosslinked scaffold was about 50 times higher than the non-crosslinked one. After 21 days, the 3D matrix submitted to hydrolytic degradation loses above 40% of its weight. MSC were collected from rat bone marrow and seeded in chitosan-gelatin-chondroitin 3D scaffolds and in 2D culture plates as well. MSC were differentiated into osteoblasts for 21 days. Cell proliferation and alkaline phosphatase activity were followed weekly during the osteogenic process. The osteogenic differentiation of MSC was improved in 3D culture as shown by MTT assay and alkaline phosphatase activity. On the 21st day, bone markers, osteopontin and osteocalcin, were detected by the PCR analysis. This study shows that the chitosan-gelatin-chondroitin 3D structure provides a good environment for the osteogenic process and enhances cellular proliferation.

  2. 3D chitosan-gelatin-chondroitin porous scaffold improves osteogenic differentiation of mesenchymal stem cells.

    Science.gov (United States)

    Machado, C B; Ventura, J M G; Lemos, A F; Ferreira, J M F; Leite, M F; Goes, A M

    2007-06-01

    A porous 3D scaffold was developed to support and enhance the differentiation process of mesenchymal stem cells (MSC) into osteoblasts in vitro. The 3D scaffold was made with chitosan, gelatin and chondroitin and it was crosslinked by EDAC. The scaffold physicochemical properties were evaluated. SEM revealed the high porosity and interconnection of pores in the scaffold; rheological measurements show that the scaffold exhibits a characteristic behavior of strong gels. The elastic modulus found in compressive tests of the crosslinked scaffold was about 50 times higher than the non-crosslinked one. After 21 days, the 3D matrix submitted to hydrolytic degradation loses above 40% of its weight. MSC were collected from rat bone marrow and seeded in chitosan-gelatin-chondroitin 3D scaffolds and in 2D culture plates as well. MSC were differentiated into osteoblasts for 21 days. Cell proliferation and alkaline phosphatase activity were followed weekly during the osteogenic process. The osteogenic differentiation of MSC was improved in 3D culture as shown by MTT assay and alkaline phosphatase activity. On the 21st day, bone markers, osteopontin and osteocalcin, were detected by the PCR analysis. This study shows that the chitosan-gelatin-chondroitin 3D structure provides a good environment for the osteogenic process and enhances cellular proliferation.

  3. 3D chitosan-gelatin-chondroitin porous scaffold improves osteogenic differentiation of mesenchymal stem cells

    International Nuclear Information System (INIS)

    Machado, C B; Ventura, J M G; Lemos, A F; Ferreira, J M F; Leite, M F; Goes, A M

    2007-01-01

    A porous 3D scaffold was developed to support and enhance the differentiation process of mesenchymal stem cells (MSC) into osteoblasts in vitro. The 3D scaffold was made with chitosan, gelatin and chondroitin and it was crosslinked by EDAC. The scaffold physicochemical properties were evaluated. SEM revealed the high porosity and interconnection of pores in the scaffold; rheological measurements show that the scaffold exhibits a characteristic behavior of strong gels. The elastic modulus found in compressive tests of the crosslinked scaffold was about 50 times higher than the non-crosslinked one. After 21 days, the 3D matrix submitted to hydrolytic degradation loses above 40% of its weight. MSC were collected from rat bone marrow and seeded in chitosan-gelatin-chondroitin 3D scaffolds and in 2D culture plates as well. MSC were differentiated into osteoblasts for 21 days. Cell proliferation and alkaline phosphatase activity were followed weekly during the osteogenic process. The osteogenic differentiation of MSC was improved in 3D culture as shown by MTT assay and alkaline phosphatase activity. On the 21st day, bone markers, osteopontin and osteocalcin, were detected by the PCR analysis. This study shows that the chitosan-gelatin-chondroitin 3D structure provides a good environment for the osteogenic process and enhances cellular proliferation

  4. 3D porous chitosan scaffolds suit survival and neural differentiation of dental pulp stem cells.

    Science.gov (United States)

    Feng, Xingmei; Lu, Xiaohui; Huang, Dan; Xing, Jing; Feng, Guijuan; Jin, Guohua; Yi, Xin; Li, Liren; Lu, Yuanzhou; Nie, Dekang; Chen, Xiang; Zhang, Lei; Gu, Zhifeng; Zhang, Xinhua

    2014-08-01

    A key aspect of cell replacement therapy in brain injury treatment is construction of a suitable biomaterial scaffold that can effectively carry and transport the therapeutic cells to the target area. In the present study, we created small 3D porous chitosan scaffolds through freeze-drying, and showed that these can support and enhance the differentiation of dental pulp stem cells (DPSCs) to nerve cells in vitro. The DPSCs were collected from the dental pulp of adult human third molars. At a swelling rate of ~84.33 ± 10.92 %, the scaffold displayed high porosity and interconnectivity of pores, as revealed by SEM. Cell counting kit-8 assay established the biocompatibility of the chitosan scaffold, supporting the growth and survival of DPSCs. The successful neural differentiation of DPSCs was assayed by RT-PCR, western blotting, and immunofluorescence. We found that the scaffold-attached DPSCs showed high expression of Nestin that decreased sharply following induction of differentiation. Exposure to the differentiation media also increased the expression of neural molecular markers Microtubule-associated protein 2, glial fibrillary acidic protein, and 2',3'-cyclic nucleotide phosphodiesterase. This study demonstrates that the granular 3D chitosan scaffolds are non-cytotoxic, biocompatible, and provide a conducive and favorable micro-environment for attachment, survival, and neural differentiation of DPSCs. These scaffolds have enormous potential to facilitate future advances in treatment of brain injury.

  5. Control of cell proliferation by a porous chitosan scaffold with multiple releasing capabilities

    Science.gov (United States)

    Cai, Shu-Jyun; Li, Ching-Wen; Weihs, Daphne; Wang, Gou-Jen

    2017-01-01

    Abstract The aim of this study was to develop a porous chitosan scaffold with long-acting drug release as an artificial dressing to promote skin wound healing. The dressing was fabricated by pre-freezing at different temperatures (−20 and −80 °C) for different periods of time, followed by freeze-drying to form porous chitosan scaffolds with different pore sizes. The chitosan scaffolds were then used to investigate the effect of the controlled release of fibroblast growth factor-basic (bFGF) and transforming growth factor-β1 (TGFβ1) on mouse fibroblast cells (L929) and bovine carotid endothelial cells (BEC). The biocompatibility of the prepared chitosan scaffold was confirmed with WST-1 proliferation and viability assay, which demonstrated that the material is suitable for cell growth. The results of this study show that the pore sizes of the porous scaffolds prepared by freeze-drying can change depending on the pre-freezing temperature and time via the formation of ice crystals. In this study, the scaffolds with the largest pore size were found to be 153 ± 32 μm and scaffolds with the smallest pores to be 34 ± 9 μm. Through cell culture analysis, it was found that the concentration that increased proliferation of L929 cells for bFGF was 0.005 to 0.1 ng/mL, and the concentration for TGFβ1 was 0.005 to 1 ng/mL. The cell culture of the chitosan scaffold and growth factors shows that 3.75 ng of bFGF in scaffolds with pore sizes of 153 ± 32 μm can promote L929 cell proliferation, while 400 pg of TGFβ1 in scaffolds with pore size of 34 ± 9 μm can enhance the proliferation of L929 cells, but also inhibit BEC proliferation. It is proposed that the prepared chitosan scaffolds can form a multi-drug (bFGF and TGFβ1) release dressing that has the ability to control wound healing via regulating the proliferation of different cell types. PMID:29230255

  6. Production and characterization of chitosan/gelatin/β-TCP scaffolds for improved bone tissue regeneration

    Energy Technology Data Exchange (ETDEWEB)

    Serra, I.R.; Fradique, R.; Vallejo, M.C.S.; Correia, T.R.; Miguel, S.P.; Correia, I.J., E-mail: icorreia@ubi.pt

    2015-10-01

    Recently, bone tissue engineering emerged as a viable therapeutic alternative, comprising bone implants and new personalized scaffolds to be used in bone replacement and regeneration. In this study, biocompatible scaffolds were produced by freeze-drying, using different formulations (chitosan, chitosan/gelatin, chitosan/β-TCP and chitosan/gelatin/β-TCP) to be used as temporary templates during bone tissue regeneration. Sample characterization was performed through attenuated total reflectance-Fourier transform infrared spectroscopy, X-ray diffraction and energy dispersive spectroscopy analysis. Mechanical characterization and porosity analysis were performed through uniaxial compression test and liquid displacement method, respectively. In vitro studies were also done to evaluate the biomineralization activity and the cytotoxic profile of the scaffolds. Scanning electron and confocal microscopy analysis were used to study cell adhesion and proliferation at the scaffold surface and within their structure. Moreover, the antibacterial activity of the scaffolds was also evaluated through the agar diffusion method. Overall, the results obtained revealed that the produced scaffolds are bioactive and biocompatible, allow cell internalization and show antimicrobial activity against Staphylococcus aureus. Such, make these 3D structures as potential candidates for being used on the bone tissue regeneration, since they promote cell adhesion and proliferation and also prevent biofilm development at their surfaces, which is usually the main cause of implant failure. - Highlights: • Production of 3D scaffolds composed by chitosan/gelatin/β-TCP by freeze-drying for bone regeneration • Physicochemical characterization of the bone substitutes by SEM, FTIR, XRD and EDS • Evaluation of the cytotoxic profile and antibacterial activity of the 3D structures through in vitro assays.

  7. Production and characterization of chitosan/gelatin/β-TCP scaffolds for improved bone tissue regeneration

    International Nuclear Information System (INIS)

    Serra, I.R.; Fradique, R.; Vallejo, M.C.S.; Correia, T.R.; Miguel, S.P.; Correia, I.J.

    2015-01-01

    Recently, bone tissue engineering emerged as a viable therapeutic alternative, comprising bone implants and new personalized scaffolds to be used in bone replacement and regeneration. In this study, biocompatible scaffolds were produced by freeze-drying, using different formulations (chitosan, chitosan/gelatin, chitosan/β-TCP and chitosan/gelatin/β-TCP) to be used as temporary templates during bone tissue regeneration. Sample characterization was performed through attenuated total reflectance-Fourier transform infrared spectroscopy, X-ray diffraction and energy dispersive spectroscopy analysis. Mechanical characterization and porosity analysis were performed through uniaxial compression test and liquid displacement method, respectively. In vitro studies were also done to evaluate the biomineralization activity and the cytotoxic profile of the scaffolds. Scanning electron and confocal microscopy analysis were used to study cell adhesion and proliferation at the scaffold surface and within their structure. Moreover, the antibacterial activity of the scaffolds was also evaluated through the agar diffusion method. Overall, the results obtained revealed that the produced scaffolds are bioactive and biocompatible, allow cell internalization and show antimicrobial activity against Staphylococcus aureus. Such, make these 3D structures as potential candidates for being used on the bone tissue regeneration, since they promote cell adhesion and proliferation and also prevent biofilm development at their surfaces, which is usually the main cause of implant failure. - Highlights: • Production of 3D scaffolds composed by chitosan/gelatin/β-TCP by freeze-drying for bone regeneration • Physicochemical characterization of the bone substitutes by SEM, FTIR, XRD and EDS • Evaluation of the cytotoxic profile and antibacterial activity of the 3D structures through in vitro assays

  8. Development of a Novel Scaffold of Chitosan, Type IV Collagen and Integrin α3β1 As Alternative Scaffold for Primary Culture of Podocytes

    Directory of Open Access Journals (Sweden)

    Diana Ginette Zárate-Triviño

    2018-06-01

    Full Text Available Loss of podocytes has been a main pathology present in renal diseases; the leak of these specialized cells increases the permeability of the glomerular basal membrane (GMB and protein release affecting the glomeruli, the main structure of the kidney. The study of different physiopathology mechanism has been a challenge because of the short lifetime of podocytes in vitro. We obtained and characterized composites based on Chitosan (CTS, polyvinyl alcohol (PVA, type IV collagen and integrin α3β1 as a possible application in primary culture of podocytes. Podocytes were extracted from the urine of patients with Idiopathic Nephrotic Syndrome (INS. To evaluate biocompatibility, we assessed cell viability through the lactate dehydrogenase assay. Immunohistochemical staining was used to detect the expression of specific proteins from podocytes such as podocin, and podocalyxin and CD80, a marker of cellular stress. The results showed that our synthesis method promotes the copolymerization of the components in the scaffold. Due to its reactivity, the amine group of chitosan made links with type IV collagen and integrin α3β1. The swelling and degradation tests allowed us to select the material with the best mechanical properties for cellular culture. The expression of podocin and podocalyxin remains the same in the culture of podocytes on the scaffold; in contrast, CD80 expression increased. The viability of podocytes cultured on the CTS/PVA/type IV collagen/integrin α3β1 scaffold increased in comparison to the culture control.

  9. Biocomposite scaffolds for bone regeneration: Role of chitosan and hydroxyapatite within poly-3-hydroxybutyrate-co-3-hydroxyvalerate on mechanical properties and in vitro evaluation.

    Science.gov (United States)

    Zhang, Sai; Prabhakaran, Molamma P; Qin, Xiaohong; Ramakrishna, Seeram

    2015-11-01

    Bio-engineered scaffolds for bone tissue regeneration is an exploding area of research mainly because they can satisfy the essential demands and current challenges in bone replacement therapies, by imitating the extracellular matrix (ECM) of the native bone. We fabricated bio-composite nanofibrous scaffolds with a blend of poly-3-hydroxybutyrate-co-3-hydroxyvalerate (PHBV), chitosan (CTS) and hydroxyapatite (HA) during this study. Morphological evaluation confirmed the fiber diameters of PHBV, PHBV/CTS (90:10), PHBV/CTS/HA4 (85.5:9.5:5) and PHBV/CTS/HA8 (81:9:10) as 405 ± 74 nm, 334 ± 82 nm, 316 ± 103 nm and 256 ± 110 nm, respectively. The PHBV/CTS/HA4 and PHBV/CTS/HA8 scaffolds were capable of enduring the long term culture of human fetal osteoblasts (hFOB) with ultimate tensile strength of 3.55 ± 0.22 MPa and 4.19 ± 0.19 MPa, respectively. The proliferation of osteoblasts on PHBV/CTS/HA8 scaffold was found 34.10% higher than that on PHBV scaffold on day 20. Cell maturation identified by alkaline phosphatase activity on day 20 was significantly higher on PHBV/CTS/HA8 scaffold than that on PHBV scaffold. The cells on PHBV/CTS/HA8 scaffold also acquired higher mineral deposition (25.79%) than the mineral deposition on PHBV scaffold by day 20, confirmed by EDX analysis. Based on the results, we concluded that the electrospun PHBV/CTS/HA8 scaffolds hold great potential to promote the regeneration of bone tissue due to the synergistic effect of chitosan and HA, whereby chitosan provided cell recognition sites while HA acted as a chelating agent for organizing the apatite-like mineralization. Copyright © 2015 Elsevier Ltd. All rights reserved.

  10. Incorporation of zinc oxide nanoparticles into chitosan-collagen 3D porous scaffolds: Effect on morphology, mechanical properties and cytocompatibility of 3D porous scaffolds.

    Science.gov (United States)

    Ullah, Saleem; Zainol, Ismail; Idrus, Ruszymah Hj

    2017-11-01

    The zinc oxide nanoparticles (particles size chitosan-collagen 3D porous scaffolds and investigated the effect of zinc oxide nanoparticles incorporation on microstructure, mechanical properties, biodegradation and cytocompatibility of 3D porous scaffolds. The 0.5%, 1.0%, 2.0% and 4.0% zinc oxide nanoparticles chitosan-collagen 3D porous scaffolds were fabricated via freeze-drying technique. The zinc oxide nanoparticles incorporation effects consisting in chitosan-collagen 3D porous scaffolds were investigated by mechanical and swelling tests, and effect on the morphology of scaffolds examined microscopically. The biodegradation and cytocompatibility tests were used to investigate the effects of zinc oxide nanoparticles incorporation on the ability of scaffolds to use for tissue engineering application. The mean pore size and swelling ratio of scaffolds were decreased upon incorporation of zinc oxide nanoparticles however, the porosity, tensile modulus and biodegradation rate were increased upon incorporation of zinc oxide nanoparticles. In vitro culture of human fibroblasts and keratinocytes showed that the zinc oxide nanoparticles facilitated cell adhesion, proliferation and infiltration of chitosan-collagen 3D porous scaffolds. It was found that the zinc oxide nanoparticles incorporation enhanced porosity, tensile modulus and cytocompatibility of chitosan-collagen 3D porous scaffolds. Copyright © 2017 Elsevier B.V. All rights reserved.

  11. Chitosan-Based Hyaluronic Acid Hybrid Polymer Fibers as a Scaffold Biomaterial for Cartilage Tissue Engineering

    Directory of Open Access Journals (Sweden)

    Shintarou Yamane

    2010-12-01

    Full Text Available An ideal scaffold material is one that closely mimics the natural environment in the tissue-specific extracellular matrix (ECM. Therefore, we have applied hyaluronic acid (HA, which is a main component of the cartilage ECM, to chitosan as a fundamental material for cartilage regeneration. To mimic the structural environment of cartilage ECM, the fundamental structure of a scaffold should be a three-dimensional (3D system with adequate mechanical strength. We structurally developed novel polymer chitosan-based HA hybrid fibers as a biomaterial to easily fabricate 3D scaffolds. This review presents the potential of a 3D fabricated scaffold based on these novel hybrid polymer fibers for cartilage tissue engineering.

  12. Biocompatibility assessment of porous chitosan-Nafion and chitosan-PTFE composites in vivo.

    Science.gov (United States)

    Liu, Bo-Ji; Ma, Li-Nan; Su, Juan; Jing, Wei-Wei; Wei, Min-Jie; Sha, Xian-Zheng

    2014-06-01

    Chitosan (CS) is widely used as a scaffold material in tissue engineering. The objective of this study was to test whether porous chitosan membrane (PCSM) coating for Nafion used in implantable sensor reduced fibrous capsule (FC) density and promoted superior vascularization compared with PCSM coating for polytetrafluoroethylene (PTFE). PCSM was fabricated with solvent casting/particulate leaching method using silica gel as porogen and characterized in vitro. Then, PCSM-Nafion and PCSM-PTFE composites were assembled with hydrated PCSM and implanted subcutaneously in rats. The histological analysis was performed in comparison with Nafion and PTFE. Implants were explanted 35, 65, and 100 days after the implantation. Histological assessments indicated that both composites achieved presumed effects of porous coatings on decreasing collagen deposition and promoting angiogenesis. PCSM-PTFE exerted higher collagen deposition by area ratio, both within and outside, compared with that of PCSM-Nafion. Angiogenesis within and outside the PCSM-Nafion both increased over time, but that of the PCSM-PTFE within decreased. Copyright © 2013 Wiley Periodicals, Inc.

  13. Fabrication and evaluation of a sustained-release chitosan-based scaffold embedded with PLGA microspheres.

    Science.gov (United States)

    Song, Kedong; Liu, Yingchao; Macedo, Hugo M; Jiang, Lili; Li, Chao; Mei, Guanyu; Liu, Tianqing

    2013-04-01

    Nutrient depletion within three-dimensional (3D) scaffolds is one of the major hurdles in the use of this technology to grow cells for applications in tissue engineering. In order to help in addressing it, we herein propose to use the controlled release of encapsulated nutrients within polymer microspheres into chitosan-based 3D scaffolds, wherein the microspheres are embedded. This method has allowed maintaining a stable concentration of nutrients within the scaffolds over the long term. The polymer microspheres were prepared using multiple emulsions (w/o/w), in which bovine serum albumin (BSA) and poly (lactic-co-glycolic) acid (PLGA) were regarded as the protein pattern and the exoperidium material, respectively. These were then mixed with a chitosan solution in order to form the scaffolds by cryo-desiccation. The release of BSA, entrapped within the embedded microspheres, was monitored with time using a BCA kit. The morphology and structure of the PLGA microspheres containing BSA before and after embedding within the scaffold were observed under a scanning electron microscope (SEM). These had a round shape with diameters in the range of 27-55 μm, whereas the chitosan-based scaffolds had a uniform porous structure with the microspheres uniformly dispersed within their 3D structure and without any morphological change. In addition, the porosity, water absorption and degradation rate at 37 °C in an aqueous environment of 1% chitosan-based scaffolds were (92.99±2.51) %, (89.66±0.66) % and (73.77±3.21) %, respectively. The studies of BSA release from the embedded microspheres have shown a sustained and cumulative tendency with little initial burst, with (20.24±0.83) % of the initial amount released after 168 h (an average rate of 0.12%/h). The protein concentration within the chitosan-based scaffolds after 168 h was found to be (11.44±1.81)×10(-2) mg/mL. This novel chitosan-based scaffold embedded with PLGA microspheres has proven to be a promising technique

  14. Biomimetic properties of an injectable chitosan/nano-hydroxyapatite/collagen composite

    Energy Technology Data Exchange (ETDEWEB)

    Huang Zhi [Laboratory of Advanced Materials, Department of Materials Science and Engineering, Tsinghua University, Beijing 100084 (China); Feng Qingling, E-mail: biomater@mail.tsinghua.edu.cn [Laboratory of Advanced Materials, Department of Materials Science and Engineering, Tsinghua University, Beijing 100084 (China); Yu Bo; Li Songjian [Department of Orthopedics, Zhujiang Hospital of Southern Medical University, Guangzhou 510282 (China)

    2011-04-08

    To meet the challenges of designing an injectable scaffold and regenerating bone with complex three-dimensional (3D) structures, a biomimetic and injectable hydrogel scaffold based on nano-hydroxyapatite (HA), collagen (Col) and chitosan (Chi) is synthesized. The chitosan/nano-hydroxyapatite/collagen (Chi/HA/Col) solution rapidly forms a stable gel at body temperature. It shows some features of natural bone both in main composition and microstructure. The Chi/HA/Col system can be expected as a candidate for workable systemic minimally invasive scaffolds with surface properties similar to physiological bone based on scanning electron microscopic (SEM), transmission electron microscopy (TEM), X-ray photoelectron spectroscopy (XPS), X-ray diffraction (XRD) and Fourier transform infrared spectroscopy (FT-IR) results.

  15. Biomimetic properties of an injectable chitosan/nano-hydroxyapatite/collagen composite

    International Nuclear Information System (INIS)

    Huang Zhi; Feng Qingling; Yu Bo; Li Songjian

    2011-01-01

    To meet the challenges of designing an injectable scaffold and regenerating bone with complex three-dimensional (3D) structures, a biomimetic and injectable hydrogel scaffold based on nano-hydroxyapatite (HA), collagen (Col) and chitosan (Chi) is synthesized. The chitosan/nano-hydroxyapatite/collagen (Chi/HA/Col) solution rapidly forms a stable gel at body temperature. It shows some features of natural bone both in main composition and microstructure. The Chi/HA/Col system can be expected as a candidate for workable systemic minimally invasive scaffolds with surface properties similar to physiological bone based on scanning electron microscopic (SEM), transmission electron microscopy (TEM), X-ray photoelectron spectroscopy (XPS), X-ray diffraction (XRD) and Fourier transform infrared spectroscopy (FT-IR) results.

  16. Fabrication and characterization of hydrothermal cross-linked chitosan porous scaffolds for cartilage tissue engineering applications.

    Science.gov (United States)

    Shamekhi, Mohammad Amin; Rabiee, Ahmad; Mirzadeh, Hamid; Mahdavi, Hamid; Mohebbi-Kalhori, Davod; Baghaban Eslaminejad, Mohamadreza

    2017-11-01

    The use of various chemical cross-linking agents for the improvement of scaffolds physical and mechanical properties is a common practical method, which is limited by cytotoxicity effects. Due to exerting contract type forces, chondrocytes are known to implement shrinkage on the tissue engineered constructs, which can be avoided by the scaffold cross-linking. In the this research, chitosan scaffolds are cross-linked with hydrothermal treatment with autoclave sterilization time of 0, 10, 20 and 30min, to avoid the application of the traditional chemical toxic materials. The optimization studies with gel content and crosslink density measurements indicate that for 20min sterilization time, the gel content approaches to ~80%. The scaffolds are fully characterized by the conventional techniques such as SEM, porosity and permeability, XRD, compression, thermal analysis and dynamic mechanical thermal analysis (DMTA). FT-IR studies shows that autoclave inter-chain cross-linking reduces the amine group absorption at 1560cm -1 and increase the absorption of N-acetylated groups at 1629cm -1 . It is anticipated, that this observation evidenced by chitosan scaffold browning upon autoclave cross-linking is an indication of the familiar maillard reaction between amine moieties and carbonyl groups. The biodegradation rate analysis shows that chitosan scaffolds with lower concentrations, possess suitable degradation rate for cartilage tissue engineering applications. In addition, cytotoxicity analysis shows that fabricated scaffolds are biocompatible. The human articular chondrocytes seeding into 3D cross-linked scaffolds shows a higher viability and proliferation in comparison with the uncross-linked samples and 2D controls. Investigation of cell morphology on the scaffolds by SEM, shows a more spherical morphology of chondrocytes on the cross-linked scaffolds for 21days of in vitro culture. Copyright © 2017. Published by Elsevier B.V.

  17. Effects of Neutralization and Crosslinking Agents on the Morphology of Chitosan Electrospun Scaffolds

    Directory of Open Access Journals (Sweden)

    Maryam Mashayekhi

    2017-01-01

    Full Text Available Chitosan, a natural polymer derived from chitin by deacetylation process of chitin, has gained an enormous interest in tissue engineering due to its unique features such as antibacterial activity and wound healing properties. Electrospinning of acidified chitosan solution is one of the most widely-used approaches in fabrication of 3D scaffolds. Although there are some reports addressing morphology tailoring of the chitosan nanofibers through solution electrospinning, there is no comparative report concerning the neutralization and stabilization conditions of chitosan electrospun fibers. Therefore, this article compares the effects of different neutralizing agents such as aqueous solutions of sodium carbonate (Na2CO3 and potassium carbonate (K2CO3, and crosslinking reagents including glutaraldehyde (GA and genipin on morphology of electrospun chitosan fibers. After neutralization and stabilization processes, Fourier transform infrared spectroscopy (FTIR was employed to investigate the morphology of fibers. Furthermore, the influence of the aforementioned parameters on stability of fibers was probed using scanning electron microscopy. SEM images illustrated that the scaffold resulting from electrospinning of 4 wt% chitosan solution in a mixture of trifluoroacetic acid (TFA and dichloromethane (DCM possessed a well-formed nanofibrous structure. Afterwards, different methods for neutralization and stabilization of the electrospun chitosan nanofiber mats were performed. In this respect, aqueous solutions of both Na2CO3 and K2CO3 salts (1M were employed as neutralization agents and GA and genipin were used as two different crosslinking agents. Based on SEM analysis, the chitosan fibers, crosslinked with genipin, showed better morphology than a scaffold which was crosslinked with glutaraldehyde

  18. Protein growth factors loaded highly porous chitosan scaffold: A comparison of bone healing properties

    Energy Technology Data Exchange (ETDEWEB)

    Nandi, Samit K., E-mail: samitnandi1967@gmail.com [Department of Veterinary Surgery and Radiology, West Bengal University of Animal and Fishery Sciences, Kolkata (India); Kundu, Biswanath, E-mail: biswa_kundu@rediffmail.com [Bioceramics and Coating Division, CSIR—Central Glass and Ceramic Research Institute, Kolkata (India); Basu, Debabrata [Bioceramics and Coating Division, CSIR—Central Glass and Ceramic Research Institute, Kolkata (India)

    2013-04-01

    Present study aimed to investigate and compare effectiveness of porous chitosan alone and in combination with insulin like growth factor-1 (IGF-1) and bone morphogenetic protein-2 (BMP-2) in bone healing. Highly porous (85 ± 2%) with wide distribution of macroporous (70–900 μm) chitosan scaffolds were fabricated as bone substitutes by employing a simple liquid hardening method using 2% (w/v) chitosan suspension. IGF-1 and BMP-2 were infiltrated using vacuum infiltration with freeze drying method. Adsorption efficiency was found to be 87 ± 2 and 90 ± 2% for BMP-2 and IGF-1 respectively. After thorough material characterization (pore details, FTIR and SEM), samples were used for subsequent in vivo animal trial. Eighteen rabbit models were used to evaluate and compare control (chitosan) (group A), chitosan with IGF-1 (group B) and chitosan with BMP-2 (group C) in the repair of critical size bone defect in tibia. Radiologically, there was evidence of radiodensity in defect area from 60th day (initiated on 30th day) in groups B and C as compared to group A and attaining nearly bony density in most of the part at day 90. Histological results depicted well developed osteoblastic proliferation around haversian canal along with proliferating fibroblast, vascularization and reticular network which was more pronounced in group B followed by groups C and A. Fluorochrome labeling and SEM studies in all groups showed similar outcome. Hence, porous chitosan alone and in combination with growth factors (GFs) can be successfully used for bone defect healing with slight advantage of IGF-1 in chitosan samples. - Highlights: ► Fabrication and characterization of porous chitosan with or without IGF-1 and BMP-2 ► Highly porous growth factor loaded chitosan studied in animal subjects for 3 months ► Parameters studied: histopathology, radiology and fluorochrome labeling ► IGF-1 loaded porous chitosan found to be very effective for bone defect healing.

  19. Protein growth factors loaded highly porous chitosan scaffold: A comparison of bone healing properties

    International Nuclear Information System (INIS)

    Nandi, Samit K.; Kundu, Biswanath; Basu, Debabrata

    2013-01-01

    Present study aimed to investigate and compare effectiveness of porous chitosan alone and in combination with insulin like growth factor-1 (IGF-1) and bone morphogenetic protein-2 (BMP-2) in bone healing. Highly porous (85 ± 2%) with wide distribution of macroporous (70–900 μm) chitosan scaffolds were fabricated as bone substitutes by employing a simple liquid hardening method using 2% (w/v) chitosan suspension. IGF-1 and BMP-2 were infiltrated using vacuum infiltration with freeze drying method. Adsorption efficiency was found to be 87 ± 2 and 90 ± 2% for BMP-2 and IGF-1 respectively. After thorough material characterization (pore details, FTIR and SEM), samples were used for subsequent in vivo animal trial. Eighteen rabbit models were used to evaluate and compare control (chitosan) (group A), chitosan with IGF-1 (group B) and chitosan with BMP-2 (group C) in the repair of critical size bone defect in tibia. Radiologically, there was evidence of radiodensity in defect area from 60th day (initiated on 30th day) in groups B and C as compared to group A and attaining nearly bony density in most of the part at day 90. Histological results depicted well developed osteoblastic proliferation around haversian canal along with proliferating fibroblast, vascularization and reticular network which was more pronounced in group B followed by groups C and A. Fluorochrome labeling and SEM studies in all groups showed similar outcome. Hence, porous chitosan alone and in combination with growth factors (GFs) can be successfully used for bone defect healing with slight advantage of IGF-1 in chitosan samples. - Highlights: ► Fabrication and characterization of porous chitosan with or without IGF-1 and BMP-2 ► Highly porous growth factor loaded chitosan studied in animal subjects for 3 months ► Parameters studied: histopathology, radiology and fluorochrome labeling ► IGF-1 loaded porous chitosan found to be very effective for bone defect healing

  20. Chitosan-based scaffolds for the support of smooth muscle constructs in intestinal tissue engineering

    Science.gov (United States)

    Zakhem, Elie; Raghavan, Shreya; Gilmont, Robert R; Bitar, Khalil N

    2012-01-01

    Intestinal tissue engineering is an emerging field due to a growing demand for intestinal lengthening and replacement procedures secondary to massive resections of the bowel. Here, we demonstrate the potential use of a chitosan/collagen scaffold as a 3D matrix to support the bioengineered circular muscle constructs maintain their physiological functionality. We investigated the biocompatibility of chitosan by growing rabbit colonic circular smooth muscle cells (RCSMCs) on chitosan-coated plates. The cells maintained their spindle-like morphology and preserved their smooth muscle phenotypic markers. We manufactured tubular scaffolds with central openings composed of chitosan and collagen in a 1:1 ratio. Concentrically-aligned 3D circular muscle constructs were bioengineered using fibrin-based hydrogel seeded with RCSMCs. The constructs were placed around the scaffold for 2 weeks, after which they were taken off and tested for their physiological functionality. The muscle constructs contracted in response to Acetylcholine (Ach) and potassium chloride (KCl) and they relaxed in response to vasoactive intestinal peptide (VIP). These results demonstrate that chitosan is a biomaterial possibly suitable for intestinal tissue engineering applications. PMID:22483012

  1. Biofabrication of chitosan-silver composite SERS substrates enabling quantification of adenine by a spectroscopic shift

    International Nuclear Information System (INIS)

    Luo, X L; Bentley, W E; Buckhout-White, S; Rubloff, G W

    2011-01-01

    Surface-enhanced Raman scattering (SERS) has grown dramatically as an analytical tool for the sensitive and selective detection of molecules adsorbed on nano-roughened noble metal structures. Quantification with SERS based on signal intensity remains challenging due to the complicated fabrication process to obtain well-dispersed nanoparticles and well-ordered substrates. We report a new biofabrication strategy of SERS substrates that enable quantification through a newly discovered spectroscopic shift resulting from the chitosan-analyte interactions in solution. We demonstrate this phenomenon by the quantification of adenine, which is an essential part of the nucleic acid structure and a key component in pathways which generate signal molecules for bacterial communications. The SERS substrates were fabricated simply by sequential electrodeposition of chitosan on patterned gold electrodes and electroplating of a silver nitrate solution through the chitosan scaffold to form a chitosan-silver nanoparticle composite. Active SERS signals of adenine solutions were obtained in real time from the chitosan-silver composite substrates with a significant concentration-dependent spectroscopic shift. The Lorentzian curve fitting of the dominant peaks suggests the presence of two separate peaks with a concentration-dependent area percentage of the separated peaks. The chitosan-mediated composite SERS substrates can be easily biofabricated on predefined electrodes within microfluidic channels for real-time detection in microsystems.

  2. Mechanical Properties, Cytocompatibility and Manufacturability of Chitosan:PEGDA Hybrid-Gel Scaffolds by Stereolithography.

    Science.gov (United States)

    Morris, Viola B; Nimbalkar, Siddharth; Younesi, Mousa; McClellan, Phillip; Akkus, Ozan

    2017-01-01

    Extracellular matrix mimetic hydrogels which hybridize synthetic and natural polymers offer molecularly-tailored, bioactive properties and tunable mechanical strength. In addition, 3D bioprinting by stereolithography allows fabrication of internal pores and defined macroscopic shapes. In this study, we formulated a hybrid biocompatible resin using natural and synthetic polymers (chitosan and polyethylene glycol diacrylate (PEGDA), respectively) by controlling molecular weight of chitosan, feed-ratios, and photo-initiator concentration. Ear-shaped, hybrid scaffolds were fabricated by a stereolithographic method using a 405 nm laser. Hybrid hydrogel scaffolds of chitosan (50-190 kDa) and PEGDA (575 Da) were mixed at varying feed-ratios. Some of the cationic, amino groups of chitosan were neutralized by dialysis in acidic solution containing chitosan in excess of sodium acetate solution to inhibit quenching of newly formed photoradicals. A feed-ratio of 1:7.5 was found to be the most appropriate of the formulations considered in this study in terms of mechanical properties, cell adhesion, and printability. The biofabricated hybrid scaffold showed interconnected, homogeneous pores with a nominal pore size of 50 µm and an elastic modulus of ~400 kPa. Moreover, long-term cell viability and cell spreading was observed via actin filament staining. Printability of the biocompatible resin was confirmed by printing thresholded MR images of an ear and the feed ratio of 1:7.5 provided the most faithful reproduction of the shape. To the best of our knowledge, this is the first report of stereolithographic printing hybridizing cell-adhesive properties of chitosan with mechanical robustness of PEG in scaffolds suitable for repair of complex tissue geometries, such as those of the human ear.

  3. The effect of hydroxyapatite addition on the mechanical properties of polyvinyl alcohol/chitosan biomaterials for bone scaffolds application

    Science.gov (United States)

    Ramahdita, Ghiska; Puspita, Debie Maya; Albab, Muh Fadhil; Alfata, Rowi; Sofyan, Nofrijon; Yuwono, Akhmad Herman

    2018-02-01

    The increasing number of bone fracture incident in Indonesia from year to year needs an urgent problem solving of the limited bone substitute which can meet the necessary criteria for that purpose. Motivated by this, therefore, the current study is focusing on the optimization of material properties used as bone scaffold. A biomaterial of polyvinyl alcohol (PVA)/chitosan-hydroxyapatite (HA) composite was successfully made by wet chemistry method, followed by freeze thawing and freeze drying. For comparison purposes, the percentage of HA has been varied from 0, 25 and 40 % (wt/v). The resulting composites were characterized by Fourier transform infrared (FTIR) spectroscopy, scanning electron microscope (SEM), compressive test, and swelling behavior. The results showed that the addition of HA up to 40% (wt/v) has yielded a porous structure with an average pore size of 42.39 µm. In addition, the compressive modulus was enhanced from 14 MPa for 0% HA to 143, and 191 MPa for composites with the addition of HA from 25 to 40% (wt/v). The addition of HA has also reduced the swelling ratio from 296% for the sample without HA to 85 and 78 % for sample with addition of HA from 25 to 40 (wt/v), respectively. The obtained results show that PVA/chitosan-HA in this study is potential to be used as scaffold in bone tissue engineering.

  4. Biodegradability and Biocompatibility Study of Poly(Chitosan-g-lactic Acid Scaffolds

    Directory of Open Access Journals (Sweden)

    Zhe Zhang

    2012-03-01

    Full Text Available A biodegradable, biocompatible poly(chitosan-g-lactic acid (PCLA scaffold was prepared and evaluated in vitro and in vivo. The PCLA scaffold was obtained by grafting lactic acid (LA onto the amino groups on chitosan (CS without a catalyst. The PCLA scaffolds were characterized by Fourier Transform infrared spectroscopy (FT-IR and scanning electron microscopy (SEM. The biodegradabilty was determined by mass loss in vitro, and degradation in vivo as a function of feed ratio of LA/CS. Bone marrow mesenchymal stem cell (BMSC culture experiments and histological examination were performed to evaluate the PCLA scaffolds’ biocompatibility. The results indicated that PCLA was promising for tissue engineering application.

  5. New type of chitosan/2-hydroxypropyl-β-cyclodextrin composite membrane for gallic acid encapsulation and controlled release.

    Science.gov (United States)

    Paun, Gabriela; Neagu, Elena; Tache, Andreia; Radu, G L

    2014-01-01

    A new type of chitosan/2-hydroxypropyl-β-cyclodextrin composite membrane have been developed for the encapsulation and controlled release of gallic acid. The morphology of the composite membrane was investigated by infrared spectroscopy (FT-IR) and scanning electron microscopy (SEM), whereas swelling gallic acid and release properties were investigated by UV-visible spectroscopy. The release behavior with pH changes was also explored. The composite membrane based on chitosan/2-hydroxypropyl-β-cyclodextrin with gallic acid included showed improved antioxidant capacities compared to plain chitosan membrane. The information obtained in this study will facilitate the design and preparation of composite membrane based on chitosan and could open a wide range of applications, particularly its use as an antioxidant in food, food packaging, biomedical (biodegradable soft porous scaffolds for enhance the surrounding tissue regeneration), pharmaceutical and cosmetics industries.

  6. Functional 3-D cardiac co-culture model using bioactive chitosan nanofiber scaffolds.

    Science.gov (United States)

    Hussain, Ali; Collins, George; Yip, Derek; Cho, Cheul H

    2013-02-01

    The in vitro generation of a three-dimensional (3-D) myocardial tissue-like construct employing cells, biomaterials, and biomolecules is a promising strategy in cardiac tissue regeneration, drug testing, and tissue engineering applications. Despite significant progress in this field, current cardiac tissue models are not yet able to stably maintain functional characteristics of cardiomyocytes for long-term culture and therapeutic purposes. The objective of this study was to fabricate bioactive 3-D chitosan nanofiber scaffolds using an electrospinning technique and exploring its potential for long-term cardiac function in the 3-D co-culture model. Chitosan is a natural polysaccharide biomaterial that is biocompatible, biodegradable, non-toxic, and cost effective. Electrospun chitosan was utilized to provide structural scaffolding characterized by scale and architectural resemblance to the extracellular matrix (ECM) in vivo. The chitosan fibers were coated with fibronectin via adsorption in order to enhance cellular adhesion to the fibers and migration into the interfibrous milieu. Ventricular cardiomyocytes were harvested from neonatal rats and studied in various culture conditions (i.e., mono- and co-cultures) for their viability and function. Cellular morphology and functionality were examined using immunofluorescent staining for alpha-sarcomeric actin (SM-actin) and gap junction protein, Connexin-43 (Cx43). Scanning electron microscopy (SEM) and light microscopy were used to investigate cellular morphology, spatial organization, and contractions. Calcium indicator was used to monitor calcium ion flux of beating cardiomyocytes. The results demonstrate that the chitosan nanofibers retained their cylindrical morphology in long-term cell cultures and exhibited good cellular attachment and spreading in the presence of adhesion molecule, fibronectin. Cardiomyocyte mono-cultures resulted in loss of cardiomyocyte polarity and islands of non-coherent contractions. However

  7. Scaffold of chitosan-sodium alginate and hydroxyapatite with application potential for bone regeneration

    International Nuclear Information System (INIS)

    Rebelo, Marcia de A.; Alves, Thais F.R.; Lopes, Francielly C.C.N; Oliveira Junior, Jose Martins de; Pontes, Katiusca S.; Fogaca, Bruna A.C.; Chaud, Marco V.

    2015-01-01

    Scaffold for organic tissue regeneration are architectural, three-dimensional, porous, biocompatible and biodegradable devices. The first challenges to be met in the development of these devices to mimic the biomechanical properties of the target tissue. The aim of this study was to develop and to characterize scaffolds composed of chitosan (Ch), sodium alginate (SA), hydroxyapatite (HA). The scaffolds were obtained by lyophilization. HA has been incorporated into the polymer dispersion in Ch-AS concentration of 20 and 60%. The mechanical properties of the scaffold were determined by tensile and compression tests. Swelling capacity was assessed in the presence of simulated saliva, purified water, HCl 0.01M, NaOH 0.01M. The calcium content was quantified using fluorescence X-rays. Analysis of the results indicates that the Qt-AS-HA-60% scaffold obtained by lyophilization meets promising properties for bone tissue regeneration. (author)

  8. A novel squid pen chitosan/hydroxyapatite/β-tricalcium phosphate composite for bone tissue engineering

    Energy Technology Data Exchange (ETDEWEB)

    Shavandi, Amin, E-mail: amin.shavandi@postgrad.otago.ac.nz [Department of Food Sciences, University of Otago, Dunedin (New Zealand); Department of Applied Sciences, University of Otago, Dunedin (New Zealand); Bekhit, Alaa El-Din A. [Department of Food Sciences, University of Otago, Dunedin (New Zealand); Sun, Zhifa; Ali, Azam [Department of Physics, University of Otago, Dunedin (New Zealand); Gould, Maree [Department of Anatomy, University of Otago, Dunedin (New Zealand)

    2015-10-01

    Squid pen chitosan was used in the fabrication of biocomposite scaffolds for bone tissue engineering. Hydroxyapatite (HA) and beta-tricalcium phosphate (β-TCP) obtained from waste mussel shells were used as the calcium phosphate source. The composite was prepared using 2.5% tripolyphosphate (TPP) and 1% glycerol as a cross-linker and plasticizer, respectively. The weight percent (wt.%) ratios of the ceramic components in the composite were 20/10/70, 30/20/50 and 40/30/30 (HA/β-TCP/Chi). The biodegradation rate and structural properties of the scaffolds were investigated. Scanning electron microscopy (SEM) and microCT(μCT) results indicated that the composites have a well defined lamellar structure with an average pore size of 200 μm. The porosity of the composites decreased from 88 to 56% by increasing the ratio of HA/β-TCP from 30 to 70%. After 28 days of incubation in a physiological solution, the scaffolds were degraded by approximately 30%. In vitro investigations showed that the composites were cytocompatible and supported the growth of L929 and Saos-2 cells. The obtained data suggests that the squid pen chitosan composites are potential candidates for bone regeneration. - Highlights: • Biocomposite scaffolds were made from mussel shells HA and β-TCP, and squid pin chitosan. • The porosity of the composites decreased with an increase in HA/β-TCP ratio. • Composites were cytocompatible and supported the growth of L929 and Saos-2 cells. • Composite containing 50% HA and β-TCP had the best mechanical properties.

  9. Fabrication of Chitosan/Poly (vinyl alcohol/Carbon Nanotube/Bioactive Glass Nanocomposite Scaffolds for Neural Tissue Engineering

    Directory of Open Access Journals (Sweden)

    S. Nikbakht Katouli

    2016-06-01

    5 and 10 wt% incorporated electrospun chitosan (CS/polyvinyl alcohol (PVA nanofibers for potential neural tissue engineering applications.The morphology, structure, and mechanical properties of the formed electrospun fibrous mats were characterized using scanning electron microscopy (SEM and mechanical testing, respectively. In vitro cell culture of embryonal carcinoma stem cells (P19 were seeded onto the electrospun scaffolds. The results showed that the incorporation of CNTs and BG nanoparticles did not appreciably affect the morphology of the CS/PVA nanofibers. The maximum tensile strength (7.9 MPa was observed in the composite sample with 5 %wt bioactive glass nanoparticles. The results suggest that BG and CNT-incorporated CS/PVA nanofibrous scaffolds with small diameters, high porosity, and promoted mechanical properties can potentially provide many possibilities for applications in the fields of neural tissue engineering and regenerative medicine.

  10. Highly defined 3D printed chitosan scaffolds featuring improved cell growth.

    Science.gov (United States)

    Elviri, Lisa; Foresti, Ruben; Bergonzi, Carlo; Zimetti, Francesca; Marchi, Cinzia; Bianchera, Annalisa; Bernini, Franco; Silvestri, Marco; Bettini, Ruggero

    2017-07-12

    The augmented demand for medical devices devoted to tissue regeneration and possessing a controlled micro-architecture means there is a need for industrial scale-up in the production of hydrogels. A new 3D printing technique was applied to the automation of a freeze-gelation method for the preparation of chitosan scaffolds with controlled porosity. For this aim, a dedicated 3D printer was built in-house: a preliminary effort has been necessary to explore the printing parameter space to optimize the printing results in terms of geometry, tolerances and mechanical properties of the product. Analysed parameters included viscosity of the starting chitosan solution, which was measured with a Brookfield viscometer, and temperature of deposition, which was determined by filming the process with a cryocooled sensor thermal camera. Optimized parameters were applied to the production of scaffolds from solutions of chitosan alone or with the addition of raffinose as a viscosity modifier. Resulting hydrogels were characterized in terms of morphology and porosity. In vitro cell culture studies comparing 3D printed scaffolds with their homologous produced by solution casting evidenced an improvement in biocompatibility deriving from the production technique as well as from the solid state modification of chitosan stemming from the addition of the viscosity modifier.

  11. Electrostatic flocking of chitosan fibres leads to highly porous, elastic and fully biodegradable anisotropic scaffolds.

    Science.gov (United States)

    Gossla, Elke; Tonndorf, Robert; Bernhardt, Anne; Kirsten, Martin; Hund, Rolf-Dieter; Aibibu, Dilibar; Cherif, Chokri; Gelinsky, Michael

    2016-10-15

    Electrostatic flocking - a common textile technology which has been applied in industry for decades - is based on the deposition of short polymer fibres in a parallel aligned fashion on flat or curved substrates, covered with a layer of a suitable adhesive. Due to their highly anisotropic properties the resulting velvet-like structures can be utilised as scaffolds for tissue engineering applications in which the space between the fibres can be defined as pores. In the present study we have developed a fully resorbable compression elastic flock scaffold from a single material system based on chitosan. The fibres and the resulting scaffolds were analysed concerning their structural and mechanical properties and the biocompatibility was tested in vitro. The tensile strength and Young's modulus of the chitosan fibres were analysed as a function of the applied sterilisation technique (ethanol, supercritical carbon dioxide, γ-irradiation and autoclaving). All sterilisation methods decreased the Young's modulus (from 14GPa to 6-12GPa). The tensile strength was decreased after all treatments - except after the autoclaving of chitosan fibres submerged in water. Compressive strength of the highly porous flock scaffolds was 18±6kPa with a elastic modulus in the range of 50-100kPa. The flocked scaffolds did not show any cytotoxic effect during indirect or direct culture of human mesenchymal stem cells or the sarcoma osteogenic cell line Saos-2. Furthermore cell adhesion and proliferation of both cell types could be observed. This is the first demonstration of a fully biodegradable scaffold manufactured by electrostatic flocking. Most tissues possess anisotropic fibrous structures. In contrast, most of the commonly used scaffolds have an isotropic morphology. By utilising the textile technology of electrostatic flocking, highly porous and clearly anisotropic scaffolds can be manufactured. Flocking leads to parallel aligned short fibres, glued on the surface of a substrate

  12. In vitro degradation of chitosan composite foams for biomedical applications and effect of bioactive glass as a crosslinker

    OpenAIRE

    Martins Talita; Moreira Cheisy D. F.; Costa-Júnior Ezequiel S.; Pereira Marivalda M.

    2018-01-01

    In tissue engineering applications, 3D scaffolds with adequate structure and composition are required to provide durability that is compatiblewith the regeneration of native tissue. In the present study, the degradation of novel flexible 3D composite foams of chitosan (CH) combined with bioactive glass (BG)was evaluated, focusing on the role of BG as a physical crosslinker in the composites, and its effect on the degradation process. Highly porous CH/BG composite foams were obtained, and an e...

  13. Physicochemical properties and bioactivity of freeze-cast chitosan nanocomposite scaffolds reinforced with bioactive glass.

    Science.gov (United States)

    Pourhaghgouy, Masoud; Zamanian, Ali; Shahrezaee, Mostafa; Masouleh, Milad Pourbaghi

    2016-01-01

    Chitosan based nanocomposite scaffolds were prepared by freeze casting method through blending constant chitosan concentration with different portions of synthesized bioactive glass nanoparticles (BGNPs). Transmission Electron Microscopy (TEM) image showed that the particles size of bioactive glass (64SiO2.28CaO.8P2O5) prepared by sol-gel method was approximately less than 20 nm. Fourier Transform Infrared Spectroscopy (FT-IR) and X-ray Diffraction (XRD) analysis showed proper interfacial bonding between BGNPs and chitosan polymers. Scanning Electron Microscopy (SEM) images depicted a unidirectional structure with homogenous distribution of BGNPs among chitosan matrix associated with the absence of pure chitosan scaffold's wall pores after addition of only 10 wt.% BGNPs. As the BGNP content increased from 0 to 50 wt.%, the compressive strength and compressive module values increased from 0.034 to 0.419 MPa and 0.41 to 10.77 MPa, respectively. Biodegradation study showed that increase in BGNP content leads to growth of weight loss amount. The in vitro biomineralization studies confirmed the bioactive nature of all nanocomposites. Amount of 30 wt.% BGNPs represented the best concentration for absorption capacity and bioactivity behaviors. Copyright © 2015. Published by Elsevier B.V.

  14. Chondrogenesis of infrapatellar fat pad derived adipose stem cells in 3D printed chitosan scaffold.

    Science.gov (United States)

    Ye, Ken; Felimban, Raed; Traianedes, Kathy; Moulton, Simon E; Wallace, Gordon G; Chung, Johnson; Quigley, Anita; Choong, Peter F M; Myers, Damian E

    2014-01-01

    Infrapatellar fat pad adipose stem cells (IPFP-ASCs) have been shown to harbor chondrogenic potential. When combined with 3D polymeric structures, the stem cells provide a source of stem cells to engineer 3D tissues for cartilage repair. In this study, we have shown human IPFP-ASCs seeded onto 3D printed chitosan scaffolds can undergo chondrogenesis using TGFβ3 and BMP6. By week 4, a pearlescent, cartilage-like matrix had formed that penetrated the top layers of the chitosan scaffold forming a 'cap' on the scaffold. Chondrocytic morphology showed typical cells encased in extracellular matrix which stained positively with toluidine blue. Immunohistochemistry demonstrated positive staining for collagen type II and cartilage proteoglycans, as well as collagen type I. Real time PCR analysis showed up-regulation of collagen type II, aggrecan and SOX9 genes when IPFP-ASCs were stimulated by TGFβ3 and BMP6. Thus, IPFP-ASCs can successfully undergo chondrogenesis using TGFβ3 and BMP6 and the cartilage-like tissue that forms on the surface of 3D-printed chitosan scaffold may prove useful as an osteochondral graft.

  15. Chondrogenesis of infrapatellar fat pad derived adipose stem cells in 3D printed chitosan scaffold.

    Directory of Open Access Journals (Sweden)

    Ken Ye

    Full Text Available Infrapatellar fat pad adipose stem cells (IPFP-ASCs have been shown to harbor chondrogenic potential. When combined with 3D polymeric structures, the stem cells provide a source of stem cells to engineer 3D tissues for cartilage repair. In this study, we have shown human IPFP-ASCs seeded onto 3D printed chitosan scaffolds can undergo chondrogenesis using TGFβ3 and BMP6. By week 4, a pearlescent, cartilage-like matrix had formed that penetrated the top layers of the chitosan scaffold forming a 'cap' on the scaffold. Chondrocytic morphology showed typical cells encased in extracellular matrix which stained positively with toluidine blue. Immunohistochemistry demonstrated positive staining for collagen type II and cartilage proteoglycans, as well as collagen type I. Real time PCR analysis showed up-regulation of collagen type II, aggrecan and SOX9 genes when IPFP-ASCs were stimulated by TGFβ3 and BMP6. Thus, IPFP-ASCs can successfully undergo chondrogenesis using TGFβ3 and BMP6 and the cartilage-like tissue that forms on the surface of 3D-printed chitosan scaffold may prove useful as an osteochondral graft.

  16. Genipin-Crosslinked Chitosan Gels and Scaffolds for Tissue Engineering and Regeneration of Cartilage and Bone.

    Science.gov (United States)

    Muzzarelli, Riccardo A A; El Mehtedi, Mohamad; Bottegoni, Carlo; Aquili, Alberto; Gigante, Antonio

    2015-12-11

    The present review article intends to direct attention to the technological advances made since 2009 in the area of genipin-crosslinked chitosan (GEN-chitosan) hydrogels. After a concise introduction on the well recognized characteristics of medical grade chitosan and food grade genipin, the properties of GEN-chitosan obtained with a safe, spontaneous and irreversible chemical reaction, and the quality assessment of the gels are reviewed. The antibacterial activity of GEN-chitosan has been well assessed in the treatment of gastric infections supported by Helicobacter pylori. Therapies based on chitosan alginate crosslinked with genipin include stem cell transplantation, and development of contraction free biomaterials suitable for cartilage engineering. Collagen, gelatin and other proteins have been associated to said hydrogels in view of the regeneration of the cartilage. Viability and proliferation of fibroblasts were impressively enhanced upon addition of poly-l-lysine. The modulation of the osteocytes has been achieved in various ways by applying advanced technologies such as 3D-plotting and electrospinning of biomimetic scaffolds, with optional addition of nano hydroxyapatite to the formulations. A wealth of biotechnological advances and know-how has permitted reaching outstanding results in crucial areas such as cranio-facial surgery, orthopedics and dentistry. It is mandatory to use scaffolds fully characterized in terms of porosity, pore size, swelling, wettability, compressive strength, and degree of acetylation, if the osteogenic differentiation of human mesenchymal stem cells is sought: in fact, the novel characteristics imparted by GEN-chitosan must be simultaneously of physico-chemical and cytological nature. Owing to their high standard, the scientific publications dated 2010-2015 have met the expectations of an interdisciplinary audience.

