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Rui L. Reis - One of the best experts on this subject based on the ideXlab platform.
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Bioceramics for Osteochondral Tissue Engineering and Regeneration.
Advances in experimental medicine and biology, 2018Co-Authors: S Pina, Rita Rebelo, Vitor M. Correlo, J. Miguel Oliveira, Rui L. ReisAbstract:Considerable advances in Tissue engineering and regeneration have been accomplished over the last decade. Bioceramics have been developed to repair, reconstruct, and substitute diseased parts of the body and to promote Tissue healing as an alternative to metallic implants. Applications embrace hip, knee, and ligament repair and replacement, maxillofacial reconstruction and augmentation, spinal fusion, bone filler, and repair of periodontal diseases. Bioceramics are well-known for their superior wear resistance, high stiffness, resistance to oxidation, and low coefficient of friction. These specially designed biomaterials are grouped in natural bioceramics (e.g., coral-derived apatites), and synthetic bioceramics, namely bioinert ceramics (e.g., alumina and zirconia), bioactive glasses and glass ceramics, and bioresorbable calcium phosphates-based materials. Physicochemical, mechanical, and biological properties, as well as bioceramics applications in diverse fields of Tissue engineering are presented herein. Ongoing clinical trials using bioceramics in Osteochondral Tissue are also considered. Based on the stringent requirements for clinical applications, prospects for the development of advanced functional bioceramics for Tissue engineering are highlighted for the future.
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layered scaffolds for Osteochondral Tissue engineering
Advances in Experimental Medicine and Biology, 2018Co-Authors: D R Pereira, Rui L. Reis, Miguel J OliveiraAbstract:Despite huge efforts, Tissue engineers and orthopedic surgeons still face a great challenge to functionally repair Osteochondral (OC) defects. Nevertheless, over the past decade great progress has been made to find suitables strategies towards OC regeneration. In the clinics, some Osteochondral Tissue engineering (OCTE) strategies have already been applied although with some incongruous outcomes as OC Tissue is complex in its architecture and function. In this chapter, we have summarized current OCTE strategies that are focused on hierarchical scaffold design, mainly layered scaffolds. Most suitable candidates towards functional regeneration of OC Tissues are envisaged from monophasic to layered scaffolds. Herein is documented a variety of strategies with their intrinsic properties for further application as bare scaffolds or in combination with biologics. Both in vitro and in vivo approaches have been thoroughly studied aiming at functional OC regeneration. The most noteworthy studies in OC regeneration developed within the past 5 years are herein documented as well as some current clinical trials.
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nanoparticles based systems for Osteochondral Tissue engineering
Advances in Experimental Medicine and Biology, 2018Co-Authors: Isabel Oliveira, Miguel J Oliveira, Silvia Vieira, Rui L. ReisAbstract:Osteochondral lesions represent one of the major causes of disabilities in the world. These defects are due to degenerative or inflammatory arthritis, but both affect the articular cartilage and the underlying subchondral bone. Defects from trauma or degenerative pathology frequently cause severe pain, joint deformity, and loss of joint motion. Osteochondral defects are a significant challenge in orthopedic surgery, due to the cartilage complexity and unique structure, as well as its exposure to high pressure and motion. Although there are treatments routinely performed in the clinical practice, they present several limitations. Tissue engineering can be a suitable alternative for Osteochondral defects since bone and cartilage engineering had experienced a notable advance over the years. Allied with nanotechnology, Osteochondral Tissue engineering (OCTE) can be leveled up, being possible to create advanced structures similar to the OC Tissue. In this chapter, the current strategies using nanoparticles-based systems are overviewed. The results of the studies herein considered confirm that advanced nanomaterials will undoubtedly play a crucial role in the design of strategies for treatment of Osteochondral defects in the near future.
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the use of electrospinning technique on Osteochondral Tissue engineering
Advances in Experimental Medicine and Biology, 2018Co-Authors: Marta R Casanova, Rui L. Reis, Albino Martins, Nuno M NevesAbstract:Electrospinning, an electrostatic fiber fabrication technique, has attracted significant interest in recent years due to its versatility and ability to produce highly tunable nanofibrous meshes. These nanofibrous meshes have been investigated as promising Tissue engineering scaffolds since they mimic the scale and morphology of the native extracellular matrix. The sub-micron diameter of fibers produced by this process presents various advantages like the high surface area to volume ratio, tunable porosity, and the ability to manipulate the nanofiber composition in order to get desired properties and functionality. Electrospun fibers can be oriented or arranged randomly, giving control over both mechanical properties and the biological response to the fibrous scaffold. Moreover, bioactive molecules can be integrated with the electrospun nanofibrous scaffolds in order to improve the cellular response. This chapter presents an overview of the developments on electrospun polymer nanofibers including processing, structure, and their applications in the field of Osteochondral Tissue engineering.
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micro nano scaffolds for Osteochondral Tissue engineering
Advances in Experimental Medicine and Biology, 2018Co-Authors: Albino Martins, Rui L. Reis, Nuno M NevesAbstract:To develop an Osteochondral Tissue regeneration strategy it is extremely important to take into account the multiscale organization of the natural extracellular matrix. The structure and gradients of organic and inorganic components present in the cartilage and bone Tissues must be considered together. Another critical aspect is an efficient interface between both Tissues. So far, most of the approaches were focused on the development of multilayer or stratified scaffolds which resemble the structural composition of bone and cartilage, not considering in detail a transitional interface layer. Typically, those scaffolds have been produced by the combined use of two or more processing techniques (microtechnologies and nanotechnologies) and materials (organic and inorganic). A significant number of works was focused on either cartilage or bone, but there is a growing interest in the development of the Osteochondral interface and in Tissue engineering models of composite constructs that can mimic the cartilage/bone Tissues. The few works that give attention to the interface between cartilage and bone, as well as to the biochemical gradients observed at the Osteochondral unit, are also herein described.