  17. Genipin-Crosslinked Chitosan Gels and Scaffolds for Tissue Engineering and Regeneration of Cartilage and Bone

    Directory of Open Access Journals (Sweden)

    Riccardo A. A. Muzzarelli

    2015-12-01

    Full Text Available The present review article intends to direct attention to the technological advances made since 2009 in the area of genipin-crosslinked chitosan (GEN-chitosan hydrogels. After a concise introduction on the well recognized characteristics of medical grade chitosan and food grade genipin, the properties of GEN-chitosan obtained with a safe, spontaneous and irreversible chemical reaction, and the quality assessment of the gels are reviewed. The antibacterial activity of GEN-chitosan has been well assessed in the treatment of gastric infections supported by Helicobacter pylori. Therapies based on chitosan alginate crosslinked with genipin include stem cell transplantation, and development of contraction free biomaterials suitable for cartilage engineering. Collagen, gelatin and other proteins have been associated to said hydrogels in view of the regeneration of the cartilage. Viability and proliferation of fibroblasts were impressively enhanced upon addition of poly-l-lysine. The modulation of the osteocytes has been achieved in various ways by applying advanced technologies such as 3D-plotting and electrospinning of biomimetic scaffolds, with optional addition of nano hydroxyapatite to the formulations. A wealth of biotechnological advances and know-how has permitted reaching outstanding results in crucial areas such as cranio-facial surgery, orthopedics and dentistry. It is mandatory to use scaffolds fully characterized in terms of porosity, pore size, swelling, wettability, compressive strength, and degree of acetylation, if the osteogenic differentiation of human mesenchymal stem cells is sought: in fact, the novel characteristics imparted by GEN-chitosan must be simultaneously of physico-chemical and cytological nature. Owing to their high standard, the scientific publications dated 2010–2015 have met the expectations of an interdisciplinary audience.

  18. Genipin-Crosslinked Chitosan Gels and Scaffolds for Tissue Engineering and Regeneration of Cartilage and Bone

    Science.gov (United States)

    Muzzarelli, Riccardo A. A.; El Mehtedi, Mohamad; Bottegoni, Carlo; Aquili, Alberto; Gigante, Antonio

    2015-01-01

    The present review article intends to direct attention to the technological advances made since 2009 in the area of genipin-crosslinked chitosan (GEN-chitosan) hydrogels. After a concise introduction on the well recognized characteristics of medical grade chitosan and food grade genipin, the properties of GEN-chitosan obtained with a safe, spontaneous and irreversible chemical reaction, and the quality assessment of the gels are reviewed. The antibacterial activity of GEN-chitosan has been well assessed in the treatment of gastric infections supported by Helicobacter pylori. Therapies based on chitosan alginate crosslinked with genipin include stem cell transplantation, and development of contraction free biomaterials suitable for cartilage engineering. Collagen, gelatin and other proteins have been associated to said hydrogels in view of the regeneration of the cartilage. Viability and proliferation of fibroblasts were impressively enhanced upon addition of poly-l-lysine. The modulation of the osteocytes has been achieved in various ways by applying advanced technologies such as 3D-plotting and electrospinning of biomimetic scaffolds, with optional addition of nano hydroxyapatite to the formulations. A wealth of biotechnological advances and know-how has permitted reaching outstanding results in crucial areas such as cranio-facial surgery, orthopedics and dentistry. It is mandatory to use scaffolds fully characterized in terms of porosity, pore size, swelling, wettability, compressive strength, and degree of acetylation, if the osteogenic differentiation of human mesenchymal stem cells is sought: in fact, the novel characteristics imparted by GEN-chitosan must be simultaneously of physico-chemical and cytological nature. Owing to their high standard, the scientific publications dated 2010–2015 have met the expectations of an interdisciplinary audience. PMID:26690453

  19. An ice-templated, linearly aligned chitosan-alginate scaffold for neural tissue engineering.

    Science.gov (United States)

    Francis, Nicola L; Hunger, Philipp M; Donius, Amalie E; Riblett, Benjamin W; Zavaliangos, Antonios; Wegst, Ulrike G K; Wheatley, Margaret A

    2013-12-01

    Several strategies have been investigated to enhance axonal regeneration after spinal cord injury, however, the resulting growth can be random and disorganized. Bioengineered scaffolds provide a physical substrate for guidance of regenerating axons towards their targets, and can be produced by freeze casting. This technique involves the controlled directional solidification of an aqueous solution or suspension, resulting in a linearly aligned porous structure caused by ice templating. In this study, freeze casting was used to fabricate porous chitosan-alginate (C/A) scaffolds with longitudinally oriented channels. Chick dorsal root ganglia explants adhered to and extended neurites through the scaffold in parallel alignment with the channel direction. Surface adsorption of a polycation and laminin promoted significantly longer neurite growth than the uncoated scaffold (poly-L-ornithine + Laminin = 793.2 ± 187.2 μm; poly-L-lysine + Laminin = 768.7 ± 241.2 μm; uncoated scaffold = 22.52 ± 50.14 μm) (P < 0.001). The elastic modulus of the hydrated scaffold was determined to be 5.08 ± 0.61 kPa, comparable to reported spinal cord values. The present data suggested that this C/A scaffold is a promising candidate for use as a nerve guidance scaffold, because of its ability to support neuronal attachment and the linearly aligned growth of DRG neurites. Copyright © 2013 Wiley Periodicals, Inc., a Wiley Company.

  20. Noninvasive Evaluation of Injectable Chitosan/Nano-Hydroxyapatite/Collagen Scaffold via Ultrasound

    Directory of Open Access Journals (Sweden)

    Yan Chen

    2012-01-01

    Full Text Available To meet the challenges of designing an in situ forming scaffold and regenerating bone with complex three-dimensional (3D structures, an in situ forming hydrogel scaffold based on nano-hydroxyapatite (nHA, collagen (Col, and chitosan (CS was synthesized. Currently, only a limited number of techniques are available to mediate and visualize the injection process of the injectable biomaterials directly and noninvasively. In this study, the potential of ultrasound for the quantitative in vivo evaluation of tissue development in CS/nHAC scaffold was evaluated. The CS/nHAC scaffold was injected into rat subcutaneous tissue and evaluated for 28 days. Quantitative measurements of the gray-scale value, volume, and blood flow of the scaffold were evaluated using diagnostic technique. This study demonstrates that ultrasound can be used to noninvasively and nondestructively monitor and evaluate the in vivo characteristics of injectable bone scaffold. In comparison to the CS, the CS/nHAC scaffold showed a greater stiffness, less degradation rate, and better blood supply in the in vivo evaluation. In conclusion, the diagnostic ultrasound method is a good tool to evaluate the in vivo formation of injectable bone scaffolds and facilitates the broad use to monitor tissue development and remodeling in bone tissue engineering.

  1. Strontium hydroxyapatite/chitosan nanohybrid scaffolds with enhanced osteoinductivity for bone tissue engineering

    Energy Technology Data Exchange (ETDEWEB)

    Lei, Yong [The Education Ministry Key Lab of Resource Chemistry and Shanghai Key Laboratory of Rare Earth Functional Materials, Shanghai Normal University, Shanghai 200234 (China); Xu, Zhengliang [Department of Orthopedic Surgery, Shanghai Jiao Tong University Affiliated Sixth People' s Hospital, 600 Yishan Road, Shanghai 200233 (China); Ke, Qinfei [The Education Ministry Key Lab of Resource Chemistry and Shanghai Key Laboratory of Rare Earth Functional Materials, Shanghai Normal University, Shanghai 200234 (China); Yin, Wenjing; Chen, Yixuan [Department of Orthopedic Surgery, Shanghai Jiao Tong University Affiliated Sixth People' s Hospital, 600 Yishan Road, Shanghai 200233 (China); Zhang, Changqing, E-mail: zhangcq@sjtu.edu.cn [Department of Orthopedic Surgery, Shanghai Jiao Tong University Affiliated Sixth People' s Hospital, 600 Yishan Road, Shanghai 200233 (China); Guo, Yaping, E-mail: ypguo@shnu.edu.cn [The Education Ministry Key Lab of Resource Chemistry and Shanghai Key Laboratory of Rare Earth Functional Materials, Shanghai Normal University, Shanghai 200234 (China)

    2017-03-01

    For the clinical application of bone tissue engineering with the combination of biomaterials and mesenchymal stem cells (MSCs), bone scaffolds should possess excellent biocompatibility and osteoinductivity to accelerate the repair of bone defects. Herein, strontium hydroxyapatite [SrHAP, Ca{sub 10−x}Sr{sub x}(PO{sub 4}){sub 6}(OH){sub 2}]/chitosan (CS) nanohybrid scaffolds were fabricated by a freeze-drying method. The SrHAP nanocrystals with the different x values of 0, 1, 5 and 10 are abbreviated to HAP, Sr1HAP, Sr5HAP and Sr10HAP, respectively. With increasing x values from 0 to 10, the crystal cell volumes and axial lengths of SrHAP become gradually large because of the greater ion radius of Sr{sup 2+} than Ca{sup 2+}, while the crystal sizes of SrHAP decrease from 70.4 nm to 46.7 nm. The SrHAP/CS nanohybrid scaffolds exhibits three-dimensional (3D) interconnected macropores with pore sizes of 100–400 μm, and the SrHAP nanocrystals are uniformly dispersed within the scaffolds. In vitro cell experiments reveal that all the HAP/CS, Sr1HAP/CS, Sr5HAP/CS and Sr10HAP/CS nanohybrid scaffolds possess excellent cytocompatibility with the favorable adhesion, spreading and proliferation of human bone marrow mesenchymal stem cells (hBMSCs). The Sr5HAP nanocrystals in the scaffolds do not affect the adhesion, spreading of hBMSCs, but they contribute remarkably to cell proliferation and osteogenic differentiation. As compared with the HAP/CS nanohybrid scaffold, the released Sr{sup 2+} ions from the SrHAP/CS nanohybrid scaffolds enhance alkaline phosphatase (ALP) activity, extracellular matrix (ECM) mineralization and osteogenic-related COL-1 and ALP expression levels. Especially, the Sr5HAP/CS nanohybrid scaffolds exhibit the best osteoinductivity among four groups because of the synergetic effect between Ca{sup 2+} and Sr{sup 2+} ions. Hence, the Sr5HAP/CS nanohybrid scaffolds with excellent cytocompatibility and osteogenic property have promising application for

  2. Strontium hydroxyapatite/chitosan nanohybrid scaffolds with enhanced osteoinductivity for bone tissue engineering

    International Nuclear Information System (INIS)

    Lei, Yong; Xu, Zhengliang; Ke, Qinfei; Yin, Wenjing; Chen, Yixuan; Zhang, Changqing; Guo, Yaping

    2017-01-01

    For the clinical application of bone tissue engineering with the combination of biomaterials and mesenchymal stem cells (MSCs), bone scaffolds should possess excellent biocompatibility and osteoinductivity to accelerate the repair of bone defects. Herein, strontium hydroxyapatite [SrHAP, Ca 10−x Sr x (PO 4 ) 6 (OH) 2 ]/chitosan (CS) nanohybrid scaffolds were fabricated by a freeze-drying method. The SrHAP nanocrystals with the different x values of 0, 1, 5 and 10 are abbreviated to HAP, Sr1HAP, Sr5HAP and Sr10HAP, respectively. With increasing x values from 0 to 10, the crystal cell volumes and axial lengths of SrHAP become gradually large because of the greater ion radius of Sr 2+ than Ca 2+ , while the crystal sizes of SrHAP decrease from 70.4 nm to 46.7 nm. The SrHAP/CS nanohybrid scaffolds exhibits three-dimensional (3D) interconnected macropores with pore sizes of 100–400 μm, and the SrHAP nanocrystals are uniformly dispersed within the scaffolds. In vitro cell experiments reveal that all the HAP/CS, Sr1HAP/CS, Sr5HAP/CS and Sr10HAP/CS nanohybrid scaffolds possess excellent cytocompatibility with the favorable adhesion, spreading and proliferation of human bone marrow mesenchymal stem cells (hBMSCs). The Sr5HAP nanocrystals in the scaffolds do not affect the adhesion, spreading of hBMSCs, but they contribute remarkably to cell proliferation and osteogenic differentiation. As compared with the HAP/CS nanohybrid scaffold, the released Sr 2+ ions from the SrHAP/CS nanohybrid scaffolds enhance alkaline phosphatase (ALP) activity, extracellular matrix (ECM) mineralization and osteogenic-related COL-1 and ALP expression levels. Especially, the Sr5HAP/CS nanohybrid scaffolds exhibit the best osteoinductivity among four groups because of the synergetic effect between Ca 2+ and Sr 2+ ions. Hence, the Sr5HAP/CS nanohybrid scaffolds with excellent cytocompatibility and osteogenic property have promising application for bone tissue engineering. - Highlights: • We

  3. Preparation of zeolite-A/chitosan hybrid composites and their bioactivities and antimicrobial activities

    International Nuclear Information System (INIS)

    Yu, Liang; Gong, Jie; Zeng, Changfeng; Zhang, Lixiong

    2013-01-01

    hybrid composites were prepared by in situ transformation. • The zeolite contents could be adjusted simply. • Mechanical strength of chitosan scaffold was enhanced by incorporating zeolite. • The hybrid composites possess macroporous structures of 100–300 μm. • It exhibits superior bioactivity and antimicrobial activity

  4. Preparation of zeolite-A/chitosan hybrid composites and their bioactivities and antimicrobial activities

    Energy Technology Data Exchange (ETDEWEB)

    Yu, Liang; Gong, Jie [State Key Laboratory of Materials-Oriented Chemical Engineering, College of Chemistry and Chemical Engineering, Nanjing University of Technology, Nanjing 210009 (China); Zeng, Changfeng [College of Mechanic and Power Engineering, Nanjing University of Technology, Nanjing 210009 (China); Zhang, Lixiong, E-mail: lixiongzhang@yahoo.com [State Key Laboratory of Materials-Oriented Chemical Engineering, College of Chemistry and Chemical Engineering, Nanjing University of Technology, Nanjing 210009 (China)

    2013-10-15

    application. Highlights: • Zeolite A/chitosan hybrid composites were prepared by in situ transformation. • The zeolite contents could be adjusted simply. • Mechanical strength of chitosan scaffold was enhanced by incorporating zeolite. • The hybrid composites possess macroporous structures of 100–300 μm. • It exhibits superior bioactivity and antimicrobial activity.

  5. PLA/chitosan/keratin composites for biomedical applications

    Energy Technology Data Exchange (ETDEWEB)

    Tanase, Constantin Edi, E-mail: etanase@live.com [Faculty of Medical Bioengineering, ‘Grigore T. Popa’ University of Medicine and Pharmacy, 9-13 Kogalniceanu Street, 700454 Iasi (Romania); Spiridon, Iuliana [“Petru Poni” Institute of Macromolecular Chemistry, 41A Grigore Ghica Voda Alley, 700487 Iasi (Romania)

    2014-07-01

    Novel composites based on PLA, chitosan and keratin was obtained via blend preparation. The goal of this contribution was to evaluate mechanical and in vitro behavior of the composites. The results point out composites with improved Young modulus and decreased tensile strength, significant increase in hardness (compared to PLA) and a good uptake of the surface properties. Biological assessments using human osteosarcoma cell line on these composites indicate a good viability/proliferation outcome. Hence preliminary results regarding mechanical behavior and in vitro osteoblast response suggest that these composites might have prospective application in medical field. - Highlights: • PLA, chitosan and keratin composites are prepared by blend preparation. • PLA, chitosan and keratin composites present improved mechanical properties and water uptake compare to PLA. • PLA, chitosan and keratin composites present good in vitro behavior.

  6. PLA/chitosan/keratin composites for biomedical applications

    International Nuclear Information System (INIS)

    Tanase, Constantin Edi; Spiridon, Iuliana

    2014-01-01

    Novel composites based on PLA, chitosan and keratin was obtained via blend preparation. The goal of this contribution was to evaluate mechanical and in vitro behavior of the composites. The results point out composites with improved Young modulus and decreased tensile strength, significant increase in hardness (compared to PLA) and a good uptake of the surface properties. Biological assessments using human osteosarcoma cell line on these composites indicate a good viability/proliferation outcome. Hence preliminary results regarding mechanical behavior and in vitro osteoblast response suggest that these composites might have prospective application in medical field. - Highlights: • PLA, chitosan and keratin composites are prepared by blend preparation. • PLA, chitosan and keratin composites present improved mechanical properties and water uptake compare to PLA. • PLA, chitosan and keratin composites present good in vitro behavior

  7. Culture & differentiation of mesenchymal stem cell into osteoblast on degradable biomedical composite scaffold: In vitro study

    Directory of Open Access Journals (Sweden)

    Krishan G Jain

    2015-01-01

    Full Text Available Background & objectives: There is a significant bone tissue loss in patients from diseases and traumatic injury. The current autograft transplantation gold standard treatment has drawbacks, namely donor site morbidity and limited supply. The field of tissue engineering has emerged with a goal to provide alternative sources for transplantations to bridge this gap between the need and lack of bone graft. The aim of this study was to prepare biocomposite scaffolds based on chitosan (CHT, polycaprolactone (PCL and hydroxyapatite (HAP by freeze drying method and to assess the role of scaffolds in spatial organization, proliferation, and osteogenic differentiation of human mesenchymal stem cells (hMSCs in vitro, in order to achieve bone graft substitutes with improved physical-chemical and biological properties. Methods: Pure chitosan (100CHT and composites (40CHT/HAP, 30CHT/HAP/PCL and 25CHT/HAP/PCL scaffolds containing 40, 30, 25 parts per hundred resin (phr filler, respectively in acetic acid were freeze dried and the porous foams were studied for physicochemical and in vitro biological properties. Results: Scanning electron microscope (SEM images of the scaffolds showed porous microstructure (20-300 μm with uniform pore distribution in all compositions. Materials were tested under compressive load in wet condition (using phosphate buffered saline at pH 7.4. The in vitro studies showed that all the scaffold compositions supported mesenchymal stem cell attachment, proliferation and differentiation as visible from SEM images, [3-(4,5-dimethylthiazole-2-yl-2,5-diphenyltetrazolium bromide] (MTT assay, alkaline phosphatase (ALP assay and quantitative reverse transcription (qRT-PCR. Interpretation & conclusions: Scaffold composition 25CHT/HAP/PCL showed better biomechanical and osteoinductive properties as evident by mechanical test and alkaline phosphatase activity and osteoblast specific gene expression studies. This study suggests that this novel

  8. Novel chitosan/collagen scaffold containing transforming growth factor-β1 DNA for periodontal tissue engineering

    International Nuclear Information System (INIS)

    Zhang Yufeng; Cheng Xiangrong; Wang Jiawei; Wang Yining; Shi Bin; Huang Cui; Yang Xuechao; Liu Tongjun

    2006-01-01

    The current rapid progression in tissue engineering and local gene delivery system has enhanced our applications to periodontal tissue engineering. In this study, porous chitosan/collagen scaffolds were prepared through a freeze-drying process, and loaded with plasmid and adenoviral vector encoding human transforming growth factor-β1 (TGF-β1). These scaffolds were evaluated in vitro by analysis of microscopic structure, porosity, and cytocompatibility. Human periodontal ligament cells (HPLCs) were seeded in this scaffold, and gene transfection could be traced by green fluorescent protein (GFP). The expression of type I and type III collagen was detected with RT-PCR, and then these scaffolds were implanted subcutaneously into athymic mice. Results indicated that the pore diameter of the gene-combined scaffolds was lower than that of pure chitosan/collagen scaffold. The scaffold containing Ad-TGF-β1 exhibited the highest proliferation rate, and the expression of type I and type III collagen up-regulated in Ad-TGF-β1 scaffold. After implanted in vivo, EGFP-transfected HPLCs not only proliferated but also recruited surrounding tissue to grow in the scaffold. This study demonstrated the potential of chitosan/collagen scaffold combined Ad-TGF-β1 as a good substrate candidate in periodontal tissue engineering

  9. Balancing mechanical strength with bioactivity in chitosan-calcium phosphate 3D microsphere scaffolds for bone tissue engineering: air- vs. freeze-drying processes.

    Science.gov (United States)

    Nguyen, D T; McCanless, J D; Mecwan, M M; Noblett, A P; Haggard, W O; Smith, R A; Bumgardner, J D

    2013-01-01

    The objective of this study was to evaluate the potential benefit of 3D composite scaffolds composed of chitosan and calcium phosphate for bone tissue engineering. Additionally, incorporation of mechanically weak lyophilized microspheres within those air-dried (AD) was considered for enhanced bioactivity. AD microsphere, alone, and air- and freeze-dried microsphere (FDAD) 3D scaffolds were evaluated in vitro using a 28-day osteogenic culture model with the Saos-2 cell line. Mechanical testing, quantitative microscopy, and lysozyme-driven enzymatic degradation of the scaffolds were also studied. FDAD scaffold showed a higher concentration (p < 0.01) in cells per scaffold mass vs. AD constructs. Collagen was ∼31% greater (p < 0.01) on FDAD compared to AD scaffolds not evident in microscopy of microsphere surfaces. Alternatively, AD scaffolds demonstrated a superior threefold increase in compressive strength over FDAD (12 vs. 4 MPa) with minimal degradation. Inclusion of FD spheres within the FDAD scaffolds allowed increased cellular activity through improved seeding, proliferation, and extracellular matrix production (as collagen), although mechanical strength was sacrificed through introduction of the less stiff, porous FD spheres.

  10. Chitosan-functionalized silk fibroin 3D scaffold for keratocyte culture.

    Science.gov (United States)

    Guan, Linan; Tian, Pei; Ge, Hongyan; Tang, Xianling; Zhang, Hong; Du, Lingling; Liu, Ping

    2013-10-01

    The goal of this study was to evaluate the potential suitability of an artificial membrane composed of silk fibroin (SF) functionalized by different ratios of chitosan (CS) as a substrate for the stroma of the cornea. Keratocytes were cultured on translucent membranes made of SF and CS with different ratios. The biophysical properties of the silk fibroin and chitosan (SF/CS) membrane were examined. The SF/CS showed tensile strengths that increased as the CS concentration increased, but the physical and mechanical properties of chitosan-functionalized silk fibroin scaffolds weakened significantly compared with those of native corneas. The resulting cell scaffolds were evaluated using western blot in addition to light and electron microscopy. The cell attachment and proliferation on the scaffold were similar to those on a plastic plate. Keratocytes cultured in serum on SF/CS exhibited stellate morphology along with a marked increase in the expression of keratocan compared with identical cultures on tissue culture plastics. The biocompatibility was tested by transplanting the acellular membrane into rabbit corneal stromal pockets. There was no inflammatory complication detected at any time point on the macroscopic level. Taken together, these results indicate that SF/CS holds promise as a substrate for corneal reconstruction.

  11. Optimization strategies on the structural modeling of gelatin/chitosan scaffolds to mimic human meniscus tissue

    International Nuclear Information System (INIS)

    Sarem, Melika; Moztarzadeh, Fathollah; Mozafari, Masoud; Shastri, V. Prasad

    2013-01-01

    Meniscus lesions are frequently occurring injuries with poor ability to heal. Typical treatment procedure includes removal of damaged regions, which can lead to sub-optimal knee biomechanics and early onset of osteoarthritis. Some of the drawbacks of current treatment approach present an opportunity for a tissue engineering solution. In this study, gelatin (G)/chitosan (Cs) scaffolds were synthesized via gel casting method and cross-linked with naturally derived cross-linker, genipin, through scaffold cross-linking method. Based on the characteristics of native meniscus tissue microstructure and function, three different layers were chosen to design the macroporous multilayered scaffolds. The multi-layered scaffolds were investigated for their ability to support human-derived meniscus cells by evaluating their morphology and proliferation using MTT assay at various time points. Based on structural, mechanical and cell compatibility considerations, laminated scaffolds composed of G60/Cs40, G80/Cs20 and G40/Cs60 samples, for the first, second and third layers, respectively, could be an appropriate combination for meniscus tissue engineering applications. - Graphical abstract: The wedge shaped multilayer/multiporous G/Cs meniscus scaffolds were mimicked by MR images of anatomical knee meniscus. The layers were chosen as G60/Cs40, G80/Cs20 and G40/Cs60, according to their characteristics similar to meniscus natural tissue, as the first, second and third layers, respectively. - Highlights: • Different gelatin/chitosan systems were chosen to engineer a multilayered scaffold. • The compressive modulus increased gradually by increasing the gelatin concentration. • Further addition of gelatin showed a meaningful decrease in the water uptake degree. • The layers supported cell growth and mimicked the meniscus fibrocartilage structure

  12. Optimization strategies on the structural modeling of gelatin/chitosan scaffolds to mimic human meniscus tissue

    Energy Technology Data Exchange (ETDEWEB)

    Sarem, Melika [Sports Engineering Group, Faculty of Biomedical Engineering (Center of Excellence), Amirkabir University of Technology, P.O. Box 15875-4413, Tehran (Iran, Islamic Republic of); Institute for Macromolecular Chemistry, University of Freiburg, Hermann Staudinger Haus, Freiburg D-79104 (Germany); Helmholtz Virtual Institute: Multifunctional Biomaterials for Medicine, Freiburg (Germany); Moztarzadeh, Fathollah [Sports Engineering Group, Faculty of Biomedical Engineering (Center of Excellence), Amirkabir University of Technology, P.O. Box 15875-4413, Tehran (Iran, Islamic Republic of); Biomaterials Group, Faculty of Biomedical Engineering (Center of Excellence), Amirkabir University of Technology, P.O. Box 15875-4413, Tehran (Iran, Islamic Republic of); Mozafari, Masoud, E-mail: mozafari.masoud@gmail.com [Sports Engineering Group, Faculty of Biomedical Engineering (Center of Excellence), Amirkabir University of Technology, P.O. Box 15875-4413, Tehran (Iran, Islamic Republic of); Biomaterials Group, Faculty of Biomedical Engineering (Center of Excellence), Amirkabir University of Technology, P.O. Box 15875-4413, Tehran (Iran, Islamic Republic of); Helmerich Advanced Technology Research Center, School of Material Science and Engineering, Oklahoma State University, OK 74106 (United States); Shastri, V. Prasad [Institute for Macromolecular Chemistry, University of Freiburg, Hermann Staudinger Haus, Freiburg D-79104 (Germany); Helmholtz Virtual Institute: Multifunctional Biomaterials for Medicine, Freiburg (Germany)

    2013-12-01

    Meniscus lesions are frequently occurring injuries with poor ability to heal. Typical treatment procedure includes removal of damaged regions, which can lead to sub-optimal knee biomechanics and early onset of osteoarthritis. Some of the drawbacks of current treatment approach present an opportunity for a tissue engineering solution. In this study, gelatin (G)/chitosan (Cs) scaffolds were synthesized via gel casting method and cross-linked with naturally derived cross-linker, genipin, through scaffold cross-linking method. Based on the characteristics of native meniscus tissue microstructure and function, three different layers were chosen to design the macroporous multilayered scaffolds. The multi-layered scaffolds were investigated for their ability to support human-derived meniscus cells by evaluating their morphology and proliferation using MTT assay at various time points. Based on structural, mechanical and cell compatibility considerations, laminated scaffolds composed of G60/Cs40, G80/Cs20 and G40/Cs60 samples, for the first, second and third layers, respectively, could be an appropriate combination for meniscus tissue engineering applications. - Graphical abstract: The wedge shaped multilayer/multiporous G/Cs meniscus scaffolds were mimicked by MR images of anatomical knee meniscus. The layers were chosen as G60/Cs40, G80/Cs20 and G40/Cs60, according to their characteristics similar to meniscus natural tissue, as the first, second and third layers, respectively. - Highlights: • Different gelatin/chitosan systems were chosen to engineer a multilayered scaffold. • The compressive modulus increased gradually by increasing the gelatin concentration. • Further addition of gelatin showed a meaningful decrease in the water uptake degree. • The layers supported cell growth and mimicked the meniscus fibrocartilage structure.

  13. Chitosan composite hydrogels reinforced with natural clay nanotubes.

    Science.gov (United States)

    Huang, Biao; Liu, Mingxian; Zhou, Changren

    2017-11-01

    Here, chitosan composites hydrogels were prepared by addition of halloysite nanotubes (HNTs) in the chitosan KOH/LiOH/urea solution. The raw chitosan and chitosan/HNTs composite hydrogels were obtained by heat treatment at 60°C for 8h and then regeneration in ethanol solution. The viscosity of the composite solution is increased with HNTs content. The Fourier transform infrared spectroscopy (FT-IR) shows that the hydrogen bonds interactions exist between the HNTs and the chitosan. X-ray diffraction (XRD) results show that the crystal structure of HNT is not changed in the composite hydrogels. The compressive property test and storage modulus determination show that the mechanical properties and anti-deformation ability of the composite hydrogel significantly increase owing to the reinforcing effect of HNTs. The composites hydrogel with 66.7% HNTs can undergo 7 times compression cycles without breaking with compressive strength of 0.71MPa at 70% deformation, while pure chitosan hydrogel is broken after bearing 5 compression cycles with compressive strength of 0.14MPa and a maximum deformation of 59%. A porous structure with pore size of 100-500μm is found in the composite hydrogels by scanning electron microscopy (SEM), and the pore size and the swelling ratio in NaCl solution decrease by the addition of HNTs and the immersing of ethanol. Chitosan/HNTs composite hydrogels show low cytotoxicity towards MC3T3-E1 cells. Also, the composite hydrogels show a maximum drug entrapment efficiency of 45.7% for doxorubicin (DOX) which is much higher than that of pure chitosan hydrogel (27.5%). All the results illustrate that the chitosan/HNTs composite hydrogels show promising applications as biomaterials. Copyright © 2017 Elsevier Ltd. All rights reserved.

  14. 3D- Printed Poly(ε-caprolactone) Scaffold Integrated with Cell-laden Chitosan Hydrogels for Bone Tissue Engineering

    OpenAIRE

    Dong, Liang; Wang, Shao-Jie; Zhao, Xin-Rong; Zhu, Yu-Fang; Yu, Jia-Kuo

    2017-01-01

    Synthetic polymeric scaffolds are commonly used in bone tissue engineering (BTE) due to their biocompatibility and adequate mechanical properties. However, their hydrophobicity and the lack of specific cell recognition sites confined their practical application. In this study, to improve the cell seeding efficiency and osteoinductivity, an injectable thermo-sensitive chitosan hydrogel (CSG) was incorporated into a 3D-printed poly(ε-caprolactone) (PCL) scaffold to form a hybrid scaffold. To de...

  15. Poly(acrylonitrile)chitosan composite membranes for urease immobilization.

    Science.gov (United States)

    Gabrovska, Katya; Georgieva, Aneliya; Godjevargova, Tzonka; Stoilova, Olya; Manolova, Nevena

    2007-05-10

    (Poly)acrylonitrile/chitosan (PANCHI) composite membranes were prepared. The chitosan layer was deposited on the surface as well as on the pore walls of the base membrane. This resulted in the reduction of the pore size of the membrane and in an increase of their hydrophilicity. The pore structure of PAN and PANCHI membranes were determined by TEM and SEM analyses. It was found that the average size of the pore under a selective layer base PAN membrane is 7 microm, while the membrane coated with 0.25% chitosan shows a reduced pore size--small or equal to 5 microm and with 0.35% chitosan--about 4 microm. The amounts of the functional groups, the degree of hydrophilicity and transport characteristics of PAN/Chitosan composite membranes were determined. Urease was covalently immobilized onto all kinds of PAN/chitosan composite membranes using glutaraldehyde. Both the amount of bound protein and relative activity of immobilized urease were measured. The highest activity (94%) was measured for urease bound to PANCHI2 membranes (0.25% chitosan). The basic characteristics (pH(opt), pH(stability), T(opt), T(stability), heat inactivation and storage stability) of immobilized urease were determined. The obtained results show that the poly(acrylonitrile)chitosan composite membranes are suitable for enzyme immobilization.

  16. Chitosan-poly(lactide-co-glycolide) microsphere-based scaffolds for bone tissue engineering: in vitro degradation and in vivo bone regeneration studies.

    Science.gov (United States)

    Jiang, Tao; Nukavarapu, Syam P; Deng, Meng; Jabbarzadeh, Ehsan; Kofron, Michelle D; Doty, Stephen B; Abdel-Fattah, Wafa I; Laurencin, Cato T

    2010-09-01

    Natural polymer chitosan and synthetic polymer poly(lactide-co-glycolide) (PLAGA) have been investigated for a variety of tissue engineering applications. We have previously reported the fabrication and in vitro evaluation of a novel chitosan/PLAGA sintered microsphere scaffold for load-bearing bone tissue engineering applications. In this study, the in vitro degradation characteristics of the chitosan/PLAGA scaffold and the in vivo bone formation capacity of the chitosan/PLAGA-based scaffolds in a rabbit ulnar critical-sized-defect model were investigated. The chitosan/PLAGA scaffold showed slower degradation than the PLAGA scaffold in vitro. Although chitosan/PLAGA scaffold showed a gradual decrease in compressive properties during the 12-week degradation period, the compressive strength and compressive modulus remained in the range of human trabecular bone. Chitosan/PLAGA-based scaffolds were able to guide bone formation in a rabbit ulnar critical-sized-defect model. Microcomputed tomography analysis demonstrated that successful bridging of the critical-sized defect on the sides both adjacent to and away from the radius occurred using chitosan/PLAGA-based scaffolds. Immobilization of heparin and recombinant human bone morphogenetic protein-2 on the chitosan/PLAGA scaffold surface promoted early bone formation as evidenced by complete bridging of the defect along the radius and significantly enhanced mechanical properties when compared to the chitosan/PLAGA scaffold. Furthermore, histological analysis suggested that chitosan/PLAGA-based scaffolds supported normal bone formation via intramembranous formation. 2010 Acta Materialia Inc. Published by Elsevier Ltd. All rights reserved.

  17. Oxygen and nitrogen plasma etching of three-dimensional hydroxyapatite/chitosan scaffolds fabricated by additive manufacturing

    Science.gov (United States)

    Myung, Sung-Woon; Kim, Byung-Hoon

    2016-01-01

    Three-dimensional (3D) chitosan and hydroxyapatite (HAp)/chitosan (CH) scaffolds were fabricated by additive manufacturing, then their surfaces were etched with oxygen (O2) and nitrogen (N2) plasma. O2 and N2 plasma etching was performed to increase surface properties such as hydrophilicity, roughness, and surface chemistry on the scaffolds. After etching, hydroxyapatite was exposed on the surface of 3D HAp/CH scaffolds. The surface morphology and chemical properties were characterized by contact angle measurement, scanning electron microscopy, X-ray diffraction, and attenuated total reflection Fourier infrared spectroscopy. The cell viability of 3D chitosan scaffolds was examined by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay. The differentiation of preosteoblast cells was evaluated by alkaline phosphatase assay. The cell viability was improved by O2 and N2 plasma etching of 3D chitosan scaffolds. The present fabrication process for 3D scaffolds might be applied to a potential tool for preparing biocompatible scaffolds.

  18. 3D- Printed Poly(ε-caprolactone) Scaffold Integrated with Cell-laden Chitosan Hydrogels for Bone Tissue Engineering.

    Science.gov (United States)

    Dong, Liang; Wang, Shao-Jie; Zhao, Xin-Rong; Zhu, Yu-Fang; Yu, Jia-Kuo

    2017-10-17

    Synthetic polymeric scaffolds are commonly used in bone tissue engineering (BTE) due to their biocompatibility and adequate mechanical properties. However, their hydrophobicity and the lack of specific cell recognition sites confined their practical application. In this study, to improve the cell seeding efficiency and osteoinductivity, an injectable thermo-sensitive chitosan hydrogel (CSG) was incorporated into a 3D-printed poly(ε-caprolactone) (PCL) scaffold to form a hybrid scaffold. To demonstrate the feasibility of this hybrid system for BTE application, rabbit bone marrow mesenchymal stem cells (BMMSCs) and bone morphogenetic protein-2 (BMP-2) were encapsulated in CSG. Pure PCL scaffolds were used as controls. Cell proliferation and viability were investigated. Osteogenic gene expressions of BMMSCs in various scaffolds were determined with reverse transcription polymerase chain reaction (RT-PCR). Growth factor releasing profile and mechanical tests were performed. CCK-8 assay confirmed greater cell retention and proliferation in chitosan and hybrid groups. Confocal microscopy showed even distribution of cells in the hybrid system. After 2-week osteogenic culture in vitro, BMMSCs in hybrid and chitosan scaffolds showed stronger osteogenesis and bone-matrix formation. To conclude, chitosan/PCL hybrid scaffolds are a favorable platform for BTE due to its capacity to carry cells and drugs, and excellent mechanical strength.

  19. Combining mechanical foaming and thermally induced phase separation to generate chitosan scaffolds for soft tissue engineering.

    Science.gov (United States)

    Biswas, D P; Tran, P A; Tallon, C; O'Connor, A J

    2017-02-01

    In this paper, a novel foaming methodology consisting of turbulent mixing and thermally induced phase separation (TIPS) was used to generate scaffolds for tissue engineering. Air bubbles were mechanically introduced into a chitosan solution which forms the continuous polymer/liquid phase in the foam created. The air bubbles entrained in the foam act as a template for the macroporous architecture of the final scaffolds. Wet foams were crosslinked via glutaraldehyde and frozen at -20 °C to induce TIPS in order to limit film drainage, bubble coalescence and Ostwald ripening. The effects of production parameters, including mixing speed, surfactant concentration and chitosan concentration, on foaming are explored. Using this method, hydrogel scaffolds were successfully produced with up to 80% porosity, average pore sizes of 120 μm and readily tuneable compressive modulus in the range of 2.6 to 25 kPa relevant to soft tissue engineering applications. These scaffolds supported 3T3 fibroblast cell proliferation and penetration and therefore show significant potential for application in soft tissue engineering.

  20. Chitosan and Its Potential Use as a Scaffold for Tissue Engineering in Regenerative Medicine

    Science.gov (United States)

    Rodríguez-Vázquez, Martin; Vega-Ruiz, Brenda; Ramos-Zúñiga, Rodrigo; Saldaña-Koppel, Daniel Alexander; Quiñones-Olvera, Luis Fernando

    2015-01-01

    Tissue engineering is an important therapeutic strategy to be used in regenerative medicine in the present and in the future. Functional biomaterials research is focused on the development and improvement of scaffolding, which can be used to repair or regenerate an organ or tissue. Scaffolds are one of the crucial factors for tissue engineering. Scaffolds consisting of natural polymers have recently been developed more quickly and have gained more popularity. These include chitosan, a copolymer derived from the alkaline deacetylation of chitin. Expectations for use of these scaffolds are increasing as the knowledge regarding their chemical and biological properties expands, and new biomedical applications are investigated. Due to their different biological properties such as being biocompatible, biodegradable, and bioactive, they have given the pattern for use in tissue engineering for repair and/or regeneration of different tissues including skin, bone, cartilage, nerves, liver, and muscle. In this review, we focus on the intrinsic properties offered by chitosan and its use in tissue engineering, considering it as a promising alternative for regenerative medicine as a bioactive polymer. PMID:26504833

  1. Osteogenic differentiation and mineralization of human exfoliated deciduous teeth stem cells on modified chitosan scaffold

    Energy Technology Data Exchange (ETDEWEB)

    Su, Wen-Ta, E-mail: f10549@ntut.edu.tw [Department of Chemical Engineering and Biotechnology, National Taipei University of Technology, Taipei, Taiwan (China); Wu, Pai-Shuen [Department of Chemical Engineering and Biotechnology, National Taipei University of Technology, Taipei, Taiwan (China); Ko, Chih-Sheng [PhytoHealth Corporation, Maywufa Biopharma Group, Taipei, Taiwan (China); Huang, Te-Yang [Mackay Memorial Hospital, Taipei, Taiwan (China)

    2014-08-01

    Stem cells from human exfoliated deciduous teeth (SHEDs) have been considered as alternative sources of adult stem cells in tissue engineering because of their potential to differentiate into multiple cell lineages. Strontium has an important function in bone remodeling because it can simulate bone formation and decrease bone resorption. In this study, the effects of strontium phosphate on the osteogenic differentiation of SHEDs were investigated. Strontium phosphate was found to enhance the osteogenic differentiation of SHEDs with up-regulated osteoblast-related gene expression. The proliferation of SHEDs was slightly inhibited by chitosan scaffolds; however, type-I collagen expression, alkaline phosphatase activity, and calcium deposition on chitosan scaffolds containing strontium were significantly enhanced. Furthermore, cells seeded in a 3D scaffold under dynamic culture at an optimal fluid rate might enhance cellular differentiation than static culture in osteoblastic gene expression. This experiment might provide a useful cell resource and dynamic 3D culture for tissue engineering and bone repair. - Highlights: • SHEDs have been considered as alternative sources of adult stem cells in tissue engineering • Strontium phosphate can enhance the osteogenic differentiation of SHEDs • 3D scaffold under dynamic culture with optimal fluid rate enhance cellular differentiation.

  2. Osteogenic differentiation and mineralization of human exfoliated deciduous teeth stem cells on modified chitosan scaffold

    International Nuclear Information System (INIS)

    Su, Wen-Ta; Wu, Pai-Shuen; Ko, Chih-Sheng; Huang, Te-Yang

    2014-01-01

    Stem cells from human exfoliated deciduous teeth (SHEDs) have been considered as alternative sources of adult stem cells in tissue engineering because of their potential to differentiate into multiple cell lineages. Strontium has an important function in bone remodeling because it can simulate bone formation and decrease bone resorption. In this study, the effects of strontium phosphate on the osteogenic differentiation of SHEDs were investigated. Strontium phosphate was found to enhance the osteogenic differentiation of SHEDs with up-regulated osteoblast-related gene expression. The proliferation of SHEDs was slightly inhibited by chitosan scaffolds; however, type-I collagen expression, alkaline phosphatase activity, and calcium deposition on chitosan scaffolds containing strontium were significantly enhanced. Furthermore, cells seeded in a 3D scaffold under dynamic culture at an optimal fluid rate might enhance cellular differentiation than static culture in osteoblastic gene expression. This experiment might provide a useful cell resource and dynamic 3D culture for tissue engineering and bone repair. - Highlights: • SHEDs have been considered as alternative sources of adult stem cells in tissue engineering • Strontium phosphate can enhance the osteogenic differentiation of SHEDs • 3D scaffold under dynamic culture with optimal fluid rate enhance cellular differentiation

  3. Chitosan scaffold modified with D-(+) raffinose and enriched with thiol-modified gelatin for improved osteoblast adhesion

    International Nuclear Information System (INIS)

    Galli, C; Parisi, L; Smerieri, A; Lumetti, S; Manfredi, E; Macaluso, G M; Elviri, L; Bianchera, A; Bettini, R; Lagonegro, P

    2016-01-01

    The aim of the present study was to investigate whether chitosan-based scaffolds modified with D-(+) raffinose and enriched with thiol-modified gelatin could selectively improve osteoblast adhesion and proliferation. 2, 3 and 4.5% chitosan films were prepared. Chitosan suitability for tissue engineering was confirmed by protein adsorption assay. Scaffolds were incubated with a 2.5 mg ml −1 BSA solution and the decrease of protein content in the supernatants was measured by spectrophotometry. Chitosan films were then enriched with thiol-modified gelatin and their ability to bind BSA was also measured. Then, 2% chitosan discs with or without thiol-modified gelatin were used as culture substrates for MC3T3-E1 cells. After 72 h cells were stained with trypan blue or with calcein AM and propidium iodide for morphology, viability and proliferation assays. Moreover, cell viability was measured at 48, 72, 96 and 168 h to obtain a growth curve. Chitosan films efficiently bound and retained BSA proportionally to the concentration of chitosan discs. The amount of protein retained was higher on chitosan enriched with thiol-modified gelatin. Moreover, chitosan discs allowed the adhesion and the viability of cells, but inhibited their proliferation. The functionalization of chitosan with thiol-modified gelatin enhanced cell spreading and proliferation. Our data confirm that chitosan is a suitable material for tissue engineering. Moreover, our data show that the enrichment of chitosan with thiol-modified gelatin enhances its biological properties. (paper)

  4. Chitosan nanofiber scaffold improves bone healing via stimulating trabecular bone production due to upregulation of the Runx2/osteocalcin/alkaline phosphatase signaling pathway

    Science.gov (United States)

    Ho, Ming-Hua; Yao, Chih-Jung; Liao, Mei-Hsiu; Lin, Pei-I; Liu, Shing-Hwa; Chen, Ruei-Ming

    2015-01-01

    Osteoblasts play critical roles in bone formation. Our previous study showed that chitosan nanofibers can stimulate osteoblast proliferation and maturation. This translational study used an animal model of bone defects to evaluate the effects of chitosan nanofiber scaffolds on bone healing and the possible mechanisms. In this study, we produced uniform chitosan nanofibers with fiber diameters of approximately 200 nm. A bone defect was surgically created in the proximal femurs of male C57LB/6 mice, and then the left femur was implanted with chitosan nanofiber scaffolds for 21 days and compared with the right femur, which served as a control. Histological analyses revealed that implantation of chitosan nanofiber scaffolds did not lead to hepatotoxicity or nephrotoxicity. Instead, imaging analyses by X-ray transmission and microcomputed tomography showed that implantation of chitosan nanofiber scaffolds improved bone healing compared with the control group. In parallel, microcomputed tomography and bone histomorphometric assays further demonstrated augmentation of the production of new trabecular bone in the chitosan nanofiber-treated group. Furthermore, implantation of chitosan nanofiber scaffolds led to a significant increase in the trabecular bone thickness but a reduction in the trabecular parameter factor. As to the mechanisms, analysis by confocal microscopy showed that implantation of chitosan nanofiber scaffolds increased levels of Runt-related transcription factor 2 (Runx2), a key transcription factor that regulates osteogenesis, in the bone defect sites. Successively, amounts of alkaline phosphatase and osteocalcin, two typical biomarkers that can simulate bone maturation, were augmented following implantation of chitosan nanofiber scaffolds. Taken together, this translational study showed a beneficial effect of chitosan nanofiber scaffolds on bone healing through stimulating trabecular bone production due to upregulation of Runx2-mediated alkaline

  5. Application of chitosan scaffolds on vascular endothelial growth factor and fibroblast growth factor 2 expressions in tissue engineering principles

    Directory of Open Access Journals (Sweden)

    Ariyati Retno Pratiwi

    2015-12-01

    Full Text Available Background: Tissue engineering has given satisfactory results as biological tissue substitutes to restore, replace, or regenerate tissues that have a defect. Chitosan is an organic biomaterial often used in the biomedical field. Chitosan has biocompatible, antifungal, and antibacterial properties. Chitosan is osteoconductive, suitable for bone regeneration applications. Bone defect healing begins with inflammatory phase as a response to the presence of vascular injury, so new vascularization is required. Vascular endothelial growth factor (VEGF and basic fibroblast growth factor-2 (FGF2 are indicators of the beginning of bone regeneration process, playing an important role in angiogenesis. Purpose: This research was aimed to determine the effects of chitosan scaffold application on the expressions of VEGF and FGF2 in tissue engineering principles. Method: Chitosan was dissolved in CH3COOH and NaOH to form a gel. Chitosan gel was then printed in mould to freeze dry for 24 hours. Those rats with defected bones were divided into two groups. Group 1 was the control group which defected bones were not administrated with chitosan scaffolds. Group 2 was the treatment group which defected bones were administrated with chitosan scaffolds. Those rats were sacrificed on day 14. Tissue preparations were made, and then immunohistochemical staining was conducted. Finally, a statistical analysis was conducted using Kruskal Wallis test. Result: There was no significant difference in the expressions of VEGF and FGF2 between the control group and the treatment group (p>0.05. Conclusion: Chitosan scaffolds do not affect the expressions of VEGF and FGF2 during bone regeneration process on day 14 in tissue engineering principles

  6. Chitosan functionalized poly-ε-caprolactone electrospun fibers and 3D printed scaffolds as antibacterial materials for tissue engineering applications.

    Science.gov (United States)

    Tardajos, Myriam G; Cama, Giuseppe; Dash, Mamoni; Misseeuw, Lara; Gheysens, Tom; Gorzelanny, Christian; Coenye, Tom; Dubruel, Peter

    2018-07-01

    Tissue engineering (TE) approaches often employ polymer-based scaffolds to provide support with a view to the improved regeneration of damaged tissues. The aim of this research was to develop a surface modification method for introducing chitosan as an antibacterial agent in both electrospun membranes and 3D printed poly-ε-caprolactone (PCL) scaffolds. The scaffolds were functionalized by grafting methacrylic acid N-hydroxysuccinimide ester (NHSMA) onto the surface after Ar-plasma/air activation. Subsequently, the newly-introduced NHS groups were used to couple with chitosan of various molecular weights (Mw). High Mw chitosan exhibited a better coverage of the surface as indicated by the higher N% detected by X-ray photoelectron spectroscopy (XPS) and the observations with either scanning electron microscopy (SEM)(for fibers) or Coomassie blue staining (for 3D-printed scaffolds). A lactate dehydrogenase assay (LDH) using L929 fibroblasts demonstrated the cell-adhesion and cell-viability capacity of the modified samples. The antibacterial properties against S. aureus ATCC 6538 and S. epidermidis ET13 revealed a slower bacterial growth rate on the surface of the chitosan modified scaffolds, regardless the chitosan Mw. Copyright © 2018 Elsevier Ltd. All rights reserved.

  7. Heparin-Functionalized chitosan/ κ-carrageenan complexes as potential scaffolds for tissue engineering

    International Nuclear Information System (INIS)

    Dofeliz, Joni L.; Rojas, Nina Rosario L.

    2015-01-01

    Cell-based approaches to tissue regeneration are playing an increasingly important role in bone and cartilage repair. This is made possible through the use of growth factors, which are signaling molecules that induce a number of effects such as cell proliferation, migration and differentiation. But problems arise as these growth factors tend to be expensive, short-lived and slow moving through the extracellular matrix, making them very inefficient in their current form of introduction. One such growth factor is basic fibroblast growth factor (bFGF). The general objective of this study is to construct heparin-functionalized chitosan/κ-carrageenan scaffolds, which when bound to basic fibroblast growth factor (bFGF), may be used in the culture of mesenchymal stem cells for differentiation into cartilage. In this study, gels of semi-interpenetrating networks (semi-IPN) of chitosan and κ-carrageenan (2:4:1 blend ration) were prepared using calcium chloride as a cross-linker. The gels were then functionalized with heparin, which is known for its growth factor binding ability, using 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide (EDC) as cross-linker. The functionalized gels were then reshaped into scaffold on the wells of 48-well plates through freeze-dry method. The scaffolds were found to be realatively porous with pore sizes larger than 100 μm which satisfies the requirements for cell culture. Subsequent thermogravimetric analysis shows that the scaffolds were relatively stable with a degradation temperature of 277°C. Additionally, there was no observable separation of the individual degradation temperatures of chitosan and κ-carrageenan, confirming that a single miscible phase in the form of a semi-IPN structure was created. Results of scanning electron microscopy also showed that the scaffolds were stable under 2 hours of UV sterilization. The FTIR spectra of the heparinized PEC showed characteristic absorption bands (S=O asymetric stretch) in the area of 1160

  8. Scaffold of chitosan/poly(vinyl alcohol) blend chemically crosslinked by glutaraldehyde for tissue engineering applications

    International Nuclear Information System (INIS)

    Costa Junior, Ezequiel de S.; Laguardia-Nascimento, Mateus; Barbosa-Stancioli, Edel F.; Mansur, Herman S.

    2009-01-01

    Chitosan/PVA based films were chemically crosslinked by glutaraldehyde (GA) in order to achieve scaffolds for potential tissue engineering application. Both precursors and developed films were characterized by FTIR and XRD in order to determine the presence of chemicals groups and nanostructural order, respectively. The results have showed that the GA crosslinking have altered the crystallinity of the chitosan and the increase on the C=N bands and decreasing of NH 2 bands suggest that Chitosan/GA crosslinking has preference to occur in the carbon 2 by Schiff's base. The mechanical properties, swelling behavior, degradation rate in vitro and cellular viability were compatible with the characteristic of an epithelial tissue. The material presented a toughness range from 1.4 to 34MJ/m3, swelling from 150% to 700% in 24h, degradation rate from 20% to 75% (wt%) in 24h and cellular viability in vitro above 60% compared to the cellular control. The developed scaffolds from the films have also showed swelling and degradation in vitro properties well-matched for biomedical applications in tissue engineering (author)

  9. Genipin cross-linked electrospun chitosan-based nanofibrous mat as tissue engineering scaffold

    Directory of Open Access Journals (Sweden)

    Esmaeil Mirzaei

    2014-04-01

    Full Text Available   Objective(s: To improve water stability of electrospun chitosan/ Polyethylene oxide (PEO nanofibers, genipin, a biocompatible and nontoxic agent, was used to crosslink chitosan based nanofibers.   Materials and Methods: Different amounts of genipin were added to the chitosan/PEO solutions, chitosan/PEO weight ratio 90/10 in 80 % acetic acid, and the solutions were then electrospun to form nanofibers. The spun nanofibers were exposed to water vapor to complete crosslinking. The nanofibrous membranes were subjected to detailed analysis by scanning electron microscopy (SEM, Fourier transform infrared-attenuated total reflection (FTIR-ATR spectroscopy, swelling test, MTT cytotoxicity, and cell attachment. Results: SEM images of electrospun mats showed that genipin-crosslinked nanofibers retained their fibrous structure after immerging in PBS (pH=7.4 for 24 hours, while the uncrosslinked samples lost their fibrous structure, indicating the water stability of genipin-crosslinked nanofibers. The genipin-crosslinked mats also showed no significant change in swelling ratio in comparison with uncrosslinked ones. FTIR-ATR spectrum of uncrosslinked and genipin-crosslinked chitosan nanofibers revealed the reaction between genipin and amino groups of chitosan. Cytotoxicity of genipin-crosslinked nanofibers was examined by MTT assay on human fibroblast cells in the presence of nanofibers extraction media. The genipin-crosslinked nanofibers did not show any toxic effects on fibroblast cells at the lowest and moderate amount of genipin. The fibroblast cells also showed a good adhesion on genipin-crosslinked nanofibers. Conclusion: This electrospun matrix would be used for biomedical applications such as wound dressing and scaffold for tissue engineering without the concern of toxicity.

  10. Evaluation of the effect of the degree of acetylation on the inflammatory response to 3D porous chitosan scaffolds.

    Science.gov (United States)

    Barbosa, Judite N; Amaral, Isabel F; Aguas, Artur P; Barbosa, Mário A

    2010-04-01

    The effect of the degree of acetylation (DA) of 3D chitosan (Ch) scaffolds on the inflammatory reaction was investigated. Chitosan porous scaffolds with DAs of 4 and 15% were implanted using a subcutaneous air-pouch model of inflammation. The initial acute inflammatory response was evaluated 24 and 48 h after implantation. To characterize the initial response, the recruitment and adhesion of inflammatory cells to the implant site was studied. The fibrous capsule formation and the infiltration of inflammatory cells within the scaffolds were evaluated for longer implantation times (2 and 4 weeks). Chitosan with DA 15% attracted the highest number of leukocytes to the implant site. High numbers of adherent inflammatory cells were also observed in this material. For longer implantation periods Ch scaffolds with a DA of 15% induced the formation of a thick fibrous capsule and a high infiltration of inflammatory cells within the scaffold. Our results indicate that the biological response to implanted Ch scaffolds was influenced by the DA. Chitosan with a DA of 15% induce a more intense inflammatory response when compared with DA 4% Ch. Because inflammation and healing are interrelated, this result may provide clues for the relative importance of acetyl and amine functional groups in tissue repair and regeneration.

  11. Synthesis and characterization of chitosan-alginate scaffolds for seeding human umbilical cord derived mesenchymal stem cells.

    Science.gov (United States)

    Kumbhar, Sneha G; Pawar, S H

    2016-01-01

    Chitosan and alginate are two natural and accessible polymers that are known to be biocompatible, biodegradable and possesses good antimicrobial activity. When combined, they exhibit desirable characteristics and can be created into a scaffold for cell culture. In this study interaction of chitosan-alginate scaffolds with mesenchymal stem cells are studied. Mesenchymal stem cells were derived from human umbilical cord tissues, characterized by flow cytometry and other growth parameters studied as well. Proliferation and viability of cultured cells were studied by MTT Assay and Trypan Blue dye exclusion assay. Besides chitosan-alginate scaffold was prepared by freeze-drying method and characterized by FTIR, SEM and Rheological properties. The obtained 3D porous structure allowed very efficient seeding of hUMSCs that are able to inhabit the whole volume of the scaffold, showing good adhesion and proliferation. These materials showed desirable rheological properties for facile injection as tissue scaffolds. The results of this study demonstrated that chitosan-alginate scaffold may be promising biomaterial in the field of tissue engineering, which is currently under a great deal of examination for the development and/or restoration of tissue and organs. It combines the stem cell therapy and biomaterials.

  12. Pore architecture and cell viability on freeze dried 3D recombinant human collagen-peptide (RHC)–chitosan scaffolds

    Energy Technology Data Exchange (ETDEWEB)

    Zhang, Jing; Zhou, Aimei; Deng, Aipeng [School of Environmental and Biological Engineering, Nanjing University of Science and Technology, Nanjing 210094 (China); Yang, Yang [Faculty of Engineering, University of Nottingham, Nottingham NG7 2RD (United Kingdom); Gao, Lihu; Zhong, Zhaocai [School of Environmental and Biological Engineering, Nanjing University of Science and Technology, Nanjing 210094 (China); Yang, Shulin, E-mail: yshulin@njust.edu.cn [School of Environmental and Biological Engineering, Nanjing University of Science and Technology, Nanjing 210094 (China)

    2015-04-01

    Pore architecture of 3D scaffolds used in tissue engineering plays a critical role in the maintenance of cell survival, proliferation and further promotion of tissue regeneration. We investigated the pore size and structure, porosity, swelling as well as cell viability of a series of recombinant human collagen-peptide–chitosan (RHCC) scaffolds fabricated by lyophilization. In this paper, freezing regime containing a final temperature of freezing (T{sub f}) and cooling rates was applied to obtain scaffolds with pore size ranging from 100 μm to 120 μm. Other protocols of RHC/chitosan suspension concentration and ratio modification were studied to produce more homogenous and appropriate structural scaffolds. The mean pore size decreased along with the decline of T{sub f} at a slow cooling rate of 0.7 °C/min; a more rapid cooling rate under 5 °C/min resulted to a smaller pore size and more homogenous microstructure. High concentration could reduce pore size and lead to thick well of scaffold, while improved the ratio of RHC, lamellar and fiber structure coexisted with cellular pores. Human umbilical vein endothelial cells (HUVECs) were seeded on these manufactured scaffolds, the cell viability represented a negative correlation to the pore size. This study provides an alternative method to fabricate 3D RHC–chitosan scaffolds with appropriate pores for potential tissue engineering. - Highlights: • Fabrication of recombinant human collagen-chitosan scaffolds by freezing drying • Influence of freeze drying protocols on lyophilized scaffolds • Pore size, microstructure, porosity, swelling and cell viability were compared. • The optimized porous scaffold is suitable for cell (HUVEC) seeding.

  13. Pore architecture and cell viability on freeze dried 3D recombinant human collagen-peptide (RHC)–chitosan scaffolds

    International Nuclear Information System (INIS)

    Zhang, Jing; Zhou, Aimei; Deng, Aipeng; Yang, Yang; Gao, Lihu; Zhong, Zhaocai; Yang, Shulin

    2015-01-01

    Pore architecture of 3D scaffolds used in tissue engineering plays a critical role in the maintenance of cell survival, proliferation and further promotion of tissue regeneration. We investigated the pore size and structure, porosity, swelling as well as cell viability of a series of recombinant human collagen-peptide–chitosan (RHCC) scaffolds fabricated by lyophilization. In this paper, freezing regime containing a final temperature of freezing (T f ) and cooling rates was applied to obtain scaffolds with pore size ranging from 100 μm to 120 μm. Other protocols of RHC/chitosan suspension concentration and ratio modification were studied to produce more homogenous and appropriate structural scaffolds. The mean pore size decreased along with the decline of T f at a slow cooling rate of 0.7 °C/min; a more rapid cooling rate under 5 °C/min resulted to a smaller pore size and more homogenous microstructure. High concentration could reduce pore size and lead to thick well of scaffold, while improved the ratio of RHC, lamellar and fiber structure coexisted with cellular pores. Human umbilical vein endothelial cells (HUVECs) were seeded on these manufactured scaffolds, the cell viability represented a negative correlation to the pore size. This study provides an alternative method to fabricate 3D RHC–chitosan scaffolds with appropriate pores for potential tissue engineering. - Highlights: • Fabrication of recombinant human collagen-chitosan scaffolds by freezing drying • Influence of freeze drying protocols on lyophilized scaffolds • Pore size, microstructure, porosity, swelling and cell viability were compared. • The optimized porous scaffold is suitable for cell (HUVEC) seeding

  14. Potential Biomedical Application of Enzymatically Treated Alginate/Chitosan Hydrosols in Sponges—Biocompatible Scaffolds Inducing Chondrogenic Differentiation of Human Adipose Derived Multipotent Stromal Cells

    Directory of Open Access Journals (Sweden)

    Anna Zimoch-Korzycka

    2016-08-01

    Full Text Available Current regenerative strategies used for cartilage repair rely on biomaterial functionality as a scaffold for cells that may have potential in chondrogenic differentiation. The purpose of the research was to investigate the biocompatibility of enzymatically treated alginate/chitosan hydrosol sponges and their suitability to support chondrogenic differentiation of human adipose derived multipotent stromal cells (hASCs. The alginate/chitosan and enzyme/alginate/chitosan sponges were formed from hydrosols with various proportions and were used as a biomaterial in this study. Sponges were tested for porosity and wettability. The porosity of each sponge was higher than 80%. An equal dose of alginate and chitosan in the composition of sponges improved their swelling ability. It was found that equal concentrations of alginate and chitosan in hydrosols sponges assure high biocompatibility properties that may be further improved by enzymatic treatment. Importantly, the high biocompatibility of these biomaterials turned out to be crucial in the context of hydrosols’ pro-chondrogenic function. After exposure to the chondrogenic conditions, the hASCs in N/A/C and L/A/C sponges formed well developed nodules and revealed increased expression of collagen type II, aggrecan and decreased expression of collagen type I. Moreover, in these cultures, the reactive oxygen species level was lowered while superoxide dismutase activity increased. Based on the obtained results, we conclude that N/A/C and L/A/C sponges may have prospective application as hASCs carriers for cartilage repair.

  15. Nanostructured natural-based polyelectrolyte multilayers to agglomerate chitosan particles into scaffolds for tissue engineering.

    Science.gov (United States)

    Miranda, Emanuel Sá; Silva, Tiago H; Reis, Rui L; Mano, João F

    2011-11-01

    The layer-by-layer (LbL) deposition technique is a self-assembly process that allows the coating of material's surface with nanostructured layers of polyelectrolytes, allowing to control several surface properties. This technique presents some advantages when compared with other thin film assembly techniques, like having the possibility to coat surfaces with complex geometries in mild conditions or to incorporate active compounds. Tissue engineering (TE) involves typically the use of porous biodegradable scaffolds for the temporary support of cells. Such structures can be produced by agglomeration of microspheres that needs to be fixed into a three-dimensional (3D) structure. In this work we suggest the use of LbL to promote such mechanical fixation in free-formed microspheres assemblies and simultaneously to control the properties of its surface. For the proof of concept the biological performance of chitosan/alginate multilayers is first investigated in two-dimensional (2D) models in which the attachment and proliferation of L929 and ATDC5 cells are studied in function of the number of layers and the nature of the final layer. Scaffolds prepared by agglomeration of chitosan particles using the same multilayered system were processed and characterized; it was found that they could support the attachment and proliferation of ATDC5 cells. This study suggests that LbL can be used as a versatile methodology to prepare scaffolds by particle agglomeration that could be suitable for TE applications.

  16. Solid state synthesis of chitosan and its unsaturated derivatives for laser microfabrication of 3D scaffolds

    Science.gov (United States)

    Akopova, T. A.; Demina, T. S.; Bagratashvili, V. N.; Bardakova, K. N.; Novikov, M. M.; Selezneva, I. I.; Istomin, A. V.; Svidchenko, E. A.; Cherkaev, G. V.; Surin, N. M.; Timashev, P. S.

    2015-07-01

    Chitosans with various degrees of deacetylation and molecular weights and their allyl substituted derivatives were obtained through a solvent-free reaction under shear deformation in an extruder. Structure and physical-chemical analysis of the samples were carried out using nuclear magnetic resonance (NMR), ultraviolet (UV) and infrared radiation (IR) spectroscopy. Photosensitive materials based on the synthesized polymers were successfully used for microfabrication of 3D well-defined architectonic structures by laser stereolithography. Study on the metabolic activity of NCTC L929 cultured in the presence of the cured chitosan extracts indicates that the engineered biomaterials could support adhesion, spreading and growth of adherent-dependent cells, and thus could be considered as biocompatible scaffolds.

  17. Solid state synthesis of chitosan and its unsaturated derivatives for laser microfabrication of 3D scaffolds

    International Nuclear Information System (INIS)

    Akopova, T A; Demina, T S; Istomin, A V; Svidchenko, E A; Cherkaev, G V; Surin, N M; Bagratashvili, V N; Bardakova, K N; Novikov, M M; Selezneva, I I; Timashev, P S

    2015-01-01

    Chitosans with various degrees of deacetylation and molecular weights and their allyl substituted derivatives were obtained through a solvent-free reaction under shear deformation in an extruder. Structure and physical-chemical analysis of the samples were carried out using nuclear magnetic resonance (NMR), ultraviolet (UV) and infrared radiation (IR) spectroscopy. Photosensitive materials based on the synthesized polymers were successfully used for microfabrication of 3D well-defined architectonic structures by laser stereolithography. Study on the metabolic activity of NCTC L929 cultured in the presence of the cured chitosan extracts indicates that the engineered biomaterials could support adhesion, spreading and growth of adherent-dependent cells, and thus could be considered as biocompatible scaffolds. (paper)

  18. Long-term liver-specific functions of hepatocytes in electrospun chitosan nanofiber scaffolds coated with fibronectin.

    Science.gov (United States)

    Rajendran, Divya; Hussain, Ali; Yip, Derek; Parekh, Amit; Shrirao, Anil; Cho, Cheul H

    2017-08-01

    In this study, a new 3D liver model was developed using biomimetic nanofiber scaffolds and co-culture system consisting of hepatocytes and fibroblasts for the maintenance of long-term liver functions. The chitosan nanofiber scaffolds were fabricated by the electrospinning technique. To enhance cellular adhesion and spreading, the surfaces of the chitosan scaffolds were coated with fibronectin (FN) by adsorption and evaluated for various cell types. Cellular phenotype, protein expression, and liver-specific functions were extensively characterized by immunofluorescent and histochemical stainings, albumin enzyme-linked immunosorbent assay and Cytochrome p450 detoxification assays, and scanning electron microscopy. The electrospun chitosan scaffolds exhibited a highly porous and randomly oriented nanofibrous structure. The FN coating on the surface of the chitosan nanofibers significantly enhanced cell attachment and spreading, as expected, as surface modification with this cell adhesion molecule on the chitosan surface is important for focal adhesion formation and integrin binding. Comparison of hepatocyte mono-cultures and co-cultures in 3D culture systems indicated that the hepatocytes in co-cultures formed colonies and maintained their morphologies and functions for prolonged periods of time. The 3D liver tissue model developed in this study will provide useful tools toward the development of engineered liver tissues for drug screening and tissue engineering applications. © 2017 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 105A: 2119-2128, 2017. © 2017 Wiley Periodicals, Inc.

  19. Pore architecture and cell viability on freeze dried 3D recombinant human collagen-peptide (RHC)-chitosan scaffolds.

    Science.gov (United States)

    Zhang, Jing; Zhou, Aimei; Deng, Aipeng; Yang, Yang; Gao, Lihu; Zhong, Zhaocai; Yang, Shulin

    2015-04-01

    Pore architecture of 3D scaffolds used in tissue engineering plays a critical role in the maintenance of cell survival, proliferation and further promotion of tissue regeneration. We investigated the pore size and structure, porosity, swelling as well as cell viability of a series of recombinant human collagen-peptide-chitosan (RHCC) scaffolds fabricated by lyophilization. In this paper, freezing regime containing a final temperature of freezing (Tf) and cooling rates was applied to obtain scaffolds with pore size ranging from 100μm to 120μm. Other protocols of RHC/chitosan suspension concentration and ratio modification were studied to produce more homogenous and appropriate structural scaffolds. The mean pore size decreased along with the decline of Tf at a slow cooling rate of 0.7°C/min; a more rapid cooling rate under 5°C/min resulted to a smaller pore size and more homogenous microstructure. High concentration could reduce pore size and lead to thick well of scaffold, while improved the ratio of RHC, lamellar and fiber structure coexisted with cellular pores. Human umbilical vein endothelial cells (HUVECs) were seeded on these manufactured scaffolds, the cell viability represented a negative correlation to the pore size. This study provides an alternative method to fabricate 3D RHC-chitosan scaffolds with appropriate pores for potential tissue engineering. Copyright © 2014 Elsevier B.V. All rights reserved.

  20. Development of an injectable chitosan/marine collagen composite gel

    International Nuclear Information System (INIS)

    Wang Wei; Itoh, Soichiro; Aizawa, Tomoyasu; Demura, Makoto; Okawa, Atsushi; Sakai, Katsuyoshi; Ohkuma, Tsuneo

    2010-01-01

    A chitosan/marine-originated collagen composite has been developed. This composite gel was characterized and its biocompatibility, as well as an inflammatory reaction, was observed. The chitosan gel including N-3-carboxypropanoil-6-O-(carboxymethyl) chitosan of 3 mol%, 6-O-(carboxymethyl) chitosan of 62 mol% and 6-O-(carboxymethyl) chitin of 35 mol% was prepared and compounded with the salmon atelocollagen (SA) gel at different mixture ratios. The composite gels were injected subcutaneously in to the back of rats. The specimens were harvested for a histological survey as well as a tumor necrosis factor-alpha (TNF-α) assay by ELISA. The inflammatory cell infiltration and release of TNF-α were successively controlled low with the ratio of SA to chitosan at 10:90 or 20:80. The SA gel first, within 2 weeks, and then chitosan in the composite gel were slowly absorbed after implantation, followed by soft tissue formation. It is expected that this composite gel will be available as a carrier for tissue filler and drug delivery systems.

  1. Iota-carrageenan/chitosan/gelatin scaffold for the osteogenic differentiation of adipose-derived MSCs in vitro.

    Science.gov (United States)

    Li, Junjie; Yang, Boguang; Qian, Yufeng; Wang, Qiyu; Han, Ruijin; Hao, Tong; Shu, Yao; Zhang, Yabin; Yao, Fanglian; Wang, Changyong

    2015-10-01

    In this study, we have developed ι-carrageenan/chitosan/gelatin (CCG) scaffold containing multiple functional groups (-NH2 , -OH, -COOH, and -SO3 H) to resemble the native extracellular matrix (ECM), using the ion-shielding technology and ultrasonic dispersion method. Fourier transform infrared spectroscopy (FTIR) of the CCG scaffolds suggests that the formation of CCG network involves electrostatic interactions between ι-carrageenan (ι-CA) and chitosan/gelatin, and the covalent cross-linking among amino groups of chitosan and/or gelatin. Scanning electron microscopic (SEM) observation reveals that the porous structure of scaffolds can be modulated by the ratio of ι-CA to chitosan/gelatin. The swelling ratio of the hydrogels increases as the ι-CA contents increase. Using differential scanning calorimetry, we found that the double helix structure of ι-CA is only stabilized at low contents of ι-CA in the CCG scaffolds (e.g., 5 wt %). The scaffolds containing 5% ι-CA showed the best protein adsorption capacity (4.46 ± 0.63 μg protein/mg scaffold) and elastic modulus (5.37 ± 1.03 MPa). In addition, the CCG scaffolds exhibit excellent support for adipose-derived mesenchymal stem cells (ADMSCs) attachment and proliferation, and they can improve the osteogenic differentiation and neovascularization capacities of ADMSCs. Overall, we conclude that the CCG may represent an ideal scaffold material for bone tissue engineering. © 2014 Wiley Periodicals, Inc.

  2. Strontium hydroxyapatite/chitosan nanohybrid scaffolds with enhanced osteoinductivity for bone tissue engineering.

    Science.gov (United States)

    Lei, Yong; Xu, Zhengliang; Ke, Qinfei; Yin, Wenjing; Chen, Yixuan; Zhang, Changqing; Guo, Yaping

    2017-03-01

    For the clinical application of bone tissue engineering with the combination of biomaterials and mesenchymal stem cells (MSCs), bone scaffolds should possess excellent biocompatibility and osteoinductivity to accelerate the repair of bone defects. Herein, strontium hydroxyapatite [SrHAP, Ca 10-x Sr x (PO 4 ) 6 (OH) 2 ]/chitosan (CS) nanohybrid scaffolds were fabricated by a freeze-drying method. The SrHAP nanocrystals with the different x values of 0, 1, 5 and 10 are abbreviated to HAP, Sr1HAP, Sr5HAP and Sr10HAP, respectively. With increasing x values from 0 to 10, the crystal cell volumes and axial lengths of SrHAP become gradually large because of the greater ion radius of Sr 2+ than Ca 2+ , while the crystal sizes of SrHAP decrease from 70.4nm to 46.7nm. The SrHAP/CS nanohybrid scaffolds exhibits three-dimensional (3D) interconnected macropores with pore sizes of 100-400μm, and the SrHAP nanocrystals are uniformly dispersed within the scaffolds. In vitro cell experiments reveal that all the HAP/CS, Sr1HAP/CS, Sr5HAP/CS and Sr10HAP/CS nanohybrid scaffolds possess excellent cytocompatibility with the favorable adhesion, spreading and proliferation of human bone marrow mesenchymal stem cells (hBMSCs). The Sr5HAP nanocrystals in the scaffolds do not affect the adhesion, spreading of hBMSCs, but they contribute remarkably to cell proliferation and osteogenic differentiation. As compared with the HAP/CS nanohybrid scaffold, the released Sr 2+ ions from the SrHAP/CS nanohybrid scaffolds enhance alkaline phosphatase (ALP) activity, extracellular matrix (ECM) mineralization and osteogenic-related COL-1 and ALP expression levels. Especially, the Sr5HAP/CS nanohybrid scaffolds exhibit the best osteoinductivity among four groups because of the synergetic effect between Ca 2+ and Sr 2+ ions. Hence, the Sr5HAP/CS nanohybrid scaffolds with excellent cytocompatibility and osteogenic property have promising application for bone tissue engineering. Copyright © 2016. Published

  3. Chitosan porous 3D scaffolds embedded with resolvin D1 to improve in vivo bone healing.

    Science.gov (United States)

    Vasconcelos, Daniela P; Costa, Madalena; Neves, Nuno; Teixeira, José H; Vasconcelos, Daniel M; Santos, Susana G; Águas, Artur P; Barbosa, Mário A; Barbosa, Judite N

    2018-06-01

    The aim of this study was to investigate the effect chitosan (Ch) porous 3D scaffolds embedded with resolvin D1 (RvD1), an endogenous pro-resolving lipid mediator, on bone tissue healing. These scaffolds previous developed by us have demonstrated to have immunomodulatory properties namely in the modulation of the macrophage inflammatory phenotypic profile in an in vivo model of inflammation. Herein, results obtained in an in vivo rat femoral defect model demonstrated that two months after Ch + RvD1 scaffolds implantation, an increase in new bone formation, in bone trabecular thickness, and in collagen type I and Coll I/Coll III ratio were observed. These results suggest that Ch scaffolds embedded with RvD1 were able to lead to the formation of new bone with improvement of trabecular thickness. This study shows that the presence of RvD1 in the acute phase of the inflammatory response to the implanted biomaterial had a positive role in the subsequent bone tissue repair, thus demonstrating the importance of innovative approaches for the control of immune responses to biomedical implants in the design of advanced strategies for regenerative medicine. © 2018 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 106A: 1626-1633, 2018. © 2018 Wiley Periodicals, Inc.

  4. Preparation of 3D fibroin/chitosan blend porous scaffold for tissue engineering via a simplified method.

    Science.gov (United States)

    Ruan, Yuhui; Lin, Hong; Yao, Jinrong; Chen, Zhengrong; Shao, Zhengzhong

    2011-03-10

    In this work, we developed a simple and flexible method to manufacture a 3D porous scaffold based on the blend of regenerated silk fibroin (RSF) and chitosan (CS). No crosslinker or other toxic reagents were used in this method. The pores of resulted 3D scaffolds were connected with each other, and their sizes could be easily controlled by the concentration of the mixed solution. Compared with pure RSF scaffolds, the water absorptivities of these RSF/CS blend scaffolds with significantly enhanced mechanical properties were greatly increased. The results of MTT and RT-PCR tests indicated that the chondrocytes grew very well in these blend RSF/CS porous scaffolds. This suggested that the RSF/CS blend scaffold prepared by this new method could be a promising candidate for applications in tissue engineering. Copyright © 2011 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  5. Molecular interactions in gelatin/chitosan composite films.

    Science.gov (United States)

    Qiao, Congde; Ma, Xianguang; Zhang, Jianlong; Yao, Jinshui

    2017-11-15

    Gelatin and chitosan were mixed at different mass ratios in solution forms, and the rheological properties of these film-forming solutions, upon cooling, were studied. The results indicate that the significant interactions between gelatin and chitosan promote the formation of multiple complexes, reflected by an increase in the storage modulus of gelatin solution. Furthermore, these molecular interactions hinder the formation of gelatin networks, consequently decreasing the storage modulus of polymer gels. Both hydrogen bonds and electrostatic interactions are formed between gelatin and chitosan, as evidenced by the shift of the amide-II bands of polymers. X-ray patterns of composite films indicate that the contents of triple helices decrease with increasing chitosan content. Only one glass transition temperature (T g ) was observed in composite films with different composition ratios, and it decreases gradually with an increase in chitosan proportion, indicating that gelatin and chitosan have good miscibility and form a wide range of blends. Copyright © 2017 Elsevier Ltd. All rights reserved.

  6. Preparation, process optimization and characterization of core-shell polyurethane/chitosan nanofibers as a potential platform for bioactive scaffolds.

    Science.gov (United States)

    Maleknia, Laleh; Dilamian, Mandana; Pilehrood, Mohammad Kazemi; Sadeghi-Aliabadi, Hojjat; Hekmati, Amir Houshang

    2018-06-01

    In this paper, polyurethane (PU), chitosan (Cs)/polyethylene oxide (PEO), and core-shell PU/Cs nanofibers were produced at the optimal processing conditions using electrospinning technique. Several methods including SEM, TEM, FTIR, XRD, DSC, TGA and image analysis were utilized to characterize these nanofibrous structures. SEM images exhibited that the core-shell PU/Cs nanofibers were spun without any structural imperfections at the optimized processing conditions. TEM image confirmed the PU/Cs core-shell nanofibers were formed apparently. It that seems the inclusion of Cs/PEO to the shell, did not induce the significant variations in the crystallinity in the core-shell nanofibers. DSC analysis showed that the inclusion of Cs/PEO led to the glass temperature of the composition increased significantly compared to those of neat PU nanofibers. The thermal degradation of core-shell PU/Cs was similar to PU nanofibers degradation due to the higher PU concentration compared to other components. It was hypothesized that the core-shell PU/Cs nanofibers can be used as a potential platform for the bioactive scaffolds in tissue engineering. Further biological tests should be conducted to evaluate this platform as a three dimensional scaffold with the capabilities of releasing the bioactive molecules in a sustained manner.

  7. Strategies for neurotrophin-3 and chondroitinase ABC release from freeze-cast chitosan-alginate nerve-guidance scaffolds.

    Science.gov (United States)

    Francis, Nicola L; Hunger, Philipp M; Donius, Amalie E; Wegst, Ulrike G K; Wheatley, Margaret A

    2017-01-01

    Freeze casting, or controlled unidirectional solidification, can be used to fabricate chitosan-alginate (C-A) scaffolds with highly aligned porosity that are suitable for use as nerve-guidance channels. To augment the guidance of growth across a spinal cord injury lesion, these scaffolds are now evaluated in vitro to assess their ability to release neurotrophin-3 (NT-3) and chondroitinase ABC (chABC) in a controlled manner. Protein-loaded microcapsules were incorporated into C-A scaffolds prior to freeze casting without affecting the original scaffold architecture. In vitro protein release was not significantly different when comparing protein loaded directly into the scaffolds with release from scaffolds containing incorporated microcapsules. NT-3 was released from the C-A scaffolds for 8 weeks in vitro, while chABC was released for up to 7 weeks. Low total percentages of protein released from the scaffolds over this time period were attributed to limitation of diffusion by the interpenetrating polymer network matrix of the scaffold walls. NT-3 and chABC released from the scaffolds retained bioactivity, as determined by a neurite outgrowth assay, and the promotion of neurite growth across an inhibitory barrier of chondroitin sulphate proteoglycans. This demonstrates the potential of these multifunctional scaffolds for enhancing axonal regeneration through growth-inhibiting glial scars via the sustained release of chABC and NT-3. Copyright © 2014 John Wiley & Sons, Ltd. Copyright © 2014 John Wiley & Sons, Ltd.

  8. Preparation of bioactive porous HA/PCL composite scaffolds

    Energy Technology Data Exchange (ETDEWEB)

    Zhao, J.; Guo, L.Y.; Yang, X.B. [Key Laboratory of Advanced Technologies of Materials (Ministry of Education), School of Materials Science and Engineering, Southwest Jiaotong University, Chengdu 610031 (China); Weng, J. [Key Laboratory of Advanced Technologies of Materials (Ministry of Education), School of Materials Science and Engineering, Southwest Jiaotong University, Chengdu 610031 (China)], E-mail: jweng@swjtu.cn

    2008-12-30

    Porous hydroxyapatite (HA) bioceramic scaffold has been widely attracted the attention to act as a three-dimensional (3D) template for cell adhesion, proliferation, differentiation and thus promoting bone and cartilage regeneration because of its osteoinduction. However, the porous bioceramic scaffold is fragile so that it is not suitable to be applied in clinic for bone repair or replacement. Therefore, it is significant to improve the mechanical property of porous HA bioceramics while the interconnected structure is maintained for tissue ingrowth in vivo. In the present research, a porous composite scaffold composed of HA scaffold and polycaprolactone (PCL) lining was fabricated by the method of polymer impregnating to produce HA scaffold coated with PCL lining. Subsequently, the composite scaffolds were deposited with biomimetic coating for improving the bioactivity. The HA/PCL composite scaffolds with improved mechanical property and bioactivity is expected to be a promising bone substitute in tissue engineering applications.

  9. Preparation of bioactive porous HA/PCL composite scaffolds

    International Nuclear Information System (INIS)

    Zhao, J.; Guo, L.Y.; Yang, X.B.; Weng, J.

    2008-01-01

    Porous hydroxyapatite (HA) bioceramic scaffold has been widely attracted the attention to act as a three-dimensional (3D) template for cell adhesion, proliferation, differentiation and thus promoting bone and cartilage regeneration because of its osteoinduction. However, the porous bioceramic scaffold is fragile so that it is not suitable to be applied in clinic for bone repair or replacement. Therefore, it is significant to improve the mechanical property of porous HA bioceramics while the interconnected structure is maintained for tissue ingrowth in vivo. In the present research, a porous composite scaffold composed of HA scaffold and polycaprolactone (PCL) lining was fabricated by the method of polymer impregnating to produce HA scaffold coated with PCL lining. Subsequently, the composite scaffolds were deposited with biomimetic coating for improving the bioactivity. The HA/PCL composite scaffolds with improved mechanical property and bioactivity is expected to be a promising bone substitute in tissue engineering applications

  10. Design and characterization of a biodegradable composite scaffold for ligament tissue engineering.

    Science.gov (United States)

    Hayami, James W S; Surrao, Denver C; Waldman, Stephen D; Amsden, Brian G

    2010-03-15

    Herein we report on the development and characterization of a biodegradable composite scaffold for ligament tissue engineering based on the fundamental morphological features of the native ligament. An aligned fibrous component was used to mimic the fibrous collagen network and a hydrogel component to mimic the proteoglycan-water matrix of the ligament. The composite scaffold was constructed from cell-adherent, base-etched, electrospun poly(epsilon-caprolactone-co-D,L-lactide) (PCLDLLA) fibers embedded in a noncell-adherent photocrosslinked N-methacrylated glycol chitosan (MGC) hydrogel seeded with primary ligament fibroblasts. Base etching improved cellular adhesion to the PCLDLLA material. Cells within the MGC hydrogel remained viable (72 +/- 4%) during the 4-week culture period. Immunohistochemistry staining revealed ligament ECM markers collagen type I, collagen type III, and decorin organizing and accumulating along the PCLDLLA fibers within the composite scaffolds. On the basis of these results, it was determined that the composite scaffold design was a viable alternative to the current approaches used for ligament tissue engineering and merits further study. (c) 2009 Wiley Periodicals, Inc.

  11. Electrophoretic deposition of composite hydroxyapatite-chitosan coatings

    International Nuclear Information System (INIS)

    Pang Xin; Zhitomirsky, Igor

    2007-01-01

    Cathodic electrophoretic deposition has been utilized for the fabrication of composite hydroxyapatite-chitosan coatings on 316L stainless steel substrates. The addition of chitosan to the hydroxyapatite suspensions promoted the electrophoretic deposition of the hydroxyapatite nanoparticles and resulted in the formation of composite coatings. The obtained coatings were investigated by X-ray diffraction, thermogravimetric and differential thermal analysis, scanning and transmission electron microscopy, potentiodynamic polarization measurements, and electrochemical impedance spectroscopy. It was shown that the deposit composition can be changed by a variation of the chitosan or hydroxyapatite concentration in the solutions. Experimental conditions were developed for the fabrication of hydroxyapatite-chitosan nanocomposites containing 40.9-89.8 wt.% hydroxyapatite. The method enabled the formation of adherent and uniform coatings of thicknesses up to 60 μm. X-ray studies revealed that the preferred orientation of the hydroxyapatite nanoparticles in the chitosan matrix increases with decreasing hydroxyapatite content in the composite coatings. The obtained coatings provided the corrosion protection for the 316L stainless steel substrates

  12. Antimicrobial Properties of Chitosan-Alumina/f-MWCNT Nano composites

    International Nuclear Information System (INIS)

    Masheane, M.; Nthunya, L.; Malinga, S.; Masheane, M.; Nthunya, L.; Nxumalo, E.; Mhlanga, S.; Barnard, T.

    2016-01-01

    Antimicrobial chitosan-alumina/functionalized-multi walled carbon nano tube (f-MWCNT) nano composites were prepared by a simple phase inversion method. Scanning electron microscopy (SEM) analyses showed the change in the internal morphology of the composites and energy dispersive spectroscopy (EDS) confirmed the presence of alumina and f-MWCNTs in the chitosan polymer matrix. Fourier transform infrared (FTIR) spectroscopy showed the appearance of new functional groups from both alumina and f-MWCNTs, and thermogravimetric analysis (TGA) revealed that the addition of alumina and f-MWCNTs improved the thermal stability of the chitosan polymer. The presence of alumina and f-MWCNTs in the polymer matrix was found to improve the thermal stability and reduced the solubility of chitosan polymer. The prepared chitosan-alumina/f-MWCNT nano composites showed inhibition of twelve strains of bacterial strains that were tested. Thus, the nano composites show a potential for use as a biocides in water treatment for the removal of bacteria at different environmental conditions.

  13. Influence of chitosan-chitin nanofiber composites on cytoskeleton structure and the proliferation of rat bone marrow stromal cells.

    Science.gov (United States)

    Kiroshka, Victoria V; Petrova, Valentina A; Chernyakov, Daniil D; Bozhkova, Yulia O; Kiroshka, Katerina V; Baklagina, Yulia G; Romanov, Dmitry P; Kremnev, Roman V; Skorik, Yury A

    2017-01-01

    Chitosan scaffolds have gained much attention in various tissue engineering applications, but the effect of their microstructure on cell-material spatial interactions remains unclear. Our objective was to evaluate the effect of chitosan-based matrices doping with chitin nano-whiskers (CNW) on adhesion, spreading, cytoskeleton structure, and proliferation of rat bone marrow stromal cells (BMSCs). The behavior of BMSCs during culture on chitosan-CNW films was determined by the molecular mass, hydrophobicity, porosity, crosslinking degree, protonation degree and molecular structure of the composite chitosan-CNW films. The shape, spreading area, cytoskeleton structure, and proliferation of BMSCs on chitosan matrices with a crystalline structure and high porosity were similar to that observed for BMSCs cultured on polystyrene tissue culture plates. The amorphous polymer structure and high swelling led to a decrease in the spreading area and cell proliferation. Thus, we can control the behavior of cells in culture (adhesion, spreading, and proliferation) by changing the physico-chemical properties of the chitosan-CNW films.

  14. Tailoring the properties and functions of phosphate/silk/Ag/chitosan scaffolds

    International Nuclear Information System (INIS)

    Abdel-Fattah, Wafa I.; Sallam, Abdel Sattar M.; Diab, A.M.; Ali, Ghareib W.

    2015-01-01

    Two novel silk composites of phosphatic phases with nanosilver/chitosan having enhanced biocompatibility were achieved. Hydroxyapatite and octa calcium phosphates were synthesized in situ within silk fibroin/chitosan/nanosilver composites recently studied. Thermo-gravimetric analysis (TGA) and Differential Scanning Calorimetry (DSC) verified their thermal behavior. The structural aspects were characterized applying X-ray diffraction analysis (XRD), transmission electron microscopy (TEM) and field emission scanning electron microscope (FESEM) with EDAX. Additionally X-ray Photoelectron Spectroscopy (XPS) and Fourier Transform Infrared spectroscopy (FTIR) were applied. Mercury porosimeter was used to verify the pore size distribution. The in vitro degradation was followed in D-MEM for 48 h in a cumulative manner for five successive periods. Biochemical analyses of Ca, P and total protein using relevant chemical kits and atomic absorption for silver were performed. ANOVA statistics was carried out. Phosphatic crystalline phases along with the presence of silk, chitosan and nano-silver were developed. The diameters of hydroxyapatite and octa calcium phosphate particles were ~ 8–17 nm and 15–22 nm respectively. Comparatively higher degradation of Octa composite possessing higher porosity proved in turn more osteoinduction with in situ apatitic development. - Highlights: • A bottom–up approach controlled the achieved in situ configurations. • The calculated (CI) XDR and (CI) FTIR for both phases are highly conformable. • Post-immersion trimodal pore system was achieved in OCP composite. • Silver nanoparticle inclusion expected to enhance composite bactericidal activity

  15. Chitosan-alginate 3D scaffolds as a mimic of the glioma tumor microenvironment.

    Science.gov (United States)

    Kievit, Forrest M; Florczyk, Stephen J; Leung, Matthew C; Veiseh, Omid; Park, James O; Disis, Mary L; Zhang, Miqin

    2010-08-01

    Despite recent advances in the understanding of its cell biology, glioma remains highly lethal. Development of effective therapies requires a cost-effective in vitro tumor model that more accurately resembles the in vivo tumor microenvironment as standard two-dimensional (2D) tissue culture conditions do so poorly. Here we report on the use of a three-dimensional (3D) chitosan-alginate (CA) scaffold to serve as an extracellular matrix that promotes the conversion of cultured cancer cells to a more malignant in vivo-like phenotype. Human U-87 MG and U-118 MG glioma cells and rat C6 glioma cells were chosen for the study. In vitro tumor cell proliferation and secretion of factors that promote tumor malignancy, including VEGF, MMP-2, fibronectin, and laminin, were assessed. The scaffolds pre-cultured with U-87 MG and C6 cells were then implanted into nude mice to evaluate tumor growth and blood vessel recruitment compared to the standard 2D cell culture and 3D Matrigel matrix xenograft controls. Our results indicate that while the behavior of C6 cells showed minimal differences due to their highly malignant and invasive nature, U-87 MG and U-118 MG cells exhibited notably higher malignancy when cultured in CA scaffolds. CA scaffolds provide a 3D microenvironment for glioma cells that is more representative of the in vivo tumor, thus can serve as a more effective platform for development and study of anticancer therapeutics. This unique CA scaffold platform may offer a valuable alternative strategy to the time-consuming and costly animal studies for a wide variety of experimental designs. Copyright 2010 Elsevier Ltd. All rights reserved.

  16. Tumor cell culture on collagen–chitosan scaffolds as three-dimensional tumor model: A suitable model for tumor studies

    Directory of Open Access Journals (Sweden)

    Aziz Mahmoudzadeh

    2016-07-01

    Full Text Available Tumor cells naturally live in three-dimensional (3D microenvironments, while common laboratory tests and evaluations are done in two-dimensional (2D plates. This study examined the impact of cultured 4T1 cancer cells in a 3D collagen–chitosan scaffold compared with 2D plate cultures. Collagen–chitosan scaffolds were provided and passed confirmatory tests. 4T1 tumor cells were cultured on scaffolds and then tumor cells growth rate, resistance to X-ray radiation, and cyclophosphamide as a chemotherapy drug were analyzed. Furthermore, 4T1 cells were extracted from the scaffold model and were injected into the mice. Tumor growth rate, survival rate, and systemic immune responses were evaluated. Our results showed that 4T1 cells infiltrated the scaffolds pores and constructed a 3D microenvironment. Furthermore, 3D cultured tumor cells showed a slower proliferation rate, increased levels of survival to the X-ray irradiation, and enhanced resistance to chemotherapy drugs in comparison with 2D plate cultures. Transfer of extracted cells to the mice caused enhanced tumor volume and decreased life span. This study indicated that collagen–chitosan nanoscaffolds provide a suitable model of tumor that would be appropriate for tumor studies.

  17. Tumor cell culture on collagen-chitosan scaffolds as three-dimensional tumor model: A suitable model for tumor studies.

    Science.gov (United States)

    Mahmoudzadeh, Aziz; Mohammadpour, Hemn

    2016-07-01

    Tumor cells naturally live in three-dimensional (3D) microenvironments, while common laboratory tests and evaluations are done in two-dimensional (2D) plates. This study examined the impact of cultured 4T1 cancer cells in a 3D collagen-chitosan scaffold compared with 2D plate cultures. Collagen-chitosan scaffolds were provided and passed confirmatory tests. 4T1 tumor cells were cultured on scaffolds and then tumor cells growth rate, resistance to X-ray radiation, and cyclophosphamide as a chemotherapy drug were analyzed. Furthermore, 4T1 cells were extracted from the scaffold model and were injected into the mice. Tumor growth rate, survival rate, and systemic immune responses were evaluated. Our results showed that 4T1 cells infiltrated the scaffolds pores and constructed a 3D microenvironment. Furthermore, 3D cultured tumor cells showed a slower proliferation rate, increased levels of survival to the X-ray irradiation, and enhanced resistance to chemotherapy drugs in comparison with 2D plate cultures. Transfer of extracted cells to the mice caused enhanced tumor volume and decreased life span. This study indicated that collagen-chitosan nanoscaffolds provide a suitable model of tumor that would be appropriate for tumor studies. Copyright © 2016. Published by Elsevier B.V.

  18. Preparation, characterization and biological test of 3D-scaffolds based on chitosan, fibroin and hydroxyapatite for bone tissue engineering.

    Science.gov (United States)

    Lima, Paulo Autran Leite; Resende, Cristiane Xavier; Soares, Glória Dulce de Almeida; Anselme, Karine; Almeida, Luís Eduardo

    2013-08-01

    This work describes the preparation and characterization of porous 3D-scaffolds based on chitosan (CHI), chitosan/silk fibroin (CHI/SF) and chitosan/silk fibroin/hydroxyapatite (CHI/SF/HA) by freeze drying. The biomaterials were characterized by X-ray diffraction, attenuated total reflection Fourier transform infrared spectroscopy, thermogravimetric analysis, differential scanning calorimetry, scanning electron microscopy and energy dispersive spectroscopy. In addition, studies of porosity, pore size, contact angle and biological response of SaOs-2osteoblastic cells were performed. The CHI scaffolds have a porosity of 94.2±0.9%, which is statistically higher than the one presented by CHI/SF/HA scaffolds, 89.7±2.6%. Although all scaffolds were able to promote adhesion, growth and maintenance of osteogenic differentiation of SaOs-2 cells, the new 3D-scaffold based on CHI/SF/HA showed a significantly higher cell growth at 7 days and 21 days and the level of alkaline phosphatase at 14 and 21 days was statistically superior compared to other tested materials. Copyright © 2013 Elsevier B.V. All rights reserved.

  19. Modeling and Reconstruction of Micro-structured 3D Chitosan/Gelatin Porous Scaffolds Using Micro-CT

    Science.gov (United States)

    Gong, Haibo; Li, Dichen; He, Jiankang; Liu, Yaxiong; Lian, Qin; Zhao, Jinna

    2008-09-01

    Three dimensional (3D) channel networks are the key to promise the uniform distribution of nutrients inside 3D hepatic tissue engineering scaffolds and prompt elimination of metabolic products out of the scaffolds. 3D chitosan/gelatin porous scaffolds with predefined internal channels were fabricated and a combination of light microscope, laser confocal microscopy and micro-CT were employed to characterize the structure of porous scaffolds. In order to evaluate the flow field distribution inside the micro-structured 3D scaffolds, a computer reconstructing method based on Micro-CT was proposed. According to this evaluating method, a contrast between 3D porous scaffolds with and without predefined internal channels was also performed to assess scaffolds' fluid characters. Results showed that the internal channel of the 3D scaffolds formed the 3D fluid channel network; the uniformity of flow field distribution of the scaffolds fabricated in this paper was better than the simple porous scaffold without micro-fluid channels.

  20. Gentamicin-Loaded Thermosetting Hydrogel and Moldable Composite Scaffold: Formulation Study and Biologic Evaluation.

    Science.gov (United States)

    Dorati, Rossella; De Trizio, Antonella; Genta, Ida; Merelli, Alessia; Modena, Tiziana; Conti, Bice

    2017-06-01

    The aim was to design biodegradable drug delivery systems for gentamicin local delivery, meanwhile acting as scaffold for bone regeneration. Gentamicin-loaded thermosetting composite hydrogels were prepared combining chitosan with bovine bone substitutes (Orthoss® granules), beta-glycerophosphate as cross-linker, and lyophilized to obtain moldable composite scaffolds (moldable composite scaffold loaded with gentamicin [mCSG]). Diverse techniques for gentamicin loading into mCS were investigated by drug incorporation during hydrogel preparation or drug absorption on preformed mCS. Rheologic hydrogel characterization was performed. mCSGs were characterized for porosity, stability (water retention, water uptake), gentamicin release, cell seeding and proliferation, and antimicrobial effect on Escherichia coli ATCC 10356. Results show suitable gentamicin loadings were 4 mg in 1 mL thermosetting composite hydrogel starting solution, irreversible hydrogel thermosetting behavior, and cosolute effect of gentamicin on sol-gel transition. Positive results in terms of porosity (80%-86%), scaffold water uptake, and retention capability were obtained. Antibiotic in vitro release was completed in 4 h. Good cell seeding results were observed for mCSG1-5; mCSG3 and mCSG5 resulted the best as cell proliferation results. mCSG exerted bactericidal effect for 24 h, with superimposition of chitosan bacteriostatic effect in the first 4 h. The results lead to consider the drug delivery for reducing infection risk during bone open surgeries. Copyright © 2017 American Pharmacists Association®. Published by Elsevier Inc. All rights reserved.

  1. In vitro degradation of chitosan composite foams for biomedical applications and effect of bioactive glass as a crosslinker

    Directory of Open Access Journals (Sweden)

    Martins Talita

    2018-02-01

    Full Text Available In tissue engineering applications, 3D scaffolds with adequate structure and composition are required to provide durability that is compatiblewith the regeneration of native tissue. In the present study, the degradation of novel flexible 3D composite foams of chitosan (CH combined with bioactive glass (BGwas evaluated, focusing on the role of BG as a physical crosslinker in the composites, and its effect on the degradation process. Highly porous CH/BG composite foams were obtained, and an elevated degradation temperature and lower degradation rate compared with pure chitosan were observed, probably as a result of greater intermolecular interaction between CH and BG. The Fourier transform infrared spectroscopy (FTIR data suggest that hydrogen bonds were responsible for the physical crosslinking between CH and BG. The results confirm that CH/BG foams can combine controllable bioactivity and degradation behavior and, therefore, could be useful for tissue regeneration matrices.

  2. Construction of a fluorescent nanostructured chitosan-hydroxyapatite scaffold by nanocrystallon induced biomimetic mineralization and its cell biocompatibility.

    Science.gov (United States)

    Wang, Guancong; Zheng, Lin; Zhao, Hongshi; Miao, Junying; Sun, Chunhui; Liu, Hong; Huang, Zhen; Yu, Xiaoqiang; Wang, Jiyang; Tao, Xutang

    2011-05-01

    Biomaterial surfaces and their nanostructures can significantly influence cell growth and viability. Thus, manipulating surface characteristics of scaffolds can be a potential strategy to control cell functions for stem cell tissue engineering. In this study, in order to construct a hydroxyapatite (HAp) coated genipin-chitosan conjugation scaffold (HGCCS) with a well-defined HAp nanostructured surface, we have developed a simple and controllable approach that allows construction of a two-level, three-dimensional (3D) networked structure to provide sufficient calcium source and achieve desired mechanical function and mass transport (permeability and diffusion) properties. Using a nontoxic cross-linker (genipin) and a nanocrystallon induced biomimetic mineralization method, we first assembled a layer of HAp network-like nanostructure on a 3D porous chitosan-based framework. X-ray diffraction (XRD) and high resolution transmission electron microscopy (HRTEM) analysis confirm that the continuous network-like nanostructure on the channel surface of the HGCCS is composed of crystalline HAp. Compressive testing demonstrated that the strength of the HGCCS is apparently enhanced because of the strong cross-linking of genipin and the resulting reinforcement of the HAp nanonetwork. The fluorescence properties of genipin-chitosan conjugation for convenient monitoring of the 3D porous scaffold biodegradability and cell localization in the scaffold was specifically explored using confocal laser scanning microscopy (CLSM). Furthermore, through scanning electron microscope (SEM) observation and immunofluorescence measurements of F-actin, we found that the HAp network-like nanostructure on the surface of the HGCCS can influence the morphology and integrin-mediated cytoskeleton organization of rat bone marrow-derived mesenchymal stem cells (BMSCs). Based on cell proliferation assays, rat BMSCs tend to have higher viability on HGCCS in vitro. The results of this study suggest that

  3. Fluorescent composite scaffolds made of nanodiamonds/polycaprolactone

    Science.gov (United States)

    Cao, Li; Hou, Yanwen; Lafdi, Khalid; Urmey, Kirk

    2015-11-01

    Polycaprolactone (PCL) has been widely studied for biological applications. Biodegradable PCL fibrous scaffold can work as an appropriate substrate for tissue regeneration. In this letter, fluorescent nanodiamonds (FNDs) were prepared after surface passivation with octadecylamine. The FNDs were then mixed with PCL polymer and subsequently electrospun into FNDs/PCL fibrous scaffolds. The obtained scaffolds not only exhibited photoluminescence, but also showed reinforced mechanical strength. Toxicity study indicated FNDs/PCL scaffolds were nontoxic. This biocompatible fluorescent composite fibrous scaffold can support in vitro cell growth and also has the potential to act as an optical probe for tissue engineering application in vitro and in vivo.

  4. POTENTIAL ANTISTATIC PROPERTIES OF A CEMENT COMPOSITION MODIFIED BY CHITOSAN

    Directory of Open Access Journals (Sweden)

    Darchiya Valentina Ivanovna

    2012-10-01

    Full Text Available Environmental compatibility of construction materials and their impact onto the human organism and the environment are the essential factors to be taken account of in the course of construction. Therefore, natural renewable biological polymers arouse interest. Polysaccharide chitin takes a special position among them. It represents one of the most widely spread biological polymers; it is extracted from 100% renewable materials. It is part of the external skeleton of crustaceans and insects, and it also part of cell walls of mushrooms and algae. Any research of potential materials to be generated from chitin and its derivative chitosan may involve a practical implementation. The research of the antistatic properties followed the introduction of 1% of chitosan into the cement composition. Electrostatic field intensity was measured by Electrostatic Field Intensity Meter ST-01. The electrostatic property of the sample modified by chitosan turned out to be lower than the one of the benchmark sample by 5.6 times. The presence of chitosan in the cement composition makes no impact on strength-related properties of the construction material. The cement composition modified by chitosan may be used in the manufacturing of antistatic self-leveling floors.

  5. 2-N, 6-O-sulfated chitosan-assisted BMP-2 immobilization of PCL scaffolds for enhanced osteoinduction

    Energy Technology Data Exchange (ETDEWEB)

    Cao, Lingyan [Key Laboratory for Ultrafine Materials of Ministry of Education, East China University of Science and Technology, Shanghai 200237 (China); Engineering Research Center for Biomedical Materials of Ministry of Education, East China University of Science and Technology, Shanghai 200237 (China); CSIRO Manufacturing, Bayview Avenue, Clayton, Victoria 3168 (Australia); Department of Prosthodontics, College of Stomatology, Ninth People' s Hospital, School of Medicine, Shanghai Jiao Tong University, 639 Zhizaoju Road, Shanghai 200011 (China); Yu, Yuanman [Key Laboratory for Ultrafine Materials of Ministry of Education, East China University of Science and Technology, Shanghai 200237 (China); Engineering Research Center for Biomedical Materials of Ministry of Education, East China University of Science and Technology, Shanghai 200237 (China); Wang, Jing, E-mail: biomatwj@163.com [Key Laboratory for Ultrafine Materials of Ministry of Education, East China University of Science and Technology, Shanghai 200237 (China); Engineering Research Center for Biomedical Materials of Ministry of Education, East China University of Science and Technology, Shanghai 200237 (China); Werkmeister, Jerome A [CSIRO Manufacturing, Bayview Avenue, Clayton, Victoria 3168 (Australia); McLean, Keith M, E-mail: Keith.McLean@csiro.au [CSIRO Manufacturing, Bayview Avenue, Clayton, Victoria 3168 (Australia); Liu, Changsheng, E-mail: liucs@ecust.edu.cn [Key Laboratory for Ultrafine Materials of Ministry of Education, East China University of Science and Technology, Shanghai 200237 (China); Engineering Research Center for Biomedical Materials of Ministry of Education, East China University of Science and Technology, Shanghai 200237 (China)

    2017-05-01

    The aim of this study was to develop a 2-N, 6-O-sulfated chitosan (26SCS) modified electrospun fibrous PCL scaffold for bone morphogenetic protein-2 (BMP-2) delivery to improve osteoinduction. The PCL scaffold was modified by an aminolysis reaction using ethylenediamine (ED) and 26SCS was immobilized via electrostatic interactions (PCL-N-S). Scaffolds were characterized by scanning electron microscopy (SEM), atomic force microscopy (AFM), X-ray photoelectron spectroscopy (XPS) and contact angle measurements. In vitro BMP-2 adsorption and release kinetics indicated that modified PCL-N-S scaffolds showed higher levels of binding of BMP-2 (about 30–100 times), moderative burst release (about one third), and prolonged releasing time compared to the unmodified PCL scaffold. The bioactivity of released BMP-2 determined by alkaline phosphatase (ALP) activity assay was maintained and improved 8– 12 times with increasing concentration of immobilized 26SCS on the scaffolds. In vitro studies demonstrated that bone marrow mesenchymal stem cells (BMSCs) attached more readily to the PCL-N-S scaffolds with increased spreading. In conclusion, 26SCS modified PCL scaffolds can be a potent system for the sustained and bioactive delivery of BMP-2. - Graphical abstract: Limited self-regenerating capacity of human body makes the reconstruction of critical size bone defect a significant challenge. Although bone morphogenetic protein-2 (BMP-2) is an important differentiation factor inducing bone regeneration, it's short half-life in vivo and potent side effect at high dosage still show lots of concerns in the clinical use. Herein, modification of electrospun PCL scaffolds was presented through immobilizing of sulfated chitosan (26SCS). The modified scaffolds effectively improve the binding capacity of BMP-2 and exhibited an enhanced bioactivity and sustained release in vitro. Thus, the use of 26SCS modified PCL scaffolds combined with BMP-2 could be a useful scaffold for tissue

  6. Super-paramagnetic responsive silk fibroin/chitosan/magnetite scaffolds with tunable pore structures for bone tissue engineering applications

    Energy Technology Data Exchange (ETDEWEB)

    Aliramaji, Shamsa; Zamanian, Ali, E-mail: a-zamanian@merc.ac.ir; Mozafari, Masoud

    2017-01-01

    Tissue engineering is a promising approach in repairing damaged tissues. During the last few years, magnetic nanoparticles have been of great interest in this field of study due to their controlled responsive characteristics in specific external magnetic fields. In this study, after synthesizing iron oxide (magnetite) nanoparticles through a reverse coprecipitation method, silk fibroin/chitosan-based magnetic scaffolds were prepared using different amounts of magnetite nanoparticles (0, 0.5, 1 and 2%) by freeze-casting method. The physicochemical activity of the scaffolds was monitored in phosphate-buffered saline (PBS) solution to determine the biodegradation and swelling behaviors. The stability of the magnetite nanoparticles in the fabricated scaffolds was determined by atomic absorption spectroscopy (AAS). Moreover, the cellular activity of the magnetic scaffolds was examined under a static magnetic field. The results showed that the lamellar structured scaffolds having MNPs in the walls could not affect the final structure and deteriorate the biological characteristics of the scaffolds, while the ability of magnetic responsivity was added to the scaffolds. This study warrants further pre-clinical and clinical evaluations. - Highlights: • Based on TEM micrograph and Rietveld refinement the particle size of MNPs was approximately 12 nm. • The water absorption of silk scaffolds increases by the addition of chitosan content. • Addition of 0.5 wt% MNPs led to decrease in scaffolds degradation and number of living cells. • By increasing the MNPs from 0.5 to 1 and 2, the degradation rate and living cells increased. • In scaffolds with 2 wt% MNPs cell attachment is slightly better than those of 0.5 wt%.

  7. Preparation and characterization of malonic acid cross-linked chitosan and collagen 3D scaffolds: an approach on non-covalent interactions.

    Science.gov (United States)

    Mitra, Tapas; Sailakshmi, G; Gnanamani, A; Mandal, A B

    2012-05-01

    The present study emphasizes the influence of non-covalent interactions on the mechanical and thermal properties of the scaffolds of chitosan/collagen origin. Malonic acid (MA), a bifuncitonal diacid was chosen to offer non-covalent cross-linking. Three dimensional scaffolds was prepared using chitosan at 1.0% (w/v) and MA at 0.2% (w/v), similarly collagen 0.5% (w/v) and MA 0.2% (w/v) and characterized. Results on FT-IR, TGA, DSC, SEM and mechanical properties (tensile strength, stiffness, Young's modulus, etc.) assessment demonstrated the existence of non-covalent interaction between MA and chitosan/collagen, which offered flexibility and high strength to the scaffolds suitable for tissue engineering research. Studies using NIH 3T3 fibroblast cells suggested biocompatibility nature of the scaffolds. Docking simulation study further supports the intermolecular hydrogen bonding interactions between MA and chitosan/collagen.

  8. Enriched fluoride sorption using alumina/chitosan composite

    Energy Technology Data Exchange (ETDEWEB)

    Viswanathan, Natrayasamy, E-mail: natrayasamy_viswanathan@rediffmail.com [Department of Chemistry, Anna University Tiruchirappalli - Dindigul Campus, Dindigul 624 622, Tamil Nadu (India); Meenakshi, S., E-mail: drs_meena@rediffmail.com [Department of Chemistry, Gandhigram Rural University, Gandhigram 624 302, Tamil Nadu (India)

    2010-06-15

    Alumina possesses an appreciable defluoridation capacity (DC) of 1566 mg F{sup -}/kg. In order to improve its DC, it is aimed to prepare alumina polymeric composites using the chitosan. Alumina/chitosan (AlCs) composite was prepared by incorporating alumina particles in the chitosan polymeric matrix, which can be made into any desired form viz., beads, candles and membranes. AlCs composite displayed a maximum DC of 3809 mg F{sup -}/kg than the alumina and chitosan (52 mg F{sup -}/kg). The fluoride removal studies were carried out in batch mode to optimize the equilibrium parameters viz., contact time, pH, co-anions and temperature. The equilibrium data was fitted with Freundlich and Langmuir isotherms to find the best fit for the sorption process. The calculated values of thermodynamic parameters indicate the nature of sorption. The surface characterisation of the sorbent was performed by FTIR, AFM and SEM with EDAX analysis. A possible mechanism of fluoride sorption by AlCs composite has been proposed. Suitability of AlCs composite at field conditions was tested with a field sample taken from a nearby fluoride-endemic village. This work provides a potential platform for the development of defluoridation technology.

  9. Influence of processing parameters on pore structure of 3D porous chitosan-alginate polyelectrolyte complex scaffolds.

    Science.gov (United States)

    Florczyk, Stephen J; Kim, Dae-Joon; Wood, David L; Zhang, Miqin

    2011-09-15

    Fabrication of porous polymeric scaffolds with controlled structure can be challenging. In this study, we investigated the influence of key experimental parameters on the structures and mechanical properties of resultant porous chitosan-alginate (CA) polyelectrolyte complex (PEC) scaffolds, and on proliferation of MG-63 osteoblast-like cells, targeted at bone tissue engineering. We demonstrated that the porous structure is largely affected by the solution viscosity, which can be regulated by the acetic acid and alginate concentrations. We found that the CA PEC solutions with viscosity below 300 Pa.s yielded scaffolds of uniform pore structure and that more neutral pH promoted more complete complexation of chitosan and alginate, yielding stiffer scaffolds. CA PEC scaffolds produced from solutions with viscosities below 300 Pa.s also showed enhanced cell proliferation compared with other samples. By controlling the key experimental parameters identified in this study, CA PEC scaffolds of different structures can be made to suit various tissue engineering applications. Copyright © 2011 Wiley Periodicals, Inc.

  10. Effects of chitosan-coated fibers as a scaffold for three-dimensional cultures of rabbit fibroblasts for ligament tissue engineering.

    Science.gov (United States)

    Sarukawa, Junichiro; Takahashi, Masaaki; Abe, Masashi; Suzuki, Daisuke; Tokura, Seiichi; Furuike, Tetsuya; Tamura, Hiroshi

    2011-01-01

    Material selection in tissue-engineering scaffolds is one of the primary factors defining cellular response and matrix formation. In this study, we fabricated chitosan-coated poly(lactic acid) (PLA) fiber scaffolds to test our hypothesis that PLA fibers coated with chitosan highly promoted cell supporting properties compared to those without chitosan. Both PLA fibers (PLA group) and chitosan-coated PLA fibers (PLA-chitosan group) were fabricated for this study. Anterior cruciate ligament (ACL) fibroblasts were isolated from Japanese white rabbits and cultured on scaffolds consisting of each type of fiber. The effects of cell adhesivity, proliferation, and synthesis of the extracellular matrix (ECM) for each fiber were analyzed by cell counting, hydroxyproline assay, scanning electron microscopy and quantitative RT-PCR. Cell adhesivity, proliferation, hydroxyproline content and the expression of type-I collagen mRNA were significantly higher in the PLA-chitosan group than in the PLA group. Scanning electron microscopic observation showed that fibroblasts proliferated with a high level of ECM synthesis around the cells. Chitosan coating improved ACL fibroblast adhesion and proliferation, and had a positive effect on matrix production. Thus, the advantages of chitosan-coated PLA fibers show them to be a suitable biomaterial for ACL tissue-engineering scaffolds.

  11. Biomedical potential of chitosan/HA and chitosan/β-1,3-glucan/HA biomaterials as scaffolds for bone regeneration — A comparative study

    Energy Technology Data Exchange (ETDEWEB)

    Przekora, Agata, E-mail: agata.przekora@umlub.pl [Department of Biochemistry and Biotechnology, Medical University of Lublin, Chodzki 1, 20-093 Lublin (Poland); Palka, Krzysztof [Department of Materials Engineering, Lublin University of Technology, Nadbystrzycka 36, 20-618 Lublin (Poland); Ginalska, Grazyna [Department of Biochemistry and Biotechnology, Medical University of Lublin, Chodzki 1, 20-093 Lublin (Poland)

    2016-01-01

    The aim of this work was to compare biomedical potential of chitosan/hydroxyapatite (chit/HA) and novel chitosan/β-1,3-glucan/hydroxyapatite (chit/glu/HA) materials as scaffolds for bone regeneration via characterization of their biocompatibility, porosity, mechanical properties, and water uptake behaviour. Biocompatibility of the scaffolds was assessed in direct-contact with the materials using normal human foetal osteoblast cell line. Cytotoxicity and osteoblast proliferation rate were evaluated. Porosity was assessed using computed microtomography analysis and mechanical properties were determined by compression testing. Obtained results demonstrated that chit/HA scaffold possessed significantly better mechanical properties (compressive strength: 1.23 MPa, Young's modulus: 0.46 MPa) than chit/glu/HA material (compressive strength: 0.26 MPa, Young's modulus: 0.25 MPa). However, addition of bacterial β-1,3-glucan to the chit/HA scaffold improved its flexibility and porosity. Moreover, chit/glu/HA scaffold revealed significantly higher water uptake capability (52.6% after 24 h of soaking) compared to the chit/HA (30.7%) and thus can serve as a very good drug delivery carrier. Chit/glu/HA scaffold was also more favourable to osteoblast survival (near 100% viability after 24-h culture), proliferation, and spreading compared to the chit/HA (63% viability). The chit/glu/HA possesses better biomedical potential than chit/HA scaffold. Nevertheless, poor mechanical properties of the chit/glu/HA limit its application to non-load bearing implantation area. - Highlights: • Chitosan/HA and chit/β-1,3-glucan/HA scaffolds for bone regeneration were compared. • Chit/HA significantly reduced osteoblast viability to 63% compared to chit/glu/HA. • Unlike chit/HA, chit/glu/HA favoured cell adhesion, spreading, and proliferation. • Chit/HA had better compressive strength and Young's modulus than chit/glu/HA. • Chit/glu/HA revealed significantly higher

  12. Biocompatible nanocomposite scaffolds based on copolymer-grafted chitosan for bone tissue engineering with drug delivery capability

    International Nuclear Information System (INIS)

    Saber-Samandari, Samaneh; Saber-Samandari, Saeed

    2017-01-01

    Significant efforts have been made to develop a suitable biocompatible scaffold for bone tissue engineering. In this work, a chitosan-graft-poly(acrylic acid-co-acrylamide)/hydroxyapatite nanocomposite scaffold was synthesized through a novel multi-step route. The prepared scaffolds were characterized for crystallinity, morphology, elemental analysis, chemical bonds, and pores size in their structure. The mechanical properties (i.e. compressive strength and elastic modulus) of the scaffolds were examined. Further, the biocompatibility of scaffolds was determined by MTT assays on HUGU cells. The result of cell culture experiments demonstrated that the prepared scaffolds have good cytocompatibility without any cytotoxicity, and with the incorporation of hydroxyapatite in their structure improves cell viability and proliferation. Finally, celecoxib as a model drug was efficiently loaded into the prepared scaffolds because of the large specific surface area. The in vitro release of the drug displayed a biphasic pattern with a low initial burst and a sustained release of up to 14 days. Furthermore, different release kinetic models were employed for the description of the release process. The results suggested that the prepared cytocompatible and non-toxic nanocomposite scaffolds might be efficient implants and drug carriers in bone-tissue engineering. - Highlights: • A series of biocompatible scaffolds were synthesized through a novel multi-step route. • The mechanical properties of the scaffolds were found close to those of trabecular bone. • The prepared scaffolds were able to load celecoxib efficiently as a model drug. • The celecoxib release was mainly controlled by a Fickian diffusion process. • The scaffold can be efficient as an implant for tissue engineering and drug delivery.

  13. Biocompatible nanocomposite scaffolds based on copolymer-grafted chitosan for bone tissue engineering with drug delivery capability

    Energy Technology Data Exchange (ETDEWEB)

    Saber-Samandari, Samaneh, E-mail: samaneh.saber@gmail.com [Department of Chemistry, Eastern Mediterranean University, Gazimagusa, TRNC via Mersin 10 (Turkey); Saber-Samandari, Saeed, E-mail: saeedss@aut.ac.ir [New Technologies Research Center, Amirkabir University of Technology, Tehran (Iran, Islamic Republic of)

    2017-06-01

    Significant efforts have been made to develop a suitable biocompatible scaffold for bone tissue engineering. In this work, a chitosan-graft-poly(acrylic acid-co-acrylamide)/hydroxyapatite nanocomposite scaffold was synthesized through a novel multi-step route. The prepared scaffolds were characterized for crystallinity, morphology, elemental analysis, chemical bonds, and pores size in their structure. The mechanical properties (i.e. compressive strength and elastic modulus) of the scaffolds were examined. Further, the biocompatibility of scaffolds was determined by MTT assays on HUGU cells. The result of cell culture experiments demonstrated that the prepared scaffolds have good cytocompatibility without any cytotoxicity, and with the incorporation of hydroxyapatite in their structure improves cell viability and proliferation. Finally, celecoxib as a model drug was efficiently loaded into the prepared scaffolds because of the large specific surface area. The in vitro release of the drug displayed a biphasic pattern with a low initial burst and a sustained release of up to 14 days. Furthermore, different release kinetic models were employed for the description of the release process. The results suggested that the prepared cytocompatible and non-toxic nanocomposite scaffolds might be efficient implants and drug carriers in bone-tissue engineering. - Highlights: • A series of biocompatible scaffolds were synthesized through a novel multi-step route. • The mechanical properties of the scaffolds were found close to those of trabecular bone. • The prepared scaffolds were able to load celecoxib efficiently as a model drug. • The celecoxib release was mainly controlled by a Fickian diffusion process. • The scaffold can be efficient as an implant for tissue engineering and drug delivery.

  14. Chitosan-based films composites for wound healing purposes

    International Nuclear Information System (INIS)

    Alves, Natali de O.; Silva, Gabriela T. da; Schulz, Gracelie A.S.; Fajardo, Andre R.

    2015-01-01

    Chitosan has been extensively applied in the developing of biomaterials due to its desirable good physico-chemical and biological properties. According to this, here films composite of chitosan, poly(vinyl alcohol) and bovine bone powder were prepared by casting willing to be applied in wound healing purposes. Moreover, the first step was the developing of a suitable method to obtain bovine bone powder, which was utilized here as filler. All the materials and films were fully characterized by FTIR, DRX and thermal analysis. Water uptake capacity was measured by swelling assays. (author)

  15. Novel development of carbonate apatite-chitosan scaffolds based on lyophilization technique for bone tissue engineering

    Directory of Open Access Journals (Sweden)

    Maretaningtias Dwi Ariani

    2012-09-01

    Full Text Available Background: The natural biopolymer chitosan (Ch is currently regarded as a candidate for bone tissue engineering. However, Ch is poor for cell adhesion and low bone formation ability. In order to enhance cell adhesion and bone formation ability, combination of Ch with carbonate apatite (CA was developed. Purpose: The aim of this study was to make carbonate apatite-chitosan scaffolds (CAChSs and evaluate its osteoconductivity in terms of cell proliferation. Methods: Chitosan scaffolds (ChSs were made by the following procedure. Twenty-five, 50, 100, 200 and 400 mg Ch was dissolved into 5 ml of 2% acetic acid (CH3COOH, shaked for 15 min and neutralized with 15 ml of 0.1 M sodium hydroxide (NaOH solution. After centrifugation, Ch gel was packed into the molds then frozen at -80°C for 2h and dried in a freeze dry machine for 24h. The sponges were subjected to UV radiation for 2h. To make CA-ChSs, 200 mg Ch was selected. After neutralization, 50 mg of 0.06 M CA were added into the 200 mg Ch gel. The structure of CA-ChSs was observed by scanning electron microscope (SEM. Mouse osteoblast-like cell (MC3T3-E1 proliferation in these scaffolds was investigated at 1, 7, 14 and 21 days. Results: Three dimensional porous structures of CA-ChSs were clearly observed by SEM. Proliferated cell numbers in CA-ChSs was significantly higher than those in ChSs (control at each stage (p<0.05. Conclusion: It can be concluded that newly developed CA-ChSs had three-dimensional interconnected porous structure, good handling property and supporting ability of proliferation of osteoblasts. It is suggested that newly developed CA-ChSs could be considered as a scaffolds material for bone tissue enginearing.Latar belakang: Kitosan yang merupakan biopolimer alami dianggap sebagai salah satu kandidat untuk rekayasa jaringan tulang. Namun, kitosan memiliki kelemahan terhadap adhesi sel dan kurang mampu membentuk tulang yang cukup. Untuk meningkatkan adhesi sel dan kemampuan

  16. PMAA-stabilized ferrofluid/chitosan/yeast composite for bioapplications

    International Nuclear Information System (INIS)

    Baldikova, Eva; Prochazkova, Jitka; Stepanek, Miroslav; Hajduova, Jana; Pospiskova, Kristyna; Safarikova, Mirka; Safarik, Ivo

    2017-01-01

    A simple, one-pot process for the preparation of magnetically responsive yeast-based biocatalysts was developed. Saccharomyces cerevisiae, Candida utilis and Kluyveromyces lactis cells were successfully incorporated into chitosan gel magnetically modified with poly(methacrylic acid)-stabilized magnetic fluid (PMAA-FF) during its formation. Magnetic PMAA-FF/chitosan/yeast composites were efficiently employed for invert sugar production. The dependence of invertase activity on used yeast, amount of magnetic biocatalyst, agitation time and after reuse was studied in detail. The tested magnetic biocatalysts retained at least 69% of their initial activity after 8 reuse cycles. - Highlights: • New types of magnetically responsive yeast biocomposites were prepared. • Recently developed PMAA-stabilized magnetic fluid was used. • Three yeast species were entrapped into magnetic chitosan gel during its formation. • All biocatalysts were efficiently employed for invert sugar formation.

  17. PMAA-stabilized ferrofluid/chitosan/yeast composite for bioapplications

    Energy Technology Data Exchange (ETDEWEB)

    Baldikova, Eva, E-mail: baldie@email.cz [Global Change Research Institute, CAS, Na Sadkach 7, 370 05 Ceske Budejovice (Czech Republic); Department of Applied Chemistry, Faculty of Agriculture, University of South Bohemia, Branisovska 1457, 370 05 Ceske Budejovice (Czech Republic); Prochazkova, Jitka [Global Change Research Institute, CAS, Na Sadkach 7, 370 05 Ceske Budejovice (Czech Republic); Stepanek, Miroslav; Hajduova, Jana [Department of Physical and Macromolecular Chemistry, Faculty of Science, Charles University, Hlavova 2030, 128 40 Prague 2 (Czech Republic); Pospiskova, Kristyna [Regional Centre of Advanced Technologies and Materials, Palacky University, Slechtitelu 27, 783 71 Olomouc (Czech Republic); Safarikova, Mirka [Global Change Research Institute, CAS, Na Sadkach 7, 370 05 Ceske Budejovice (Czech Republic); Department of Nanobiotechnology, Biology Centre, CAS, ISB, Na Sadkach 7, 370 05 Ceske Budejovice (Czech Republic); Safarik, Ivo [Global Change Research Institute, CAS, Na Sadkach 7, 370 05 Ceske Budejovice (Czech Republic); Regional Centre of Advanced Technologies and Materials, Palacky University, Slechtitelu 27, 783 71 Olomouc (Czech Republic); Department of Nanobiotechnology, Biology Centre, CAS, ISB, Na Sadkach 7, 370 05 Ceske Budejovice (Czech Republic)

    2017-04-01

    A simple, one-pot process for the preparation of magnetically responsive yeast-based biocatalysts was developed. Saccharomyces cerevisiae, Candida utilis and Kluyveromyces lactis cells were successfully incorporated into chitosan gel magnetically modified with poly(methacrylic acid)-stabilized magnetic fluid (PMAA-FF) during its formation. Magnetic PMAA-FF/chitosan/yeast composites were efficiently employed for invert sugar production. The dependence of invertase activity on used yeast, amount of magnetic biocatalyst, agitation time and after reuse was studied in detail. The tested magnetic biocatalysts retained at least 69% of their initial activity after 8 reuse cycles. - Highlights: • New types of magnetically responsive yeast biocomposites were prepared. • Recently developed PMAA-stabilized magnetic fluid was used. • Three yeast species were entrapped into magnetic chitosan gel during its formation. • All biocatalysts were efficiently employed for invert sugar formation.

  18. 3D nanocomposite chitosan/bioactive glass scaffolds obtained using two different routes: an evaluation of the porous structure and mechanical properties

    Directory of Open Access Journals (Sweden)

    Elke M. F. Lemos

    2016-05-01

    Full Text Available Porous synthetic substrates are developed through tissue engineering technologies to grow new tissue, restoring the function of tissue or an organ. For bone regeneration, these scaffolds must support the dynamic load exerted on this tissue, achieved primarily by increasing their compression strength, as established in the literature. The aim of this paper was to incorporate an inorganic composite bioactive glass (60%SiO2 - 36%CaO - 4%P2O5 as a reinforcing agent in mechanical 3D scaffolds that must remain porous. Two strategies were adopted: a co-precipitation method to obtain a nanoparticulate dispersion of bioactive glass (BGNP and a sol-gel method to combine a bioactive glass solution (BG with a previously prepared chitosan polymer solution. Moreover, a lyophilization process was also used, generating highly porous scaffolds. Various aspects of the scaffold were evaluated, including the morphology, orientation and size of the pores, and mechanical strength, as obtained using the two synthetic methods. The data for compressive strength revealed increased strength after the incorporation of bioactive glass, which was more pronounced when utilizing the nanoscale bioactive glass.

  19. In vitro cartilage construct generation from silk fibroin- chitosan porous scaffold and umbilical cord blood derived human mesenchymal stem cells in dynamic culture condition.

    Science.gov (United States)

    Agrawal, Parinita; Pramanik, Krishna; Biswas, Amit; Ku Patra, Ranjan

    2018-02-01

    Cartilage construct generation includes a scaffold with appropriate composition to mimic matrix of the damaged tissue on which the stem cells grow and differentiate. In this study, umbilical cord blood (UCB) derived human mesenchymal stem cells (hMSCs) were seeded on freeze dried porous silk-fibroin (SF)/chitosan (CS) scaffolds. Influence of static and dynamic (spinner flask bioreactor) culture conditions on the developing cartilage construct were studied by in-vitro characterization for viability, proliferation, distribution, and chondrogenic differentiation of hMSCs over the scaffold. Constructs developed in spinner flask consisted of 62% live cells, and exhibited 543% more cell density at the core than constructs cultured in static system. Quantification of DNA and glycosaminoglycans accumulation after 21 days showed the progression of chondrogenic differentiation of hMSCs was higher in dynamic culture compared to static one. In constructs generated under dynamic condition, histology staining for proteoglycan matrix, and fluorescence staining for collagen-II and aggrecan showed positive correlation between early and late stage chondrogenic markers, which was further confirmed by quantitative PCR analysis, showing low collagen-I expression and highly expressed Sox9, collagen-II and aggrecan. The present study demonstrated that construct generated by combining 3D SF/CS scaffold with UCB-hMSCs under dynamic condition using spinner flask bioreactor can be used for cartilage tissue regeneration for future medical treatments. © 2017 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 106A: 397-407, 2018. © 2017 Wiley Periodicals, Inc.

  20. Enhanced biological properties of biomimetic apatite fabricated polycaprolactone/chitosan nanofibrous bio-composite for tendon and ligament regeneration.

    Science.gov (United States)

    Wu, Geng; Deng, Xuefeng; Song, Jinqi; Chen, Feiqiang

    2018-01-01

    The development of tailored nanofibrous scaffolds for tendon and ligament tissue engineering has been a goal of clinical research for current researchers. Here, we establish a formation of novel nanofibrous matrix with significant mechanical and biological properties by electro-spinning process. The fine fibrous morphology of the nanostructured hydroxyapatite (HAp) dispersed in the polycaprolactone/chitosan (HAp-PCL/CS) nanofibrous matrix was exhibited by microscopic (SEM and TEM) techniques. The favorable mechanical properties (load and modulus) were achieved. The load and modulus of the HAp-PCL/CS composite fibers was 250.1N and 215.5MPa, which is very similar to that of standard value of the human tendon and ligament tissues. The cellular responses and biocompatibility of HAp-PCL/CS nanofibrous scaffolds were investigated with human osteoblast (HOS) cells for tendon regeneration and examined the primary osteoblast mechanism by in vitro method. The morphological (FE-SEM and fluorescence) microscopic images clearly exhibited that HOS cells are well attached and flatted on the nanofibrous composites. The HAp dispersed PCL/CS nanofibrous scaffolds promoted higher adhesion and proliferation of HOS cells comparable to the nanofibrous scaffolds without HAp nanoparticles. The physic-chemical and biological properties of the synthesized nanofibrous scaffold were very close to that of normal ligament and tendon in human body. Over all, these studied results confirmed that the prepared nanofibrous scaffolds will be effective biomaterial of tendon ligament regeneration applications. Copyright © 2017 Elsevier B.V. All rights reserved.

  1. Skin derived precursor Schwann cell-generated acellular matrix modified chitosan/silk scaffolds for bridging rat sciatic nerve gap.

    Science.gov (United States)

    Zhu, Changlai; Huang, Jing; Xue, Chengbin; Wang, Yaxian; Wang, Shengran; Bao, Shuangxi; Chen, Ruyue; Li, Yuan; Gu, Yun

    2017-12-27

    Extracellular/acellular matrix has been attracted much research interests for its unique biological characteristics, and ACM modified neural scaffolds shows the remarkable role of promoting peripheral nerve regeneration. In this study, skin-derived precursors pre-differentiated into Schwann cells (SKP-SCs) were used as parent cells to generate acellular(ACM) for constructing a ACM-modified neural scaffold. SKP-SCs were co-cultured with chitosan nerve guidance conduits (NGC) and silk fibroin filamentous fillers, followed by decellularization to stimulate ACM deposition. This NGC-based, SKP-SC-derived ACM-modified neural scaffold was used for bridging a 10 mm long rat sciatic nerve gap. Histological and functional evaluation after grafting demonstrated that regenerative outcomes achieved by this engineered neural scaffold were better than those achieved by a plain chitosan-silk fibroin scaffold, and suggested the benefits of SKP-SC-derived ACM for peripheral nerve repair. Copyright © 2017 Elsevier Ireland Ltd and Japan Neuroscience Society. All rights reserved.

  2. Nanofibrous Chitosan-Polyethylene Oxide Engineered Scaffolds: A Comparative Study between Simulated Structural Characteristics and Cells Viability

    Directory of Open Access Journals (Sweden)

    Mohammad Kazemi Pilehrood

    2014-01-01

    Full Text Available 3D nanofibrous chitosan-polyethylene oxide (PEO scaffolds were fabricated by electrospinning at different processing parameters. The structural characteristics, such as pore size, overall porosity, pore interconnectivity, and scaffold percolative efficiency (SPE, were simulated by a robust image analysis. Mouse fibroblast cells (L929 were cultured in RPMI for 2 days in the presence of various samples of nanofibrous chitosan/PEO scaffolds. Cell attachments and corresponding mean viability were enhanced from 50% to 110% compared to that belonging to a control even at packed morphologies of scaffolds constituted from pores with nanoscale diameter. To elucidate the correlation between structural characteristics within the depth of the scaffolds’ profile and cell viability, a comparative analysis was proposed. This analysis revealed that larger fiber diameters and pore sizes can enhance cell viability. On the contrary, increasing the other structural elements such as overall porosity and interconnectivity due to a simultaneous reduction in fiber diameter and pore size through the electrospinning process can reduce the viability of cells. In addition, it was found that manipulation of the processing parameters in electrospinning can compensate for the effects of packed morphologies of nanofibrous scaffolds and can thus potentially improve the infiltration and viability of cells.

  3. Sphere-shaped nano-hydroxyapatite/chitosan/gelatin 3D porous scaffolds increase proliferation and osteogenic differentiation of human induced pluripotent stem cells from gingival fibroblasts

    International Nuclear Information System (INIS)

    Ji, Jun; Tong, Xin; Huang, Xiaofeng; Wang, Tiancong; Lin, Zitong; Cao, Yazhou; Qin, Haiyan; Hu, Qingang; Zhang, Junfeng; Dong, Lei

    2015-01-01

    Hydroxyapatite (HA) is an important component of human bone and bone tissue engineering scaffolds. A plethora of bone tissue engineering scaffolds have been synthesized so far, including nano-HA/chitosan/gelatin (nHA/CG) scaffolds; and for seeding cells, stem cells, especially induced pluripotent stem cells (iPSCs), have been a promising cell source for bone tissue engineering recently. However, the influence of different HA nano-particle morphologies on the osteogenic differentiation of human iPSCs (hiPSCs) from human gingival fibroblasts (hGFs) is unknown. The purpose of this study was to investigate the osteogenic differentiation of hiPSCs from hGFs seeded on nHA/CG scaffolds with 2 shapes (rod and sphere) of nHA particles. Firstly, hGFs isolated from discarded normal gingival tissues were reprogrammed into hiPSCs. Secondly, hiPSCs were seeded on rod-like nHA/CG (rod-nHA/CG) and sphere-shaped nHA/CG (sphere-nHA/CG) scaffolds respectively and then cell/scaffold complexes were cultured in vitro. Scanning electron microscope, hematoxyline and eosin (HE) staining, Masson’s staining, and quantitative real-time polymerase chain reaction techniques were used to examine hiPSC morphology, proliferation, and differentiation on rod-nHA/CG and sphere-nHA/CG scaffolds. Finally, hiPSCs composited with 2 kinds of nHA/CG were transplanted in vivo in a subcutaneous implantation model for 12 weeks; pure scaffolds were also transplanted as a blank control. HE, Masson’s, and immunohistochemistry staining were applied to detect new bone regeneration ability. The results showed that sphere-nHA/CG significantly increased hiPSCs from hGF proliferation and osteogenic differentiation in vitro. hiPSCs and sphere-nHA/CG composities generated large bone, whereas hiPSCs and rod-nHA/CG composities produced tiny bone in vivo. Moreover, pure scaffolds without cells almost produced no bone. In conclusion, our work provided a potential innovative bone tissue engineering approach using

  4. Sphere-shaped nano-hydroxyapatite/chitosan/gelatin 3D porous scaffolds increase proliferation and osteogenic differentiation of human induced pluripotent stem cells from gingival fibroblasts.

    Science.gov (United States)

    Ji, Jun; Tong, Xin; Huang, Xiaofeng; Wang, Tiancong; Lin, Zitong; Cao, Yazhou; Zhang, Junfeng; Dong, Lei; Qin, Haiyan; Hu, Qingang

    2015-07-08

    Hydroxyapatite (HA) is an important component of human bone and bone tissue engineering scaffolds. A plethora of bone tissue engineering scaffolds have been synthesized so far, including nano-HA/chitosan/gelatin (nHA/CG) scaffolds; and for seeding cells, stem cells, especially induced pluripotent stem cells (iPSCs), have been a promising cell source for bone tissue engineering recently. However, the influence of different HA nano-particle morphologies on the osteogenic differentiation of human iPSCs (hiPSCs) from human gingival fibroblasts (hGFs) is unknown. The purpose of this study was to investigate the osteogenic differentiation of hiPSCs from hGFs seeded on nHA/CG scaffolds with 2 shapes (rod and sphere) of nHA particles. Firstly, hGFs isolated from discarded normal gingival tissues were reprogrammed into hiPSCs. Secondly, hiPSCs were seeded on rod-like nHA/CG (rod-nHA/CG) and sphere-shaped nHA/CG (sphere-nHA/CG) scaffolds respectively and then cell/scaffold complexes were cultured in vitro. Scanning electron microscope, hematoxyline and eosin (HE) staining, Masson's staining, and quantitative real-time polymerase chain reaction techniques were used to examine hiPSC morphology, proliferation, and differentiation on rod-nHA/CG and sphere-nHA/CG scaffolds. Finally, hiPSCs composited with 2 kinds of nHA/CG were transplanted in vivo in a subcutaneous implantation model for 12 weeks; pure scaffolds were also transplanted as a blank control. HE, Masson's, and immunohistochemistry staining were applied to detect new bone regeneration ability. The results showed that sphere-nHA/CG significantly increased hiPSCs from hGF proliferation and osteogenic differentiation in vitro. hiPSCs and sphere-nHA/CG composities generated large bone, whereas hiPSCs and rod-nHA/CG composities produced tiny bone in vivo. Moreover, pure scaffolds without cells almost produced no bone. In conclusion, our work provided a potential innovative bone tissue engineering approach using

  5. Poly(N-isopropylacrylamide) hydrogel/chitosan scaffold hybrid for three-dimensional stem cell culture and cartilage tissue engineering.

    Science.gov (United States)

    Mellati, Amir; Kiamahalleh, Meisam Valizadeh; Madani, S Hadi; Dai, Sheng; Bi, Jingxiu; Jin, Bo; Zhang, Hu

    2016-11-01

    Providing a controllable and definable three-dimensional (3D) microenvironment for chondrogenic differentiation of mesenchymal stem cells (MSCs) remains a great challenge for cartilage tissue engineering. In this work, poly(N-isopropylacrylamide) (PNIPAAm) polymers with the degrees of polymerization of 100 and 400 (NI100 and NI400) were prepared and the polymer solutions were introduced into the preprepared chitosan porous scaffolds (CS) to form hybrids (CSNI100 and CSNI400, respectively). SEM images indicated that the PNIPAAm gel partially occupied chitosan pores while the interconnected porous structure of chitosan was preserved. MSCs were incorporated within the hybrid and cell proliferation and chondrogenic differentiation were monitored. After 7-day incubation of the cell-laden constructs in a growth medium, the cell viability in CSNI100 and CSNI400 were 54 and 108% higher than that in CS alone, respectively. Glycosaminoglycan and total collagen contents increased 2.6- and 2.5-fold after 28-day culture of cell-laden CSNI400 in the chondrogenic medium. These results suggest that the hybrid structure composed of the chitosan porous scaffold and the well-defined PNIPAAm hydrogel, in particular CSNI400, is suitable for 3D stem cell culture and cartilage tissue engineering. © 2016 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 104A: 2764-2774, 2016. © 2016 Wiley Periodicals, Inc.

  6. Compatibility of Porous Chitosan Scaffold with the Attachment and Proliferation of human Adipose-Derived Stem Cells In Vitro

    Directory of Open Access Journals (Sweden)

    Gomathysankar S

    2016-11-01

    Full Text Available Adipose-derived stem cells (ASCs have potential applications in the repair and regeneration of various tissues and organs. The use of various scaffold materials as an excellent template for mimicking the extracellular matrix to induce the attachment and proliferation of different cell types has always been of interest in the field of tissue engineering because ideal biomaterials are in great demand. Chitosan, a marine polysaccharide, have wide clinical applications and it acts as a promising scaffold for cell migration and proliferation. ASCs, with their multi-differentiation potential, and chitosan, with its great biocompatibility with ASCs, were investigated in the present study. ASCs were isolated and were characterized by two different methods: immunocytochemistry and flow cytometry, using the mesenchymal stem cell markers CD90, CD105, CD73 and CD29. The ASCs were then induced to differentiate into adipogenic, osteogenic and chondrogenic lineages. These ASCs were incorporated into a porous chitosan scaffold (PCS, and their structural morphology was studied using a scanning electron microscope and hematoxylin and eosin staining. The proliferation rate of the ASCs on the PCS was assessed using a PrestoBlue viability assay. The results indicated that the PCS provides an excellent template for the adhesion and proliferation of ASCs. Thus, this study revealed that PCS is a promising biomaterial for inducing the proliferation of ASCs, which could lead to successful tissue reconstruction in the field of tissue engineering.

  7. Use of synovium-derived stromal cells and chitosan/collagen type I scaffolds for cartilage tissue engineering

    Energy Technology Data Exchange (ETDEWEB)

    Gong Zhongcheng; Lin Zhaoquan [Department of Oral and Maxillofacial Surgery, First Affiliated Hospital of Xinjiang Medical University, Urumqi, Xinjiang 830054 (China); Xiong Hui; Long Xing; Wei Lili; Li Jian; Wu Yang, E-mail: xinglong1957@yahoo.com.c [State Key Laboratory Breeding Base of Basic Science of Stomatology (Hubei-MOST) and Key Laboratory of Oral Biomedicine, Ministry of Education, School and Hospital of Stomatology, Wuhan University, Wuhan, Hubei 430079 (China)

    2010-10-01

    The objective was to investigate synovium-derived stromal cells (SDSCs) coupled with chitosan/collagen type I (CS/COL-I) scaffolds for cartilage engineering. CS/COL-I scaffolds were fabricated through freeze-drying and cross-linked by 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide. SDSCs were isolated from synovium and cultured onto CS/COL-I scaffolds, constructs of which were incubated in serum-free chondrogenic medium with sequential application of TGF-{beta}1 and bFGF for up to 21 days and then implanted into nude mice. The physical characteristics of the scaffolds were examined. The quality of the in vitro constructs was assessed in terms of DNA content by PicoGreen assay and cartilaginous matrix by histological examination. The implants of the constructs were evaluated by histological and immunohistochemical examinations and reverse transcription PCR. Results indicated that the CS/COL-I scaffold showed porous structures, and the DNA content of SDSCs in CS/COL-I scaffolds increased at 1 week culture time. Both of the constructs in vitro and the implants were examined with positive stained GAGs histologically and the implants with positive collagen type II immunohistochemically. RT-PCR of the implants indicated that aggrecan and collagen type II expressed. It suggested that SDSCs coupled with CS/COL-I scaffolds treated sequentially with TGF-{beta}1 and bFGF in vitro were highly competent for engineered cartilage formation in vitro and in vivo.

  8. Enhanced Healing of Rat Calvarial Defects with MSCs Loaded on BMP-2 Releasing Chitosan/Alginate/Hydroxyapatite Scaffolds

    Science.gov (United States)

    He, Xiaoning; Liu, Yang; Yuan, Xue; Lu, Li

    2014-01-01

    In this study, we designed a chitosan/alginate/hydroxyapatite scaffold as a carrier for recombinant BMP-2 (CAH/B2), and evaluated the release kinetics of BMP-2. We evaluated the effect of the CAH/B2 scaffold on the viability and differentiation of bone marrow mesenchymal stem cells (MSCs) by scanning electron microscopy, MTS, ALP assay, alizarin-red staining and qRT-PCR. Moreover, MSCs were seeded on scaffolds and used in a 8 mm rat calvarial defect model. New bone formation was assessed by radiology, hematoxylin and eosin staining 12 weeks postoperatively. We found the release kinetics of BMP-2 from the CAH/B2 scaffold were delayed compared with those from collagen gel, which is widely used for BMP-2 delivery. The BMP-2 released from the scaffold increased MSC differentiation and did not show any cytotoxicity. MSCs exhibited greater ALP activity as well as stronger calcium mineral deposition, and the bone-related markers Col1α, osteopontin, and osteocalcin were upregulated. Analysis of in vivo bone formation showed that the CAH/B2 scaffold induced more bone formation than other groups. This study demonstrates that CAH/B2 scaffolds might be useful for delivering osteogenic BMP-2 protein and present a promising bone regeneration strategy. PMID:25084008

  9. Defluoridation of water using magnesia/chitosan composite

    Energy Technology Data Exchange (ETDEWEB)

    Sairam Sundaram, C. [Department of Science and Humanities, Karaikal Polytechnic College, Karaikal 609609, Puducherry (India)], E-mail: sairam_adithya@yahoo.com; Viswanathan, Natrayasamy [Department of Chemistry, Gandhigram Rural University, Gandhigram 624302, Tamilnadu (India)], E-mail: natrayasamy_viswanathan@rediffmail.com; Meenakshi, S. [Department of Chemistry, Gandhigram Rural University, Gandhigram 624302, Tamilnadu (India)], E-mail: drs_meena@rediffmail.com

    2009-04-30

    Magnesia (MgO) is a well-known adsorbent showing extremely high defluoridation capacity (DC). In order to over come the limitations of MgO for field applications, an attempt has been made to modify magnesia with abundant biomaterial chitosan to form magnesia/chitosan (MgOC) composite in a usable form and its merits over conventional magnesia and raw chitosan is established. Removal of fluoride from aqueous solution with MgO and MgOC composite was studied with batch equilibrium experiments. At equilibrium, MgOC composite has a DC of 4440 mg F{sup -}/kg while for magnesia it is only 2175 mg F{sup -}/kg. The physicochemical properties of the synthesised MgOC composite were analyzed with FTIR and SEM with EDAX studies. The equilibrium data were fitted with isotherm and kinetic models. Thermodynamic parameters viz, {delta}G{sup o}, {delta}H{sup o} and {delta}S{sup o} were calculated to understand the nature of sorption. Field studies were carried out to find the suitability of these sorbents at field conditions.

  10. THE USE OF A NOVEL ALDEHYDE-FUNCTIONALIZED CHITOSAN HYDROGEL TO PREPARE POROUS TUBULAR SCAFFOLDS FOR VASCULAR TISSUE ENGINEERING APPLICATIONS

    Directory of Open Access Journals (Sweden)

    Eduardo P. Azevedo

    Full Text Available In this work, porous tubular scaffolds were prepared from a novel water soluble aldehyde-functionalized chitosan (ALDCHIT hydrogel, which was obtained by dissolving this chitosan derivative in water and using oxidized dextrose (OXDEXT as the crosslinking agent at different ALDCHIT:OXDEXT mole ratios (10:1, 10:2 and 10:4. By increasing the amount of OXDEXT in respect to ALDCHIT the hydrogels became more rigid and could absorb more than 200% of its weight in water. Since the ALDCHIT:OXDEXT 10:4 was the most stable hydrogel, its ability to form porous tubular scaffolds was investigated. The tubular scaffolds were prepared by the lyophilization method, where the orientation of the pores was controlled by exposing either the internal or the external surface of the frozen hydrogel during the sublimation step. When only the inner surface of the frozen hydrogel was exposed, tubular scaffolds with a highly porous lumen and a sealed outer surface were obtained, where the orientation of the pores, their sizes and interconnectivity seem to be optimum for vascular tissue engineering application.

  11. Preparation, characteristics and assessment of a novel gelatin-chitosan sponge scaffold as skin tissue engineering material.

    Science.gov (United States)

    Han, Fei; Dong, Yang; Su, Zhen; Yin, Ran; Song, Aihua; Li, Sanming

    2014-12-10

    In order to develop a skin tissue engineering material for wound dressing application, a novel gelatin-chitosan sponge scaffold was designed and studied. The effect of chitosan and gelatin ratio on the morphology, pore size, porosity, water uptake capacity, water retention capacity and the degradation behavior were evaluated. Biocompatibility was investigated by both MTT method and AO/EB staining method. Antibacterial assessment and in vivo pharmacodynamic was also studied to evaluate the potential for wound healing. Results showed the sponge scaffold have uniform porous structure with pore size range between 120 and 140 μm, high porosity (>90%), high water uptake capacity (>1500%), high water retention capacity (>400%), and degradation percent in 28 days between 38.3 and 53.9%. Biocompatibility results showed that the activity of cells could not be affected by the nature of the sponge and it was suitable for cell adhesion and proliferation for 21 days. In vivo evaluation indicated that the sponge scaffold could offer effective support and attachment to cells for skin wound healing. In conclusion, the developed sponge scaffold was a potential skin tissue engineering material with appropriate physical properties and good biocompatibility. Copyright © 2014 Elsevier B.V. All rights reserved.

  12. Reinforced chitosan-based heart valve scaffold and utility of bone marrow-derived mesenchymal stem cells for cardiovascular tissue engineering

    Science.gov (United States)

    Albanna, Mohammad Zaki

    Recent research has demonstrated a strong correlation between the differentiation profile of mesenchymal stem cells (MSCs) and scaffold stiffness. Chitosan is being widely studied for tissue engineering applications due to its biocompatibility and biodegradability. However, its use in load-bearing applications is limited due to moderate to low mechanical properties. In this study, we investigated the effectiveness of a fiber reinforcement method for enhancing the mechanical properties of chitosan scaffolds. Chitosan fibers were fabricated using a solution extrusion and neutralization method and incorporated into porous chitosan scaffolds. The effects of different fiber/scaffold mass ratios, fiber mechanical properties and fiber lengths on scaffold mechanical properties were studied. The results showed that incorporating fibers improved scaffold strength and stiffness in proportion to the fiber/scaffold mass ratio. A fiber-reinforced heart valve leaflet scaffold achieved strength values comparable to the radial values of human pulmonary and aortic valves. Additionally, the effects of shorter fibers (2 mm) were found to be up to 3-fold greater than longer fibers (10 mm). Despite this reduction in fiber mechanical properties caused by heparin crosslinking, the heparin-modified fibers still improved the mechanical properties of the reinforced scaffolds, but to a lesser extent than the unmodified fibers. The results demonstrate that chitosan fiber-reinforcement can be used to generate tissue-matching mechanical properties in porous chitosan scaffolds and that fiber length and mechanical properties are important parameters in defining the degree of mechanical improvement. We further studied various chemical and physical treatments to improve the mechanical properties of chitosan fibers. With combination of chemical and physical treatments, fiber stiffness improved 40fold compared to unmodified fibers. We also isolated ovine bone marrow-derived MSCs and evaluated their

  13. Antibacterial chitosan/silk sericin 3D porous scaffolds as a wound dressing material.

    Science.gov (United States)

    Karahaliloglu, Zeynep; Kilicay, Ebru; Denkbas, Emir Baki

    2017-09-01

    Antimicrobial mixed dressings have traditionally been used to minimize bacterial infection of burns and other wounds. This study presents the advancement of biocompatible chitosan/silk sericin (CHT/SS) scaffolds combined with lauric acid (LA) and zinc oxide nanoparticles (nZnO) for the successful wound dressing applications. Antibacterial assay results showed that the diameters of the inhibition zone increased from 2 ± 0.4 to 7 ± 0.1 mm for Escherichia coli, as well as from 2.5 ± 0.2 to 6 ± 0.4 mm for Staphylococcus aureus while CHTS/SS/100nZnO compared to CHT/SS/0.01LA. The results not only showed excellent inhibition against Gram-positive and Gram-negative bacterial growth but also revealed improved proliferation and extended viability for HaCaT cells.

  14. Evaluation of structural and mechanical properties of electrospun nano-micro hybrid of poly hydroxybutyrate-chitosan/silk scaffold for cartilage tissue engineering.

    Science.gov (United States)

    Karbasi, Saeed; Fekrat, Farnoosh; Semnani, Daryoush; Razavi, Shahnaz; Zargar, Elham Naghash

    2016-01-01

    One of the new methods of scaffold fabrication is a nano-micro hybrid structure in which the properties of the scaffold are improved by introducing nanometer and micrometer structures. This method could be suitable for scaffold designing if some features improve. In this study, electrospun nanofibers of 9% weight solution of poly (3-hydroxybutyrate) (P3HB) and a 15% weight of chitosan by trifluoroacetic acid were coated on both the surface of a silk knitted substrate in the optimum condition to improve the mechanical properties of scaffolds for cartilage tissue engineering application. These hybrid nano-micro fibrous scaffolds were characterized by structural and mechanical evaluation methods. Scanning electron microscopy values and porosity analysis showed that average diameter of nanofibers was 584.94 nm in electrospinning part and general porosity was more than 80%. Fourier transform infrared spectroscopy results indicated the presence of all elements without pollution. The tensile test also stated that by electrospinning, as well as adding chitosan, both maximum strength and maximum elongation increased to 187 N and 10 mm. It means that the microfibrous part of scaffold could affect mechanical properties of nano part of the hybrid scaffold, significantly. It could be concluded that P3HB-chitosan/silk hybrid scaffolds can be a good candidate for cartilage tissue engineering.

  15. Alveolar bone tissue engineering using composite scaffolds for drug delivery

    Directory of Open Access Journals (Sweden)

    Tomonori Matsuno

    2010-08-01

    Full Text Available For many years, bone graft substitutes have been used to reconstruct bone defects in orthopedic and dental fields. However, synthetic bone substitutes such as hydroxyapatite or β-tricalcium phosphate have no osteoinductive or osteogenic abilities. Bone tissue engineering has also been promoted as an alternative approach to regenerating bone tissue. To succeed in bone tissue engineering, osteoconductive scaffolding biomaterials should provide a suitable environment for osteogenic cells and provide local controlled release of osteogenic growth factors. In addition, the scaffold for the bone graft substitute should biodegrade to replace the newly formed bone. Recent advances in bone tissue engineering have allowed the creation of composite scaffolds with tailored functional properties. This review focuses on composite scaffolds that consist of synthetic ceramics and natural polymers as drug delivery carriers for alveolar bone tissue engineering.

  16. Preparation and characterization of photocured poly (ε-caprolactone) diacrylate/poly (ethylene glycol) diacrylate/chitosan for photopolymerization-type 3D printing tissue engineering scaffold application.

    Science.gov (United States)

    Cheng, Yih-Lin; Chen, Freeman

    2017-12-01

    Because of its biocompatible, biodegradable and antimicrobial properties, chitosan is an attractive biomaterial for use in tissue engineering scaffolds. This work builds on previous research by incorporating 95% DD chitosan into a visible-light curable resin which is compatible with a digital light processing (DLP™) projection additive manufacturing (3D printing) system. Different concentrations of chitosan were added to a poly (ε-caprolactone)-diacrylate/poly (ethylene glycol)-diacrylate baseline resin and the samples were extensively characterized. Thermal and mechanical analysis conformed to established scaffold requirements. L929 cells were cultured on the photo-crosslinked films and MTT assays were performed at 1, 3, and 5days to assess cytocompatibility of the resins. Data and SEM images verified a correlation between the concentration of chitosan in the photocurable resin and the adhesion, proliferation, and viability of cell cultures. Finally, the processability of the resins with the dynamic masking DLP system was demonstrated by constructing multi-layer scaffolds with actual measurements that were consistent with the CAD models. These findings encourage the use of chitosan as an additive in visible-light curable resins to improve desired properties in tissue engineering scaffolds. Copyright © 2017 Elsevier B.V. All rights reserved.

  17. HPLC detection of loss rate and cell migration of HUVECs in a proanthocyanidin cross-linked recombinant human collagen-peptide (RHC)–chitosan scaffold

    Energy Technology Data Exchange (ETDEWEB)

    Zhang, Jing; Deng, Aipeng [School of Environmental and Biological Engineering, Nanjing University of Science and Technology, Nanjing 210094 (China); Yang, Yang [Faculty of Engineering, University of Nottingham, Nottingham NG7 2RD (United Kingdom); Gao, Lihu; Xu, Na; Liu, Xin; Hu, Lunxiang; Chen, Junhua [School of Environmental and Biological Engineering, Nanjing University of Science and Technology, Nanjing 210094 (China); Yang, Shulin, E-mail: yshulin@njust.edu.cn [School of Environmental and Biological Engineering, Nanjing University of Science and Technology, Nanjing 210094 (China)

    2015-11-01

    Porous scaffolds with appropriate pore structure, biocompatibility, mechanical property and processability play an important role in tissue engineering. In this paper, we fabricated a recombinant human collagen-peptide (RHC)–chitosan scaffold cross-linked by premixing 30% proanthocyanidin (PA) in one-step freeze-drying. To remove the residual acetic acid, optimized 0.2 M phosphate buffer of pH 6.24 with 30% ethanol (PBSE) was selected to neutralize the lyophilized scaffold followed by three times deionized water rinse. Ninhydrin assay was used to characterize the components loss during the fabrication process. To detect the exact RHC loss under optimized neutralization condition, high performance liquid chromatography (HPLC) equipped size exclusion chromatography column was used and the total RHC loss rate through PBSE rinse was 19.5 ± 5.08%. Fourier transform infrared spectroscopy (FT-IR) indicated hydrogen bonding among RHC, chitosan and PA, it also presented a probative but not strong hydrophobic interaction between phenyl rings of polyphenols and pyrrolidine rings of proline in RHC. Further, human umbilical vein endothelial cell (HUVEC) viability analyzed by a scanning electron microscope (SEM) and acridine orange/ethidium bromide (AO/EB) fluorescence staining exhibited that this scaffold could not only promote cell proliferation on scaffold surface but also permit cells migration into the scaffold. qRT-PCR exhibited that the optimized scaffold could stimulate angiogenesis associated genes VEGF and CD31 expression. These characterizations indicated that this scaffold can be considered as an ideal candidate for tissue engineering. - Highlights: • PA cross-linked recombinant human collagen–chitosan scaffold. • Fabrication in one-step lyophilization with neutralization. • HPLC detection of RHC loss rate • HUVEC proliferation and migration in scaffold • Angiogenesis associated gene expressions were increased in scaffold cell culturing.

  18. Preparation of composite hydroxybutyl chitosan sponge and its role in promoting wound healing.

    Science.gov (United States)

    Hu, Shihao; Bi, Shichao; Yan, Dong; Zhou, Zhongzheng; Sun, Guohui; Cheng, Xiaojie; Chen, Xiguang

    2018-03-15

    In this work, a composite sponge was produced by physically mixing hydroxybutyl chitosan with chitosan to form a porous spongy material through vacuum freeze-drying. Hydrophilic and macroporous composite hydroxybutyl chitosan sponge was developed via the incorporation of chitosan into hydroxybutyl chitosan. The composite sponge showed higher porosity (about 85%), greater water absorption (about 25 times), better softness and lower blood-clotting index (BCI) than those of chitosan sponge and hydroxybutyl chitosan sponge. The composite sponge with good hydrophilic could absorb the moisture in the blood to increase blood concentration and viscosity, and become a semi-swelling viscous colloid to clog the capillaries. Cytocompatibility tests with L929 cells and HUVEC cells demonstrated that composite sponge were no cytotoxicity, and could promote the growth of fibroblasts. It made up for the shortcomings of hydroxybutyl chitosan with unfavorable antibacterial effect to achieve a higher level of antibacterial (>99.99% reduction). Eventually, the vivo evaluations in Sprague-Dawley rats revealed that epithelial cells attached to the composite sponge and penetrated into the interior, in addition to this, it was also proved that the composite sponge (HC-1) had a better ability to promote wound healing and helped for faster formation of skin glands and re-epithelialization. The obtained data encourage the use of this composite sponge for wound dressings. Copyright © 2017. Published by Elsevier Ltd.

  19. Macro- and micro-designed chitosan-alginate scaffold architecture by three-dimensional printing and directional freezing

    International Nuclear Information System (INIS)

    Reed, Stephanie; Wu, Benjamin M; Lau, Grace; Delattre, Benjamin; Lopez, David Don; Tomsia, Antoni P

    2016-01-01

    While many tissue-engineered constructs aim to treat cartilage defects, most involve chondrocyte or stem cell seeding on scaffolds. The clinical application of cell-based techniques is limited due to the cost of maintaining cellular constructs on the shelf, potential immune response to allogeneic cell lines, and autologous chondrocyte sources requiring biopsy from already diseased or injured, scarce tissue. An acellular scaffold that can induce endogenous influx and homogeneous distribution of native stem cells from bone marrow holds great promise for cartilage regeneration. This study aims to develop such an acellular scaffold using designed, channeled architecture that simultaneously models the native zones of articular cartilage and subchondral bone. Highly porous, hydrophilic chitosan-alginate (Ch-Al) scaffolds were fabricated in three-dimensionally printed (3DP) molds designed to create millimeter scale macro-channels. Different polymer preform casting techniques were employed to produce scaffolds from both negative and positive 3DP molds. Macro-channeled scaffolds improved cell suspension distribution and uptake overly randomly porous scaffolds, with a wicking volumetric flow rate of 445.6 ± 30.3 mm 3 s −1 for aqueous solutions and 177 ± 16 mm 3 s −1 for blood. Additionally, directional freezing was applied to Ch-Al scaffolds, resulting in lamellar pores measuring 300 μm and 50 μm on the long and short axes, thus creating micrometer scale micro-channels. After directionally freezing Ch-Al solution cast in 3DP molds, the combined macro- and micro-channeled scaffold architecture enhanced cell suspension uptake beyond either macro- or micro-channels alone, reaching a volumetric flow rate of 1782.1 ± 48 mm 3 s −1 for aqueous solutions and 440.9 ± 0.5 mm 3 s −1 for blood. By combining 3DP and directional freezing, we can control the micro- and macro-architecture of Ch-Al to drastically improve cell influx into and distribution within the

  20. Sorption of technetium on composite chitosan-hydroxyapatite from aqueous solutions

    International Nuclear Information System (INIS)

    Pivarciova, L.; Rosskopfova, O.; Galambos, M.; Rajec, P.

    2013-01-01

    Biomaterials such as natural polymers (chitosan) and hydroxyapatite have an important application in material for bone replacement. Most of chitosan/hydroxyapatite composites are prepared by mixing hydroxyapatite particles with chitosan matrices. Another method of preparation of chitosan/hydroxyapatite composite is in-situ generation of nano-hydroxyapatite in chitosan matrix. The most common biomaterial used in the past years in hard tissue regeneration was hydroxyapatite, owing to its properties as biocompatibility, bioactivity, non-toxicity, non-immunogenicity etc. Chitosan is a polyaminosacharide, partially deacetylated product of chitin. Chitosan can be used in combination with other materials to enhance bone growth such as bone filling paste. The aims of this work were: the influence of the contact time on sorption of pertechnate anions on chitosan/hydroxyapatite composites; the effect of pH on sorption of pertechnate anions on chitosan/hydroxyapatite composites; the effect of foreign ions on sorption of pertechnate anions on chitosan/hydroxyapatite composites. The author concluded: the percentage of technetium sorption after 1 hour of contact time was > 97 %. In the initial pH range of 2.9-10.2, the percentage of technetium sorption on chitosan/hydroxyapatite composites CH/HA(A), CH/HA(B), CH/HA 30:70, ZCH was > 98 % and on CH/HA 50:50 was > 94%. The competition effect of Fe 2+ towards TcO 4 :- sorption is stronger than competition effect of other observed cations for all examined composites with the same weight ratio. The percentage of the technetium sorption was the same for all composites with the weight ratio of 30:70. (authors)

  1. Effects of chitosan and bioactive glass modifications of knitted and rolled polylactide-based 96/4 L/D scaffolds on chondrogenic differentiation of adipose stem cells.

    Science.gov (United States)

    Ahtiainen, Katja; Sippola, Laura; Nurminen, Manu; Mannerström, Bettina; Haimi, Suvi; Suuronen, Riitta; Hyttinen, Jari; Ylikomi, Timo; Kellomäki, Minna; Miettinen, Susanna

    2015-01-01

    The performance of biodegradable knitted and rolled 3-dimensional (3D) polylactide-based 96/4 scaffolds modified with bioactive glass (BaG) 13-93, chitosan and both was compared with regard to the viability, proliferation and chondrogenic differentiation of rabbit adipose stem cells (ASCs). Scaffold porosities were determined by micro-computed tomography (μCT). Water absorption and degradation of scaffolds were studied during 28-day hydrolysis in Tris-buffer. Viability, number and differentiation of ASCs in PLA96/4 scaffolds were examined in vitro. The dimensions of the scaffolds were maintained during hydrolysis and mass loss was detected only in the BaG13-93 containing scaffolds. ASCs adhered and proliferated on each scaffold type. Cell aggregation and expression of chondral matrix components improved in all scaffold types in chondrogenic medium. Signs of hypertrophy were detected in the modified scaffolds but not in the plain PLA96/4 scaffold. Chondrogenic differentiation was most enhanced in the presence of chitosan. These findings indicate that the plain P scaffold provided a good 3D-matrix for ASC proliferation whereas the addition of chitosan to the PLA96/4 scaffold induced chondrogenic differentiation independent of the medium. Accordingly, a PLA96/4 scaffold modified by chitosan could provide a functional and bioactive basis for tissue-engineered chondral implants. Copyright © 2012 John Wiley & Sons, Ltd.

  2. Porous Lactose-Modified Chitosan Scaffold for Liver Tissue Engineering: Influence of Galactose Moieties on Cell Attachment and Mechanical Stability

    Directory of Open Access Journals (Sweden)

    Birong Wang

    2016-01-01

    Full Text Available Galactosylated chitosan (CTS has been widely applied in liver tissue engineering as scaffold. However, the influence of degree of substitution (DS of galactose moieties on cell attachment and mechanical stability is not clear. In this study, we synthesized the lactose-modified chitosan (Lact-CTS with various DS of galactose moieties by Schiff base reaction and reducing action of NaBH4, characterized by FTIR. The DS of Lact-CTS-1, Lact-CTS-2, and Lact-CTS-3 was 19.66%, 48.62%, and 66.21% through the method of potentiometric titration. The cell attachment of hepatocytes on the CTS and Lact-CTS films was enhanced accompanied with the increase of galactose moieties on CTS chain because of the galactose ligand-receptor recognition; however, the mechanical stability of Lact-CTS-3 was reduced contributing to the extravagant hydrophilicity, which was proved using the sessile drop method. Then, the three-dimensional Lact-CTS scaffolds were fabricated by freezing-drying technique. The SEM images revealed the homogeneous pore bearing the favorable connectivity and the pore sizes of scaffolds with majority of 100 μm; however, the extract solution of Lact-CTS-3 scaffold significantly damaged red blood cells by hemolysis assay, indicating that exorbitant DS of Lact-CTS-3 decreased the mechanical stability and increased the toxicity. To sum up, the Lact-CTS-2 with 48.62% of galactose moieties could facilitate the cell attachment and possess great biocompatibility and mechanical stability, indicating that Lact-CTS-2 was a promising material for liver tissue engineering.

  3. Fabrication of Novel Porous Chitosan Matrices as Scaffolds for Bone Tissue Engineering

    National Research Council Canada - National Science Library

    Jiang, Tao; Pilane, Cyril M; Laurencin, Cato T

    2005-01-01

    .... Chitosan, a natural polymer obtained from chitin, which forms a major component of crustacean exoskeleton, is a potential candidate for bone tissue engineering due to its excellent osteocompatibility...

  4. Proliferation and enrichment of CD133(+) glioblastoma cancer stem cells on 3D chitosan-alginate scaffolds.

    Science.gov (United States)

    Kievit, Forrest M; Florczyk, Stephen J; Leung, Matthew C; Wang, Kui; Wu, Jennifer D; Silber, John R; Ellenbogen, Richard G; Lee, Jerry S H; Zhang, Miqin

    2014-11-01

    Emerging evidence implicates cancer stem cells (CSCs) as primary determinants of the clinical behavior of human cancers, representing an ideal target for next-generation anti-cancer therapies. However CSCs are difficult to propagate in vitro, severely limiting the study of CSC biology and drug development. Here we report that growing cells from glioblastoma (GBM) cell lines on three dimensional (3D) porous chitosan-alginate (CA) scaffolds dramatically promotes the proliferation and enrichment of cells possessing the hallmarks of CSCs. CA scaffold-grown cells were found more tumorigenic in nude mouse xenografts than cells grown from monolayers. Growing in CA scaffolds rapidly promoted expression of genes involved in the epithelial-to-mesenchymal transition that has been implicated in the genesis of CSCs. Our results indicate that CA scaffolds have utility as a simple and inexpensive means to cultivate CSCs in vitro in support of studies to understand CSC biology and develop more effective anti-cancer therapies. Copyright © 2014 Elsevier Ltd. All rights reserved.

  5. Synthesis and Study the Effect of HNTs on PVA/Chitosan Composite Material

    OpenAIRE

    Malek Ali

    2016-01-01

    Composites materials of Poly (vinyl alcohol) (PVA)/Chitosan (CS) have been synthesized and characterized successfully. HNTs have been added to composites to enhance the mechanical and degradation properties by hydrogen bonding interactions, compatibility, and chemical crosslink between HNTs and PVA. PVA/CS/HNTs composites prepared with different concentration ratio. SEM micrographs of composites surface showed that more agglomeration with more chitosan ratio. Mechanical and degradation proper...

  6. Functional chitosan-based grapefruit seed extract composite films for applications in food packaging technology

    International Nuclear Information System (INIS)

    Tan, Y.M.; Lim, S.H.; Tay, B.Y.; Lee, M.W.; Thian, E.S.

    2015-01-01

    Highlights: • Chitosan-based grapefruit seed extract (GFSE) films were solution casted. • GFSE was uniformly dispersed within all chitosan film matrices. • All chitosan-based composite films showed remarkable transparency. • Increasing amounts of GFSE incorporated increased the elongation at break of films. • Chitosan-based GFSE composite films inhibited the proliferation of fungal growth. - Abstract: Chitosan-based composite films with different amounts of grapefruit seed extract (GFSE) (0.5, 1.0 and 1.5% v/v) were fabricated via solution casting technique. Experimental results showed that GFSE was uniformly dispersed within all chitosan film matrices. The presence of GFSE made the films more amorphous and tensile strength decreased, while elongation at break values increased as GFSE content increased. Results from the measurement of light transmission revealed that increasing amounts of GFSE (from 0.5 to 1.5% v/v) did not affect transparency of the films. Furthermore, packaging of bread samples with chitosan-based GFSE composite films inhibited the proliferation of fungal growth as compared to control samples. Hence, chitosan-based GFSE composite films have the potential to be a useful material in the area of food technology

  7. Functional chitosan-based grapefruit seed extract composite films for applications in food packaging technology

    Energy Technology Data Exchange (ETDEWEB)

    Tan, Y.M. [Department of Mechanical Engineering, National University of Singapore (Singapore); Lim, S.H.; Tay, B.Y. [Forming Technology Group, Singapore Institute of Manufacturing Technology (Singapore); Lee, M.W. [Food Innovation and Resource Centre, Singapore Polytechnic (Singapore); Thian, E.S., E-mail: mpetes@nus.edu.sg [Department of Mechanical Engineering, National University of Singapore (Singapore)

    2015-09-15

    Highlights: • Chitosan-based grapefruit seed extract (GFSE) films were solution casted. • GFSE was uniformly dispersed within all chitosan film matrices. • All chitosan-based composite films showed remarkable transparency. • Increasing amounts of GFSE incorporated increased the elongation at break of films. • Chitosan-based GFSE composite films inhibited the proliferation of fungal growth. - Abstract: Chitosan-based composite films with different amounts of grapefruit seed extract (GFSE) (0.5, 1.0 and 1.5% v/v) were fabricated via solution casting technique. Experimental results showed that GFSE was uniformly dispersed within all chitosan film matrices. The presence of GFSE made the films more amorphous and tensile strength decreased, while elongation at break values increased as GFSE content increased. Results from the measurement of light transmission revealed that increasing amounts of GFSE (from 0.5 to 1.5% v/v) did not affect transparency of the films. Furthermore, packaging of bread samples with chitosan-based GFSE composite films inhibited the proliferation of fungal growth as compared to control samples. Hence, chitosan-based GFSE composite films have the potential to be a useful material in the area of food technology.

  8. In Vitro Degradation of PHBV Scaffolds and nHA/PHBV Composite Scaffolds Containing Hydroxyapatite Nanoparticles for Bone Tissue Engineering

    Directory of Open Access Journals (Sweden)

    Naznin Sultana

    2012-01-01

    Full Text Available This paper investigated the long-term in vitro degradation properties of scaffolds based on biodegradable polymers and osteoconductive bioceramic/polymer composite materials for the application of bone tissue engineering. The three-dimensional porous scaffolds were fabricated using emulsion-freezing/freeze-drying technique using poly(hydroxybutyrate-co-hydroxyvalerate (PHBV which is a natural biodegradable and biocompatible polymer. Nanosized hydroxyapatite (nHA particles were successfully incorporated into the PHBV scaffolds to render the scaffolds osteoconductive. The PHBV and nHA/PHBV scaffolds were systematically evaluated using various techniques in terms of mechanical strength, porosity, porous morphology, and in vitro degradation. PHBV and nHA/PHBV scaffolds degraded over time in phosphate-buffered saline at 37°C. PHBV polymer scaffolds exhibited slow molecular weight loss and weight loss in the in vitro physiological environment. Accelerated weight loss was observed in nHA incorporated PHBV composite scaffolds. An increasing trend of crystallinity was observed during the initial period of degradation time. The compressive properties decreased more than 40% after 5-month in vitro degradation. Together with interconnected pores, high porosity, suitable mechanical properties, and slow degradation profile obtained from long-term degradation studies, the PHBV scaffolds and osteoconductive nHA/PHBV composite scaffolds showed promises for bone tissue engineering application.

  9. The interplay between nanostructured carbon-grafted chitosan scaffolds and protein adsorption on the cellular response of osteoblasts: structure-function property relationship.

    Science.gov (United States)

    Depan, D; Misra, R D K

    2013-04-01

    The rapid adsorption of proteins occurs during the early stages of biomedical device implantation into physiological systems. In this regard, the adsorption of proteins is a strong function of the nature of a biomedical device, which ultimately governs the biological functions. The objective of this study was to elucidate the interplay between nanostructured carbon-modified (graphene oxide and single-walled carbon nanohorn) chitosan scaffolds and consequent protein adsorption and biological function (osteoblast function). We compare and contrast the footprint of protein adsorption on unmodified chitosan and nanostructured carbon-modified chitosan. A comparative analysis of cell-substrate interactions using an osteoblast cell line (MC3T3-E1) implied that biological functions were significantly enhanced in the presence of nanostructured carbon, compared with unmodified chitosan. The difference in their respective behaviors is related to the degree and topography of protein adsorption on the scaffolds. Furthermore, there was a synergistic effect of nanostructured carbon and protein adsorption in terms of favorably modulating biological functions, including cell attachment, proliferation and viability, with the effect being greater on nanostructured carbon-modified scaffolds. The study also underscores that protein adsorption is favored in nanostructured carbon-modified scaffolds such that bioactivity and biological function are promoted. Copyright © 2012 Acta Materialia Inc. Published by Elsevier Ltd. All rights reserved.

  10. Chitosan/γ-poly(glutamic acid) scaffolds with surface-modified albumin, elastin and poly-l-lysine for cartilage tissue engineering.

    Science.gov (United States)

    Kuo, Yung-Chih; Ku, Hao-Fu; Rajesh, Rajendiran

    2017-09-01

    Cartilage has limited ability to self-repair due to the absence of blood vessels and nerves. The application of biomaterial scaffolds using biomimetic extracellular matrix (ECM)-related polymers has become an effective approach to production of engineered cartilage. Chitosan/γ-poly(glutamic acid) (γ-PGA) scaffolds with different mass ratios were prepared using genipin as a cross-linker and a freeze-drying method, and their surfaces were modified with elastin, human serum albumin (HSA) and poly-l-lysine (PLL). The scaffolds were formed through a complex between NH 3 + of chitosan and COO - of γ-PGA, confirmed by Fourier transform infrared spectroscopy, and exhibited an interconnected porous morphology in field emission scanning electron microscopy analysis. The prepared chitosan/γ-PGA scaffolds, at a 3:1 ratio, obtained the required porosity (90%), pore size (≥100μm), mechanical strength (compressive strength>4MPa, Young's modulus>4MPa) and biodegradation (30-60%) for articular cartilage tissue engineering applications. Surface modification of the scaffolds showed positive indications with improved activity toward cell proliferation (deoxyribonucleic acid), cell adhesion and ECM (glycoaminoglycans and type II collagen) secretion of bovine knee chondrocytes compared with unmodified scaffolds. In caspase-3 detection, elastin had a higher inhibitory effect on chondrocyte apoptosis in vitro, followed by HSA, and then PLL. We concluded that utilizing chitosan/γ-PGA scaffolds with surface active biomolecules, including elastin, HSA and PLL, can effectively promote the growth of chondrocytes, secrete ECM and improve the regenerative ability of cartilaginous tissues. Copyright © 2017 Elsevier B.V. All rights reserved.

  11. Developing bioactive composite scaffolds for bone tissue engineering

    Science.gov (United States)

    Chen, Yun

    Poly(L-lactic acid) (PLLA) films were fabricated using the method of dissolving and evaporation. PLLA scaffold was prepared by solid-liquid phase separation of polymer solutions and subsequent sublimation of solvent. Bonelike apatite coating was formed on PLLA films, PLLA scaffolds and poly(glycolic acid) (PGA) scaffolds in 24 hours through an accelerated biomimetic process. The ion concentrations in the simulated body fluid (SBF) were nearly 5 times of those in human blood plasma. The apatite formed was characterized using scanning electron microscopy (SEM), energy dispersive X-ray spectroscopy (EDX), X-ray diffraction (XRD), and Fourier transform infrared spectroscopy (FTIR). The apatite formed in 5SBF was similar in morphology and composition to that formed in the classical biomimetic process employing SBF or 1.5SBF, and similar to that of natural bone. This indicated that the biomimetic apatite coating process could be accelerated by using concentrated simulated body fluid at 37°C. Besides saving time, the accelerated biomimetic process is particularly significant to biodegradable polymers. Some polymers which degrade too fast to be coated with apatite by a classical biomimetic process, for example PGA, could be coated with bone-like apatite in an accelerated biomimetic process. Collagen and apatite were co-precipitated as a composite coating on poly(L-lactic acid) (PLLA) in an accelerated biomimetic process. The incubation solution contained collagen (1g/L) and simulated body fluid (SBF) with 5 times inorganic ionic concentrations as human blood plasma. The coating formed on PLLA films and scaffolds after 24 hours incubation was characterized using EDX, XRD, FTIR, and SEM. It was shown that the coating contained carbonated bone-like apatite and collagen, the primary constituents of natural bone. SEM showed a complex composite coating of submicron bone-like apatite particulates combined with collagen fibrils. This work provided an efficient process to obtain

  12. Hard tissue compatibility of natural hydroxyapatite/chitosan composite

    International Nuclear Information System (INIS)

    Tang Xiaojun; Gui Lai; Lue Xiaoying

    2008-01-01

    The natural hydroxyapatite/chitosan (NHC) composite is a new synthesized material. The aim of this experiment was to assess the bone tissue compatibility of this NHC composite in vivo. Twenty-four healthy New Zealand rabbits were included in this study. Of those, 20 were used as the experimental group and four as the control group. In the experimental group, animals receive a cranium defect procedure and NHC composite repair. In the control group, animals underwent the cranium defect procedure without NHC composite repair. At 1, 4, 12, 24, and 40 weeks after surgery, the animals were sacrificed and samples were taken and assessed by gross observation, three-dimensional (3D) computerized tomographic (CT) reconstruction, histology and scanning electron microscope. Our results showed that at 1 week after repairing the bone defect with the NHC composite in the experimental group, new bone appeared around the composite and matured gradually. At 24 weeks after surgery, there were little collagenous tissues present between the material and surrounding bones. At 40 weeks after surgery, new bone had grown into the mature bone and total osseointegration had occurred. In the control group, however, no bone defect healing was observed at 40 weeks after surgery. All these results of the present in vivo work suggest that the NHC composite has a good hard tissue biocompatibility and an excellent osteoconductivity. It is suitable for artificial bone implants and frame materials of tissue engineering.

  13. Hard tissue compatibility of natural hydroxyapatite/chitosan composite

    Energy Technology Data Exchange (ETDEWEB)

    Tang Xiaojun; Gui Lai [Department of Cranio-maxillofacial Surgery, Plastic Surgery Hospital, Chinese Academy of Medical Sciences and Peking Union Medical College, 33 Ba-Da-Chu Road, Beijing, 100144 (China); Lue Xiaoying [State Key Laboratory of Bioelectronics, School of Biological Science and Medical Engineering, Southeast University, Nanjing, 210096 (China)], E-mail: laiguiplastic@tom.com, E-mail: luxy@seu.edu.cn

    2008-12-15

    The natural hydroxyapatite/chitosan (NHC) composite is a new synthesized material. The aim of this experiment was to assess the bone tissue compatibility of this NHC composite in vivo. Twenty-four healthy New Zealand rabbits were included in this study. Of those, 20 were used as the experimental group and four as the control group. In the experimental group, animals receive a cranium defect procedure and NHC composite repair. In the control group, animals underwent the cranium defect procedure without NHC composite repair. At 1, 4, 12, 24, and 40 weeks after surgery, the animals were sacrificed and samples were taken and assessed by gross observation, three-dimensional (3D) computerized tomographic (CT) reconstruction, histology and scanning electron microscope. Our results showed that at 1 week after repairing the bone defect with the NHC composite in the experimental group, new bone appeared around the composite and matured gradually. At 24 weeks after surgery, there were little collagenous tissues present between the material and surrounding bones. At 40 weeks after surgery, new bone had grown into the mature bone and total osseointegration had occurred. In the control group, however, no bone defect healing was observed at 40 weeks after surgery. All these results of the present in vivo work suggest that the NHC composite has a good hard tissue biocompatibility and an excellent osteoconductivity. It is suitable for artificial bone implants and frame materials of tissue engineering.

  14. The molecular understanding of interfacial interactions of functionalized graphene and chitosan

    International Nuclear Information System (INIS)

    Zhang, Hong-ping; Luo, Xue-gang; Lin, Xiao-yan; Lu, Xiong; Tang, Youhong

    2016-01-01

    Graphical abstract: The type of the functional groups can be used to modulating interactions between graphene sheet and chitosan. - Highlights: • Investigate interfacial interactions between chitosan and functionalized graphene by DFT. • Observe covalent linkages between COOH-modified graphene and chitosan units. • Multi-functionalized graphene regulates the interfacial interactions with chitosan. • It is useful for guiding the preparation of graphene/chitosan composites. - Abstract: Graphene-reinforced chitosan scaffolds have been extensively studied for several years as promising hard tissue replacements. However, the interfacial interactions between graphene and chitosan are strongly related to the solubility, processability, and mechanical properties of graphene-reinforced chitosan (G–C) composites. The functionalization of graphene is regarded as the most effective way to improve the abovementioned properties of the G–C composite. In this study, the interfacial interactions between chitosan and functionalized graphene sheets with carboxylization (COOH-), amination (NH 2 -), and hydroxylation (OH-) groups were systematically studied at the electronic level using the method of ab initio simulations based on quantum mechanics theory and the observations were compared with reported experimental results. The covalent linkages between COOH-modified graphene and the chitosan units were demonstrated and the combination of multi-functionalization on graphene could regulate the interfacial interactions between graphene and the chitosan. The interfacial interactions between chitosan and properly functionalized graphene are critical for the preparation of G–C-based composites for tissue engineering scaffolds and other applications.

  15. Curcumin-Loaded Chitosan/Gelatin Composite Sponge for Wound Healing Application

    Directory of Open Access Journals (Sweden)

    Van Cuong Nguyen

    2013-01-01

    Full Text Available Three composite sponges were made with 10% of curcumin and by using polymers, namely, chitosan and gelatin with various ratios. The chemical structure and morphology were evaluated by FTIR and SEM. These sponges were evaluated for water absorption capacity, antibacterial activity, in vitro drug release, and in vivo wound healing studies by excision wound model using rabbits. The in vivo study presented a greater wound closure in wounds treated with curcumin-composite sponge than those with composite sponge without curcumin and untreated group. These obtained results showed that combination of curcumin, chitosan and gelatin could improve the wound healing activity in comparison to chitosan, and gelatin without curcumin.

  16. Three-dimensional dynamic fabrication of engineered cartilage based on chitosan/gelatin hybrid hydrogel scaffold in a spinner flask with a special designed steel frame.

    Science.gov (United States)

    Song, Kedong; Li, Liying; Li, Wenfang; Zhu, Yanxia; Jiao, Zeren; Lim, Mayasari; Fang, Meiyun; Shi, Fangxin; Wang, Ling; Liu, Tianqing

    2015-10-01

    Cartilage transplantation using in vitro tissue engineered cartilage is considered a promising treatment for articular cartilage defects. In this study, we assessed the advantages of adipose derived stem cells (ADSCs) combined with chitosan/gelatin hybrid hydrogel scaffolds, which acted as a cartilage biomimetic scaffold, to fabricate a tissue engineered cartilage dynamically in vitro and compared this with traditional static culture. Physical properties of the hydrogel scaffolds were evaluated and ADSCs were inoculated into the hydrogel at a density of 1×10(7) cells/mL and cultured in a spinner flask with a special designed steel framework and feed with chondrogenic inductive media for two weeks. The results showed that the average pore size, porosity, swelling rate and elasticity modulus of hybrid scaffolds with good biocompatibility were 118.25±19.51 μm, 82.60±2.34%, 361.28±0.47% and 61.2±0.16 kPa, respectively. ADSCs grew well in chitosan/gelatin hybrid scaffold and successfully differentiated into chondrocytes, showing that the scaffolds were suitable for tissue engineering applications in cartilage regeneration. Induced cells cultivated in a dynamic spinner flask with a special designed steel frame expressed more proteoglycans and the cell distribution was much more uniform with the scaffold being filled mostly with extracellular matrix produced by cells. A spinner flask with framework promoted proliferation and chondrogenic differentiation of ADSCs within chitosan/gelatin hybrid scaffolds and accelerated dynamic fabrication of cell-hydrogel constructs, which could be a selective and good method to construct tissue engineered cartilage in vitro. Copyright © 2015 Elsevier B.V. All rights reserved.

  17. Scaffold of chitosan-sodium alginate and hydroxyapatite with application potential for bone regeneration; Scaffold de quitosana-alginato de sodio e hidroxiapatita com potencial de aplicacao para regeneracao ossea

    Energy Technology Data Exchange (ETDEWEB)

    Rebelo, Marcia de A.; Alves, Thais F.R.; Lopes, Francielly C.C.N; Oliveira Junior, Jose Martins de; Pontes, Katiusca S.; Fogaca, Bruna A.C.; Chaud, Marco V., E-mail: marco.chaud@prof.uniso.br [Universidade de Sorocaba (LABNUS/UNISO), Sorocaba, SP (Brazil). Laboratorio de Biomateriais e Nanotecnologia

    2015-07-01

    Scaffold for organic tissue regeneration are architectural, three-dimensional, porous, biocompatible and biodegradable devices. The first challenges to be met in the development of these devices to mimic the biomechanical properties of the target tissue. The aim of this study was to develop and to characterize scaffolds composed of chitosan (Ch), sodium alginate (SA), hydroxyapatite (HA). The scaffolds were obtained by lyophilization. HA has been incorporated into the polymer dispersion in Ch-AS concentration of 20 and 60%. The mechanical properties of the scaffold were determined by tensile and compression tests. Swelling capacity was assessed in the presence of simulated saliva, purified water, HCl 0.01M, NaOH 0.01M. The calcium content was quantified using fluorescence X-rays. Analysis of the results indicates that the Qt-AS-HA-60% scaffold obtained by lyophilization meets promising properties for bone tissue regeneration. (author)

  18. Hyaluronic acid doped-poly(3,4-ethylenedioxythiophene)/chitosan/gelatin (PEDOT-HA/Cs/Gel) porous conductive scaffold for nerve regeneration

    International Nuclear Information System (INIS)

    Wang, Shuping; Guan, Shui; Zhu, Zhibo; Li, Wenfang; Liu, Tianqing; Ma, Xuehu

    2017-01-01

    Conducting polymer, as a “smart” biomaterial, has been increasingly used to construct tissue engineered scaffold for nerve tissue regeneration. In this study, a novel porous conductive scaffold was prepared by incorporating conductive hyaluronic acid (HA) doped-poly(3,4-ethylenedioxythiophene) (PEDOT-HA) nanoparticles into a chitosan/gelatin (Cs/Gel) matrix. The physicochemical characteristics of Cs/Gel scaffold with 0–10 wt% PEDOT-HA were analyzed and the results indicated that the incorporation of PEDOT-HA into scaffold increased the electrical and mechanical properties while decreasing the porosity and water absorption. Moreover, in vitro biodegradation of scaffold displayed a declining trend with the PEDOT-HA content increased. About the biocompatibility of conductive scaffold, neuron-like rat phaeochromocytoma (PC12) cells were cultured in scaffold to evaluate cell adhesion and growth. 8% PEDOT-HA/Cs/Gel scaffold had a higher cell adhesive efficiency and cell viability than the other conductive scaffolds. Furthermore, cells in the scaffold with 8 wt% PEDOT-HA expressed higher synapse growth gene of GAP43 and SYP compared with Cs/Gel control group. These results suggest that 8%PEDOT-HA/Cs/Gel scaffold is an attractive cell culture conductive substrate which could support cell adhesion, survival, proliferation, and synapse growth for the application in nerve tissue regeneration. - Highlights: • A porous conductive scaffold was prepared by freeze-drying method. • Conductive scaffold could support PC12 cells adhesion, survival, and proliferation. • Cells in conductive scaffold expressed high synapse growth gene of GAP43 and SYP.

  19. Hyaluronic acid doped-poly(3,4-ethylenedioxythiophene)/chitosan/gelatin (PEDOT-HA/Cs/Gel) porous conductive scaffold for nerve regeneration

    Energy Technology Data Exchange (ETDEWEB)

    Wang, Shuping; Guan, Shui, E-mail: guanshui@dlut.edu.cn; Zhu, Zhibo; Li, Wenfang; Liu, Tianqing; Ma, Xuehu

    2017-02-01

    Conducting polymer, as a “smart” biomaterial, has been increasingly used to construct tissue engineered scaffold for nerve tissue regeneration. In this study, a novel porous conductive scaffold was prepared by incorporating conductive hyaluronic acid (HA) doped-poly(3,4-ethylenedioxythiophene) (PEDOT-HA) nanoparticles into a chitosan/gelatin (Cs/Gel) matrix. The physicochemical characteristics of Cs/Gel scaffold with 0–10 wt% PEDOT-HA were analyzed and the results indicated that the incorporation of PEDOT-HA into scaffold increased the electrical and mechanical properties while decreasing the porosity and water absorption. Moreover, in vitro biodegradation of scaffold displayed a declining trend with the PEDOT-HA content increased. About the biocompatibility of conductive scaffold, neuron-like rat phaeochromocytoma (PC12) cells were cultured in scaffold to evaluate cell adhesion and growth. 8% PEDOT-HA/Cs/Gel scaffold had a higher cell adhesive efficiency and cell viability than the other conductive scaffolds. Furthermore, cells in the scaffold with 8 wt% PEDOT-HA expressed higher synapse growth gene of GAP43 and SYP compared with Cs/Gel control group. These results suggest that 8%PEDOT-HA/Cs/Gel scaffold is an attractive cell culture conductive substrate which could support cell adhesion, survival, proliferation, and synapse growth for the application in nerve tissue regeneration. - Highlights: • A porous conductive scaffold was prepared by freeze-drying method. • Conductive scaffold could support PC12 cells adhesion, survival, and proliferation. • Cells in conductive scaffold expressed high synapse growth gene of GAP43 and SYP.

  20. Chitin Fiber and Chitosan 3D Composite Rods

    International Nuclear Information System (INIS)

    Wang, Z.; Hu, Q.; Cai, L.

    2010-01-01

    Chitin fiber (CHF) and chitosan (CS) 3D composite rods with layer-by-layer structure were constructed by in situ precipitation method. CHF could not be dissolved in acetic acid aqueous solution, but CS could be dissolved due to the different deacetylation degree (D.D) between CHF and CS. CHF with undulate surfaces could be observed using SEM to demonstrate that the sufficiently rough surfaces and edges of the fiber could enhance the mechanical combining stress between fiber and matrix. XRD indicated that the crystallinity of CHF/CS composites decreased and CS crystal plane d-spacing of CHF/CS composites became larger than that of pure CS rod. TG analysis showed that mixing a little amount of CHF could enhance thermal stability of CS rod, but when the content of CHF was higher than the optimum amount, its thermal stability decreased. When 0.5% CHF was added into CS matrix, the bending strength and bending modulus of the composite rods arrived at 114.2 MPa and 5.2 GPa, respectively, increased by 23.6% and 26.8% compared with pure CS rods, indicating that CHF/CS composite rods could be a better candidate for bone fracture internal fixation.

  1. Chitin Fiber and Chitosan 3D Composite Rods

    Directory of Open Access Journals (Sweden)

    Zhengke Wang

    2010-01-01

    Full Text Available Chitin fiber (CHF and chitosan (CS 3D composite rods with layer-by-layer structure were constructed by in situ precipitation method. CHF could not be dissolved in acetic acid aqueous solution, but CS could be dissolved due to the different deacetylation degree (D.D between CHF and CS. CHF with undulate surfaces could be observed using SEM to demonstrate that the sufficiently rough surfaces and edges of the fiber could enhance the mechanical combining stress between fiber and matrix. XRD indicated that the crystallinity of CHF/CS composites decreased and CS crystal plane d-spacing of CHF/CS composites became larger than that of pure CS rod. TG analysis showed that mixing a little amount of CHF could enhance thermal stability of CS rod, but when the content of CHF was higher than the optimum amount, its thermal stability decreased. When 0.5% CHF was added into CS matrix, the bending strength and bending modulus of the composite rods arrived at 114.2 MPa and 5.2 GPa, respectively, increased by 23.6% and 26.8% compared with pure CS rods, indicating that CHF/CS composite rods could be a better candidate for bone fracture internal fixation.

  2. Investigation of particle-functionalized tissue engineering scaffolds using X-ray tomographic microscopy

    DEFF Research Database (Denmark)

    Nygaard, J V; Andersen, M Ø; Howard, K A

    2008-01-01

    A low-density, porous chitosan/poly-(dl-lactide-co-glycolide) (PLGA) microparticle composite scaffold was produced by thermally induced phase separation followed by lyophilization, to provide a bicontinuous microstructure potentially suitable for tissue engineering and locally controlled drug...

  3. New composite materials prepared by calcium phosphate precipitation in chitosan/collagen/hyaluronic acid sponge cross-linked by EDC/NHS.

    Science.gov (United States)

    Kaczmarek, B; Sionkowska, A; Kozlowska, J; Osyczka, A M

    2018-02-01

    Nowadays, fabrication of composite materials based on biopolymers is a rising field due to potential for bone repair and tissue engineering application. Blending of different biopolymers and incorporation of inorganic particles in the blend can lead to new materials with improved physicochemical properties and biocompatibility. In this work 3D porous structures called scaffolds based on chitosan, collagen and hyaluronic acid were obtained through the lyophilization process. Scaffolds were cross-linked by EDC/NHS. Infrared spectra for the materials were made, the percentage of swelling, scaffolds porosity and density, mechanical parameters, thermal stability were studied. Moreover, the scaffolds were used as matrixes for the calcium phosphate in situ precipitation. SEM images were taken and EDX analysis was carried out for calcium and phosphorous content determination in the scaffold. In addition, the adhesion and proliferation of human osteosarcoma SaOS-2 cells was examined on obtained scaffolds. The results showed that the properties of 3D composites cross-linked by EDC/NHS were altered after the addition of 1, 2 and 5% hyaluronic acid. Mechanical parameters, thermal stability and porosity of scaffolds were improved. Moreover, calcium and phosphorous were found in each kind of scaffold. SEM images showed that the precipitation was homogeneously carried in the whole volume of samples. Attachment of SaOS-2 cells to all modified materials was better compared to unmodified control and proliferation of these cells was markedly increased on scaffolds with precipitated calcium phosphate. Obtained materials can provide the support useful in tissue engineering and regenerative medicine. Copyright © 2017 Elsevier B.V. All rights reserved.

  4. Preparation and characterization of chitosan-natural nano hydroxyapatite-fucoidan nanocomposites for bone tissue engineering.

    Science.gov (United States)

    Lowe, Baboucarr; Venkatesan, Jayachandran; Anil, Sukumaran; Shim, Min Suk; Kim, Se-Kwon

    2016-12-01

    Solid three dimensional (3D) composite scaffolds for bone tissue engineering were prepared using the freeze-drying method. The scaffolds were composed of chitosan, natural nano-hydroxyapatite (nHA) and fucoidan in the following combinations: chitosan, chitosan-fucoidan, chitosan-nHA, and chitosan-nHA-fucoidan. Fourier transform infrared spectroscopy (FT-IR), thermal gravimetric analysis (TGA), X-ray diffraction analysis (XRD), scanning electron microscopy (SEM), and optical microscopy (OM) were used to determine the physiochemical constituents and the morphology of the scaffolds. The addition of nHA into the chitosan-fucoidan composite scaffold reduced the water uptake and water retention. FT-IR analysis confirmed the presence of a phosphate group in the chitosan-nHA-fucoidan scaffold. This group is present because of the presence of nHA (isolated via alkaline hydrolysis from salmon fish bones). Microscopic results indicated that the dispersion of nHA and fucoidan in the chitosan matrix was uniform with a pore size of 10-400μm. The composite demonstrated a suitable micro architecture for cell growth and nutrient supplementation. This compatibility was further elucidated in vitro using periosteum-derived mesenchymal stem cells (PMSCs). The cells demonstrated high biocompatibility and excellent mineralization for the chitosan-nHA-fucoidan scaffold. We believe that a chitosan-nHA-fucoidan composite is a promising biomaterial for the scaffold that can be used for bone tissue regeneration. Copyright © 2016 Elsevier B.V. All rights reserved.

  5. Surface-enrichment with hydroxyapatite nanoparticles in stereolithography-fabricated composite polymer scaffolds promotes bone repair

    NARCIS (Netherlands)

    Guillaume, O.; Geven, M. A.; Sprecher, C. M.; Stadelmann, V. A.; Grijpma, D. W.; Tang, T.T.; Qin, L.; Lai, Y.; Alini, M.; de Bruijn, J. D.; Yuan, H.; Richards, R.G.; Eglin, D.

    2017-01-01

    Fabrication of composite scaffolds using stereolithography (SLA) for bone tissue engineering has shown great promises. However, in order to trigger effective bone formation and implant integration, exogenous growth factors are commonly combined to scaffold materials. In this study, we fabricated

  6. A clinical and histologic evaluation of gingival fibroblasts seeding on a chitosan-based scaffold and its effect on the width of keratinized gingiva in dogs.

    Science.gov (United States)

    Lotfi, Ghogha; Shokrgozar, Mohammad Ali; Mofid, Rasoul; Abbas, Fatemeh Mashhadi; Ghanavati, Farzin; Bagheban, Alireza Akbarzadeh; Shariati, Ramin Pajoum

    2011-09-01

    Finding biocompatible matrix materials capable of enhancing the procedures of gingival augmentation is a major concern in periodontal research. This has prompted the investigation of a safe grafting technique by means of synthetic or natural polymers. The objective of this study is to examine the effect of a gingival fibroblast cultured on a naturally derived (i.e., chitosan-based) scaffold on the width of keratinized gingiva in dogs. Gingival fibroblasts were cultured from a small portion of hard palates of five dogs. A bilayered chitosan scaffold was seeded with the gingival fibroblasts and transferred to dogs. Surgery was performed bilaterally, and the regions were randomly divided into two groups: chitosan only (control site) and chitosan + fibroblast (test site). Periodontal parameters, including probing depth and width of keratinized and attached gingiva, were measured at baseline and 3 months after surgery. A histologic evaluation was also performed on the healed grafted sites. Comparison of width of keratinized and attached gingiva in control and test sites showed that the mean width of keratinized and attached gingiva increased in each group after surgery. However, the difference between control and test groups was not statistically significant. Concerning the existence of the keratinized epithelium, exocytosis, and epithelium thickness, no significant difference was observed in test and control sites. The difference was significant in relation to rete ridge formation. The tissue-engineered graft consisting of chitosan + fibroblast was applied to gingival augmentation procedures and generated keratinized tissue without any complications usually associated with donor-site surgery.

  7. Silk fibroin/chitosan scaffold with tunable properties and low inflammatory response assists the differentiation of bone marrow mesenchymal stem cells.

    Science.gov (United States)

    Li, Da-Wei; Lei, Xiaohua; He, Feng-Li; He, Jin; Liu, Ya-Li; Ye, Ya-Jing; Deng, Xudong; Duan, Enkui; Yin, Da-Chuan

    2017-12-01

    The physical and chemical properties of the scaffold are known to play important roles in three-dimensional (3D) cell culture, which always determine the cellular fate or the results of implantation. To control these properties becomes necessary for meeting the requirements of a variety of tissue engineering applications. In this study, a series of silk fibroin/chitosan (SF/CS) scaffolds with tunable properties were prepared using freeze-drying method, and the rat bone marrow-derived mesenchymal stem cells (BM-MSCs) were seeded in these scaffolds to evaluate their availability of use in tissue engineering. The 3D structure, mechanical properties and degradation ability of SF/CS scaffold can be tuned by changing the total concentration of the precursor solution and the blending ratio between SF and CS. BM-MSCs cultured in the SF/CS scaffold exhibited excellent proliferation and multiple morphologies. The induction of osteogenic and adipogenic differentiation of BM-MSCs were successful in this scaffold when cultured in vitro. Subcutaneous implantation of the SF/CS scaffolds did not cause any inflammatory response within four weeks, which revealed good compatibility. Moreover, the implanted scaffold allowed host cells to invade, adhere, grow and form new blood vessels. With these excellent performance, SF/CS scaffold has great potential in preparing implants for tissue engineering applications. Copyright © 2017. Published by Elsevier B.V.

  8. Effect of degree of deacetylation of chitosan on adsorption capacity and reusability of chitosan/polyvinyl alcohol/TiO2 nano composite.

    Science.gov (United States)

    Habiba, Umma; Joo, Tan Chin; Siddique, Tawsif A; Salleh, Areisman; Ang, Bee Chin; Afifi, Amalina M

    2017-11-01

    The chitosan/polyvinyl alcohol/TiO 2 composite was synthesized. Two different degrees of deacetylation of chitosan were prepared by hydrolysis to compare the effectiveness of them. The composite was analyzed via field emission scanning electron microscopy, Fourier transform infrared, X-ray diffraction, thermal gravimetric analysis, weight loss test and adsorption study. The FTIR and XRD results proved the interaction among chitosan, PVA and TiO 2 without any chemical reaction. It was found that, chitosan with higher degree of deacetylation has better stability. Furthermore, it also showed that higher DD of chitosan required less time to reach equilibrium for methyl orange. The adsorption followed the pseudo-second-order kinetic model. The Langmuir and Freundlich isotherm models were fitted well for isotherm study. Adsorption capacity was higher for the composite containing chitosan with higher DD. The dye removal rate was independent of the dye's initial concentration. The adsorption capacity was increased with temperature and it was found from reusability test that the composite containing chitosan with higher DD is more reusable. It was notable that adsorption capacity was even after 15 runs. Therefore, chitosan/PVA/TiO 2 composite can be a very useful material for dye removal. Copyright © 2017 Elsevier B.V. All rights reserved.

  9. Gelatin–PMVE/MA composite scaffold promotes expansion of embryonic stem cells

    International Nuclear Information System (INIS)

    Chhabra, Hemlata; Gupta, Priyanka; Verma, Paul J.; Jadhav, Sameer; Bellare, Jayesh R.

    2014-01-01

    We introduce a new composite scaffold of gelatin and polymethyl vinyl ether-alt-maleic anhydride (PMVE/MA) for expansion of embryonic stem cells (ESCs) in an in vitro environment. To optimize the scaffold, we prepared a gelatin scaffold (G) and three composite scaffolds namely GP-1, GP-2, and GP-3 with varying PMVE/MA concentrations (0.2–1%) and characterized them by scanning electron microscopy (SEM), swelling study, compression testing and FTIR. SEM micrographs revealed interconnected porous structure in all the scaffolds. The permissible hemolysis ratio and activation of platelets by scaffolds confirmed the hemocompatibility of scaffolds. Initial biocompatibility assessment of scaffolds was conducted using hepatocarcinoma (Hep G2) cells and adhesion, proliferation and infiltration of Hep G2 cells in depth of scaffolds were observed, proving the scaffold's biocompatibility. Further Oct4B2 mouse embryonic stem cells (mESCs), which harbor a green fluorescence protein transgene under regulatory control of the Oct4 promotor, were examined for expansion on scaffolds with MTT assay. The GP-2 scaffold demonstrated the best cell proliferation and was further explored for ESC adherence and infiltration in depth (SEM and confocal), and pluripotent state of mESCs was assessed with the expression of Oct4-GFP and stage-specific embryonic antigen-1 (SSEA-1). This study reports the first demonstration of biocompatibility of gelatin–PMVE/MA composite scaffold and presents this scaffold as a promising candidate for embryonic stem cell based tissue engineering. - Highlights: • Composite scaffolds of gelatin and PMVE/MA were prepared by freeze-drying method. • SEM micrographs showed porous structure in all scaffolds of varying pore dimension. • GP-2 composite exhibited better cellular response in comparison to other scaffolds. • mESCs proliferated and expressed Oct-4 and SSEA-1, when cultured on GP-2 scaffold

  10. Gelatin–PMVE/MA composite scaffold promotes expansion of embryonic stem cells

    Energy Technology Data Exchange (ETDEWEB)

    Chhabra, Hemlata [Department of Chemical Engineering, Indian Institute of Technology Bombay, Powai, Mumbai (India); Gupta, Priyanka [Department of Chemical Engineering, Indian Institute of Technology Bombay, Powai, Mumbai (India); IITB-Monash Research Academy, Mumbai (India); Department of Chemical Engineering, Monash University, Melbourne (Australia); Verma, Paul J. [Turretfield Research Centre, South Australian Research and Development Institute, Rosedale, South Australia (Australia); Jadhav, Sameer; Bellare, Jayesh R. [Department of Chemical Engineering, Indian Institute of Technology Bombay, Powai, Mumbai (India)

    2014-04-01

    We introduce a new composite scaffold of gelatin and polymethyl vinyl ether-alt-maleic anhydride (PMVE/MA) for expansion of embryonic stem cells (ESCs) in an in vitro environment. To optimize the scaffold, we prepared a gelatin scaffold (G) and three composite scaffolds namely GP-1, GP-2, and GP-3 with varying PMVE/MA concentrations (0.2–1%) and characterized them by scanning electron microscopy (SEM), swelling study, compression testing and FTIR. SEM micrographs revealed interconnected porous structure in all the scaffolds. The permissible hemolysis ratio and activation of platelets by scaffolds confirmed the hemocompatibility of scaffolds. Initial biocompatibility assessment of scaffolds was conducted using hepatocarcinoma (Hep G2) cells and adhesion, proliferation and infiltration of Hep G2 cells in depth of scaffolds were observed, proving the scaffold's biocompatibility. Further Oct4B2 mouse embryonic stem cells (mESCs), which harbor a green fluorescence protein transgene under regulatory control of the Oct4 promotor, were examined for expansion on scaffolds with MTT assay. The GP-2 scaffold demonstrated the best cell proliferation and was further explored for ESC adherence and infiltration in depth (SEM and confocal), and pluripotent state of mESCs was assessed with the expression of Oct4-GFP and stage-specific embryonic antigen-1 (SSEA-1). This study reports the first demonstration of biocompatibility of gelatin–PMVE/MA composite scaffold and presents this scaffold as a promising candidate for embryonic stem cell based tissue engineering. - Highlights: • Composite scaffolds of gelatin and PMVE/MA were prepared by freeze-drying method. • SEM micrographs showed porous structure in all scaffolds of varying pore dimension. • GP-2 composite exhibited better cellular response in comparison to other scaffolds. • mESCs proliferated and expressed Oct-4 and SSEA-1, when cultured on GP-2 scaffold.

  11. Polymer-Ceramic Composite Scaffolds: The Effect of Hydroxyapatite and β-tri-Calcium Phosphate

    OpenAIRE

    Boyang Huang; Guilherme Caetano; Cian Vyas; Jonny James Blaker; Carl Diver; Paulo Bártolo

    2018-01-01

    The design of bioactive scaffolds with improved mechanical and biological properties is an important topic of research. This paper investigates the use of polymer-ceramic composite scaffolds for bone tissue engineering. Different ceramic materials (hydroxyapatite (HA) and β-tri-calcium phosphate (TCP)) were mixed with poly-ε-caprolactone (PCL). Scaffolds with different material compositions were produced using an extrusion-based additive manufacturing system. The produced scaffolds were physi...

  12. 3D-macroporous chitosan-based scaffolds with in situ formed Pd and Pt nanoparticles for nitrophenol reduction.

    Science.gov (United States)

    Berillo, Dmitriy; Cundy, Andrew

    2018-07-15

    3D-macroporous chitosan-based scaffolds (cryogels) were produced via growth of metal-polymer coordinated complexes and electrostatic interactions between oppositely charged groups of chitosan and metal ions under subzero temperatures. A mechanism of reduction of noble metal complexes inside the cryogel walls by glutaraldehyde is proposed, which produces discrete and dispersed noble metal nanoparticles. 3D-macroporous scaffolds prepared under different conditions were characterised using TGA, FTIR, nitrogen adsorption, SEM, EDX and TEM, and the distribution of platinum nanoparticles (PtNPs) and palladium nanoparticles (PdNPs) in the material assessed. The catalytic activity of the in situ synthesised PdNPs, at 2.6, 12.5 and 21.0 μg total mass, respectively, was studied utilising a model system of 4-nitrophenol reduction. The kinetics of the reaction under different conditions (temperature, concentration of catalyst) were examined, and a decrease of catalytic activity was not observed over 17 treatment cycles. Increasing the temperature of the catalytic reaction from 10 to 22 and 35 °C by PdNPs supported within the cryogel increased the kinetic rate by 44 and 126%, respectively. Turnover number and turnover frequency of the PdNPs catalysts at room temperature were in the range 0.20-0.53 h -1 . The conversion degree of 4-nitrophenol at room temperature reached 98.9% (21.0 μg PdNPs). Significantly less mass of palladium nanoparticles (by 30-40 times) was needed compared to published data to obtain comparable rates of reduction of 4-nitrophenol. Copyright © 2018 Elsevier Ltd. All rights reserved.

  13. Microwave Irradiation Assisted Preparation of Chitosan Composite Microsphere for Dye Adsorption

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    Xiaoyu Chen

    2017-01-01

    Full Text Available Chitosan-activated carbon composite microspheres were prepared by emulsion cross-linking method and its adsorption properties for methyl orange were studied. Chitosan solution was mixed with activated carbon powder and then chitosan was cross-linked by epichlorohydrin under microwave irradiation. SEM photos show that the composite microspheres have diameters of 200–400 μm and activated carbon powder dispersed on the surface of composite microsphere. FTIR spectrum indicates chitosan is successfully cross-linked. Microwave irradiation can effectively shorten the cross-linking time. Composite microspheres have enhanced dye adsorption capacity for methyl orange compared to chitosan microspheres. Kinetic studies showed that the adsorption followed a pseudo-second-order model. Isotherm studies show that the isotherm adsorption equilibrium is better described by Freundlich isotherm. Regeneration results show that adsorption capacity of composite microsphere decreased about 5.51% after being reused for three times. These results indicated that chitosan-activated carbon composite microsphere has potential application in the removal of dye from wastewaters.

  14. Microwave Absorbent Packaging Material from Composites Chitosan-Polyvinyl Alcohol Polymer

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    Bambang - Riyanto

    2014-11-01

    Full Text Available Microwave absorbent packaging materials currently tend to biomaterial. Chitosan is a dielectric biomaterial with polycationic properties. The aim of this study was to analyze characteristics of microwave absorbing packaging material made from composite chitosan-polyvinyl alcohol (PVA polymer. The ability of the packaging material to absorb microwave was determined by reflection loss measurement. Formed packaging prototype resembles as a thin transparent yellowish plastic with thickness (0.11-0.22 mm and the tensile strength (106.33±2.82-143.00±2.59 kPa. SEM analysis showed homogenous structure characterized by interaction between chitosan and PVA. Optimum absorption value was obtained from chitosan concentration of 1%, with average value of reflection loss was (-31.9289±4.0094 dB.Keywords: chitosan, material packaging, microwave, reflection loss

  15. Microwave Absorbent Packaging Material from Composites Chitosan-Polyvinyl Alcohol Polymer

    Directory of Open Access Journals (Sweden)

    Bambang - Riyanto

    2015-07-01

    Full Text Available Microwave absorbent packaging materials currently tend to biomaterial. Chitosan is a dielectric biomaterial with polycationic properties. The aim of this study was to analyze characteristics of microwave absorbing packaging material made from composite chitosan-polyvinyl alcohol (PVA polymer. The ability of the packaging material to absorb microwave was determined by reflection loss measurement. Formed packaging prototype resembles as a thin transparent yellowish plastic with thickness (0.11-0.22 mm and the tensile strength (106.33±2.82-143.00±2.59 kPa. SEM analysis showed homogenous structure characterized by interaction between chitosan and PVA. Optimum absorption value was obtained from chitosan concentration of 1%, with average value of reflection loss was (-31.9289±4.0094 dB.Keywords: chitosan, material packaging, microwave, reflection loss

  16. Physico-mechanical properties of silanized-montmorillonite reinforced chitosan-co-poly(maleic anhydride) composites

    Science.gov (United States)

    Saputra, O. A.; Fajrin, A.; Nauqinida, M.; Suryanti, V.; Pramono, E.

    2017-07-01

    To solve the problems of dependence on petroleum as starting material in the manufacturing of plastics in Indonesia, green plastic from biopolymer like chitosan to be one of promising options and alternative to replace the conventional plastics. However, to overcome the mechanical and physical properties of chitosan, the addition of reinforcement agent was introduced. In this study, silanized-montmorillonite (sMMt) has been prepared as a reinforcement agent in the chitosan-co-poly(maleic anhydride) (referred as Cs-MAH) matrix. Silanizing of montmorillonite is one of strategy to improve the interaction between montmorillonite and chitosan, consequently, the mechanical properties, tensile strength of composites contained 6 phr of sMMt improved 56.5% to chitosan. Moreover, the presence both MAH and sMMt on the comosites also reduced swelling degree and swelling area by 20.6% and 26.7%.

  17. Inhibitory effect on Streptococcus mutans and mechanical properties of the chitosan containing composite resin

    Directory of Open Access Journals (Sweden)

    Ji-Sun Kim

    2013-02-01

    Full Text Available Objectives This study evaluated the antibacterial effect and mechanical properties of composite resins (LCR, MCR, HCR incorporating chitosan with three different molecular weights (L, Low; M, Medium; H, High. Materials and Methods Streptococcus (S. mutans 100 mL and each chitosan powder were inoculated in sterilized 10 mL Brain-Heart Infusion (BHI solution, and was centrifuged for 12 hr. Absorbance of the supernatent was measured at OD660 to estimate the antibacterial activities of chitosan. After S. mutans was inoculated in the disc shaped chitosan-containing composite resins, the disc was cleansed with BHI and diluted with serial dilution method. S. mutans was spread on Mitis-salivarius bacitracin agar. After then, colony forming unit (CFU was measured to verify the inhibitory effect on S. mutans biofilm. To ascertain the effect on the mechanical properties of composite resin, 3-point bending and Vickers hardness tests were done after 1 and 3 wk water storage, respectively. Using 2-way analysis of variance (ANOVA and Scheffe test, statistical analysis was done with 95% significance level. Results All chitosan powder showed inhibition effect against S. mutans. CFU number in chitosan-containing composite resins was smaller than that of control resin without chitosan. The chitosan containing composite resins did not show any significant difference in flexural strength and Vickers hardness in comparison with the control resin. However, the composite resin, MCR showed a slightly decreased flexural strength and the maximum load than those of control and the other composite resins HCR and LCR. Conclusions LCR and HCR would be recommended as a feasible antibacterial restorative due to its antibacterial nature and mechanical properties.

  18. Mechanical Properties of Chitosan-Starch Composite Filled Hydroxyapatite Micro- and Nanopowders

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    Jafar Ai

    2011-01-01

    Full Text Available Hydroxyapatite is a biocompatible ceramic and reinforcing material for bone implantations. In this study, Starch-chitosan hydrogel was produced using the oxidation of starch solution and subsequently cross-linked with chitosan via reductive alkylation method (weight ratio (starch/chitosan: 0.38. The hydroxyapatite micropowders and nanopowders synthesized by sol-gel method (10, 20, 30, 40 %W were composited to hydrogels and were investigated by mechanical analysis. The results of SEM images and Zetasizer experiments for synthesized nanopowders showed an average size of 100 nm. The nanoparticles distributed as uniform in the chitosan-starch film. The tensile modulus increased for composites containing hydroxyapatite nano-(size particle: 100 nanometer powders than composites containing micro-(size particle: 100 micrometer powders. The swelling percentage decreased for samples containing hydroxyapatite nanopowder than the micropowders. These nanocomposites could be applied for hard-tissue engineering.

  19. Development of Biomimetic Hybrid Porous Scaffold of Chitosan/Polyvinyl Alcohol/Carboxymethyl Cellulose by Freeze-Dried and Salt Leached Technique.

    Science.gov (United States)

    Kanimozhi, K; Basha, S Khaleel; Kumari, V Sugantha; Kaviyarasu, K

    2018-07-01

    Freeze drying and salt leaching methods were applied to fabricate Chitosan/Poly(vinyl alcohol)/Carboxymethyl cellulose (CPCMC) biomimetic porous scaffolds for soft tissue engineering. The properties of these scaffolds were investigated and compared to those by freeze drying and salt leaching methods respectively. The salt-leached CS/PVA/CMC scaffolds were easily formed into desired shapes with a uniformly distributed and interconnected pore structure with an average pore size. The mechanical strength of the scaffolds increased with the porosity, and were easily modulated by the addition of carboxymethyl cellulose. The morphology of the porous scaffolds observed using a SEM exhibited good porosity and interconnectivity of pores. MTT assay using L929 fibroblast cells demonstrated that the cell viability of the porous scaffold was good. Scaffolds prepared by salt leached method show larger swelling capacity, and mechanical strength, potent antibacterial activity and more cell viability than freeze dried method. It is found that salt leaching method has distinguished characteristics of simple, efficient, feasible and less economic than freeze dried scaffolds.

  20. In vitro evaluation of alginate/halloysite nanotube composite scaffolds for tissue engineering

    International Nuclear Information System (INIS)

    Liu, Mingxian; Dai, Libing; Shi, Huizhe; Xiong, Sheng; Zhou, Changren

    2015-01-01

    In this study, a series of alginate/halloysite nanotube (HNTs) composite scaffolds were prepared by solution-mixing and freeze-drying method. HNTs are incorporated into alginate to improve both the mechanical and cell-attachment properties of the scaffolds. The interfacial interactions between alginate and HNTs were confirmed by the atomic force microscope (AFM), transmission electron microscope (TEM) and FTIR spectroscopy. The mechanical, morphological, and physico-chemical properties of the composite scaffolds were investigated. The composite scaffolds exhibit significant enhancement in compressive strength and compressive modulus compared with pure alginate scaffold both in dry and wet states. A well-interconnected porous structure with size in the range of 100–200 μm and over 96% porosity is found in the composite scaffolds. X-ray diffraction (XRD) result shows that HNTs are uniformly dispersed and partly oriented in the composite scaffolds. The incorporation of HNTs leads to increase in the scaffold density and decrease in the water swelling ratio of alginate. HNTs improve the stability of alginate scaffolds against enzymatic degradation in PBS solution. Thermogravimetrica analysis (TGA) shows that HNTs can improve the thermal stability of the alginate. The mouse fibroblast cells display better attachment to the alginate/HNT composite than those to the pure alginate, suggesting the good cytocompatibility of the composite scaffolds. Alginate/HNT composite scaffolds exhibit great potential for applications in tissue engineering. - Highlights: • We fabricated HNTs reinforced alginate composite scaffolds for biomedical applications. • The hydrogen bond interactions between HNTs and alginate are confirmed. • HNTs can significantly enhance the mechanical properties of alginate scaffold. • The scaffolds exhibit a highly porous structure with interconnected pores. • HNTs can improve the cell attachment and proliferation on alginate

  1. In vitro evaluation of alginate/halloysite nanotube composite scaffolds for tissue engineering

    Energy Technology Data Exchange (ETDEWEB)

    Liu, Mingxian [Department of Materials Science and Engineering, Jinan University, Guangzhou 510632 (China); Dai, Libing [Guangzhou Institute of Traumatic Surgery, Guangzhou Red Cross Hospital Medical College, Jinan University, Guangzhou 510220 (China); Shi, Huizhe; Xiong, Sheng [Institute of Biomedicine, National Engineering Research Center of Genetic Medicine, Jinan University, Guangzhou 510632 (China); Zhou, Changren, E-mail: tcrz9@jnu.edu.cn [Department of Materials Science and Engineering, Jinan University, Guangzhou 510632 (China)

    2015-04-01

    In this study, a series of alginate/halloysite nanotube (HNTs) composite scaffolds were prepared by solution-mixing and freeze-drying method. HNTs are incorporated into alginate to improve both the mechanical and cell-attachment properties of the scaffolds. The interfacial interactions between alginate and HNTs were confirmed by the atomic force microscope (AFM), transmission electron microscope (TEM) and FTIR spectroscopy. The mechanical, morphological, and physico-chemical properties of the composite scaffolds were investigated. The composite scaffolds exhibit significant enhancement in compressive strength and compressive modulus compared with pure alginate scaffold both in dry and wet states. A well-interconnected porous structure with size in the range of 100–200 μm and over 96% porosity is found in the composite scaffolds. X-ray diffraction (XRD) result shows that HNTs are uniformly dispersed and partly oriented in the composite scaffolds. The incorporation of HNTs leads to increase in the scaffold density and decrease in the water swelling ratio of alginate. HNTs improve the stability of alginate scaffolds against enzymatic degradation in PBS solution. Thermogravimetrica analysis (TGA) shows that HNTs can improve the thermal stability of the alginate. The mouse fibroblast cells display better attachment to the alginate/HNT composite than those to the pure alginate, suggesting the good cytocompatibility of the composite scaffolds. Alginate/HNT composite scaffolds exhibit great potential for applications in tissue engineering. - Highlights: • We fabricated HNTs reinforced alginate composite scaffolds for biomedical applications. • The hydrogen bond interactions between HNTs and alginate are confirmed. • HNTs can significantly enhance the mechanical properties of alginate scaffold. • The scaffolds exhibit a highly porous structure with interconnected pores. • HNTs can improve the cell attachment and proliferation on alginate.

  2. Chitosan-Graphene Oxide 3D scaffolds as Promising Tools for Bone Regeneration in Critical-Size Mouse Calvarial Defects.

    Science.gov (United States)

    Hermenean, Anca; Codreanu, Ada; Herman, Hildegard; Balta, Cornel; Rosu, Marcel; Mihali, Ciprian Valentin; Ivan, Alexandra; Dinescu, Sorina; Ionita, Mariana; Costache, Marieta

    2017-11-30

    Limited self-regenerating capacity of human skeleton makes the reconstruction of critical size bone defect a significant challenge for clinical practice. Aimed for regenerating bone tissues, this study was designed to investigate osteogenic differentiation, along with bone repair capacity of 3D chitosan (CHT) scaffolds enriched with graphene oxide (GO) in critical-sized mouse calvarial defect. Histopathological/histomorphometry and scanning electron microscopy(SEM) analysis of the implants revealed larger amount of new bone in the CHT/GO-filled defects compared with CHT alone (p < 0.001). When combined with GO, CHT scaffolds synergistically promoted the increase of alkaline phosphatase activity both in vitro and in vivo experiments. This enhanced osteogenesis was corroborated with increased expression of bone morphogenetic protein (BMP) and Runx-2 up to week 4 post-implantation, which showed that GO facilitates the differentiation of osteoprogenitor cells. Meanwhile, osteogenesis was promoted by GO at the late stage as well, as indicated by the up-regulation of osteopontin and osteocalcin at week 8 and overexpressed at week 18, for both markers. Our data suggest that CHT/GO biomaterial could represent a promising tool for the reconstruction of large bone defects, without using exogenous living cells or growth factors.

  3. Biomimetic composite coating on rapid prototyped scaffolds for bone tissue engineering.

    Science.gov (United States)

    Arafat, M Tarik; Lam, Christopher X F; Ekaputra, Andrew K; Wong, Siew Yee; Li, Xu; Gibson, Ian

    2011-02-01

    The objective of this present study was to improve the functional performance of rapid prototyped scaffolds for bone tissue engineering through biomimetic composite coating. Rapid prototyped poly(ε-caprolactone)/tri-calcium phosphate (PCL/TCP) scaffolds were fabricated using the screw extrusion system (SES). The fabricated PCL/TCP scaffolds were coated with a carbonated hydroxyapatite (CHA)-gelatin composite via biomimetic co-precipitation. The structure of the prepared CHA-gelatin composite coating was studied by scanning electron microscopy (SEM), X-ray photoelectron spectroscopy and Fourier transform infrared spectroscopy. Compressive mechanical testing revealed that the coating process did not have any detrimental effect on the mechanical properties of the scaffolds. The cell-scaffold interaction was studied by culturing porcine bone marrow stromal cells (BMSCs) on the scaffolds and assessing the proliferation and bone-related gene and protein expression capabilities of the cells. Confocal laser microscopy and SEM images of the cell-scaffold constructs showed a uniformly distributed cell sheet and accumulation of extracellular matrix in the interior of CHA-gelatin composite-coated PCL/TCP scaffolds. The proliferation rate of BMSCs on CHA-gelatin composite-coated PCL/TCP scaffolds was about 2.3 and 1.7 times higher than that on PCL/TCP scaffolds and CHA-coated PCL/TCP scaffolds, respectively, by day 10. Furthermore, reverse transcription polymerase chain reaction and Western blot analysis revealed that CHA-gelatin composite-coated PCL/TCP scaffolds stimulate osteogenic differentiation of BMSCs the most, compared with PCL/TCP scaffolds and CHA-coated PCL/TCP scaffolds. These results demonstrate that CHA-gelatin composite-coated rapid prototyped PCL/TCP scaffolds are promising for bone tissue engineering. Copyright © 2010 Acta Materialia Inc. Published by Elsevier Ltd. All rights reserved.

  4. Polycaprolactone foam functionalized with chitosan microparticles – a suitable scaffold for cartilage regeneration.

    Czech Academy of Sciences Publication Activity Database

    Filová, Eva; Jakubcová, B.; Danilová, I.; Kuželová Košťáková, E.; Jarošíková, T.; Chernyavskiy, Oleksandr; Hejda, J.; Handl, M.; Beznoska, J.; Nečas, A.; Rosina, J.; Amler, Evžen

    2016-01-01

    Roč. 65, č. 1 (2016), s. 121-131 ISSN 0862-8408 Institutional support: RVO:68378041 ; RVO:67985823 Keywords : poly-epsilon-caprolactone * chitosan * cartilage regeneration * foam Subject RIV: FI - Traumatology, Orthopedics Impact factor: 1.461, year: 2016

  5. Pore size and LbL chitosan coating influence mesenchymal stem cell in vitro fibrosis and biomineralization in 3D porous poly(epsilon-caprolactone) scaffolds.

    Science.gov (United States)

    Mehr, Nima Ghavidel; Li, Xian; Chen, Gaoping; Favis, Basil D; Hoemann, Caroline D

    2015-07-01

    Poly(epsilon-caprolactone) (PCL) is a hydrophobic bioplastic under development for bone tissue engineering applications. Limited information is available on the role of internal geometry and cell-surface attachment on osseous integration potential. We tested the hypothesis that human bone marrow mesenchymal stem cells (MSCs) deposit more mineral inside porous 3D PCL scaffolds with fully interconnected 84 or 141 µm pores, when the surfaces are coated with chitosan via Layer-by-Layer (LbL)-deposited polyelectrolytes. Freshly trypsinized MSCs were seeded on PCL 3D cylinders using a novel static cold seeding method in 2% serum to optimally populate all depths of the scaffold discs, followed by 10 days of culture in proliferation medium and 21 additional days in osteogenic medium. MSCs were observed by SEM and histology to spread faster and to proliferate more on chitosan-coated pore surfaces. Most pores, with or without chitosan, became filled by collagen networks sparsely populated with fibroblast-like cells. After 21 days of culture in osteogenic medium, sporadic matrix mineralization was detected histologically and by micro-CT in highly cellular surface layers that enveloped all scaffolds and in cell aggregates in 141 µm pores near the edges. LbL-chitosan promoted punctate mineral deposition on the surfaces of 84 µm pores (p chitosan coatings are sufficient to promote MSC attachment to PCL but only enhance mineral formation in 84 µm pores, suggesting a potential inhibitory role for MSC-derived fibroblasts in osteoblast terminal differentiation. © 2014 Wiley Periodicals, Inc.

  6. A gelatin composite scaffold strengthened by drug-loaded halloysite nanotubes.

    Science.gov (United States)

    Ji, Lijun; Qiao, Wei; Zhang, Yuheng; Wu, Huayu; Miao, Shiyong; Cheng, Zhilin; Gong, Qianming; Liang, Ji; Zhu, Aiping

    2017-09-01

    Mechanical properties and anti-infection are two of the most concerned issues for artificial bone grafting materials. Bone regeneration porous scaffolds with sustained drug release were developed by freeze-drying the mixture of nanosized drug-loaded halloysite nanotubes (HNTs) and gelatin. The scaffolds showed porous structure and excellent biocompatibility. The mechanical properties of the obtained composite scaffolds were enhanced significantly by HNTs to >300%, comparing to those of gelatin scaffold, and match to those of natural cancellous bones. The ibuprofen-loaded HNTs incorporated in the scaffolds allowed extended drug release over 100h, comparing to 8h when directly mixed the drug into the gelatin scaffold. The biological properties of the composite scaffolds were investigated by culturing MG63 cells on them. The HNTs/gelatin scaffolds with excellent mechanical properties and sustained drug release could be a promising artificial bone grating material. Copyright © 2017 Elsevier B.V. All rights reserved.

  7. Development of chitosan-pullulan composite nanoparticles for nasal delivery of vaccines: in vivo studies.

    Science.gov (United States)

    Cevher, Erdal; Salomon, Stefan K; Somavarapu, Satyanarayana; Brocchini, Steve; Alpar, H Oya

    2015-01-01

    Here, we aimed at developing chitosan/pullulan composite nanoparticles and testing their potential as novel systems for the nasal delivery of diphtheria toxoid (DT). All the chitosan derivatives [N-trimethyl (TMC), chloride and glutamate] and carboxymethyl pullulan (CMP) were synthesised and antigen-loaded composites were prepared by polyion complexation of chitosan and pullulan derivatives (particle size: 239-405 nm; surface charge: +18 and +27 mV). Their immunological effects after intranasal administration to mice were compared to intramuscular route. Composite nanoparticles induced higher levels of IgG responses than particles formed with chitosan derivative and antigen. Nasally administered TMC-pullulan composites showed higher DT serum IgG titre when compared with the other composites. Co-encapsulation of CpG ODN within TMC-CMP-DT nanoparticles resulted in a balanced Th1/Th2 response. TMC/pullulan composite nanoparticles also induced highest cytokine levels compared to those of chitosan salts. These findings demonstrated that TMC-CMP-DT composite nanoparticles are promising delivery system for nasal vaccination.

  8. Effects of 3-dimensional culture conditions (collagen-chitosan nano-scaffolds) on maturation of dendritic cells and their capacity to interact with T-lymphocytes.

    Science.gov (United States)

    Daneshmandi, Saeed; Dibazar, Shaghayegh Pishkhan; Fateh, Shirin

    2016-01-01

    In the body, there is a natural three-dimensional (3D) microenvironment in which immune cells, including dendritic cells (DC), play their functions. This study evaluated the impact of using collagen-chitosan 3D nano-scaffolds in comparisons to routine 2D culture plates on DC phenotype and functions. Bone marrow-derived DC were cultured on scaffolds and plates and then stimulated with lipopolysaccharide (LPS) or chitosan-based nanoparticles (NP) for 24 h. Thereafter, DC viability, expression of maturation markers and levels of cytokines secretion were evaluated. In another set of studies, the DC were co-cultured with allogenic T-lymphocytes in both the 2D and 3D systems and effects on DC-induction of T-lymphocyte proliferation and cytokine release were analyzed. The results indicated that CD40, CD86 and MHC II marker expression and interleukin (IL)-12, IL-6 and tumor necrosis factor (TNF)-α secretion by DC were enhanced in 3D cultures in comparison to by cells maintained in the 2D states. The data also showed that DNA/chitosan NP activated DC more than LPS in the 3D system. T-Lymphocyte proliferation was induced to a greater extent by DNA/NP-treated DC when both cell types were maintained on the scaffolds. Interestingly, while DC induction of T-lymphocyte interferon (IFN)-γ and IL-4 release was enhanced in the 3D system (relative to controls), there was a suppression of transforming growth factor (TGF)-β production; effects on IL-10 secretion were variable. The results here suggested that collagen-chitosan scaffolds could provide a pro-inflammatory and activator environment to perform studies to analyze effects of exogenous agents on the induction of DC maturation, NP uptake and/or cytokines release, as well as for the ability of these cells to potentially interact with other immune system cells in vitro.

  9. Novel High-Viscosity Polyacrylamidated Chitosan for Neural Tissue Engineering: Fabrication of Anisotropic Neurodurable Scaffold via Molecular Disposition of Persulfate-Mediated Polymer Slicing and Complexation

    Directory of Open Access Journals (Sweden)

    Viness Pillay

    2012-10-01

    Full Text Available Macroporous polyacrylamide-grafted-chitosan scaffolds for neural tissue engineering were fabricated with varied synthetic and viscosity profiles. A novel approach and mechanism was utilized for polyacrylamide grafting onto chitosan using potassium persulfate (KPS mediated degradation of both polymers under a thermally controlled environment. Commercially available high molecular mass polyacrylamide was used instead of the acrylamide monomer for graft copolymerization. This grafting strategy yielded an enhanced grafting efficiency (GE = 92%, grafting ratio (GR = 263%, intrinsic viscosity (IV = 5.231 dL/g and viscometric average molecular mass (MW = 1.63 × 106 Da compared with known acrylamide that has a GE = 83%, GR = 178%, IV = 3.901 dL/g and MW = 1.22 × 106 Da. Image processing analysis of SEM images of the newly grafted neurodurable scaffold was undertaken based on the polymer-pore threshold. Attenuated Total Reflectance-FTIR spectral analyses in conjugation with DSC were used for the characterization and comparison of the newly grafted copolymers. Static Lattice Atomistic Simulations were employed to investigate and elucidate the copolymeric assembly and reaction mechanism by exploring the spatial disposition of chitosan and polyacrylamide with respect to the reactional profile of potassium persulfate. Interestingly, potassium persulfate, a peroxide, was found to play a dual role initially degrading the polymers—“polymer slicing”—thereby initiating the formation of free radicals and subsequently leading to synthesis of the high molecular mass polyacrylamide-grafted-chitosan (PAAm-g-CHT—“polymer complexation”. Furthermore, the applicability of the uniquely grafted scaffold for neural tissue engineering was evaluated via PC12 neuronal cell seeding. The novel PAAm-g-CHT exhibited superior neurocompatibility in terms of cell infiltration owing to the anisotropic porous architecture, high molecular mass mediated robustness

  10. Effects of Genipin Concentration on Cross-Linked Chitosan Scaffolds for Bone Tissue Engineering: Structural Characterization and Evidence of Biocompatibility Features

    Directory of Open Access Journals (Sweden)

    Simona Dimida

    2017-01-01

    Full Text Available Genipin (GN is a natural molecule extracted from the fruit of Gardenia jasminoides Ellis according to modern microbiological processes. Genipin is considered as a favorable cross-linking agent due to its low cytotoxicity compared to widely used cross-linkers; it cross-links compounds with primary amine groups such as proteins, collagen, and chitosan. Chitosan is a biocompatible polymer that is currently studied in bone tissue engineering for its capacity to promote growth and mineral-rich matrix deposition by osteoblasts in culture. In this work, two genipin cross-linked chitosan scaffolds for bone repair and regeneration were prepared with different GN concentrations, and their chemical, physical, and biological properties were explored. Scanning electron microscopy and mechanical tests revealed that nonremarkable changes in morphology, porosity, and mechanical strength of scaffolds are induced by increasing the cross-linking degree. Also, the degradation rate was shown to decrease while increasing the cross-linking degree, with the high cross-linking density of the scaffold disabling the hydrolysis activity. Finally, basic biocompatibility was investigated in vitro, by evaluating proliferation of two human-derived cell lines, namely, the MG63 (human immortalized osteosarcoma and the hMSCs (human mesenchymal stem cells, as suitable cell models for bone tissue engineering applications of biomaterials.

  11. Culture on 3D Chitosan-Hyaluronic Acid Scaffolds Enhances Stem Cell Marker Expression and Drug Resistance in Human Glioblastoma Cancer Stem Cells.

    Science.gov (United States)

    Wang, Kui; Kievit, Forrest M; Erickson, Ariane E; Silber, John R; Ellenbogen, Richard G; Zhang, Miqin

    2016-12-01

    The lack of in vitro models that support the growth of glioblastoma (GBM) stem cells (GSCs) that underlie clinical aggressiveness hinders developing new, effective therapies for GBM. While orthotopic patient-derived xenograft models of GBM best reflect in vivo tumor behavior, establishing xenografts is a time consuming, costly, and frequently unsuccessful endeavor. To address these limitations, a 3D porous scaffold composed of chitosan and hyaluronic acid (CHA) is synthesized. Growth and expression of the cancer stem cell (CSC) phenotype of the GSC GBM6 taken directly from fresh xenogratfs grown on scaffolds or as adherent monolayers is compared. While 2D adherent cultures grow as monolayers of flat epitheliod cells, GBM6 cells proliferate within pores of CHA scaffolds as clusters of self-adherent ovoid cells. Growth on scaffolds is accompanied by greater expression of genes that mediate epithelial-mesenchymal transition and maintain a primitive, undifferentiated phenotype, hallmarks of CSCs. Scaffold-grown cells also display higher expression of genes that promote resistance to hypoxia-induced oxidative stress. In accord, scaffold-grown cells show markedly greater resistance to clinically utilized alkylating agents compared to adherent cells. These findings suggest that our CHA scaffolds better mimic in vivo biological and clinical behavior and provide insights for developing novel individualized treatments. © 2016 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  12. 3D Porous Chitosan-Alginate Scaffolds as an In Vitro Model for Evaluating Nanoparticle-Mediated Tumor Targeting and Gene Delivery to Prostate Cancer.

    Science.gov (United States)

    Wang, Kui; Kievit, Forrest M; Florczyk, Stephen J; Stephen, Zachary R; Zhang, Miqin

    2015-10-12

    Cationic nanoparticles (NPs) for targeted gene delivery are conventionally evaluated using 2D in vitro cultures. However, this does not translate well to corresponding in vivo studies because of the marked difference in NP behavior in the presence of the tumor microenvironment. In this study, we investigated whether prostate cancer (PCa) cells cultured in three-dimensional (3D) chitosan-alginate (CA) porous scaffolds could model cationic NP-mediated gene targeted delivery to tumors in vitro. We assessed in vitro tumor cell proliferation, formation of tumor spheroids, and expression of marker genes that promote tumor malignancy in CA scaffolds. The efficacy of NP-targeted gene delivery was evaluated in PCa cells in 2D cultures, PCa tumor spheroids grown in CA scaffolds, and PCa tumors in a mouse TRAMP-C2 flank tumor model. PCa cells cultured in CA scaffolds grew into tumor spheroids and displayed characteristics of higher malignancy as compared to those in 2D cultures. Significantly, targeted gene delivery was only observed in cells cultured in CA scaffolds, whereas cells cultured on 2D plates showed no difference in gene delivery between targeted and nontarget control NPs. In vivo NP evaluation confirmed targeted gene delivery, indicating that only CA scaffolds correctly modeled NP-mediated targeted delivery in vivo. These findings suggest that CA scaffolds serve as a better in vitro platform than 2D cultures for evaluation of NP-mediated targeted gene delivery to PCa.

  13. Three-dimensional dynamic fabrication of engineered cartilage based on chitosan/gelatin hybrid hydrogel scaffold in a spinner flask with a special designed steel frame

    International Nuclear Information System (INIS)

    Song, Kedong; Li, Liying; Li, Wenfang; Zhu, Yanxia; Jiao, Zeren; Lim, Mayasari; Fang, Meiyun; Shi, Fangxin; Wang, Ling; Liu, Tianqing

    2015-01-01

    Cartilage transplantation using in vitro tissue engineered cartilage is considered a promising treatment for articular cartilage defects. In this study, we assessed the advantages of adipose derived stem cells (ADSCs) combined with chitosan/gelatin hybrid hydrogel scaffolds, which acted as a cartilage biomimetic scaffold, to fabricate a tissue engineered cartilage dynamically in vitro and compared this with traditional static culture. Physical properties of the hydrogel scaffolds were evaluated and ADSCs were inoculated into the hydrogel at a density of 1 × 10 7 cells/mL and cultured in a spinner flask with a special designed steel framework and feed with chondrogenic inductive media for two weeks. The results showed that the average pore size, porosity, swelling rate and elasticity modulus of hybrid scaffolds with good biocompatibility were 118.25 ± 19.51 μm, 82.60 ± 2.34%, 361.28 ± 0.47% and 61.2 ± 0.16 kPa, respectively. ADSCs grew well in chitosan/gelatin hybrid scaffold and successfully differentiated into chondrocytes, showing that the scaffolds were suitable for tissue engineering applications in cartilage regeneration. Induced cells cultivated in a dynamic spinner flask with a special designed steel frame expressed more proteoglycans and the cell distribution was much more uniform with the scaffold being filled mostly with extracellular matrix produced by cells. A spinner flask with framework promoted proliferation and chondrogenic differentiation of ADSCs within chitosan/gelatin hybrid scaffolds and accelerated dynamic fabrication of cell–hydrogel constructs, which could be a selective and good method to construct tissue engineered cartilage in vitro. - Highlights: • ADSCs/hybrid scaffold constructs are dynamically fabricated in a spinner flask with a special framework. • Inside convection in spinner flask made enough supplement of oxygen and nutrients far beyond the depth of passive diffusion. • 3D culture environment accelerated mass

  14. Three-dimensional dynamic fabrication of engineered cartilage based on chitosan/gelatin hybrid hydrogel scaffold in a spinner flask with a special designed steel frame

    Energy Technology Data Exchange (ETDEWEB)

    Song, Kedong, E-mail: kedongsong@dlut.edu.cn [State Key Laboratory of Fine Chemicals, Dalian R& D Center for Stem Cell and Tissue Engineering, Dalian University of Technology, Dalian 116024 (China); Li, Liying; Li, Wenfang [State Key Laboratory of Fine Chemicals, Dalian R& D Center for Stem Cell and Tissue Engineering, Dalian University of Technology, Dalian 116024 (China); Zhu, Yanxia [Anti-Ageing and Regenerative Medicine Centre, Shenzhen University, 3688 Nanhai Avenue, Shenzhen 518060 Guangdong (China); Jiao, Zeren [State Key Laboratory of Fine Chemicals, Dalian R& D Center for Stem Cell and Tissue Engineering, Dalian University of Technology, Dalian 116024 (China); Lim, Mayasari [Division of Bioengineering, School of Chemical and Biomedical Engineering, Nanyang Technological University, Singapore 637457 (Singapore); Fang, Meiyun [Department of Hematology, First Affiliated Hospital, Dalian Medical University, Dalian 116011 (China); Shi, Fangxin [Department of Oncology, First Affiliated Hospital of Dalian Medical University, Dalian 116011 (China); Wang, Ling, E-mail: whwl@hotmail.com [Department of Obstetrics and Gynecology, First Affiliated Hospital, Dalian Medical University, Dalian 116011 (China); Liu, Tianqing, E-mail: liutq@dlut.edu.cn [State Key Laboratory of Fine Chemicals, Dalian R& D Center for Stem Cell and Tissue Engineering, Dalian University of Technology, Dalian 116024 (China)

    2015-10-01

    Cartilage transplantation using in vitro tissue engineered cartilage is considered a promising treatment for articular cartilage defects. In this study, we assessed the advantages of adipose derived stem cells (ADSCs) combined with chitosan/gelatin hybrid hydrogel scaffolds, which acted as a cartilage biomimetic scaffold, to fabricate a tissue engineered cartilage dynamically in vitro and compared this with traditional static culture. Physical properties of the hydrogel scaffolds were evaluated and ADSCs were inoculated into the hydrogel at a density of 1 × 10{sup 7} cells/mL and cultured in a spinner flask with a special designed steel framework and feed with chondrogenic inductive media for two weeks. The results showed that the average pore size, porosity, swelling rate and elasticity modulus of hybrid scaffolds with good biocompatibility were 118.25 ± 19.51 μm, 82.60 ± 2.34%, 361.28 ± 0.47% and 61.2 ± 0.16 kPa, respectively. ADSCs grew well in chitosan/gelatin hybrid scaffold and successfully differentiated into chondrocytes, showing that the scaffolds were suitable for tissue engineering applications in cartilage regeneration. Induced cells cultivated in a dynamic spinner flask with a special designed steel frame expressed more proteoglycans and the cell distribution was much more uniform with the scaffold being filled mostly with extracellular matrix produced by cells. A spinner flask with framework promoted proliferation and chondrogenic differentiation of ADSCs within chitosan/gelatin hybrid scaffolds and accelerated dynamic fabrication of cell–hydrogel constructs, which could be a selective and good method to construct tissue engineered cartilage in vitro. - Highlights: • ADSCs/hybrid scaffold constructs are dynamically fabricated in a spinner flask with a special framework. • Inside convection in spinner flask made enough supplement of oxygen and nutrients far beyond the depth of passive diffusion. • 3D culture environment accelerated mass

  15. Developing multi-cellular tumor spheroid model (MCTS) in the chitosan/collagen/alginate (CCA) fibrous scaffold for anticancer drug screening

    Energy Technology Data Exchange (ETDEWEB)

    Wang, Jian-Zheng, E-mail: wppzheng@126.com [Laboratory of Biomedical Material Engineering, Dalian Institute of Chemical Physics, Chinese Academy of Sciences, 457 Zhongshan Road, Dalian 116023 (China); Affiliated General Hospital, Tianguan Group Co., Ltd, Nanyang 473000 (China); Testing Center of Henan Tianguan Group Co., Ltd, Nanyang 473000 (China); Zhu, Yu-Xia [Laboratory of Biomedical Material Engineering, Dalian Institute of Chemical Physics, Chinese Academy of Sciences, 457 Zhongshan Road, Dalian 116023 (China); Affiliated General Hospital, Tianguan Group Co., Ltd, Nanyang 473000 (China); Testing Center of Henan Tianguan Group Co., Ltd, Nanyang 473000 (China); Ma, Hui-Chao; Chen, Si-Nan; Chao, Ji-Ye; Ruan, Wen-Ding; Wang, Duo; Du, Feng-guang [Affiliated General Hospital, Tianguan Group Co., Ltd, Nanyang 473000 (China); Testing Center of Henan Tianguan Group Co., Ltd, Nanyang 473000 (China); Meng, Yue-Zhong [State Key Laboratory of Optoelectronic Materials and Technologies, Sun Yat-sen University, Guangzhou 510275 (China)

    2016-05-01

    In this work, a 3D MCTS-CCA system was constructed by culturing multi-cellular tumor spheroid (MCTS) in the chitosan/collagen/alginate (CCA) fibrous scaffold for anticancer drug screening. The CCA scaffolds were fabricated by spray-spinning. The interactions between the components of the spray-spun fibers were evidenced by methods of Coomassie Blue stain, X-ray diffraction (XRD) and Fourier transform-infrared spectroscopy (FTIR). Co-culture indicated that MCF-7 cells showed a spatial growth pattern of multi-cellular tumor spheroid (MCTS) in the CCA fibrous scaffold with increased proliferation rate and drug-resistance to MMC, ADM and 5-Aza comparing with the 2D culture cells. Significant increases of total viable cells were found in 3D MCTS groups after drug administration by method of apoptotic analysis. Glucose–lactate analysis indicated that the metabolism of MCTS in CCA scaffold was closer to the tumor issue in vivo than the monolayer cells. In addition, MCTS showed the characteristic of epithelial mesenchymal transition (EMT) which is subverted by carcinoma cells to facilitate metastatic spread. These results demonstrated that MCTS in CCA scaffold possessed a more conservative phenotype of tumor than monolayer cells, and anticancer drug screening in 3D MCTS-CCA system might be superior to the 2D culture system. - Highlights: • Chitosan/collagen/alginate (CCA) scaffolds were fabricated by spray-spinning. • MCF-7 cells presented a multi-cellular tumor spheroid model (MCTS) in CCA scaffold. • MCTS in CCA possessed a more conservative phenotype of tumor than monolayer cells. • Anticancer drug screening in MCTS-CCA system is superior to 2D culture system.

  16. Developing multi-cellular tumor spheroid model (MCTS) in the chitosan/collagen/alginate (CCA) fibrous scaffold for anticancer drug screening

    International Nuclear Information System (INIS)

    Wang, Jian-Zheng; Zhu, Yu-Xia; Ma, Hui-Chao; Chen, Si-Nan; Chao, Ji-Ye; Ruan, Wen-Ding; Wang, Duo; Du, Feng-guang; Meng, Yue-Zhong

    2016-01-01

    In this work, a 3D MCTS-CCA system was constructed by culturing multi-cellular tumor spheroid (MCTS) in the chitosan/collagen/alginate (CCA) fibrous scaffold for anticancer drug screening. The CCA scaffolds were fabricated by spray-spinning. The interactions between the components of the spray-spun fibers were evidenced by methods of Coomassie Blue stain, X-ray diffraction (XRD) and Fourier transform-infrared spectroscopy (FTIR). Co-culture indicated that MCF-7 cells showed a spatial growth pattern of multi-cellular tumor spheroid (MCTS) in the CCA fibrous scaffold with increased proliferation rate and drug-resistance to MMC, ADM and 5-Aza comparing with the 2D culture cells. Significant increases of total viable cells were found in 3D MCTS groups after drug administration by method of apoptotic analysis. Glucose–lactate analysis indicated that the metabolism of MCTS in CCA scaffold was closer to the tumor issue in vivo than the monolayer cells. In addition, MCTS showed the characteristic of epithelial mesenchymal transition (EMT) which is subverted by carcinoma cells to facilitate metastatic spread. These results demonstrated that MCTS in CCA scaffold possessed a more conservative phenotype of tumor than monolayer cells, and anticancer drug screening in 3D MCTS-CCA system might be superior to the 2D culture system. - Highlights: • Chitosan/collagen/alginate (CCA) scaffolds were fabricated by spray-spinning. • MCF-7 cells presented a multi-cellular tumor spheroid model (MCTS) in CCA scaffold. • MCTS in CCA possessed a more conservative phenotype of tumor than monolayer cells. • Anticancer drug screening in MCTS-CCA system is superior to 2D culture system.

  17. Surface biofunctionalization of three-dimensional porous poly(lactic acid) scaffold using chitosan/OGP coating for bone tissue engineering.

    Science.gov (United States)

    Zeng, Sen; Ye, Jianhua; Cui, Zhixiang; Si, Junhui; Wang, Qianting; Wang, Xiaofeng; Peng, Kaiping; Chen, Wenzhe

    2017-08-01

    As one of the stimulators on bone formation, osteogenic growth peptide (OGP) improves both proliferation and differentiation of the bone cells in vitro and in vivo. The aim of this work was the preparation of three dimensional porous poly(lactic acid) (PLA) scaffold with high porosity from PLA-dioxane-water ternary system with the use of vacuum-assisted solvent casting, phase separation, solvent extraction and particle leaching methods. Then, by surface coating of PLA scaffold with chitosan (CS)/OGP solution, biofunctionalization of PLA scaffold had been completed for application in bone regeneration. The effects of frozen temperature (-20, -50, -80°C) and PLA solution concentration (10, 12, 14wt%) on the microstructure, water absorption, porosity, hydrophilicity, mechanical properties, and biocompatibility of PLA and CS/OGP/PLA scaffold were investigated. Results showed that both PLA and CS/OGP/PLA scaffolds have an interconnected network structure and a porosity of up to 96.1% and 91.5%, respectively. The CS/OGP/PLA scaffold exhibited better hydrophilicity and mechanical properties than that of uncoated PLA scaffold. Moreover, the results of cell culture test showed that CS/OGP coating could stimulate the proliferation and growth of osteoblast cells on CS/OGP/PLA scaffold. These finding suggested that the surface biofunctionalization by CS/OGP coating layer could be an effective method on enhancing cell adhesion to synthetic polymer-based scaffolds in tissue engineering application and the developed porous CS/OGP/PLA scaffold should be considered as alternative biomaterials for bone regeneration. Copyright © 2017 Elsevier B.V. All rights reserved.

  18. Biodegradable polycaprolactone-chitosan three-dimensional scaffolds fabricated by melt stretching and multilayer deposition for bone tissue engineering: assessment of the physical properties and cellular response

    International Nuclear Information System (INIS)

    Thuaksuban, Nuttawut; Nuntanaranont, Thongchai; Suttapreyasri, Srisurang; Pattanachot, Wachirapan; Cheung, Lim Kwong

    2011-01-01

    Fabrication of polycaprolactone (PCL)-chitosan (CS) three-dimensional (3D) scaffolds using the novel technique of melt stretching and multilayer deposition was introduced. In brief, firstly, the PCL-CS monofilaments containing 0% (pure PCL), 10%, 20% and 30% CS by weight were fabricated by melting and stretching processes. Secondly, the desired multilayer (3D) scaffolds were fabricated by arranging and depositing the filaments. Physical properties of the filaments and the scaffolds were evaluated. MC3T3-E1 cell lines were seeded on the scaffolds to assess their proliferation. A typical micro-groove pattern was found on the surfaces of pure PCL filaments due to stretching. The filaments of PCL-30%CS had the highest tendency of fracture during stretching and could not be used to form the scaffold. Increasing CS proportions tended to reduce the micro-groove pattern, surface roughness, tensile strength and elasticity of the filaments, whilst compressive strength of the PCL-CS scaffolds was not affected. The average pore size and porosity of the scaffolds were 536.90 ± 17.91 μm and 45.99 ± 2.8% respectively. Over 60 days, degradation of the scaffolds gradually increased (p > 0.05). The more CS containing scaffolds were found to increase in water uptake, but decrease in degradation rate. During the culture period, the growth of the cells in PCL-CS groups was significantly higher than in the pure PCL group (p < 0.05). On culture-day 21, the growth in the PCL-20%CS group was significantly higher than the other groups (p < 0.05). In conclusion, the PCL-20%CS scaffolds obtained the optimum results in terms of physical properties and cellular response.

  19. Biodegradable polycaprolactone-chitosan three-dimensional scaffolds fabricated by melt stretching and multilayer deposition for bone tissue engineering: assessment of the physical properties and cellular response

    Energy Technology Data Exchange (ETDEWEB)

    Thuaksuban, Nuttawut; Nuntanaranont, Thongchai; Suttapreyasri, Srisurang [Department of Oral and Maxillofacial Surgery, Faculty of Dentistry, Prince of Songkla University, Kanjanavanij Road, Hatyai, Songkhla, 90112 (Thailand); Pattanachot, Wachirapan [Polymer Science Program, Faculty of Science, Prince of Songkla University, Kanjanavanij Road, Hatyai, Songkhla, 90112 (Thailand); Cheung, Lim Kwong, E-mail: nuttawut.t@psu.ac.t [Discipline of Oral and Maxillofacial Surgery, Faculty of Dentistry, the University of Hong Kong, Hong Kong (China)

    2011-02-15

    Fabrication of polycaprolactone (PCL)-chitosan (CS) three-dimensional (3D) scaffolds using the novel technique of melt stretching and multilayer deposition was introduced. In brief, firstly, the PCL-CS monofilaments containing 0% (pure PCL), 10%, 20% and 30% CS by weight were fabricated by melting and stretching processes. Secondly, the desired multilayer (3D) scaffolds were fabricated by arranging and depositing the filaments. Physical properties of the filaments and the scaffolds were evaluated. MC3T3-E1 cell lines were seeded on the scaffolds to assess their proliferation. A typical micro-groove pattern was found on the surfaces of pure PCL filaments due to stretching. The filaments of PCL-30%CS had the highest tendency of fracture during stretching and could not be used to form the scaffold. Increasing CS proportions tended to reduce the micro-groove pattern, surface roughness, tensile strength and elasticity of the filaments, whilst compressive strength of the PCL-CS scaffolds was not affected. The average pore size and porosity of the scaffolds were 536.90 {+-} 17.91 {mu}m and 45.99 {+-} 2.8% respectively. Over 60 days, degradation of the scaffolds gradually increased (p > 0.05). The more CS containing scaffolds were found to increase in water uptake, but decrease in degradation rate. During the culture period, the growth of the cells in PCL-CS groups was significantly higher than in the pure PCL group (p < 0.05). On culture-day 21, the growth in the PCL-20%CS group was significantly higher than the other groups (p < 0.05). In conclusion, the PCL-20%CS scaffolds obtained the optimum results in terms of physical properties and cellular response.

  20. The osteoinductive effect of chitosan-collagen composites around pure titanium implant surfaces in rats.

    Science.gov (United States)

    Kung, S; Devlin, H; Fu, E; Ho, K-Y; Liang, S-Y; Hsieh, Y-D

    2011-02-01

    The enhancing effects of chitosan on activation of platelets and differentiation of osteoprogenitor cells have been demonstrated in vitro. The purpose of this study was to evaluate the in vivo osteoinductive effect of chitosan-collagen composites around pure titanium implant surfaces. Chitosan-collagen composites containing chitosan of different molecular weights (450 and 750 kDa) were wrapped onto titanium implants and embedded into the subcutaneous area on the back of 15 Sprague-Dawley rats. The control consisted of implants wrapped with plain collagen type I membranes. Implants and surrounding tissues were retrieved 6 wks after surgery and identified by Alizarin red and Alcian blue whole mount staining. The newly formed structures in the test groups were further analyzed by Toluidine blue and Masson-Goldner trichrome staining, and immunohistochemical staining with osteopontin and alkaline phosphotase. The bone formation parameters of the new bone in the two test groups were measured and compared. New bone formed ectopically in both chitosan-collagen groups, whereas no bone induction occurred in the negative control group. These newly formed bone-like structures were further confirmed by immunohistochemical staining. Comparison of bone parameters of the newly induced bone revealed no statistically significant differences between the 450 and 750 kDa chitosan-collagen groups. Our results demonstrated that chitosan-collagen composites might induce in vivo new bone formation around pure titanium implant surfaces. Different molecular weights of chitosan did not show significantly different effects on the osteoinductive potential of the test materials. © 2010 John Wiley & Sons A/S.

  1. Direct electrochemistry and electrocatalysis of hemoglobin protein entrapped in graphene and chitosan composite film.

    Science.gov (United States)

    Xu, Huifeng; Dai, Hong; Chen, Guonan

    2010-04-15

    A novel, biocompatible sensing strategy based on graphene and chitosan composite film for immobilizing the hemoglobin protein was firstly adopted. The direct electron transfer and bioelectrocatalytic activity of hemoglobin after incorporation into the composite film were investigated. A pair of reversible redox waves of hemoglobin was appeared, and hemoglobin could exhibit its bioelectrocatalytic activity toward H(2)O(2) in a long term. Such results indicated that graphene and chitosan composite could be a friendly biocompatible interface for immobilizing biomolecules and keeping their native structure. Furthermore, the appearance of graphene in the composite film could facilitate the electron transfer between matrix and the electroactive center of hemoglobin. Hence, this graphene and chitosan based protocol would be a promising platform for protein immobilization and biosensor preparation. (c) 2010 Elsevier B.V. All rights reserved.

  2. Tailoring Functional Chitosan-based Composites for Food Applications.

    Science.gov (United States)

    Nunes, Cláudia; Coimbra, Manuel A; Ferreira, Paula

    2018-03-08

    Chitosan-based functional materials are emerging for food applications. The covalent bonding of molecular entities demonstrates to enhance resistance to the typical acidity of food assigning mechanical and moisture/gas barrier properties. Moreover, the grafting to chitosan of some functional molecules, like phenolic compounds or essential oils, gives antioxidant, antimicrobial, among others properties to chitosan. The addition of nanofillers to chitosan and other biopolymers improves the already mentioned required properties for food applications and can attribute electrical conductivity and magnetic properties for active and intelligent packaging. Electrical conductivity is a required property for the processing of food at low temperature using electric fields or for sensors application. © 2018 The Chemical Society of Japan & Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim.

  3. Preparation, physicochemical properties and biocompatibility of PBLG/PLGA/bioglass composite scaffolds

    Energy Technology Data Exchange (ETDEWEB)

    Cui, Ning [State Key Laboratory for Mechanical Behavior of Materials, Xi' an Jiaotong University, Xi' an 710049 (China); Qian, Junmin, E-mail: jmqian@mail.xjtu.edu.cn [State Key Laboratory for Mechanical Behavior of Materials, Xi' an Jiaotong University, Xi' an 710049 (China); Wang, Jinlei [State Key Laboratory for Mechanical Behavior of Materials, Xi' an Jiaotong University, Xi' an 710049 (China); Ji, Chuanlei [The Orthopaedic Department, XiJing Hospital Affiliated to the Fourth Military Medical University, Xi' an 710032 (China); Xu, Weijun; Wang, Hongjie [State Key Laboratory for Mechanical Behavior of Materials, Xi' an Jiaotong University, Xi' an 710049 (China)

    2017-02-01

    In this study, novel poly(γ-benzyl L-glutamate)/poly(lactic-co-glycolic acid)/bioglass (PBLG/PLGA/BG) composite scaffolds with different weight ratios were fabricated using a negative NaCl-templating method. The morphology, compression modulus and degradation kinetics of the scaffolds were characterized. The results showed that the PBLG/PLGA/BG composite scaffolds with a weight ratio of 5:5:1, namely PBLG5PLGA5BG composite scaffolds, displayed a pore size range of 50–500 μm, high compressive modulus (566.6 ± 8.8 kPa), suitable glass transition temperature (46.8 ± 0.2 °C) and low degradation rate (> 8 weeks). The in vitro biocompatibility of the scaffolds was evaluated with MC3T3-E1 cells by live-dead staining, MTT and ALP activity assays. The obtained results indicated that the PBLG5PLGA5BG composite scaffolds were more conducive to the adhesion, proliferation and osteoblastic differentiation of MC3T3-E1 cells than PBLG and PBLG/PLGA composite scaffolds. The in vivo biocompatibility of the scaffolds was evaluated in both SD rat subcutaneous model and rabbit tibia defect model. The results of H&E, Masson's trichrome and CD34 staining assays demonstrated that the PBLG5PLGA5BG composite scaffolds allowed the ingrowth of tissue and microvessels more effectively than PBLG/PLGA composite scaffolds. The results of digital radiography confirmed that the PBLG5PLGA5BG composite scaffolds significantly improved in vivo osteogenesis. Collectively, the PBLG5PLGA5BG composite scaffolds could be a promising candidate for tissue engineering applications. - Highlights: • Foamy PBLG/PLGA/bioglass composite scaffolds were fabricated by negative templating. • PBLG/PLGA/bioglass composite scaffolds displayed tunable physicochemical properties. • PBLG/PLGA/bioglass composite scaffolds had good biocompatibility in vitro and in vivo. • PBLG/PLGA/bioglass composite scaffolds could promote the healing of bone defects.

  4. Structure-property-processing correlations in freeze-cast composite scaffolds.

    Science.gov (United States)

    Hunger, Philipp M; Donius, Amalie E; Wegst, Ulrike G K

    2013-05-01

    Surprisingly few reports have been published, to date, on the structure-property-processing correlations observed in freeze-cast materials directionally solidified from polymer solutions, or ceramic or metal slurries. The studies that exist focus on properties of sintered ceramics, that is materials whose structure was altered by further processing. In this contribution, we report first results on correlations observed in alumina-chitosan-gelatin composites, which were chosen as a model system to test and compare the effect of particle size and processing parameters on their mechanical properties at a specific composition. Our study reveals that highly porous (>90%) hybrid materials can be manufactured by freeze casting, through the self-assembly of a polymer and a ceramic phase that occurs during directional solidification, without the need of additional processing steps such as sintering or infiltration. It further illustrates that the properties of freeze-cast hybrid materials can independently be tailored at two levels of their structural hierarchy, allowing for the simultaneous optimization of both mechanical and structural requirements. An increase in freezing rate resulted in decreases in lamellar spacing, cell wall thickness, pore aspect ratio and cross-sectional area, as well as increases in both Young's modulus and compressive yield strength. The mechanical properties of the composite scaffolds increased with an increasing particle size. The results show that both structure and mechanical properties of the freeze-cast composites can be custom-designed and that they are thus ideally suited for a large variety of applications that require high porosity at low or medium load-bearing capacity. Copyright © 2013 Acta Materialia Inc. Published by Elsevier Ltd. All rights reserved.

  5. Laminated electrospun nHA/PHB-composite scaffolds mimicking bone extracellular matrix for bone tissue engineering

    Energy Technology Data Exchange (ETDEWEB)

    Chen, Zhuoyue [Lab of Tissue Engineering, Faculty of Life Science, Northwest University, 229 TaiBai North Road, Xi' an, Shaanxi Province 710069 (China); Provincial Key Laboratory of Biotechnology of Shaanxi, Northwest University, 229 TaiBai North Road, Xi' an, Shaanxi Province 710069 (China); Song, Yue [Lab of Tissue Engineering, Faculty of Life Science, Northwest University, 229 TaiBai North Road, Xi' an, Shaanxi Province 710069 (China); Zhang, Jing [Lab of Tissue Engineering, Faculty of Life Science, Northwest University, 229 TaiBai North Road, Xi' an, Shaanxi Province 710069 (China); Provincial Key Laboratory of Biotechnology of Shaanxi, Northwest University, 229 TaiBai North Road, Xi' an, Shaanxi Province 710069 (China); Key Laboratory of Resource Biology and Modern Biotechnology in Western China, Ministry of Education, Northwest University, 229 TaiBai North Road, Xi' an, Shaanxi Province, 710069 (China); Liu, Wei [Lab of Tissue Engineering, Faculty of Life Science, Northwest University, 229 TaiBai North Road, Xi' an, Shaanxi Province 710069 (China); Cui, Jihong, E-mail: cjh@nwu.edu.cn [Lab of Tissue Engineering, Faculty of Life Science, Northwest University, 229 TaiBai North Road, Xi' an, Shaanxi Province 710069 (China); Provincial Key Laboratory of Biotechnology of Shaanxi, Northwest University, 229 TaiBai North Road, Xi' an, Shaanxi Province 710069 (China); Key Laboratory of Resource Biology and Modern Biotechnology in Western China, Ministry of Education, Northwest University, 229 TaiBai North Road, Xi' an, Shaanxi Province, 710069 (China); and others

    2017-03-01

    Electrospinning is an effective means to generate nano- to micro-scale polymer fibers resembling native extracellular matrix for tissue engineering. However, a major problem of electrospun materials is that limited pore size and porosity may prevent adequate cellular infiltration and tissue ingrowth. In this study, we first prepared thin layers of hydroxyapatite nanoparticle (nHA)/poly-hydroxybutyrate (PHB) via electrospinning. We then laminated the nHA/PHB thin layers to obtain a scaffold for cell seeding and bone tissue engineering. The results demonstrated that the laminated scaffold possessed optimized cell-loading capacity. Bone marrow mesenchymal stem cells (MSCs) exhibited better adherence, proliferation and osteogenic phenotypes on nHA/PHB scaffolds than on PHB scaffolds. Thereafter, we seeded MSCs onto nHA/PHB scaffolds to fabricate bone grafts. Histological observation showed osteoid tissue formation throughout the scaffold, with most of the scaffold absorbed in the specimens 2 months after implantation, and blood vessels ingrowth into the graft could be observed in the graft. We concluded that electrospun and laminated nanoscaled biocomposite scaffolds hold great therapeutic potential for bone regeneration. - Highlights: • We laminated the nHA/PHB layers to obtain a scaffold for bone tissue engineering. • The laminated scaffold performed optimized cell-loading capacity. • MSCs exhibited osteogenic phenotypes on the laminated scaffold. • Osteoid tissue formed throughout the laminated scaffold after 2 months in vivo. The laminated bio-composite scaffolds can be applied to bone regeneration.

  6. Laminated electrospun nHA/PHB-composite scaffolds mimicking bone extracellular matrix for bone tissue engineering

    International Nuclear Information System (INIS)

    Chen, Zhuoyue; Song, Yue; Zhang, Jing; Liu, Wei; Cui, Jihong

    2017-01-01

    Electrospinning is an effective means to generate nano- to micro-scale polymer fibers resembling native extracellular matrix for tissue engineering. However, a major problem of electrospun materials is that limited pore size and porosity may prevent adequate cellular infiltration and tissue ingrowth. In this study, we first prepared thin layers of hydroxyapatite nanoparticle (nHA)/poly-hydroxybutyrate (PHB) via electrospinning. We then laminated the nHA/PHB thin layers to obtain a scaffold for cell seeding and bone tissue engineering. The results demonstrated that the laminated scaffold possessed optimized cell-loading capacity. Bone marrow mesenchymal stem cells (MSCs) exhibited better adherence, proliferation and osteogenic phenotypes on nHA/PHB scaffolds than on PHB scaffolds. Thereafter, we seeded MSCs onto nHA/PHB scaffolds to fabricate bone grafts. Histological observation showed osteoid tissue formation throughout the scaffold, with most of the scaffold absorbed in the specimens 2 months after implantation, and blood vessels ingrowth into the graft could be observed in the graft. We concluded that electrospun and laminated nanoscaled biocomposite scaffolds hold great therapeutic potential for bone regeneration. - Highlights: • We laminated the nHA/PHB layers to obtain a scaffold for bone tissue engineering. • The laminated scaffold performed optimized cell-loading capacity. • MSCs exhibited osteogenic phenotypes on the laminated scaffold. • Osteoid tissue formed throughout the laminated scaffold after 2 months in vivo. The laminated bio-composite scaffolds can be applied to bone regeneration.

  7. Polymer-Ceramic Composite Scaffolds: The Effect of Hydroxyapatite and β-tri-Calcium Phosphate

    Directory of Open Access Journals (Sweden)

    Boyang Huang

    2018-01-01

    Full Text Available The design of bioactive scaffolds with improved mechanical and biological properties is an important topic of research. This paper investigates the use of polymer-ceramic composite scaffolds for bone tissue engineering. Different ceramic materials (hydroxyapatite (HA and β-tri-calcium phosphate (TCP were mixed with poly-ε-caprolactone (PCL. Scaffolds with different material compositions were produced using an extrusion-based additive manufacturing system. The produced scaffolds were physically and chemically assessed, considering mechanical, wettability, scanning electron microscopy and thermal gravimetric tests. Cell viability, attachment and proliferation tests were performed using human adipose derived stem cells (hADSCs. Results show that scaffolds containing HA present better biological properties and TCP scaffolds present improved mechanical properties. It was also possible to observe that the addition of ceramic particles had no effect on the wettability of the scaffolds.

  8. Polymer-Ceramic Composite Scaffolds: The Effect of Hydroxyapatite and β-tri-Calcium Phosphate.

    Science.gov (United States)

    Huang, Boyang; Caetano, Guilherme; Vyas, Cian; Blaker, Jonny James; Diver, Carl; Bártolo, Paulo

    2018-01-14

    The design of bioactive scaffolds with improved mechanical and biological properties is an important topic of research. This paper investigates the use of polymer-ceramic composite scaffolds for bone tissue engineering. Different ceramic materials (hydroxyapatite (HA) and β-tri-calcium phosphate (TCP)) were mixed with poly-ε-caprolactone (PCL). Scaffolds with different material compositions were produced using an extrusion-based additive manufacturing system. The produced scaffolds were physically and chemically assessed, considering mechanical, wettability, scanning electron microscopy and thermal gravimetric tests. Cell viability, attachment and proliferation tests were performed using human adipose derived stem cells (hADSCs). Results show that scaffolds containing HA present better biological properties and TCP scaffolds present improved mechanical properties. It was also possible to observe that the addition of ceramic particles had no effect on the wettability of the scaffolds.

  9. Collagen/silk fibroin composite scaffold incorporated with PLGA microsphere for cartilage repair

    Energy Technology Data Exchange (ETDEWEB)

    Wang, Jianhua; Yang, Qiu; Cheng, Niangmei [Institute of Biomedical and Pharmaceutical Technology, Fuzhou University, Fuzhou 350002 (China); Tao, Xiaojun [Department of Pharmacy, School of Medicine, Hunan Normal University, Changsha, 410013, Hunan (China); Zhang, Zhihua; Sun, Xiaomin [Institute of Biomedical and Pharmaceutical Technology, Fuzhou University, Fuzhou 350002 (China); Zhang, Qiqing, E-mail: zhangqiq@126.com [Institute of Biomedical and Pharmaceutical Technology, Fuzhou University, Fuzhou 350002 (China); Key Laboratory of Biomedical Materials of Tianjin, Institute of Biomedical Engineering, Chinese Academy of Medical Science & Peking Union Medical College, Tianjin 300192 (China)

    2016-04-01

    For cartilage repair, ideal scaffolds should mimic natural extracellular matrix (ECM) exhibiting excellent characteristics, such as biocompatibility, suitable porosity, and good cell affinity. This study aimed to prepare a collagen/silk fibroin composite scaffold incorporated with poly-lactic-co-glycolic acid (PLGA) microsphere that can be applied in repairing cartilage. To obtain optimum conditions for manufacturing a composite scaffold, a scaffold composed of different collagen-to-silk fibroin ratios was evaluated by determining porosity, water absorption, loss rate in hot water, and cell proliferation. Results suggested that the optimal ratio of collagen and silk fibroin composite scaffold was 7:3. The microstructure and morphological characteristics of the obtained scaffold were also examined through scanning electron microscopy and Fourier transform infrared spectroscopy. The results of in vitro fluorescence staining of bone marrow stromal cells revealed that collagen/silk fibroin composite scaffold enhanced cell proliferation without eliciting side effects. The prepared composite scaffold incorporated with PLGA microsphere was implanted in fully thick articular cartilage defects in rabbits. Collagen/silk fibroin composite scaffold with PLGA microspheres could enhance articular cartilage regeneration and integration between the repaired cartilage and the surrounding cartilage. Therefore, this composite will be a promising material for cartilage repair and regeneration. - Highlights: • Collagen/silk fibroin composite scaffold incorporated with PLGA microsphere proposed for cartilage repair was created. • In vivo, scaffold could enhance cartilage regeneration and integration between the repaired and surrounding cartilage. • In vitro, scaffold exhibits excellent characteristics, such as, improved porosity water absorption and good cell affinity.

  10. Preparation of chitosan-collagen-alginate composite dressing and its promoting effects on wound healing.

    Science.gov (United States)

    Xie, Haixia; Chen, Xiuli; Shen, Xianrong; He, Ying; Chen, Wei; Luo, Qun; Ge, Weihong; Yuan, Weihong; Tang, Xue; Hou, Dengyong; Jiang, Dingwen; Wang, Qingrong; Liu, Yuming; Liu, Qiong; Li, Kexian

    2018-02-01

    The present study aimed to prepare a composite dressing composed of collagen, chitosan, and alginate, which may promote wound healing and prevent from seawater immersion. Chitosan-collagen-alginate (CCA) cushion was prepared by paintcoat and freeze-drying, and it was attached to a polyurethane to compose CCA composite dressing. The swelling, porosity, degradation, and mechanical properties of CCA cushion were evaluated. The effects on wound healing and seawater prevention of CCA composite dressing were tested by rat wound model. Preliminary biosecurity was tested by cytotoxicity and hemocompatibility. The results revealed that CCA cushion had good water absorption and mechanical properties. A higher wound healing ratio was observed in CCA composite dressing treated rats than in gauze or chitosan treated ones. On the fifth day, the healing rates of CCA composite dressing, gauze, and chitosan were 48.49%±1.07%, 28.02%±6.4%, and 38.97%±8.53%, respectively. More fibroblast and intact re-epithelialization were observed in histological images of CCA composite dressing treated rats, and the expressions of EGF, bFGF, TGF-β, and CD31 increased significantly. CCA composite dressing showed no significant cytotoxicity, and favorable hemocompatibility. These results suggested that CCA composite dressing could prevent against seawater immersion and promote wound healing while having a good biosecurity. Copyright © 2017. Published by Elsevier B.V.

  11. Diatomite as a novel composite ingredient for chitosan film with enhanced physicochemical properties.

    Science.gov (United States)

    Akyuz, Lalehan; Kaya, Murat; Koc, Behlul; Mujtaba, Muhammad; Ilk, Sedef; Labidi, Jalel; Salaberria, Asier M; Cakmak, Yavuz Selim; Yildiz, Aysegul

    2017-12-01

    Practical applications of biopolymers in different industries are gaining considerable increase day by day. But still, these biopolymers lack important properties in order to meet the industrial demands. In the same regard, in the current study, chitosan composite films are produced by incorporating diatomite soil at two different concentrations. In order to obtain a homogeneous film, glutaraldehyde was supplemented to chitosan solution as a cross-linker. Compositing diatomaceous earth to chitosan film resulted in improvement of various important physicochemical properties compared to control such as; enhanced film wettability, increase elongation at break and improved thermal stability (264-277°C). The microstructure of the film was observed to haveconsisted of homogeneously distributed blister-shaped structures arised due to the incorporation of diatomite. The incorporation of diatomite did not influence the overall antioxidant activity of the composite films, which can be ascribe to the difficulty radicals formation. Chitosan film incorporated with increasing fraction of diatomite revealed a notable enhancement in the antimicrobial activity. Additionally with the present study, for the first time possible interactions between chitosan/diatomite were determined via quantum chemical calculations. Current study will be helpful in giving a new biotechnological perspective to diatom in terms of its successful application in hydrophobic composite film production. Copyright © 2017 Elsevier B.V. All rights reserved.

  12. Engineered 3D-scaffolds of photocrosslinked chitosan-gelatin hydrogel hybrids for chronic wound dressings and regeneration.

    Science.gov (United States)

    Carvalho, Isadora C; Mansur, Herman S

    2017-09-01

    Wound repair is one of the most complex biological processes in human life. To date, no ideal biomaterial solution has been identified, which that encompasses all functions and properties of real skin tissue. Thus, this study focused on the synthesis of new biocompatible hybrid hydrogel scaffolds based on methacrylate-functionalized high molecular mass chitosan with gelatin-A photocrosslinked with UV radiation to tailor matrix network properties. These hybrid hydrogels were produced via freeze-drying and were extensively characterized by swelling and degradation measurements, Fourier transform infrared spectroscopy (FTIR), UV-visible spectroscopy (UV-Vis), scanning electron microscopy (SEM-EDS), and micro-computed tomography (micro-CT). The results demonstrated that hydrogels were produced with broadly designed swelling degrees typically ranging from 500% to 2000%, which were significantly dependent on the relative concentration of polymers and irradiation time for crosslinking. Analogously, degradation was reduced with increased photocrosslinking of the network. Moreover, insights into the mechanism of photochemical crosslinking were suggested based on FTIR and UV-Vis analyses of the characteristic functional groups involved in the reactions. SEM analysis associated with micro-CT imaging of the hybrid scaffolds showed uniformly interconnected 3D porous structures, with architectural features affected by the crosslinking of the network. These hydrogels were biocompatible, with live cell viability responses of human embryonic kidney (HEK293T) cells being above 95%. Hence, novel hybrid hydrogels were designed and produced with tunable properties through photocrosslinking and with a biocompatible response suitable for use in wound dressing and skin tissue repair applications. Copyright © 2017 Elsevier B.V. All rights reserved.

  13. Morphological Effects of HA on the Cell Compatibility of Electrospun HA/PLGA Composite Nanofiber Scaffolds

    Directory of Open Access Journals (Sweden)

    Adnan Haider

    2014-01-01

    Full Text Available Tissue engineering is faced with an uphill challenge to design a platform with appropriate topography and suitable surface chemistry, which could encourage desired cellular activities and guide bone tissue regeneration. To develop such scaffolds, composite nanofiber scaffolds of nHA and sHA with PLGA were fabricated using electrospinning technique. nHA was synthesized using precipitation method, whereas sHA was purchased. The nHA and sHA were suspended in PLGA solution separately and electrospun at optimized electrospinning parameters. The composite nanofiber scaffolds were characterized by FE-SEM, EDX analysis, TEM, XRD analysis, FTIR, and X-ray photoelectron. The potential of the HA/PLGA composite nanofiber as bone scaffolds in terms of their bioactivity and biocompatibility was assessed by culturing the osteoblastic cells onto the composite nanofiber scaffolds. The results from in vitro studies revealed that the nHA/PLGA composite nanofiber scaffolds showed higher cellular adhesion, proliferation, and enhanced osteogenesis performance, along with increased Ca+2 ions release compared to the sHA/PLGA composite nanofiber scaffolds and pristine PLGA nanofiber scaffold. The results show that the structural dependent property of HA might affect its potential as bone scaffold and implantable materials in regenerative medicine and clinical tissue engineering.

  14. Nano-hydroxyapatite/poly ε-caprolactone composite 3D scaffolds for mastoid obliteration

    International Nuclear Information System (INIS)

    Kim, S E; Yun, H S; Hyun, Y T; Shin, J W; Song, J J

    2009-01-01

    The aim of this study is to evaluate the use of our nano-HA/PCL composite 3D scaffolds as graft materials for mastoid cavity obliteration in an animal model. Nano-HA particles were synthesized by chemical precipitation technique and mixed them with PCL solution to make composite paste. 3D scaffolds were fabricated by a paste extruding deposition process. The nano-HA/PCL 3D scaffolds showed good in vivo bone regeneration behaviour in a rabbit model after 4 and 8 week implantation. To characterize the 3D scaffolds as a grafting material for mastoid obliteration, mastoid cavities were introduced in rats and implanted the scaffolds. After two week implantation, histological examination showed good tissue ingrowth and new bone formation behaviour. It can be argued that our nano-HA/PCL composite 3D scaffold is a promising alternative material for mastoid obliteration.

  15. In-Vitro Enzymatic Degradation of γ-irradiated Porous Chitosan Scaffold: Crystallinity and degree of deacetylation

    International Nuclear Information System (INIS)

    Ismail Zainol; Azreena Mastor; Suhaida Md Ghani; Ahmad Fuad Yahya; Hazizan Md Akil

    2009-01-01

    Full text: Enzymatic degradation behavior of porous chitosan membrane was carried out in vitro by using enzymatic hydrolysis of chitosan in lysozyme solution. Chitosan was first modified by reducing its molecular weight by gamma (γ) radiation in the solid state. The chitosan powder was irradiated with gamma Co 60 source with various doses of 10, 25, 50 and 100 kGy. The molecular weight of irradiated chitosan was measured using visco metric method. The modified chitosan was transform into a porous membrane by lyophilization method. Degree of deacetylation (DD) and crystallinity of samples were measured using FTIR and XRD respectively on both gamma irradiated and enzymatic degradation samples. The results suggested that the irradiated chitosan become less crystalline without changes in DD. The enzymatic degradation of chitosan however shows an increment in DD and crystallinity. (author)

  16. Injectable alginate-O-carboxymethyl chitosan/nano fibrin composite hydrogels for adipose tissue engineering.

    Science.gov (United States)

    Jaikumar, Dhanya; Sajesh, K M; Soumya, S; Nimal, T R; Chennazhi, K P; Nair, Shantikumar V; Jayakumar, R

    2015-03-01

    Injectable, biodegradable scaffolds are required for soft tissue reconstruction owing to its minimally invasive approach. Such a scaffold can mimic the native extracellular matrix (ECM), provide uniform distribution of cells and overcome limitations like donor site morbidity, volume loss, etc. So, here we report two classes of biocompatible and biodegradable hydrogel blend systems namely, Alginate/O-carboxymethyl chitosan (O-CMC) and Alginate/poly (vinyl alcohol) (PVA) with the inclusion of fibrin nanoparticles in each. The hydrogels were prepared by ionic cross-linking method. The developed hydrogels were compared in terms of its swelling ratio, degradation profile, compressive strength and elastic moduli. From these preliminary findings, it was concluded that Alginate/O-CMC formed a better blend for tissue engineering applications. The potential of the formed hydrogel as an injectable scaffold was revealed by the survival of adipose derived stem cells (ADSCs) on the scaffold by its adhesion, proliferation and differentiation into adipocytes. Cell differentiation studies of fibrin incorporated hydrogel scaffolds showed better differentiation was confirmed by Oil Red O staining technique. These injectable gels have potential in soft tissue regeneration. Copyright © 2014 Elsevier B.V. All rights reserved.

  17. Development of a PVAl/chitosan composite membrane compatible with the dermo-epidermic system

    International Nuclear Information System (INIS)

    Almeida, Tiago Luiz de

    2009-03-01

    Due to the frequent incidence of people with skin lesions such as burns and ulcers and the lack of available donors, biomaterials with the capacity to mimic skin must be developed. In order to develop these biomaterials, polymers are used in the attempt to achieve characteristics which are closer to the target organ. In this direction, for several years our group has been developing dermo-epidermic substitutes, specifically biodegradable and biocompatible membranes made up of PVAl and chitosan. PVAl, a synthetic polymer, was used to imitate part of the human dermis and chitosan, a polymer of organic origin, was used in this study to stimulate growth and maintenance of the epidermis. Due to the variations of these commercially obtained polymers, the objective of this study was to characterize their physical and chemical properties, comparing them with the membrane previously obtained by our group with the intention of confirming the hypotheses of interferences put forward in this study. The PVAl membranes in the study (PVAl MP) that obtained characteristics most similar to the standard were those irradiated with 13 and 15 kGy; this last was chosen because it was the minimum dose necessary to achieve sterility. These membranes were also those which had the largest percentage of pores between 70 and 100 μm. For chitosan, the principal characteristics studied were the degree of acetylation (DA) and average molecular weight, both results demonstrated different characteristics than commercially indicated. Various membrane preparation protocols were carried out from the chitosan solution (2%). The membrane composed of the solution of chitosan homogenized with glycerol (20%) and dried at room temperature had the best interaction with keratinocytes. To finalize the study, this chitosan solution was poured over a PVAl membrane, lyophilized and impregnated with chitosan (2%) solution and the compound was kept at room temperature until a chitosan film formed on the upper

  18. In-situ birth of MSCs multicellular spheroids in poly(L-glutamic acid)/chitosan scaffold for hyaline-like cartilage regeneration.

    Science.gov (United States)

    Zhang, Kunxi; Yan, Shifeng; Li, Guifei; Cui, Lei; Yin, Jingbo

    2015-12-01

    The success of mesenchymal stem cells (MSCs) based articular cartilage tissue engineering is limited by the presence of fibrous tissue in generated cartilage, which is associated with the current scaffold strategy that promotes cellular adhesion and spreading. Here we design a non-fouling scaffold based on amide bonded poly(l-glutamic acid) (PLGA) and chitosan (CS) to drive adipose stem cells (ASCs) to aggregate to form multicellular spheroids with diameter of 80-110 μm in-situ. To illustrate the advantage of the present scaffolds, a cellular adhesive scaffold based on the same amide bonded PLGA and CS was created through a combination of air-drying and freeze-drying to limit the hydration effect while also achieving porous structure. Compared to ASCs spreading along the surface of pores within scaffold, the dense mass of aggregated ASCs in PLGA/CS scaffold exhibited enhanced chondrogenic differentiation capacity, as determined by up-regulated GAGs and COL II expression, and greatly decreased COL I deposition during in vitro chondrogenesis. Furthermore, after 12 weeks of implantation, neo-cartilages generated by ASCs adhered on scaffold significantly presented fibrous matrix which was characterized by high levels of COL I deposition. However, neo-cartilage at 12 weeks post-implantation generated by PLGA/CS scaffold carrying ASC spheroids possessed similar high level of GAGs and COL II and low level of COL I as that in normal cartilage. The in vitro and in vivo results indicated the present strategy could not only promote chondrogenesis of ASCs, but also facilitate hyaline-like cartilage regeneration with reduced fibrous tissue formation which may attenuate cartilage degradation in future long-term follow-up. Copyright © 2015 Elsevier Ltd. All rights reserved.

  19. Characterization of Three-Dimensional Printed Composite Scaffolds Prepared with Different Fabrication Methods

    Directory of Open Access Journals (Sweden)

    Szlązak K.

    2016-06-01

    Full Text Available An optimal method for composites preparation as an input to rapid prototyping fabrication of scaffolds with potential application in osteochondral tissue engineering is still needed. Scaffolds in tissue engineering applications play a role of constructs providing appropriate mechanical support with defined porosity to assist regeneration of tissue. The aim of the presented study was to analyze the influence of composite fabrication methods on scaffolds mechanical properties. The evaluation was performed on polycaprolactone (PCL with 5 wt% beta-tricalcium phosphate (TCP scaffolds fabricated using fused deposition modeling (FDM. Three different methods of PCL-TCP composite preparation: solution casting, particles milling, extrusion and injection were used to provide material for scaffold fabrication. The obtained scaffolds were investigated by means of scanning electron microscope, x-ray micro computed tomography, thermal gravimetric analysis and static material testing machine. All of the scaffolds had the same geometry (cylinder, 4×6 mm and fiber orientation (0/60/120°. There were some differences in the TCP distribution and formation of the ceramic agglomerates in the scaffolds. They depended on fabrication method. The use of composites prepared by solution casting method resulted in scaffolds with the best combination of compressive strength (5.7±0.2 MPa and porosity (48.5±2.7 %, both within the range of trabecular bone.

  20. Chitosan/bioactive glass nanoparticle composite membranes for periodontal regeneration

    NARCIS (Netherlands)

    Mota, J.; Yu, N.; Caridade, S.G.; Luz, G.M.; Gomes, M.E.R.; Reis, R.L.; Jansen, J.A.; Walboomers, X.F.; Mano, J.F.

    2012-01-01

    Barrier membranes are used in periodontal applications with the aim of supporting periodontal regeneration by physically blocking migration of epithelial cells. The present work proposes a combination of chitosan (CHT) with bioactive glass nanoparticles (BG-NPs) in order to produce a novel guided

  1. Developing multi-cellular tumor spheroid model (MCTS) in the chitosan/collagen/alginate (CCA) fibrous scaffold for anticancer drug screening.

    Science.gov (United States)

    Wang, Jian-Zheng; Zhu, Yu-Xia; Ma, Hui-Chao; Chen, Si-Nan; Chao, Ji-Ye; Ruan, Wen-Ding; Wang, Duo; Du, Feng-guang; Meng, Yue-Zhong

    2016-05-01

    In this work, a 3D MCTS-CCA system was constructed by culturing multi-cellular tumor spheroid (MCTS) in the chitosan/collagen/alginate (CCA) fibrous scaffold for anticancer drug screening. The CCA scaffolds were fabricated by spray-spinning. The interactions between the components of the spray-spun fibers were evidenced by methods of Coomassie Blue stain, X-ray diffraction (XRD) and Fourier transform-infrared spectroscopy (FTIR). Co-culture indicated that MCF-7 cells showed a spatial growth pattern of multi-cellular tumor spheroid (MCTS) in the CCA fibrous scaffold with increased proliferation rate and drug-resistance to MMC, ADM and 5-Aza comparing with the 2D culture cells. Significant increases of total viable cells were found in 3D MCTS groups after drug administration by method of apoptotic analysis. Glucose-lactate analysis indicated that the metabolism of MCTS in CCA scaffold was closer to the tumor issue in vivo than the monolayer cells. In addition, MCTS showed the characteristic of epithelial mesenchymal transition (EMT) which is subverted by carcinoma cells to facilitate metastatic spread. These results demonstrated that MCTS in CCA scaffold possessed a more conservative phenotype of tumor than monolayer cells, and anticancer drug screening in 3D MCTS-CCA system might be superior to the 2D culture system. Copyright © 2016 Elsevier B.V. All rights reserved.

  2. Composite particles formed by complexation of poly(methacrylic acid) - stabilized magnetic fluid with chitosan: Magnetic material for bioapplications.

    Science.gov (United States)

    Safarik, Ivo; Stepanek, Miroslav; Uchman, Mariusz; Slouf, Miroslav; Baldikova, Eva; Nydlova, Leona; Pospiskova, Kristyna; Safarikova, Mirka

    2016-10-01

    A simple procedure for the synthesis of magnetic fluid (ferrofluid) stabilized by poly(methacrylic acid) has been developed. This ferrofluid was used to prepare a novel type of magnetically responsive chitosan-based composite material. Both ferrofluid and magnetic chitosan composite were characterized by a combination of microscopy (optical microscopy, TEM, SEM), scattering (static and dynamic light scattering, SANS) and spectroscopy (FTIR) techniques. Magnetic chitosan was found to be a perspective material for various bioapplications, especially as a magnetic carrier for immobilization of enzymes and cells. Lipase from Candida rugosa was covalently attached after cross-linking and activation of chitosan using glutaraldehyde. Baker's yeast cells (Saccharomyces cerevisiae) were incorporated into the chitosan composite during its preparation; both biocatalysts were active after reaction with appropriate substrates. Copyright © 2016 Elsevier B.V. All rights reserved.

  3. The roles of knitted mesh-reinforced collagen-chitosan hybrid scaffold in the one-step repair of full-thickness skin defects in rats.

    Science.gov (United States)

    Wang, Xingang; You, Chuangang; Hu, Xinlei; Zheng, Yurong; Li, Qiyin; Feng, Zhanzeng; Sun, Huafeng; Gao, Changyou; Han, Chunmao

    2013-08-01

    Full-thickness skin defects represent a significant and urgent clinical problem. Dermal substitutes serving as a regenerative template to induce dermal reconstruction provide a promising method to treat serious skin defects. Although collagen-chitosan dermal scaffolds display good biocompatibility and a suitable porous structure for angiogenesis and tissue regeneration, their poor mechanical properties compromise their application. To develop a well-supported dermal substitute, a poly(l-lactide-co-glycolide) (PLGA) knitted mesh was fabricated and integrated with collagen-chitosan scaffold (CCS) to obtain a PLGA knitted mesh-reinforced CCS (PLGAm/CCS). The morphology of this PLGAm/CCS was investigated in vitro. To characterize the tissue response, specifically angiogenesis and tissue regeneration, the PLGAm/CCS was transplanted in combination with thin split-thickness autografts to repair full-thickness skin wounds using a one-step surgical procedure in Sprague-Dawley rats. These results were then compared with CCSs. At weeks 2, 4 and 8 after the operation, the healing wounds were imaged to analyse wound changes, and tissue specimens were harvested for histology, immunohistochemistry, real-time quantitative polymerase chain reaction and Western blot analysis. The results demonstrated that collagen-chitosan sponge in the PLGAm/CCS remained porous, interconnected and occupied the openings of PLGA mesh, and the incorporation of the PLGA knitted mesh into CCS improved the mechanical strength with little influence on its mean pore size and porosity. Following transplantation, PLGAm/CCS inhibited wound contraction, and effectively promoted neotissue formation and blood vessel ingrowth. In conclusion, the mechanical strength of the scaffolds plays an important role in the process of tissue regeneration and vascularization. The ability of PLGAm/CCS to promote angiogenesis and induce in situ tissue regeneration demonstrates its potential in skin tissue engineering. Copyright

  4. Chitosan/chitin nanowhiskers composites: effect of plasticisers on the mechanical behaviour

    Czech Academy of Sciences Publication Activity Database

    Kelnar, Ivan; Kovářová, Jana; Tishchenko, Galina; Kaprálková, Ludmila; Pavlova, Ewa; Carezzi, F.; Morganti, P.

    2015-01-01

    Roč. 22, č. 2 (2015), 5_1-5_6 ISSN 1022-9760 R&D Projects: GA ČR(CZ) GA13-15255S EU Projects: European Commission(XE) 315233 - N-CHITOPACK Institutional support: RVO:61389013 Keywords : chitosan * chitin nanowhiskers * composite Subject RIV: CD - Macromolecular Chemistry Impact factor: 1.969, year: 2015

  5. Magnetic responsive hydroxyapatite composite scaffolds construction for bone defect reparation.

    Science.gov (United States)

    Zeng, Xiao Bo; Hu, Hao; Xie, Li Qin; Lan, Fang; Jiang, Wen; Wu, Yao; Gu, Zhong Wei

    2012-01-01

    In recent years, interest in magnetic biomimetic scaffolds for tissue engineering has increased considerably. A type of magnetic scaffold composed of magnetic nanoparticles (MNPs) and hydroxyapatite (HA) for bone repair has been developed by our research group. In this study, to investigate the influence of the MNP content (in the scaffolds) on the cell behaviors and the interactions between the magnetic scaffold and the exterior magnetic field, a series of MNP-HA magnetic scaffolds with different MNP contents (from 0.2% to 2%) were fabricated by immersing HA scaffold into MNP colloid. ROS 17/2.8 and MC3T3-E1 cells were cultured on the scaffolds in vitro, with and without an exterior magnetic field, respectively. The cell adhesion, proliferation and differentiation were evaluated via scanning electron microscopy; confocal laser scanning microscopy; and 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT), alkaline phosphatase, and bone gla protein activity tests. The results demonstrated the positive influence of the magnetic scaffolds on cell adhesion, proliferation, and differentiation. Further, a higher amount of MNPs on the magnetic scaffolds led to more significant stimulation. The magnetic scaffold can respond to the exterior magnetic field and engender some synergistic effect to intensify the stimulating effect of a magnetic field to the proliferation and differentiation of cells.

  6. Utilizing two-photon fluorescence and second harmonic generation microscopy to study human bone marrow mesenchymal stem cell morphogenesis in chitosan scaffold

    Science.gov (United States)

    Su, Ping-Jung; Huang, Chi-Hsiu; Huang, Yi-You; Lee, Hsuan-Sue; Dong, Chen-Yuan

    2008-02-01

    A major goal of tissue engineering is to cultivate the cartilage in vitro. One approach is to implant the human bone marrow mesenchymal stem cells into the three dimensional biocompatible and biodegradable material. Through the action of the chondrogenic factor TGF-β3, the stem cells can be induced to secrete collagen. In this study, mesenchymal stem cells are implanted on the chitosan scaffold and TGF-β3 was added to produce the cartilage tissue and TP autofluorescence and SHG microscopy was used to image the process of chondrogenesis. With additional development, multiphoton microscopy can be developed into an effective tool for evaluating the quality of tissue engineering products.

  7. Elaboration of chitosan/activated carbon composites for the removal of organic micropollutants from waters.

    Science.gov (United States)

    Venault, A; Vachoud, L; Pochat, C; Bouyer, D; Faur, C

    2008-12-01

    Composite hydrogels were prepared by a wet-casting process by blending a biopolymer, chitosan, with activated carbon (AC) for use in water treatment. Adsorption properties of the composite gels for an organic micro-pollutant (phenol) which may be encountered in wastewaters was studied with an experimental design approach as a function of: - the concentration of raw materials and thus the AC weight within the chitosan matrix. - the accessibility of AC in the polymeric matrix, which is assumed to be related to the coating and thus to the pH of the immersion bath. ESEM observations showed that at a higher pH of gelation (pH = 14), AC particles were entrapped at the surface of the polymer matrix because of a faster gelation kinetic than at a lower pH (13.3). Adsorption kinetic tests showed that phenol adsorption occurred according to two mechanisms. During the first step, phenol molecules were adsorbed by the AC particles located at the surface. The second step corresponded to a slow diffusion through chitosan chains leading to an adsorption by AC particles entrapped within the polymeric matrix coupled to an adsorption on to the chitosan. A mass transfer model was used to describe this two-step adsorption phenomenon. However, due to a heterogeneous coating of AC by chitosan, this phenomenon was not supported by experimental design results: the initial kinetic coefficients were associated with a high experimental error which didn't allow for an analysis of the influence of elaboration parameters on kinetic coefficients. Regardling equilibrium adsorption properties, it was shown that composite gels were good adsorbents for phenol with removal ranging from 94% to 98% corresponding to adsorption capacities from 30 to 41 mg g(-1). The pH of the immersion bath had no influence on equilibrium adsorption properties, contrary to the AC weight within the chitosan matrix which wasdemonstrated to influence significantly adsorption capacities. Because carbon particles may improve

  8. Thermogel-Coated Poly(ε-Caprolactone Composite Scaffold for Enhanced Cartilage Tissue Engineering

    Directory of Open Access Journals (Sweden)

    Shao-Jie Wang

    2016-05-01

    Full Text Available A three-dimensional (3D composite scaffold was prepared for enhanced cartilage tissue engineering, which was composed of a poly(ε-caprolactone (PCL backbone network and a poly(lactide-co-glycolide-block-poly(ethylene glycol-block-poly(lactide-co-glycolide (PLGA–PEG–PLGA thermogel surface. The composite scaffold not only possessed adequate mechanical strength similar to native osteochondral tissue as a benefit of the PCL backbone, but also maintained cell-friendly microenvironment of the hydrogel. The PCL network with homogeneously-controlled pore size and total pore interconnectivity was fabricated by fused deposition modeling (FDM, and was impregnated into the PLGA–PEG–PLGA solution at low temperature (e.g., 4 °C. The PCL/Gel composite scaffold was obtained after gelation induced by incubation at body temperature (i.e., 37 °C. The composite scaffold showed a greater number of cell retention and proliferation in comparison to the PCL platform. In addition, the composite scaffold promoted the encapsulated mesenchymal stromal cells (MSCs to differentiate chondrogenically with a greater amount of cartilage-specific matrix production compared to the PCL scaffold or thermogel. Therefore, the 3D PCL/Gel composite scaffold may exhibit great potential for in vivo cartilage regeneration.

  9. The effect of hydroxylation on CNT to form Chitosan-CNT composites: A DFT study

    International Nuclear Information System (INIS)

    Yu, Rui; Ran, Maofei; Wen, Jie; Sun, Wenjing; Chu, Wei; Jiang, Chengfa; He, Zhiwei

    2015-01-01

    Graphical abstract: - Highlights: • The effect of hydroxylation on CNT to form Chitosan-CNT composites was studied. • The adsorption of Chitosan on CNTs is very weak by electrostatic interactions. • Chitosan loads onto CNT-OH_n via hydrogen-bond interactions. • Chitosan transfers electron to CNT-OH_n and thus improves the reactivity of CNT. - Abstract: The effect of types of CNTs (pristine and hydroxylated) on the synthesis of Chitosan-CNT (CS-CNT) composites was investigated theoretically. The adsorption energy (E_a_d_s) of CS on the pristine CNT and hydroxylated CNTs (CNT-OH_n, n = 1–6) as well as the structural and electronic properties of said composites have been investigated. Results show that the adsorption of CS on CNT and CNT-OH_n is thermodynamically favored. The E_a_d_s of CS on CNTs was calculated to be −20.387 kcal/mol from electrostatic interactions. For CS adsorbed into CNT-OH_n, E_a_d_s ranges from −20.612 to −37.567 kcal/mol. Hydroxyl groups on CNT are the main adsorption sites for CS loading onto CNT-OH_n via hydrogen-bond interactions. The CS-CNT-OH_3 is the most sable composite among tested complexes. The energy gap (ΔE_g_a_p) of CS-CNT-OH_3 was calculated less than pristine CNT and CNT-OH_3, indicative of the composites being more reactive than that of pristine CNTs and CNT-OH_3. It was proved that CS can transfer electron to the hydroxylated CNTs, thus overcoming the drawbacks of CNTs being chemically inert.

  10. The effect of hydroxylation on CNT to form Chitosan-CNT composites: A DFT study

    Energy Technology Data Exchange (ETDEWEB)

    Yu, Rui [China-America Cancer Research Institute, Key Laboratory for Medical Molecular Diagnostics of Guangdong Province, Guangdong Medical University, Dongguan, Guangdong 523808 (China); Department of Chemical Engineering, Sichuan University, Chengdu 610065 (China); Ran, Maofei [College of Chemistry & Environment Protection Engineering, Southwest University for Nationalities, Chengdu 610041, Sichuan (China); Wen, Jie [College of Chemistry and Chemical Engineering, Southwest Petroleum University, Chengdu 610500, Sichuan (China); Sun, Wenjing, E-mail: swj_gdmc@163.com [China-America Cancer Research Institute, Key Laboratory for Medical Molecular Diagnostics of Guangdong Province, Guangdong Medical University, Dongguan, Guangdong 523808 (China); Chu, Wei; Jiang, Chengfa [Department of Chemical Engineering, Sichuan University, Chengdu 610065 (China); He, Zhiwei, E-mail: zhiweihe688@yahoo.com [China-America Cancer Research Institute, Key Laboratory for Medical Molecular Diagnostics of Guangdong Province, Guangdong Medical University, Dongguan, Guangdong 523808 (China)

    2015-12-30

    Graphical abstract: - Highlights: • The effect of hydroxylation on CNT to form Chitosan-CNT composites was studied. • The adsorption of Chitosan on CNTs is very weak by electrostatic interactions. • Chitosan loads onto CNT-OH{sub n} via hydrogen-bond interactions. • Chitosan transfers electron to CNT-OH{sub n} and thus improves the reactivity of CNT. - Abstract: The effect of types of CNTs (pristine and hydroxylated) on the synthesis of Chitosan-CNT (CS-CNT) composites was investigated theoretically. The adsorption energy (E{sub ads}) of CS on the pristine CNT and hydroxylated CNTs (CNT-OH{sub n}, n = 1–6) as well as the structural and electronic properties of said composites have been investigated. Results show that the adsorption of CS on CNT and CNT-OH{sub n} is thermodynamically favored. The E{sub ads} of CS on CNTs was calculated to be −20.387 kcal/mol from electrostatic interactions. For CS adsorbed into CNT-OH{sub n}, E{sub ads} ranges from −20.612 to −37.567 kcal/mol. Hydroxyl groups on CNT are the main adsorption sites for CS loading onto CNT-OH{sub n} via hydrogen-bond interactions. The CS-CNT-OH{sub 3} is the most sable composite among tested complexes. The energy gap (ΔE{sub gap}) of CS-CNT-OH{sub 3} was calculated less than pristine CNT and CNT-OH{sub 3}, indicative of the composites being more reactive than that of pristine CNTs and CNT-OH{sub 3}. It was proved that CS can transfer electron to the hydroxylated CNTs, thus overcoming the drawbacks of CNTs being chemically inert.

  11. Ceramic Identity Contributes to Mechanical Properties and Osteoblast Behavior on Macroporous Composite Scaffolds

    Directory of Open Access Journals (Sweden)

    J. Kent Leach

    2012-05-01

    Full Text Available Implants formed of metals, bioceramics, or polymers may provide an alternative to autografts for treating large bone defects. However, limitations to each material motivate the examination of composites to capitalize on the beneficial aspects of individual components and to address the need for conferring bioactive behavior to the polymer matrix. We hypothesized that the inclusion of different bioceramics in a ceramic-polymer composite would alter the physical properties of the implant and the cellular osteogenic response. To test this, composite scaffolds formed from poly(lactide-co-glycolide (PLG and either hydroxyapatite (HA, β-tricalcium phosphate (TCP, or bioactive glass (Bioglass 45S®, BG were fabricated, and the physical properties of each scaffold were examined. We quantified cell proliferation by DNA content, osteogenic response of human osteoblasts (NHOsts to composite scaffolds by alkaline phosphatase (ALP activity, and changes in gene expression by qPCR. Compared to BG-PLG scaffolds, HA-PLG and TCP-PLG composite scaffolds possessed greater compressive moduli. NHOsts on BG-PLG substrates exhibited higher ALP activity than those on control, HA-, or TCP-PLG scaffolds after 21 days, and cells on composites exhibited a 3-fold increase in ALP activity between 7 and 21 days versus a minimal increase on control scaffolds. Compared to cells on PLG controls, RUNX2 expression in NHOsts on composite scaffolds was lower at both 7 and 21 days, while expression of genes encoding for bone matrix proteins (COL1A1 and SPARC was higher on BG-PLG scaffolds at both time points. These data demonstrate the importance of selecting a ceramic when fabricating composites applied for bone healing.

  12. Preparation of zeolite-A/chitosan hybrid composites and their bioactivities and antimicrobial activities.

    Science.gov (United States)

    Yu, Liang; Gong, Jie; Zeng, Changfeng; Zhang, Lixiong

    2013-10-01

    Zeolite-A/chitosan hybrid composites with zeolite contents of 20-55 wt.% were prepared by in situ transformation of silica/chitosan mixtures in a sodium aluminate alkaline solution through impregnation-gelation-hydrothermal synthesis. The products were characterized by X-ray diffraction, diffuse reflectance infrared Fourier transform spectroscopy, scanning electron microscopy, thermogravimetric analysis, and mercury penetration porosimetry. Their in vitro bioactivities were examined using as-synthesized and Ca(2+)-exchanged hybrid composites in simulated body fluid (SBF) for hydroxyapatite (HAP) growth. Their antimicrobial activities for Escherichia coli (E. coli) in trypticase soy broth (TSB) were evaluated using Ag(+)-exchanged hybrid composites. The zeolite-A/chitosan hybrid composites could be prepared as various shapes, including cylinders, plates and thin films. They possessed macropores with pore sizes ranging from 100 to 300 μm and showed compressive mechanical strength as high as 3.2 MPa when the zeolite content was 35 wt.%. Fast growth on the Ca(2+)-exchanged hybrid composites was observed with the highest weight gain of 51.4% in 30 days. The 35 wt.% Ag(+)-exchanged hybrid composite showed the highest antimicrobial activity, which could reduce the 9×10(6) CFU mL(-1)E. coli concentration to zero within 4h of incubation time with the Ag(+)-exchanged hybrid composite amount of 0.4 g L(-1). The bioactivity and antimicrobial activity could be combined by ion-exchanging the composites first with Ca(2+) and then with Ag(+). These zeolite-A/chitosan hybrid composites have potential applications on tissue engineering and antimicrobial food packaging. Copyright © 2013 Elsevier B.V. All rights reserved.

  13. Atmospheric plasma surface modifications of electrospun PCL/chitosan/PCL hybrid scaffolds by nozzle type plasma jets for usage of cell cultivation

    Energy Technology Data Exchange (ETDEWEB)

    Surucu, Seda [Department of Metallurgical and Materials Engineering, Atilim University, Incek, Golbasi, 06836, Ankara (Turkey); Masur, Kai [Leibniz Institute for Plasma Science and Technology (Germany); Turkoglu Sasmazel, Hilal, E-mail: hilal.sasmazel@atilim.edu.tr [Department of Metallurgical and Materials Engineering, Atilim University, Incek, Golbasi, 06836, Ankara (Turkey); Von Woedtke, Thomas; Weltmann, Klaus Dieter [Leibniz Institute for Plasma Science and Technology (Germany)

    2016-11-01

    Highlights: • Electrospun PCL/chitosan/PCL scaffolds introduced to the literature by us were modified with atmospheric pressure plasma jets. • Plasma was fed into the system with different gas flow rates, time and distances. • Topographical and functional changes were examined by various characterization methods. • Optimum plasma treatment parameters for enhanced topography and functionality were determined. • Electrospun hybrid plasma surface modified samples showed the increased biocompatibility performance of L929 fibroblast cells. - Abstract: This paper reports Ar gas, Ar + O{sub 2}, Ar + O{sub 2} + N{sub 2} gas mixtures and dry air plasma modifications by atmospheric pressure argon driven kINPen and air driven Diener (PlasmaBeam) plasma jets to alter surface properties of three dimensional (3D), electrospun PCL/Chitosan/PCL layer by layer hybrid scaffolds to improve human fibroblast (MRC5) cell attachment and growth. The characterizations of the samples were done by contact angle (CA) measurements, scanning electron microscopy (SEM), X-Ray Photoelectron spectroscopy (XPS) analysis. The results showed that the plasma modification carried out under dry air and Ar + O{sub 2} + N{sub 2} gas mixtures were altered effectively the nanotopography and the functionality of the material surfaces. It was found that the samples treated with Ar + O{sub 2} + N{sub 2} gas mixtures for 1 min and dry air for 9 min have better hydrophilicity 78.9° ± 1.0 and 75.6° ± 0.1, respectively compared to the untreated samples (126.5°). Biocompatibility performance of the scaffolds was determined with alamarBlue (aB) assay and MTT assay methods, Giemsa staining, fluorescence microscope, confocal laser scanning microscope (CLSM) and scanning electron microscope (SEM) analyses. The results showed that plasma treated samples increased the hydrophilicity and oxygen functionality and topography of the surfaces significantly, thus affecting the cell viability and proliferation on

  14. Atmospheric plasma surface modifications of electrospun PCL/chitosan/PCL hybrid scaffolds by nozzle type plasma jets for usage of cell cultivation

    International Nuclear Information System (INIS)

    Surucu, Seda; Masur, Kai; Turkoglu Sasmazel, Hilal; Von Woedtke, Thomas; Weltmann, Klaus Dieter

    2016-01-01

    Highlights: • Electrospun PCL/chitosan/PCL scaffolds introduced to the literature by us were modified with atmospheric pressure plasma jets. • Plasma was fed into the system with different gas flow rates, time and distances. • Topographical and functional changes were examined by various characterization methods. • Optimum plasma treatment parameters for enhanced topography and functionality were determined. • Electrospun hybrid plasma surface modified samples showed the increased biocompatibility performance of L929 fibroblast cells. - Abstract: This paper reports Ar gas, Ar + O_2, Ar + O_2 + N_2 gas mixtures and dry air plasma modifications by atmospheric pressure argon driven kINPen and air driven Diener (PlasmaBeam) plasma jets to alter surface properties of three dimensional (3D), electrospun PCL/Chitosan/PCL layer by layer hybrid scaffolds to improve human fibroblast (MRC5) cell attachment and growth. The characterizations of the samples were done by contact angle (CA) measurements, scanning electron microscopy (SEM), X-Ray Photoelectron spectroscopy (XPS) analysis. The results showed that the plasma modification carried out under dry air and Ar + O_2 + N_2 gas mixtures were altered effectively the nanotopography and the functionality of the material surfaces. It was found that the samples treated with Ar + O_2 + N_2 gas mixtures for 1 min and dry air for 9 min have better hydrophilicity 78.9° ± 1.0 and 75.6° ± 0.1, respectively compared to the untreated samples (126.5°). Biocompatibility performance of the scaffolds was determined with alamarBlue (aB) assay and MTT assay methods, Giemsa staining, fluorescence microscope, confocal laser scanning microscope (CLSM) and scanning electron microscope (SEM) analyses. The results showed that plasma treated samples increased the hydrophilicity and oxygen functionality and topography of the surfaces significantly, thus affecting the cell viability and proliferation on/within scaffolds.

  15. Disinfection of water with new chitosan-modified hybrid clay composite adsorbent

    Directory of Open Access Journals (Sweden)

    Emmanuel I. Unuabonah

    2017-08-01

    Full Text Available Hybrid clay composites were prepared from Kaolinite clay and Carica papaya seeds via modification with chitosan, Alum, NaOH, and ZnCl2 in different ratios, using solvothermal and surface modification techniques. Several composite adsorbents were prepared, and the most efficient of them for the removal of gram negative enteric bacteria was the hybrid clay composite that was surface-modified with chitosan, Ch-nHYCA1:5 (Chitosan: nHYCA = 1:5. This composite adsorbent had a maximum adsorption removal value of 4.07 × 106 cfu/mL for V. cholerae after 120 min, 1.95 × 106 cfu/mL for E. coli after ∼180 min and 3.25 × 106 cfu/mL for S. typhi after 270 min. The Brouers-Sotolongo model was found to better predict the maximum adsorption capacity (qmax of Ch-nHYCA1:5 composite adsorbent for the removal of E. coli with a qmax of 103.07 mg/g (7.93 × 107 cfu/mL and V. cholerae with a qmax of 154.18 mg/g (1.19 × 108 cfu/mL while the Sips model best described S. typhi adsorption by Ch-nHYCA1:5 composite with an estimated qmax of 83.65 mg/g (6.43 × 107 cfu/mL. These efficiencies do far exceed the alert/action levels of ca. 500 cfu/mL in drinking water for these bacteria. The simplicity of the composite preparation process and the availability of raw materials used for its preparation underscore the potential of this low-cost chitosan-modified composite adsorbent (Ch-nHYCA1:5 for water treatment.

  16. Multifunctional chitosan/polyvinyl pyrrolidone/45S5 Bioglass® scaffolds for MC3T3-E1 cell stimulation and drug release

    Energy Technology Data Exchange (ETDEWEB)

    Yao, Qingqing [Institute of Advanced Materials for Nano-Bio Applications, School of Ophthalmology & Optometry, Wenzhou Medical University, 270 Xueyuan Xi Road, Wenzhou, Zhejiang 325027 (China); Li, Wei [Institute of Biomaterials, Department of Materials Science and Engineering, University of Erlangen-Nuremberg, Cauerstrasse 6, Erlangen 91058 (Germany); Yu, Shanshan; Ma, Liwei [Institute of Advanced Materials for Nano-Bio Applications, School of Ophthalmology & Optometry, Wenzhou Medical University, 270 Xueyuan Xi Road, Wenzhou, Zhejiang 325027 (China); Jin, Dayong [Institute for Biomedical Materials and Devices, Faculty of Science, University of Technology Sydney, NSW 2007 (Australia); Advanced Cytometry Labs, ARC Center of Excellence for Nanoscale BioPhotonics, Macquarie University, Sydney, NSW 2109 (Australia); Boccaccini, Aldo R., E-mail: Aldo.Boccaccini@ww.uni-erlangen.de [Institute of Biomaterials, Department of Materials Science and Engineering, University of Erlangen-Nuremberg, Cauerstrasse 6, Erlangen 91058 (Germany); Liu, Yong, E-mail: yongliu1980@hotmail.com [Institute of Advanced Materials for Nano-Bio Applications, School of Ophthalmology & Optometry, Wenzhou Medical University, 270 Xueyuan Xi Road, Wenzhou, Zhejiang 325027 (China); Advanced Cytometry Labs, ARC Center of Excellence for Nanoscale BioPhotonics, Macquarie University, Sydney, NSW 2109 (Australia)

    2015-11-01

    Novel chitosan–polyvinyl pyrrolidone/45S5 Bioglass® (CS-PVP/BG) scaffolds were prepared via foam replication and chemical cross-linking techniques. The pristine BG, CS-PVP coated BG and genipin cross-linked CS-PVP/BG (G-CS-PVP/BG) scaffolds were synthesized and characterized in terms of chemical composition, physical structure and morphology respectively. Resistance to enzymatic degradation of the scaffold is improved significantly with the use of genipin cross-linked CS-PVP. The bio-effects of scaffolds on MC3T3-E1 osteoblast-like cells were evaluated by studying cell viability, adhesion and proliferation. The CCK-8 assay shows that cell viability on the resulting G-CS-PVP/BG scaffold is improved obviously after cross-linking of genipin. Cell skeleton images exhibit that well-stretched F-actin bundles are obtained on the G-CS-PVP/BG scaffold. SEM results present significant improvement on the cell adhesion and proliferation for cells cultured on the G-CS-PVP/BG scaffold. The drug release performance on the as-synthesized scaffold was studied in a phosphate buffered saline (PBS) solution. Vancomycin is found to be released in burst fashion within 24 h from the pristine BG scaffold, however, the release period from the G-CS-PVP/BG scaffold is enhanced to 7 days, indicating improved drug release properties of the G-CS-PVP/BG scaffold. Our results suggest that the G-CS-PVP/BG scaffolds possess promising physicochemical properties, sustained drug release capability and good biocompatibility for MC3T3-E1 cells' proliferation and adhesion, suggesting their potential applications in areas such as MC3T3-E1 cell stimulation and bone tissue engineering. - Highlights: • Novel genipi–chitosan–polyvinyl pyrrolidone/45S5 Bioglass® scaffolds are prepared. • Resistance to enzymatic degradation of the scaffold is improved significantly. • The resulting scaffold shows enhanced MC3T3-E1 cell adhesion and proliferation. • Release of antibiotic vancomycin from the

  17. Multifunctional chitosan/polyvinyl pyrrolidone/45S5 Bioglass® scaffolds for MC3T3-E1 cell stimulation and drug release

    International Nuclear Information System (INIS)

    Yao, Qingqing; Li, Wei; Yu, Shanshan; Ma, Liwei; Jin, Dayong; Boccaccini, Aldo R.; Liu, Yong

    2015-01-01

    Novel chitosan–polyvinyl pyrrolidone/45S5 Bioglass® (CS-PVP/BG) scaffolds were prepared via foam replication and chemical cross-linking techniques. The pristine BG, CS-PVP coated BG and genipin cross-linked CS-PVP/BG (G-CS-PVP/BG) scaffolds were synthesized and characterized in terms of chemical composition, physical structure and morphology respectively. Resistance to enzymatic degradation of the scaffold is improved significantly with the use of genipin cross-linked CS-PVP. The bio-effects of scaffolds on MC3T3-E1 osteoblast-like cells were evaluated by studying cell viability, adhesion and proliferation. The CCK-8 assay shows that cell viability on the resulting G-CS-PVP/BG scaffold is improved obviously after cross-linking of genipin. Cell skeleton images exhibit that well-stretched F-actin bundles are obtained on the G-CS-PVP/BG scaffold. SEM results present significant improvement on the cell adhesion and proliferation for cells cultured on the G-CS-PVP/BG scaffold. The drug release performance on the as-synthesized scaffold was studied in a phosphate buffered saline (PBS) solution. Vancomycin is found to be released in burst fashion within 24 h from the pristine BG scaffold, however, the release period from the G-CS-PVP/BG scaffold is enhanced to 7 days, indicating improved drug release properties of the G-CS-PVP/BG scaffold. Our results suggest that the G-CS-PVP/BG scaffolds possess promising physicochemical properties, sustained drug release capability and good biocompatibility for MC3T3-E1 cells' proliferation and adhesion, suggesting their potential applications in areas such as MC3T3-E1 cell stimulation and bone tissue engineering. - Highlights: • Novel genipi–chitosan–polyvinyl pyrrolidone/45S5 Bioglass® scaffolds are prepared. • Resistance to enzymatic degradation of the scaffold is improved significantly. • The resulting scaffold shows enhanced MC3T3-E1 cell adhesion and proliferation. • Release of antibiotic vancomycin from the

  18. Effects of scaffold composition and architecture on human nasal chondrocyte redifferentiation and cartilaginous matrix deposition

    NARCIS (Netherlands)

    Miot, Sylvie; Woodfield, T.B.F.; Daniels, Alma U.; Suetterlin, Rosemarie; Peterschmitt, Iman; Heberer, Michael; van Blitterswijk, Clemens; Riesle, J.U.; Martin, Ivan

    2005-01-01

    We investigated whether the post-expansion redifferentiation and cartilage tissue formation capacity of adult human nasal chondrocytes can be regulated by controlled modifications of scaffold composition and architecture. As a model system, we used poly(ethylene

  19. Electrospun Hydroxyapatite-Containing Chitosan Nanofibers Crosslinked with Genipin for Bone Tissue Engineering

    Science.gov (United States)

    Frohbergh, Michael E.; Katsman, Anna; Botta, Gregory P.; Lazarovici, Phillip; Schauer, Caroline L.; Wegst, Ulrike G. K.; Lelkes, Peter I.

    2012-01-01

    Reconstruction of large bone defects remains problematic in orthopedic and craniofacial clinical practice. Autografts are limited in supply and are associated with donor site morbidity while other materials show poor integration with the host’s own bone. This lack of integration is often due to the absence of periosteum, the outer layer of bone that contains osteoprogenitor cells and is critical for the growth and remodeling of bone tissue. In this study we developed a one-step platform to electrospin nanofibrous scaffolds from chitosan, which also contain hydroxyapatite nanoparticles and are crosslinked with genipin. We hypothesized that the resulting composite scaffolds represent a microenvironment that emulates the physical, mineralized structure and mechanical properties of non-weight bearing bone extracellular matrix while promoting osteoblast differentiation and maturation similar to the periosteum. The ultrastructure and physicochemical properties of the scaffolds were studied using scanning electron microscopy and spectroscopic techniques. The average fiber diameters of the electrospun scaffolds were 227±154 nm as spun, and increased to 335±119 nm after crosslinking with genipin. Analysis by X-ray diffraction, Fourier transformed infrared spectroscopy and energy dispersive spectroscopy confirmed the presence of characteristic features of hydroxyapatite in the composite chitosan fibers. The Young’s modulus of the composite fibrous scaffolds was 142±13 MPa, which is similar to that of the natural periosteum. Both pure chitosan scaffolds and composite hydroxyapatite-containing chitosan scaffolds supported adhesion, proliferation and osteogenic differentiation of mouse 7F2 osteoblast-like cells. Expression and enzymatic activity of alkaline phosphatase, an early osteogenic marker, were higher in cells cultured on the composite scaffolds as compared to pure chitosan scaffolds, reaching a significant, 2.4 fold, difference by day 14 (phydroxyapatite

  20. Dielectric properties: A gateway to antibacterial assay-A case study of low-density polyethylene/chitosan composite films.

    Digital Repository Service at National Institute of Oceanography (India)

    Sunilkumar, M.; Gafoor, A.A.; Anas, A.; Haseena, A.P.; Sujith, A.

    anhydride and dicumyl peroxide were used as a coupling agent and a free radical initiator, respectively. The dielectric properties of the composite films were studied as a function of chitosan loading, presence of plasticizer and variable applied frequency...

  1. Removal of hexavalent chromium from wastewater using a new composite chitosan biosorbent.

    Science.gov (United States)

    Boddu, Veera M; Abburi, Krishnaiah; Talbott, Jonathan L; Smith, Edgar D

    2003-10-01

    A new composite chitosan biosorbent was prepared by coating chitosan, a glucosamine biopolymer, onto ceramic alumina. The composite bioadsorbent was characterized by high-temperature pyrolysis, porosimetry, scanning electron microscopy, and X-ray photoelectron spectroscopy. Batch isothermal equilibrium and continuous column adsorption experiments were conducted at 25 degrees C to evaluate the biosorbent for the removal of hexavalent chromium from synthetic as well as field samples obtained from chrome plating facilities. The effect of pH, sulfate, and chloride ion on adsorption was also investigated. The biosorbent loaded with Cr(VI) was regenerated using 0.1 M sodium hydroxide solution. A comparison of the results of the present investigation with those reported in the literature showed that chitosan coated on alumina exhibits greater adsorption capacity for chromium(VI). Further, experimental equilibrium data were fitted to Langmuir and Freundlich adsorption isotherms, and values of the parameters of the isotherms are reported. The ultimate capacity obtained from the Langmuir model is 153.85 mg/g chitosan.

  2. Magnetic responsive hydroxyapatite composite scaffolds construction for bone defect reparation

    Directory of Open Access Journals (Sweden)

    Zeng XB

    2012-07-01

    Full Text Available Xiao Bo Zeng, Hao Hu, Li Qin Xie, Fang Lan, Wen Jiang, Yao Wu, Zhong Wei GuNational Engineering Research Center for Biomaterials, Sichuan University, Chengdu, Sichuan, People's Republic of ChinaIntroduction: In recent years, interest in magnetic biomimetic scaffolds for tissue engineering has increased considerably. A type of magnetic scaffold composed of magnetic nanoparticles (MNPs and hydroxyapatite (HA for bone repair has been developed by our research group.Aim and methods: In this study, to investigate the influence of the MNP content (in the scaffolds on the cell behaviors and the interactions between the magnetic scaffold and the exterior magnetic field, a series of MNP-HA magnetic scaffolds with different MNP contents (from 0.2% to 2% were fabricated by immersing HA scaffold into MNP colloid. ROS 17/2.8 and MC3T3-E1 cells were cultured on the scaffolds in vitro, with and without an exterior magnetic field, respectively. The cell adhesion, proliferation and differentiation were evaluated via scanning electron microscopy; confocal laser scanning microscopy; and 3-(4,5-Dimethylthiazol-2-yl-2,5-diphenyltetrazolium bromide (MTT, alkaline phosphatase, and bone gla protein activity tests.Results: The results demonstrated the positive influence of the magnetic scaffolds on cell adhesion, proliferation, and differentiation. Further, a higher amount of MNPs on the magnetic scaffolds led to more significant stimulation.Conclusion: The magnetic scaffold can respond to the exterior magnetic field and engender some synergistic effect to intensify the stimulating effect of a magnetic field to the proliferation and differentiation of cells.Keywords: magnetic therapy, magnetic nanoparticles, bone repair, magnetic responsive

  3. Strengthening injectable thermo-sensitive NIPAAm-g-chitosan hydrogels using chemical cross-linking of disulfide bonds as scaffolds for tissue engineering.

    Science.gov (United States)

    Wu, Shu-Wei; Liu, Xifeng; Miller, A Lee; Cheng, Yu-Shiuan; Yeh, Ming-Long; Lu, Lichun

    2018-07-15

    In the present study, we fabricated non-toxic, injectable, and thermo-sensitive NIPAAm-g-chitosan (NC) hydrogels with thiol modification for introduction of disulfide cross-linking strategy. Previously, NIPAAm and chitosan copolymer has been proven to have excellent biocompatibility, biodegradability and rapid phase transition after injection, suitable to serve as cell carriers or implanted scaffolds. However, weak mechanical properties significantly limit their potential for biomedical fields. In order to overcome this issue, we incorporated thiol side chains into chitosan by covalently conjugating N-acetyl-cysteine (NAC) with carbodiimide chemistry to strengthen mechanical properties. After oxidation of thiols into disulfide bonds, modified NC hydrogels did improve the compressive modulus over 9 folds (11.4 kPa). Oscillatory frequency sweep showed a positive correlation between storage modulus and cross-liking density as well. Additionally, there was no cytotoxicity observed to mesenchymal stem cells, fibroblasts and osteoblasts. We suggested that the thiol-modified thermo-sensitive polysaccharide hydrogels are promising to be a cell-laden biomaterial for tissue regeneration. Copyright © 2018 Elsevier Ltd. All rights reserved.

  4. Long-term antibiotic delivery by chitosan-based composite coatings with bone regenerative potential

    Science.gov (United States)

    Ordikhani, F.; Simchi, A.

    2014-10-01

    Composite coatings with bone-bioactivity and drug-eluting capacity are considered as promising materials for titanium bone implants. In this work, drug-eluting chitosan-bioactive glass coatings were fabricated by a single-step electrophoretic deposition technique. Drug-loading and -releasing capacity of the composite coatings were carried out using the vancomycin antibiotic. Uniform coatings with a thickness of ∼55 μm containing 23.7 wt% bioactive glass particles and various amounts of the antibiotic (380-630 μg/cm2) were produced. The coatings were bioactive in terms of apatite-forming ability in simulated body fluid and showed favorable cell adhesion and growth. In vitro biological tests also indicated that the composite coatings had better cellular affinity than pristine chitosan coatings. The in vitro elution kinetics of the composite coating revealed an initial burst release of around 40% of the drug within the first elution step of 1 h and following by a continuous eluting over 4 weeks, revealing long-term drug-delivering potential. Antibacterial tests using survival assay against Gram-positive Staphylococcus aureus bacteria determined the effect of vancomycin release on reduction of infection risk. Almost no bacteria were survived on the coatings prepared from the EPD suspension containing ≥0.5 g/l vancomycin. The developed chitosan-based composite coatings with bone bioactivity and long-term drug-delivery ability may be potentially useful for metallic implants to reduce infection risk.

  5. Bioactive glass-poly (ε-caprolactone) composite scaffolds with 3 dimensionally hierarchical pore networks

    International Nuclear Information System (INIS)

    Yun, Hui-suk; Kim, Seung-eon; Park, Eui Kyun

    2011-01-01

    Hierarchically mesoporous-macroporous-giant-porous bioactive glass/poly ε-caprolactone (PCL) composite scaffolds were prepared using a combination of the sol-gel method, evaporation-induced self-assembly process in the presence of nonionic triblock copolymer, EO 100 PO 65 EO 100 (F127), as template, salt leaching method, and rapid prototyping techniques. F127 acts as a template, inducing the formation of mesopores, NaCl with sizes between 25 and 33 μm provides macro-pores after leaching, and rapid prototyping produces giant-pores. The structure and morphology of the scaffolds were characterized by the field emission scanning electron microscopy, transmission electron microscopy, and Hg porosimetry. The mechanical properties of the scaffolds were examined by the dynamic mechanical analysis. Their in vitro bioactivities were confirmed by immersing the scaffolds in simulated body fluid. Their biocompatibilities were also evaluated by culturing human bone marrow stromal cells on the scaffolds. The scaffolds show good molding capabilities, mechanical properties, 3 dimensionally well-interconnected pore structures, bioactivities, and biocompatibilities in vitro. Depending on the amount of NaCl, the scaffolds also show unique sponge-like properties, but still retain better mechanical properties than general salt leaching derived PCL scaffolds. All of the data provide good evidence that the obtained scaffolds possess excellent potential for applications in the fields of tissue engineering and drug storage.

  6. Nano-ceramic composite scaffolds for bioreactor-based bone engineering.

    Science.gov (United States)

    Lv, Qing; Deng, Meng; Ulery, Bret D; Nair, Lakshmi S; Laurencin, Cato T

    2013-08-01

    Composites of biodegradable polymers and bioactive ceramics are candidates for tissue-engineered scaffolds that closely match the properties of bone. We previously developed a porous, three-dimensional poly (D,L-lactide-co-glycolide) (PLAGA)/nanohydroxyapatite (n-HA) scaffold as a potential bone tissue engineering matrix suitable for high-aspect ratio vessel (HARV) bioreactor applications. However, the physical and cellular properties of this scaffold are unknown. The present study aims to evaluate the effect of n-HA in modulating PLAGA scaffold properties and human mesenchymal stem cell (HMSC) responses in a HARV bioreactor. By comparing PLAGA/n-HA and PLAGA scaffolds, we asked whether incorporation of n-HA (1) accelerates scaffold degradation and compromises mechanical integrity; (2) promotes HMSC proliferation and differentiation; and (3) enhances HMSC mineralization when cultured in HARV bioreactors. PLAGA/n-HA scaffolds (total number = 48) were loaded into HARV bioreactors for 6 weeks and monitored for mass, molecular weight, mechanical, and morphological changes. HMSCs were seeded on PLAGA/n-HA scaffolds (total number = 38) and cultured in HARV bioreactors for 28 days. Cell migration, proliferation, osteogenic differentiation, and mineralization were characterized at four selected time points. The same amount of PLAGA scaffolds were used as controls. The incorporation of n-HA did not alter the scaffold degradation pattern. PLAGA/n-HA scaffolds maintained their mechanical integrity throughout the 6 weeks in the dynamic culture environment. HMSCs seeded on PLAGA/n-HA scaffolds showed elevated proliferation, expression of osteogenic phenotypic markers, and mineral deposition as compared with cells seeded on PLAGA scaffolds. HMSCs migrated into the scaffold center with nearly uniform cell and extracellular matrix distribution in the scaffold interior. The combination of PLAGA/n-HA scaffolds with HMSCs in HARV bioreactors may allow for the generation of engineered

  7. Fabrication and in vitro biocompatibility of biomorphic PLGA/nHA composite scaffolds for bone tissue engineering

    Energy Technology Data Exchange (ETDEWEB)

    Qian, Junmin, E-mail: jmqian@mail.xjtu.edu.cn; Xu, Weijun; Yong, Xueqing; Jin, Xinxia; Zhang, Wei

    2014-03-01

    In this study, biomorphic poly(DL-lactic-co-glycolic acid)/nano-hydroxyapatite (PLGA/nHA) composite scaffolds were successfully prepared using cane as a template. The porous morphology, phase, compression characteristics and in vitro biocompatibility of the PLGA/nHA composite scaffolds and biomorphic PLGA scaffolds as control were investigated. The results showed that the biomorphic scaffolds preserved the original honeycomb-like architecture of cane and exhibited a bimodal porous structure. The average channel diameter and micropore size of the PLGA/nHA composite scaffolds were 164 ± 52 μm and 13 ± 8 μm, respectively, with a porosity of 89.3 ± 1.4%. The incorporation of nHA into PLGA decreased the degree of crystallinity of PLGA, and significantly improved the compressive modulus of biomorphic scaffolds. The in vitro biocompatibility evaluation with MC3T3-E1 cells demonstrated that the biomorphic PLGA/nHA composite scaffolds could better support cell attachment, proliferation and differentiation than the biomorphic PLGA scaffolds. The localization depth of MC3T3-E1 cells within the channels of the biomorphic PLGA/nHA composite scaffolds could reach approximately 400 μm. The results suggested that the biomorphic PLGA/nHA composite scaffolds were promising candidates for bone tissue engineering. - Highlights: • Novel biomimetic PLGA/nHA composite scaffolds were successfully prepared. • nHA addition improved elastic modulus of PLGA scaffold and decreased its crystallinity. • PLGA/nHA composite scaffolds had better biocompatibility than PLGA scaffolds. • Biomorphic PLGA/nHA composite scaffold had great potential in bone tissue engineering.

  8. Fabrication and in vitro biocompatibility of biomorphic PLGA/nHA composite scaffolds for bone tissue engineering

    International Nuclear Information System (INIS)

    Qian, Junmin; Xu, Weijun; Yong, Xueqing; Jin, Xinxia; Zhang, Wei

    2014-01-01

    In this study, biomorphic poly(DL-lactic-co-glycolic acid)/nano-hydroxyapatite (PLGA/nHA) composite scaffolds were successfully prepared using cane as a template. The porous morphology, phase, compression characteristics and in vitro biocompatibility of the PLGA/nHA composite scaffolds and biomorphic PLGA scaffolds as control were investigated. The results showed that the biomorphic scaffolds preserved the original honeycomb-like architecture of cane and exhibited a bimodal porous structure. The average channel diameter and micropore size of the PLGA/nHA composite scaffolds were 164 ± 52 μm and 13 ± 8 μm, respectively, with a porosity of 89.3 ± 1.4%. The incorporation of nHA into PLGA decreased the degree of crystallinity of PLGA, and significantly improved the compressive modulus of biomorphic scaffolds. The in vitro biocompatibility evaluation with MC3T3-E1 cells demonstrated that the biomorphic PLGA/nHA composite scaffolds could better support cell attachment, proliferation and differentiation than the biomorphic PLGA scaffolds. The localization depth of MC3T3-E1 cells within the channels of the biomorphic PLGA/nHA composite scaffolds could reach approximately 400 μm. The results suggested that the biomorphic PLGA/nHA composite scaffolds were promising candidates for bone tissue engineering. - Highlights: • Novel biomimetic PLGA/nHA composite scaffolds were successfully prepared. • nHA addition improved elastic modulus of PLGA scaffold and decreased its crystallinity. • PLGA/nHA composite scaffolds had better biocompatibility than PLGA scaffolds. • Biomorphic PLGA/nHA composite scaffold had great potential in bone tissue engineering

  9. Evaluating protein incorporation and release in electrospun composite scaffolds for bone tissue engineering applications.

    Science.gov (United States)

    Briggs, Tonye; Matos, Jeffrey; Collins, George; Arinzeh, Treena Livingston

    2015-10-01

    Electrospun polymer/ceramic composites have gained interest for use as scaffolds for bone tissue engineering applications. In this study, we investigated methods to incorporate Platelet Derived Growth Factor-BB (PDGF-BB) in electrospun polycaprolactone (PCL) or PCL prepared with polyethylene oxide (PEO), where both contained varying levels (up to 30 wt %) of ceramic composed of biphasic calcium phosphates, hydroxyapatite (HA)/β-tricalcium phosphate (TCP). Using a model protein, lysozyme, we compared two methods of protein incorporation, adsorption and emulsion electrospinning. Adsorption of lysozyme on scaffolds with ceramic resulted in minimal release of lysozyme over time. Using emulsion electrospinning, lysozyme released from scaffolds containing a high concentration of ceramic where the majority of the release occurred at later time points. We investigated the effect of reducing the electrostatic interaction between the protein and the ceramic on protein release with the addition of the cationic surfactant, cetyl trimethylammonium bromide (CTAB). In vitro release studies demonstrated that electrospun scaffolds prepared with CTAB released more lysozyme or PDGF-BB compared with scaffolds without the cationic surfactant. Human mesenchymal stem cells (MSCs) on composite scaffolds containing PDGF-BB incorporated through emulsion electrospinning expressed higher levels of osteogenic markers compared to scaffolds without PDGF-BB, indicating that the bioactivity of the growth factor was maintained. This study revealed methods for incorporating growth factors in polymer/ceramic scaffolds to promote osteoinduction and thereby facilitate bone regeneration. © 2015 Wiley Periodicals, Inc.

  10. A PEGylated platelet free plasma hydrogel based composite scaffold enables stable vascularization and targeted cell delivery for volumetric muscle loss.

    Science.gov (United States)

    Aurora, Amit; Wrice, Nicole; Walters, Thomas J; Christy, Robert J; Natesan, Shanmugasundaram

    2018-01-01

    Extracellular matrix (ECM) scaffolds are being used for the clinical repair of soft tissue injuries. Although improved functional outcomes have been reported, ECM scaffolds show limited tissue specific remodeling response with concomitant deposition of fibrotic tissue. One plausible explanation is the regression of blood vessels which may be limiting the diffusion of oxygen and nutrients across the scaffold. Herein we develop a composite scaffold as a vasculo-inductive platform by integrating PEGylated platelet free plasma (PFP) hydrogel with a muscle derived ECM scaffold (m-ECM). In vitro, adipose derived stem cells (ASCs) seeded onto the composite scaffold differentiated into two distinct morphologies, a tubular network in the hydrogel, and elongated structures along the m-ECM scaffold. The composite scaffold showed a high expression of ITGA5, ITGB1, and FN and a synergistic up-regulation of ang1 and tie-2 transcripts. The in vitro ability of the composite scaffold to provide extracellular milieu for cell adhesion and molecular cues to support vessel formation was investigated in a rodent volumetric muscle loss (VML) model. The composite scaffold delivered with ASCs supported robust and stable vascularization. Additionally, the composite scaffold supported increased localization of ASCs in the defect demonstrating its ability for localized cell delivery. Interestingly, ASCs were observed homing in the injured muscle and around the perivascular space possibly to stabilize the host vasculature. In conclusion, the composite scaffold delivered with ASCs presents a promising approach for scaffold vascularization. The versatile nature of the composite scaffold also makes it easily adaptable for the repair of soft tissue injuries. Decellularized extracellular matrix (ECM) scaffolds when used for soft tissue repair is often accompanied by deposition of fibrotic tissue possibly due to limited scaffold vascularization, which limits the diffusion of oxygen and nutrients

  11. Innovative biodegradable poly(L-lactide/collagen/hydroxyapatite composite fibrous scaffolds promote osteoblastic proliferation and differentiation

    Directory of Open Access Journals (Sweden)

    Zhou GQ

    2017-10-01

    Full Text Available Guoqiang Zhou,1–3 Sudan Liu,1 Yanyan Ma,1 Wenshi Xu,1 Wei Meng,1 Xue Lin,1 Wenying Wang,1,3 Shuxiang Wang,1–3 Jinchao Zhang1–3 1College of Chemistry and Environmental Science, 2Key Laboratory of Medicinal Chemistry and Molecular Diagnosis of Ministry of Education, 3Key Laboratory of Chemical Biology of Hebei Province, Hebei University, Baoding, Hebei, People’s Republic of China Abstract: The development of an artificial bone graft which can promote the regeneration of fractures or diseased bones is currently the most challenging aspect in bone tissue engineering. To achieve the purpose of promoting bone proliferation and differentiation, the artificial graft needs have a similar structure and composition of extracellular matrix. One-step electrospinning method of biocomposite nanofibers containing hydroxyapatite (HA nanoparticles and collagen (Coll were developed for potential application in bone tissue engineering. Nanocomposite scaffolds of poly(L-lactide (PLLA, PLLA/HA, PLLA/Coll, and PLLA/Coll/HA were fabricated by electrospinning. The morphology, diameter, elements, hydrophilicity, and biodegradability of the composite scaffolds have been investigated. The biocompatibility of different nanocomposite scaffolds was assessed using mouse osteoblasts MC3T3-E1 in vitro, and the proliferation, differentiation, and mineralization of cells on different nanofibrous scaffolds were investigated. The results showed that PLLA/Coll/HA nanofiber scaffolds enhanced cell adhesion, spreading, proliferation, differentiation, mineralization, and gene expression of osteogenic markers compared to other scaffolds. In addition, the nanofibrous scaffolds maintained a stable composition at the beginning of the degradation period and morphology wastage and weight loss were observed when incubated for up to 80 days in physiological simulated conditions. The PLLA/Coll/HA composite nanofibrous scaffolds could be a potential material for guided bone regeneration

  12. Treatment of osteomyelitis defects by a vancomycin-loaded gelatin/β-tricalcium phosphate composite scaffold

    Science.gov (United States)

    Zhou, J.; Zhou, X. G.; Wang, J. W.; Zhou, H.; Dong, J.

    2018-01-01

    Objective In the present study, we aimed to assess whether gelatin/β-tricalcium phosphate (β-TCP) composite porous scaffolds could be used as a local controlled release system for vancomycin. We also investigated the efficiency of the scaffolds in eliminating infections and repairing osteomyelitis defects in rabbits. Methods The gelatin scaffolds containing differing amounts of of β-TCP (0%, 10%, 30% and 50%) were prepared for controlled release of vancomycin and were labelled G-TCP0, G-TCP1, G-TCP3 and G-TCP5, respectively. The Kirby-Bauer method was used to examine the release profile. Chronic osteomyelitis models of rabbits were established. After thorough debridement, the osteomyelitis defects were implanted with the scaffolds. Radiographs and histological examinations were carried out to investigate the efficiency of eliminating infections and repairing bone defects. Results The prepared gelatin/β-TCP scaffolds exhibited a homogeneously interconnected 3D porous structure. The G-TCP0 scaffold exhibited the longest duration of vancomycin release with a release duration of eight weeks. With the increase of β-TCP contents, the release duration of the β-TCP-containing composite scaffolds was decreased. The complete release of vancomycin from the G-TCP5 scaffold was achieved within three weeks. In the treatment of osteomyelitis defects in rabbits, the G-TCP3 scaffold showed the most efficacious performance in eliminating infections and repairing bone defects. Conclusions The composite scaffolds could achieve local therapeutic drug levels over an extended duration. The G-TCP3 scaffold possessed the optimal porosity, interconnection and controlled release performance. Therefore, this scaffold could potentially be used in the treatment of chronic osteomyelitis defects. Cite this article: J. Zhou, X. G. Zhou, J. W. Wang, H. Zhou, J. Dong. Treatment of osteomyelitis defects by a vancomycin-loaded gelatin/β-tricalcium phosphate composite scaffold. Bone Joint Res

  13. Synthesis of magnetite/graphene oxide/chitosan composite and its application for protein adsorption

    Energy Technology Data Exchange (ETDEWEB)

    Ye, Nengsheng, E-mail: yensh@cnu.edu.cn; Xie, Yali; Shi, Pengzhi; Gao, Ting; Ma, Jichao

    2014-12-01

    In this study, a facile and novel strategy was developed to fabricate magnetite/graphene oxide/chitosan (Fe{sub 3}O{sub 4}/GO/CS) composite, and the composite was used as a magnetic adsorbent for the enrichment of protein, and followed by matrix-assisted laser desorption ionization mass spectrometry (MALDI-TOF MS) analysis. The phase composition, chemical structure and morphology of the composite were characterized by X-ray diffraction (XRD), Fourier transform infrared spectrometer (FTIR), transmission electron microscopy (TEM), scanning electronic microscope (SEM) and vibrating sample magnetometer (VSM). Protein cytochrome c was chosen as model target to evaluate the adsorptive property of Fe{sub 3}O{sub 4}/GO/CS. After enrichment procedure and magnetic separation, protein bounded with the material was analyzed by MALDI-TOF MS without desorption. The results indicated that Fe{sub 3}O{sub 4}/GO/CS composite exhibited a good adsorptive capacity for protein, and Fe{sub 3}O{sub 4}/GO/CS composite had a promising potential in magnetic separation research. - Highlights: • Magnetite/graphene oxide/chitosan composite was synthesized by novel route. • The composite was used as magnetic absorbent for protein enrichment. • The composite had excellent adsorption performance for protein enrichment.

  14. In vitro biomechanical and biocompatible evaluation of natural hydroxyapatite/chitosan composite for bone repair.

    Science.gov (United States)

    Lü, Xiaoying; Zheng, Buzhong; Tang, Xiaojun; Zhao, Lifeng; Lu, Jieyan; Zhang, Zhiwei; Zhang, Jizhong; Cui, Wei

    2011-01-01

    To evaluate the biomechanical properties and biocompatibility of natural hydroxyapatite/chitosan (HA/CS) composites. The natural HA/CS composites with a different proportion of HA and CS were prepared by the cross-linking method, and then the compressive strength, microstructure and pH values of extracts from these composites were measured by SEM and pH meter, respectively. Subsequently, the biocompatibility of the composites was evaluated by means of a series of biological tests, including MTT, acute systemic toxicity, heat source, and hemolysis tests in vitro. The chitosan content in the composites had significantly influenced the mechanical properties and microstructure of the composites. The pH value of the composite extract was approximately 7.0, which was very close to that of human plasma. Furthermore, the natural HA/CS composites showed no cytotoxicity, irritation, teratogenicity, carcinogenicity and special pyrogen. These results indicated that the natural HA/CS composite may be a potential bone repair material.

  15. Engineering of Corneal Tissue through an Aligned PVA/Collagen Composite Nanofibrous Electrospun Scaffold.

    Science.gov (United States)

    Wu, Zhengjie; Kong, Bin; Liu, Rui; Sun, Wei; Mi, Shengli

    2018-02-24

    Corneal diseases are the main reason of vision loss globally. Constructing a corneal equivalent which has a similar strength and transparency with the native cornea, seems to be a feasible way to solve the shortage of donated cornea. Electrospun collagen scaffolds are often fabricated and used as a tissue-engineered cornea, but the main drawback of poor mechanical properties make it unable to meet the requirement for surgery suture, which limits its clinical applications to a large extent. Aligned polyvinyl acetate (PVA)/collagen (PVA-COL) scaffolds were electrospun by mixing collagen and PVA to reinforce the mechanical strength of the collagen electrospun scaffold. Human keratocytes (HKs) and human corneal epithelial cells (HCECs) inoculated on aligned and random PVA-COL electrospun scaffolds adhered and proliferated well, and the aligned nanofibers induced orderly HK growth, indicating that the designed PVA-COL composite nanofibrous electrospun scaffold is suitable for application in tissue-engineered cornea.

  16. Biomimetic fabrication of calcium phosphate/chitosan nanohybrid composite in modified simulated body fluids

    Directory of Open Access Journals (Sweden)

    K. H. Park

    2017-01-01

    Full Text Available In this study, nucleation and growth of bone-like hydroxyapatite (HAp mineral in modified simulated body fluids (m-SBF were induced on chitosan (CS substrates, which were prepared by spin coating of chitosan on Ti substrate. The m-SBF showed a two fold increase in the concentrations of calcium and phosphate ions compared to SBF, and the post-NaOH treatment provided stabilization of the coatings. The calcium phosphate/chitosan composite prepared in m-SBF showed homogeneous distribution of approximately 350 nm-sized spherical clusters composed of octacalcium phosphate (OCP; Ca8H2(PO46·5H2O crystalline structure. Chitosan provided a control over the size of calcium phosphate prepared by immersion in m-SBF, and post-NaOH treatment supported the binding of calcium phosphate compound on the Ti surface. Post-NaOH treatment increased hydrophilicity and crystallinity of carbonate apatite, which increased its potential for biomedical application.

  17. Development of chitosan supported zirconium(IV) tungstophosphate composite for fluoride removal

    Energy Technology Data Exchange (ETDEWEB)

    Viswanathan, Natrayasamy, E-mail: natrayasamy_viswanathan@rediffmail.com [Department of Chemistry, Anna University Tiruchirappalli - Dindigul Campus, Dindigul 624622, Tamil Nadu (India); Meenakshi, S., E-mail: drs_meena@rediffmail.com [Department of Chemistry, Gandhigram Rural University, Gandhigram 624302, Tamil Nadu (India)

    2010-04-15

    A new biocomposite was prepared by incorporating inorganic ion exchanger namely zirconium(IV) tungstophosphate (ZrWP) into the chitosan biopolymeric matrix. The sorption behaviour of fluoride from aqueous solutions by this ZrWP/chitosan (ZrWPCs) composite has been investigated by batch technique. The fluoride sorption was studied as a function of contact time, pH, initial fluoride concentration, competing co-ions and temperature. The defluoridation capacity (DC) of the adsorbent was found to be 2025 mgF{sup -} kg{sup -1}. The composite was characterized using FTIR and SEM with EDAX analysis. The equilibrium sorption data were fitted to Freundlich and Langmuir isotherms. The kinetics of sorption was found to follow pseudo-second-order and intraparticle diffusion models. The values of thermodynamic parameters indicate the nature of sorption is spontaneous and endothermic. The biocomposite was successfully used for the removal of fluoride from the field water taken in a nearby fluoride endemic village.

  18. Synthesis and characterisation of cross-linked chitosan composites functionalised with silver and gold nanoparticles for antimicrobial applications

    Science.gov (United States)

    Ryan, Catherine; Alcock, Emma; Buttimer, Finbarr; Schmidt, Michael; Clarke, David; Pemble, Martyn; Bardosova, Maria

    2017-12-01

    We present a study of a range of cross-linked chitosan composites with potential antimicrobial applications. They were formed by cross-linking chitosan and siloxane networks and by introducing silver and gold nanoparticles (NPs). The aim was to investigate whether adding the metal NPs to the chitosan-siloxane composite would lead to a material with enhanced antimicrobial ability as compared to chitosan itself. The composites were synthesised in hydrogel form with the metal NPs embedded in the cross-linked chitosan network. Spectroscopic and microscopic techniques were employed to investigate the structural properties of the composite and the tensile strength of the structures was measured. It was found that the addition of metal NPs did not influence the mechanical strength of the composite. A crystal violet attachment assay results displayed a significant reduction in the attachment of E. coli to the cross-linked chitosan surfaces. Release profile tests suggest that the metal NPs do not contribute to the overall antimicrobial activity under neutral conditions. The contribution to the mechanical and antimicrobial properties from cross-linking with siloxane is significant, giving rise to a versatile, durable, antimicrobial material suitable for thin film formation, wound dressings or the coating of various surfaces where robustness and antimicrobial control are required.

  19. Growth of apatite on chitosan-multiwall carbon nanotube composite membranes

    Energy Technology Data Exchange (ETDEWEB)

    Yang Jun; Yao Zhiwen [State Key Laboratory of Oral Diseases, West China College of Stomatology, Sichuan University, No 14, 3rd Section South People' s Road, Chengdu 610041 (China); Tang Changyu [Department of Polymer Science and Materials, Sichuan University (China); Darvell, B.W. [Dental Materials Science, Faculty of Dentistry, University of Hong Kong (Hong Kong); Zhang Hualin; Pan Lingzhan; Liu Jingsong [State Key Laboratory of Oral Diseases, West China College of Stomatology, Sichuan University, No 14, 3rd Section South People' s Road, Chengdu 610041 (China); Chen Zhiqing, E-mail: yangj0710@gmail.com [State Key Laboratory of Oral Diseases, West China College of Stomatology, Sichuan University, No 14, 3rd Section South People' s Road, Chengdu 610041 (China)

    2009-07-30

    Bioactive membranes for guided tissue regeneration would be of value for periodontal therapy. Chitosan-multiwall carbon nanotube (CS-MWNT) composites were treated to deposit nanoscopic apatite for MWNT proportions of 0-4 mass%. Fourier-transform infrared spectroscopy, scanning electron microscopy, energy-dispersive X-ray analysis, and X-ray diffraction were used for characterization. Apatite was formed on the CS-MWNT composites at low MWNT concentrations, but the dispersion of the MWNT affects the crystallite size and the Ca/P molar ratio of the composite. The smallest crystallite size was 9 nm at 1 mass% MWNT.

  20. Synthesis and characterization of chitosan-graft-poly(acrylic acid)/rice husk ash hydrogels composites

    International Nuclear Information System (INIS)

    Rodrigues, Francisco H.A.; Lopes, Gabriel V.; Pereira, Antonio G.B.; Fajardo, Andre R.; Muniz, Edvani C.

    2011-01-01

    According to environmental concerns, super absorbent hydrogel composites were synthesized based on rice husk ash (RHA), an industrial waste, and Chitosan-graft-poly(acrylic acid). The WAXS and FTIR data confirmed the syntheses of hydrogel composites. The effect of crystalline or amorphous RHA on water uptake was investigated. It was found that the RHA in crystalline form induces higher water capacity (W eq ) of composites hydrogels due to the fact that the intra-interactions among silanol groups on RHA make available new sites in the polymer matrix, which could interact to water. (author)

  1. Polyvinyl alcohol composite nanofibres containing conjugated levofloxacin-chitosan for controlled drug release

    International Nuclear Information System (INIS)

    Jalvandi, Javid; White, Max; Gao, Yuan; Truong, Yen Bach; Padhye, Rajiv; Kyratzis, Ilias Louis

    2017-01-01

    A range of biodegradable drug-nanofibres composite mats have been reported as drug delivery systems. However, their main disadvantage is the rapid release of the drug immediately after application. This paper reports an improved system based on the incorporation of drug conjugated-chitosan into polyvinyl alcohol (PVA) nanofibers. The results showed that controlled release of levofloxacin (LVF) could be achieved by covalently binding LVF to low molecular weight chitosan (CS) via a cleavable amide bond and then blending the conjugated CS with polyvinyl alcohol (PVA) nanofibres prior to electrospinning. PVA/LVF and PVA-CS/LVF nanofibres were fabricated as controls. The conjugated CS-LVF was characterized by FTIR, DSC, TGA and 1 H NMR. Scanning electron microscopy (SEM) showed that the blended CS-PVA nanofibres had a reduced fibre diameter compared to the controls. Drug release profiles showed that burst release was decreased from 90% in the control PVA/LVF electrospun mats to 27% in the PVA/conjugated CS-LVF mats after 8 h in phosphate buffer at 37 °C. This slower release is due to the cleavable bond between LVF and CS that slowly hydrolysed over time at neutral pH. The results indicate that conjugation of the drug to the polymer backbone is an effective way of minimizing burst release behaviour and achieving sustained release of the drug, LVF. - Highlights: • A novel drug delivery system for controlled release of drug was designed. • Composite PVA/conjugated CS-LVF nanofibres was fabricated by electrospinning. • Conjugated chitosan and composite nanofibres were characterized by various techniques. • Release profiles of drug were significantly improved in composite nanofibres containing drug conjugated chitosan.

  2. Polyvinyl alcohol composite nanofibres containing conjugated levofloxacin-chitosan for controlled drug release

    Energy Technology Data Exchange (ETDEWEB)

    Jalvandi, Javid, E-mail: Javid.jlv@gmail.com [CSIRO, Manufacturing Flagship, Bayview Ave, Clayton, Victoria 3168 (Australia); School of Fashion and Textiles, College of Design and Social Context, RMIT University, 25 Dawson Street, Brunswick, Victoria 3056 (Australia); White, Max, E-mail: tamrak@bigpond.com [School of Fashion and Textiles, College of Design and Social Context, RMIT University, 25 Dawson Street, Brunswick, Victoria 3056 (Australia); Gao, Yuan, E-mail: Yuan.Gao@csiro.au [CSIRO, Manufacturing Flagship, Bayview Ave, Clayton, Victoria 3168 (Australia); Truong, Yen Bach, E-mail: Yen.truong@csiro.au [CSIRO, Manufacturing Flagship, Bayview Ave, Clayton, Victoria 3168 (Australia); Padhye, Rajiv, E-mail: rajiv.padhye@rmit.edu.au [School of Fashion and Textiles, College of Design and Social Context, RMIT University, 25 Dawson Street, Brunswick, Victoria 3056 (Australia); Kyratzis, Ilias Louis, E-mail: Louis.kyratzis@csiro.au [CSIRO, Manufacturing Flagship, Bayview Ave, Clayton, Victoria 3168 (Australia)

    2017-04-01

    A range of biodegradable drug-nanofibres composite mats have been reported as drug delivery systems. However, their main disadvantage is the rapid release of the drug immediately after application. This paper reports an improved system based on the incorporation of drug conjugated-chitosan into polyvinyl alcohol (PVA) nanofibers. The results showed that controlled release of levofloxacin (LVF) could be achieved by covalently binding LVF to low molecular weight chitosan (CS) via a cleavable amide bond and then blending the conjugated CS with polyvinyl alcohol (PVA) nanofibres prior to electrospinning. PVA/LVF and PVA-CS/LVF nanofibres were fabricated as controls. The conjugated CS-LVF was characterized by FTIR, DSC, TGA and {sup 1}H NMR. Scanning electron microscopy (SEM) showed that the blended CS-PVA nanofibres had a reduced fibre diameter compared to the controls. Drug release profiles showed that burst release was decreased from 90% in the control PVA/LVF electrospun mats to 27% in the PVA/conjugated CS-LVF mats after 8 h in phosphate buffer at 37 °C. This slower release is due to the cleavable bond between LVF and CS that slowly hydrolysed over time at neutral pH. The results indicate that conjugation of the drug to the polymer backbone is an effective way of minimizing burst release behaviour and achieving sustained release of the drug, LVF. - Highlights: • A novel drug delivery system for controlled release of drug was designed. • Composite PVA/conjugated CS-LVF nanofibres was fabricated by electrospinning. • Conjugated chitosan and composite nanofibres were characterized by various techniques. • Release profiles of drug were significantly improved in composite nanofibres containing drug conjugated chitosan.

  3. Mechanical properties, biological activity and protein controlled release by poly(vinyl alcohol)–bioglass/chitosan–collagen composite scaffolds: A bone tissue engineering applications

    Energy Technology Data Exchange (ETDEWEB)

    Pon-On, Weeraphat, E-mail: fsciwpp@ku.ac.th [Department of Physics, Faculty of Science, Kasetsart University, Bangkok 10900 (Thailand); Charoenphandhu, Narattaphol; Teerapornpuntakit, Jarinthorn; Thongbunchoo, Jirawan; Krishnamra, Nateetip [Center of Calcium and Bone Research (COCAB), Faculty of Science, Mahidol University (Thailand); Department of Physiology, Faculty of Science, Mahidol University (Thailand); Tang, I-Ming [ThEP Center, Commission of Higher Education, 328 Si Ayutthaya Rd. (Thailand); Department of Materials Science, Faculty of Science, Kasetsart University, Bangkok 10900 (Thailand)

    2014-05-01

    In the present study, composite scaffolds made with different weight ratios (0.5:1, 1:1 and 2:1) of bioactive glass (15Ca:80Si:5P) (BG)/polyvinyl alcohol (PVA) (PVABG) and chitosan (Chi)/collagen (Col) (ChiCol) were prepared by three mechanical freeze–thaw followed by freeze-drying to obtain the porous scaffolds. The mechanical properties and the in vitro biocompatibility of the composite scaffolds to simulated body fluid (SBF) and to rat osteoblast-like UMR-106 cells were investigated. The results from the studies indicated that the porosity and compressive strength were controlled by the weight ratio of PVABG:ChiCol. The highest compressive modulus of the composites made was 214.64 MPa which was for the 1:1 weight ratio PVABG:ChiCol. Mineralization study in SBF showed the formation of apatite crystals on the PVABG:ChiCol surface after 7 days of incubation. In vitro cell availability and proliferation tests confirmed the osteoblast attachment and growth on the PVABG:ChiCol surface. MTT and ALP tests on the 1:1 weight ratio PVABG:ChiCol composite indicated that the UMR-106 cells were viable. Alkaline phosphatase activity was found to increase with increasing culturing time. In addition, we showed the potential of PVABG:ChiCol drug delivery through PBS solution studies. 81.14% of BSA loading had been achieved and controlled release for over four weeks was observed. Our results indicated that the PVABG:ChiCol composites, especially the 1:1 weight ratio composite exhibited significantly improved mechanical, mineral deposition, biological properties and controlled release. This made them potential candidates for bone tissue engineering applications. - Graphical abstract: Mechanical properties, biological activity and protein controlled release by poly(vinyl alcohol)–bioglass/chitosan–collagen composite scaffolds: A bone tissue engineering applications. - Highlights: • Preparation of PVABG:ChiCol hybrid composites and their bioactivities • Mechanical

  4. Mechanical properties, biological activity and protein controlled release by poly(vinyl alcohol)–bioglass/chitosan–collagen composite scaffolds: A bone tissue engineering applications

    International Nuclear Information System (INIS)

    Pon-On, Weeraphat; Charoenphandhu, Narattaphol; Teerapornpuntakit, Jarinthorn; Thongbunchoo, Jirawan; Krishnamra, Nateetip; Tang, I-Ming

    2014-01-01

    In the present study, composite scaffolds made with different weight ratios (0.5:1, 1:1 and 2:1) of bioactive glass (15Ca:80Si:5P) (BG)/polyvinyl alcohol (PVA) (PVABG) and chitosan (Chi)/collagen (Col) (ChiCol) were prepared by three mechanical freeze–thaw followed by freeze-drying to obtain the porous scaffolds. The mechanical properties and the in vitro biocompatibility of the composite scaffolds to simulated body fluid (SBF) and to rat osteoblast-like UMR-106 cells were investigated. The results from the studies indicated that the porosity and compressive strength were controlled by the weight ratio of PVABG:ChiCol. The highest compressive modulus of the composites made was 214.64 MPa which was for the 1:1 weight ratio PVABG:ChiCol. Mineralization study in SBF showed the formation of apatite crystals on the PVABG:ChiCol surface after 7 days of incubation. In vitro cell availability and proliferation tests confirmed the osteoblast attachment and growth on the PVABG:ChiCol surface. MTT and ALP tests on the 1:1 weight ratio PVABG:ChiCol composite indicated that the UMR-106 cells were viable. Alkaline phosphatase activity was found to increase with increasing culturing time. In addition, we showed the potential of PVABG:ChiCol drug delivery through PBS solution studies. 81.14% of BSA loading had been achieved and controlled release for over four weeks was observed. Our results indicated that the PVABG:ChiCol composites, especially the 1:1 weight ratio composite exhibited significantly improved mechanical, mineral deposition, biological properties and controlled release. This made them potential candidates for bone tissue engineering applications. - Graphical abstract: Mechanical properties, biological activity and protein controlled release by poly(vinyl alcohol)–bioglass/chitosan–collagen composite scaffolds: A bone tissue engineering applications. - Highlights: • Preparation of PVABG:ChiCol hybrid composites and their bioactivities • Mechanical

  5. Enhanced electrokinetic properties and antimicrobial activities of biodegradable chitosan/organo-bentonite composites.

    Science.gov (United States)

    Cabuk, Mehmet; Alan, Yusuf; Unal, H Ibrahim

    2017-04-01

    In this study, chitosan (CS), Na + -bentonite (Na + -BNT) and chitosan/organo-bentonite (CS/O-BNT) biodegradable composites having three different compositions were investigated. Electrokinetic measurements were examined in aqueous medium by taking the effects pH, electrolytes (NaCl and BaCl 2 ), surfactants (CTAB and SDS), and temperature into account. It was noticed that the initial ζ-potential of Na + -BNT shifted from negative (ζ=-35mV) to positive region (ζ=+13mV) with increasing polycationic CS content in the composite structure as aimed. Divalent 2:1 electrolyte (BaCl 2 ) caused to shift the ζ-potentials of all the dispersions to more positive regions. While the most negative effect on ζ-potential of the composites was reached with SDS, which reduced the value of ζ-potential to -39mV for CS(1)/O-BNT composite, the most positive effect was monitored with CTAB (ζ=+40mV) for CS(3)/O-BNT composite. Further, the composites were tested against various bacterial (Gram-positive and Gram-negative) and fungal microorganisms at various concentrations and results obtained were compared with the reference antibiotics and fungicide. According to inhibition zone values accomplished, antibacterial and antifungal activities of the CS/O-BNT composites are increased with increasing CS content as proportional with their positive ζ-potential values. Copyright © 2017 Elsevier Ltd. All rights reserved.

  6. Atmospheric plasma surface modifications of electrospun PCL/chitosan/PCL hybrid scaffolds by nozzle type plasma jets for usage of cell cultivation

    Science.gov (United States)

    Surucu, Seda; Masur, Kai; Turkoglu Sasmazel, Hilal; Von Woedtke, Thomas; Weltmann, Klaus Dieter

    2016-11-01

    This paper reports Ar gas, Ar + O2, Ar + O2 + N2 gas mixtures and dry air plasma modifications by atmospheric pressure argon driven kINPen and air driven Diener (PlasmaBeam) plasma jets to alter surface properties of three dimensional (3D), electrospun PCL/Chitosan/PCL layer by layer hybrid scaffolds to improve human fibroblast (MRC5) cell attachment and growth. The characterizations of the samples were done by contact angle (CA) measurements, scanning electron microscopy (SEM), X-Ray Photoelectron spectroscopy (XPS) analysis. The results showed that the plasma modification carried out under dry air and Ar + O2 + N2 gas mixtures were altered effectively the nanotopography and the functionality of the material surfaces. It was found that the samples treated with Ar + O2 + N2 gas mixtures for 1 min and dry air for 9 min have better hydrophilicity 78.9° ± 1.0 and 75.6° ± 0.1, respectively compared to the untreated samples (126.5°). Biocompatibility performance of the scaffolds was determined with alamarBlue (aB) assay and MTT assay methods, Giemsa staining, fluorescence microscope, confocal laser scanning microscope (CLSM) and scanning electron microscope (SEM) analyses. The results showed that plasma treated samples increased the hydrophilicity and oxygen functionality and topography of the surfaces significantly, thus affecting the cell viability and proliferation on/within scaffolds.

  7. Electrospun Chitosan-Gelatin Biopolymer Composite Nanofibers for Horseradish Peroxidase Immobilization in a Hydrogen Peroxide Biosensor

    Directory of Open Access Journals (Sweden)

    Siriwan Teepoo

    2017-10-01

    Full Text Available A biosensor based on chitosan-gelatin composite biopolymers nanofibers is found to be effective for the immobilization of horseradish peroxidase to detect hydrogen peroxide. The biopolymer nanofibers were fabricated by an electrospining technique. Upon optimization of synthesis parameters, biopolymers nanofibers, an average of 80 nm in diameter, were obtained and were then modified on the working electrode surface. The effects of the concentration of enzyme, pH, and concentration of the buffer and the working potential on the current response of the nanofibers-modified electrode toward hydrogen peroxide were optimized to obtain the maximal current response. The results found that horseradish peroxidase immobilization on chitosan-gelatin composite biopolymer nanofibers had advantages of fast response, excellent reproducibility, high stability, and showed a linear response to hydrogen peroxide in the concentration range from 0.1 to 1.7 mM with a detection limit of 0.05 mM and exhibited high sensitivity of 44 µA∙mM−1∙cm−2. The developed system was evaluated for analysis of disinfectant samples and showed good agreement between the results obtained by the titration method without significant differences at the 0.05 significance level. The proposed strategy based on chitosan-gelatin composite biopolymer nanofibers for the immobilization of enzymes can be extended for the development of other enzyme-based biosensors.

  8. Protein-adsorption and Ca-phosphate formation on chitosan-bioactive glass composite coatings

    Science.gov (United States)

    Wagener, V.; Boccaccini, A. R.; Virtanen, S.

    2017-09-01

    In the last years, chitosan-bioactive glass (BG) composites have been developed and investigated as bioactive coatings for orthopedic applications. The increase of bioactivity occurs due to the stimulation of calcium-phosphate/hydroxyapatite formation on the surface while the coating is degrading. In the present work, protein adsorption and its influence on calcium-phosphate precipitation was studied for the first time on such composite coatings. The experiments involved coating of 316L stainless steel substrates with chitosan (Ch) and chitosan-bioactive glass (Ch-BG) and immersion of the coated samples in two different bovine serum albumin (BSA) containing solutions, namely DI H2O (with pH adjusted to about 7.2 with diluted NaOH) and simulated body fluid (SBF). In order to investigate the influence of protein adsorption on calcium-phosphate precipitation, samples were also immersed in DI H2O and in SBF without BSA. Samples were analyzed by scanning electron microscopy (SEM), X-ray photoelectron spectroscopy (XPS) and time-of-flight secondary ion mass spectrometry (ToF-SIMS). Surface analysis revealed that adsorption of BSA takes place on all studied samples and that protein adsorption is influenced by the presence of Ca2+ and PO43- ions. Bioactivity in the form of hydroxyapatite pre-stage formation is significantly increased on Ch-BG composite coating as compared with bare stainless steel surface. However, calcium-phosphate precipitation in SBF is reduced by the presence of BSA.

  9. Adsorption of methyl orange from aqueous solution using chitosan/diatomite composite.

    Science.gov (United States)

    Zhao, Peng; Zhang, Runhu; Wang, Jianglin

    2017-04-01

    A novel chitosan/diatomite composite was prepared by a simple mixture in the mass ratio to remove methyl orange (MO) from aqueous media in this study. The composite adsorbent was characterized by Fourier transform infrared spectroscopy and scanning electron microscopy analysis. The parameters to influence the adsorption of MO were studied under such conditions as kinetics, adsorption isotherm, pH effect, and thermodynamics. The results revealed that adsorption of MO was initially rapid and the equilibrium time was reached after 40 min. The optimal value of the pH was 5.0 for better adsorption. The equilibrium data were well fitted to the Langmuir isotherm compared to the Freundlich isotherm, and exhibited the highest capacity and a removal rate of 88.37% under an initial dye concentration of 50 mg/L. The kinetic data were well described by the pseudo-second order model. The thermodynamic calculations revealed that the sorption was viable, spontaneous, and exothermic under the conditions studied. In addition, the chitosan/diatomite composite had good adsorption and desorption performance with respect to reusability after six cycles. These results showed that the chitosan/diatomite could be considered as a potential adsorbent for the removal of MO in aqueous solution.

  10. Electrochemical monitoring of the interaction between mitomycin C and DNA at chitosan--carbon nanotube composite modified electrodes

    OpenAIRE

    CANAVAR, Pembe Ece; EKŞİN, Ece; ERDEM, Arzum

    2015-01-01

    Single-walled carbon nanotube (CNT) and chitosan composite (chitosan*CNT) based sensors were developed as DNA biosensors, and then they were applied for electrochemical investigation of the interaction between the anticancer drug mitomycin C (MC) and DNA. The oxidation signals of MC and guanine were monitored before and after the interaction process by differential pulse voltammetry (DPV). The DPV results were in good agreement with those of electrochemical impedance spectroscopy (EIS)....

  11. Development of Useful Biomaterial for Bone Tissue Engineering by Incorporating Nano-Copper-Zinc Alloy (nCuZn in Chitosan/Gelatin/Nano-Hydroxyapatite (Ch/G/nHAp Scaffold

    Directory of Open Access Journals (Sweden)

    Juan Carlos Forero

    2017-10-01

    Full Text Available Ceramic and metallic nanoparticles can improve the mechanical and biological properties of polymeric scaffolds for bone tissue engineering (BTE. In this work, nanohydroxyapatite (nHAp and nano-copper-zinc alloy (nCuZn were added to a chitosan/gelatin (Ch/G scaffold in order to investigate the effects on morphological, physical, and biocompatibility properties. Scaffolds were fabricated by a freeze-drying technique using different pre-freezing temperatures. Microstructure and morphology were studied by scanning electron microscopy (SEM, glass transition (Tg was studied using differential scanning calorimetry (DSC, cell growth was estimated by MTT assay, and biocompatibility was examined in vitro and in vivo by histochemistry analyses. Scaffolds and nanocomposite scaffolds presented interconnected pores, high porosity, and pore size appropriate for BTE. Tg of Ch/G scaffolds was diminished by nanoparticle inclusion. Mouse embryonic fibroblasts (MEFs cells loaded in the Ch/G/nHAp/nCuZn nanocomposite scaffold showed suitable behavior, based on cell adhesion, cell growth, alkaline phosphatase (ALP activity as a marker of osteogenic differentiation, and histological in vitro cross sections. In vivo subcutaneous implant showed granulation tissue formation and new tissue infiltration into the scaffold. The favorable microstructure, coupled with the ability to integrate nanoparticles into the scaffold by freeze-drying technique and the biocompatibility, indicates the potential of this new material for applications in BTE.

  12. Hemocompatibility and cytocompatibility of pristine and plasma-treated silver-zeolite-chitosan composites

    Science.gov (United States)

    Taaca, Kathrina Lois M.; Vasquez, Magdaleno R.

    2018-02-01

    Silver-exchanged zeolite-chitosan (AgZ-Ch) composites with varying AgZ content were prepared by solvent casting and modified under argon (Ar) plasma excited by a 13.56 MHz radio frequency (RF) power source. Silver (Ag) was successfully incorporated in a natural zeolite host without losing its antibacterial activity against Escherichia coli and Staphylococcus aureus. The AgZ particles were incorporated into a chitosan matrix without making significant changes in the matrix structure. The composites also exhibited antibacterial sensitivity due to the inclusion of AgZ. Plasma treatment enhanced the surface wettability of polar and nonpolar test liquids of the composites. The average increase in total surface free energy after treatment was around 49% with the polar component having a significant change. Cytocompatibility tests showed at least 87% cell viability for pristine and plasma-treated composites comparable with supplemented RPMI as positive control. Hemocompatibility tests revealed that pristine composites does not promote hemolysis and the blood clotting ability is less than 10 min. Coupled with antibacterial property, the fabricated composites have promising biomedical applications.

  13. Preparation and characterization of chitosan/Aloe Vera composite nanofibers generated by electrostatic spinning

    Energy Technology Data Exchange (ETDEWEB)

    Ibrahim, Illani; Sekak, Khairunnadim Ahmad; Hasbullah, Norazurean [Center of Physics and Material Studies, Faculty of Applied Sciences, Universiti Teknologi Mara (UiTM) 40450 Shah Alam, Selangor (Malaysia)

    2015-08-28

    Researches on the fabrication of nanostructured based membrane have attracted great attention amongst scientists due to their wide potential applications on medical application. In this work, Chitosan and Aloe Vera sol-gel solution were electrospun using 20 kV DC supply at room temperature. Morphological structure and functional group of nanofibers were characterized using field emission scanning electron microscopy (FESEM) and Fourier-transform infrared spectroscopy (FT-IR) respectively. The optimum parameter obtained at 90% concentration of acetic acid, 0.3 ml/h of solution flow rate and 10 cm distance of nozzle to collector. The fiber diameters were analyzed using the ImageJ software. Average diameters of the Chitosan/Aloe Vera composite nanofibers is 183nm in ranges of 140–260nm.

  14. Development of Bioadhesive Chitosan Superporous Hydrogel Composite Particles Based Intestinal Drug Delivery System

    Directory of Open Access Journals (Sweden)

    Hitesh Chavda

    2013-01-01

    Full Text Available Bioadhesive superporous hydrogel composite (SPHC particles were developed for an intestinal delivery of metoprolol succinate and characterized for density, porosity, swelling, morphology, and bioadhesion studies. Chitosan and HPMC were used as bioadhesive and release retardant polymers, respectively. A 32 full factorial design was applied to optimize the concentration of chitosan and HPMC. The drug loaded bioadhesive SPHC particles were filled in capsule, and the capsule was coated with cellulose acetate phthalate and evaluated for drug content, in vitro drug release, and stability studies. To ascertain the drug release kinetics, the drug release profiles were fitted for mathematical models. The prepared system remains bioadhesive up to eight hours in intestine and showed Hixson-Crowell release with anomalous nonfickian type of drug transport. The application of SPHC polymer particles as a biomaterial carrier opens a new insight into bioadhesive drug delivery system and could be a future platform for other molecules for intestinal delivery.

  15. Preparation and characterization of chitosan/Aloe Vera composite nanofibers generated by electrostatic spinning

    International Nuclear Information System (INIS)

    Ibrahim, Illani; Sekak, Khairunnadim Ahmad; Hasbullah, Norazurean

    2015-01-01

    Researches on the fabrication of nanostructured based membrane have attracted great attention amongst scientists due to their wide potential applications on medical application. In this work, Chitosan and Aloe Vera sol-gel solution were electrospun using 20 kV DC supply at room temperature. Morphological structure and functional group of nanofibers were characterized using field emission scanning electron microscopy (FESEM) and Fourier-transform infrared spectroscopy (FT-IR) respectively. The optimum parameter obtained at 90% concentration of acetic acid, 0.3 ml/h of solution flow rate and 10 cm distance of nozzle to collector. The fiber diameters were analyzed using the ImageJ software. Average diameters of the Chitosan/Aloe Vera composite nanofibers is 183nm in ranges of 140–260nm

  16. Preparation and characterization of chitosan/Aloe Vera composite nanofibers generated by electrostatic spinning

    Science.gov (United States)

    Ibrahim, Illani; Sekak, Khairunnadim Ahmad; Hasbullah, Norazurean

    2015-08-01

    Researches on the fabrication of nanostructured based membrane have attracted great attention amongst scientists due to their wide potential applications on medical application. In this work, Chitosan and Aloe Vera sol-gel solution were electrospun using 20 kV DC supply at room temperature. Morphological structure and functional group of nanofibers were characterized using field emission scanning electron microscopy (FESEM) and Fourier-transform infrared spectroscopy (FT-IR) respectively. The optimum parameter obtained at 90% concentration of acetic acid, 0.3 ml/h of solution flow rate and 10 cm distance of nozzle to collector. The fiber diameters were analyzed using the ImageJ software. Average diameters of the Chitosan/Aloe Vera composite nanofibers is 183nm in ranges of 140-260nm.

  17. Development of Composite Scaffolds for Load Bearing Segmental Bone Defects

    Science.gov (United States)

    2013-07-01

    osteoarthritis [24], generic infections, congenital deformity corrections [25], pathological degenerative bone destruction and other degenerative diseases [20...resection and reconstruction), osteoporosis, osteoarthritis , generic infections, congenital deformity corrections, pathological degenerative bone...the construct. Pore size/ Porosity: Quality of a bone scaffold to have pore size and porosity percent similar to established guidelines . Ideal

  18. Surface modification of nanofibrous polycaprolactone/gelatin composite scaffold by collagen type I grafting for skin tissue engineering

    International Nuclear Information System (INIS)

    Gautam, Sneh; Chou, Chia-Fu; Dinda, Amit K.; Potdar, Pravin D.; Mishra, Narayan C.

    2014-01-01

    In the present study, a tri-polymer polycaprolactone (PCL)/gelatin/collagen type I composite nanofibrous scaffold has been fabricated by electrospinning for skin tissue engineering and wound healing applications. Firstly, PCL/gelatin nanofibrous scaffold was fabricated by electrospinning using a low cost solvent mixture [chloroform/methanol for PCL and acetic acid (80% v/v) for gelatin], and then the nanofibrous PCL/gelatin scaffold was modified by collagen type I (0.2–1.5 wt.%) grafting. Morphology of the collagen type I-modified PCL/gelatin composite scaffold that was analyzed by field emission scanning electron microscopy (FE-SEM), showed that the fiber diameter was increased and pore size was decreased by increasing the concentration of collagen type I. Fourier transform infrared (FT-IR) spectroscopy and thermogravimetric (TG) analysis indicated the surface modification of PCL/gelatin scaffold by collagen type I immobilization on the surface of the scaffold. MTT assay demonstrated the viability and high proliferation rate of L929 mouse fibroblast cells on the collagen type I-modified composite scaffold. FE-SEM analysis of cell-scaffold construct illustrated the cell adhesion of L929 mouse fibroblasts on the surface of scaffold. Characteristic cell morphology of L929 was also observed on the nanofiber mesh of the collagen type I-modified scaffold. Above results suggest that the collagen type I-modified PCL/gelatin scaffold was successful in maintaining characteristic shape of fibroblasts, besides good cell proliferation. Therefore, the fibroblast seeded PCL/gelatin/collagen type I composite nanofibrous scaffold might be a potential candidate for wound healing and skin tissue engineering applications. - Highlights: • PCL/gelatin/collagen type I scaffold was fabricated for skin tissue engineering. • PCL/gelatin/collagen type I scaffold showed higher fibroblast growth than PCL/gelatin one. • PCL/gelatin/collagen type I might be one of the ideal scaffold for

  19. Electrospun Polyhydroxybutyrate and Poly(L-lactide-co-ε-caprolactone Composites as Nanofibrous Scaffolds

    Directory of Open Access Journals (Sweden)

    Donraporn Daranarong

    2014-01-01

    Full Text Available Electrospinning can produce nanofibrous scaffolds that mimic the architecture of the extracellular matrix and support cell attachment for tissue engineering applications. In this study, fibrous membranes of polyhydroxybutyrate (PHB with various loadings of poly(L-lactide-co-ε-caprolactone (PLCL were successfully prepared by electrospinning. In comparison to PLCL scaffolds, PLCL blends with PHB exhibited more irregular fibre diameter distributions and higher average fibre diameters but there were no significant differences in pore size. PLCL/PHB scaffolds were more hydrophilic (<120° with significantly reduced tensile strength (ca. 1 MPa compared to PLCL scaffolds (150.9±2.8∘ and 5.8±0.5 MPa. Increasing PLCL loading in PHB/PLCL scaffolds significantly increased the extension at break, (4–6-fold. PLCL/PHB scaffolds supported greater adhesion and proliferation of olfactory ensheathing cells (OECs than those exhibiting asynchronous growth on culture plates. Mitochondrial activity of cells cultivated on the electrospun blended membranes was enhanced compared to those grown on PLCL and PHB scaffolds (212, 179, and 153%, resp.. Analysis showed that PLCL/PHB nanofibrous membranes promoted cell cycle progression and reduced the onset of necrosis. Thus, electrospun PLCL/PHB composites promoted adhesion and proliferation of OECs when compared to their individual PLCL and PHB components suggesting potential in the repair and engineering of nerve tissue.

  20. Incorporating pTGF-β1/calcium phosphate nanoparticles with fibronectin into 3-dimensional collagen/chitosan scaffolds: Efficient, sustained gene delivery to stem cells for chondrogenic differentiation

    Directory of Open Access Journals (Sweden)

    X Cao

    2012-02-01

    Full Text Available The objective of this study was to prepare a 3-dimensional nanoparticle gene delivery system (3D-NGDS based on collagen/chitosan scaffolds, in which plasmid transforming growth factor beta 1 (TGF-β1/calcium phosphate nanoparticles mixed with fibronectin (FN were used to transfect mesenchymal stem cells (MSCs. Scanning electron microscopy was used to characterise the microstructure of 3-dimensional collagen/chitosan scaffolds. An analysis performed to quantify the TGF-b1 concentrations in MSC cultures revealed that the MSCs transfected with the 3D-NGDS showed remarkably high levels of TGF-b1 over long periods, retaining a concentration of TGF-b1 of approximately 10 ng/mL within two weeks, with the highest level (12.6 ng/mL being observed on the 6th day. An immunohistochemistry analysis for collagen type II revealed that much higher production of collagen II from the 9th to 15th day was observed in the 3D-NGDS-transfected MSCs than that in MSCs transfected by the Lipofectamine 2000 method. The glycosaminoglycan content of the 3D-NGDS was comparable to those treated with TGF-β1 as well as TGF-β1 plus dexamethasone, and was significantly higher than those treated with free plasmid and Lipofectamine 2000. A remarkable type I collagen expression inhibition of the 3D-NGDS at day 21 was observed via ELISA. These results suggested that transfection with the 3D-NGDS could successfully induce MSC chondrogenic differentiation in vitro without dexamethasone. In summary, the 3D-NGDS could be developed into a promising alternative method to transfer exogenous nucleic acid to MSCs in clinical trials.