Anderson Oliveira Lobo - One of the best experts on this subject based on the ideXlab platform.
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cell viability of porous poly d l lactic acid vertically aligned carbon nanotubes nanohydroxyapatite scaffolds for Osteochondral Tissue engineering
Materials, 2019Co-Authors: Thiago Domingues Stocco, Eliane Antonioli, Conceicao De Maria Vaz Elias, Bruno V M Rodrigues, Idalia A W B Siqueira, Mario Ferretti, Fernanda Roberta Marciano, Anderson Oliveira LoboAbstract:Treatment of articular cartilage lesions remains an important challenge. Frequently the bone located below the cartilage is also damaged, resulting in defects known as Osteochondral lesions. Tissue engineering has emerged as a potential approach to treat cartilage and Osteochondral defects. The principal challenge of Osteochondral Tissue engineering is to create a scaffold with potential to regenerate both cartilage and the subchondral bone involved, considering the intrinsic properties of each Tissue. Recent nanocomposites based on the incorporation of nanoscale fillers into polymer matrix have shown promising results for the treatment of Osteochondral defects. In this present study, it was performed using the recently developed methodologies (electrodeposition and immersion in simulated body fluid) to obtain porous superhydrophilic poly(d,l-lactic acid)/vertically aligned carbon nanotubes/nanohydroxyapatite (PDLLA/VACNT-O:nHAp) nanocomposite scaffolds, to analyze cell behavior and gene expression of chondrocytes, and then assess the applicability of this nanobiomaterial for Osteochondral regenerative medicine. The results demonstrate that PDLLA/VACNT-O:nHAp nanocomposite supports chondrocytes adhesion and decreases type I Collagen mRNA expression. Therefore, these findings suggest the possibility of novel nanobiomaterial as a scaffold for Osteochondral Tissue engineering applications.
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Cell Viability of Porous Poly(d,l-lactic acid)/Vertically Aligned Carbon Nanotubes/Nanohydroxyapatite Scaffolds for Osteochondral Tissue Engineering
'MDPI AG', 2019Co-Authors: Thiago Domingues Stocco, Eliane Antonioli, Conceicao De Maria Vaz Elias, Bruno V M Rodrigues, Mario Ferretti, Fernanda Roberta Marciano, Idália Aparecida Waltrick De Brito Siqueira, Anderson Oliveira LoboAbstract:Treatment of articular cartilage lesions remains an important challenge. Frequently the bone located below the cartilage is also damaged, resulting in defects known as Osteochondral lesions. Tissue engineering has emerged as a potential approach to treat cartilage and Osteochondral defects. The principal challenge of Osteochondral Tissue engineering is to create a scaffold with potential to regenerate both cartilage and the subchondral bone involved, considering the intrinsic properties of each Tissue. Recent nanocomposites based on the incorporation of nanoscale fillers into polymer matrix have shown promising results for the treatment of Osteochondral defects. In this present study, it was performed using the recently developed methodologies (electrodeposition and immersion in simulated body fluid) to obtain porous superhydrophilic poly(d,l-lactic acid)/vertically aligned carbon nanotubes/nanohydroxyapatite (PDLLA/VACNT-O:nHAp) nanocomposite scaffolds, to analyze cell behavior and gene expression of chondrocytes, and then assess the applicability of this nanobiomaterial for Osteochondral regenerative medicine. The results demonstrate that PDLLA/VACNT-O:nHAp nanocomposite supports chondrocytes adhesion and decreases type I Collagen mRNA expression. Therefore, these findings suggest the possibility of novel nanobiomaterial as a scaffold for Osteochondral Tissue engineering applications
Antonios G Mikos - One of the best experts on this subject based on the ideXlab platform.
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a rabbit femoral condyle defect model for assessment of Osteochondral Tissue regeneration
Tissue Engineering Part C-methods, 2020Co-Authors: Jason L Guo, Yu Seon Kim, Elysse A Orchard, Jeroen J J P Van Den Beucken, John A Jansen, Mark E Wong, Antonios G MikosAbstract:Osteochondral Tissue repair represents a common clinical need, with multiple approaches in Tissue engineering and regenerative medicine being investigated for the repair of defects of articular cartilage and subchondral bone. A full thickness rabbit femoral condyle defect is a clinically relevant model of an articulating and load bearing joint surface for the investigation of Osteochondral Tissue repair by various cell-, biomolecule-, and biomaterial-based implants. In this protocol, we describe the methodology and 1.5- to 2-h surgical procedure for the generation of a reproducible, full thickness defect for construct implantation in the rabbit medial femoral condyle. Furthermore, we describe a step-by-step procedure for Osteochondral Tissue collection and the assessment of Tissue formation using standardized histological, radiological, mechanical, and biochemical analytical techniques. This protocol illustrates the critical steps for reproducibility and minimally invasive surgery as well as applications to evaluate the efficacy of cartilage and bone Tissue engineering implants, with emphasis on the usage of histological and radiological measures of Tissue growth. Impact statement Although multiple surgical techniques have been developed for the treatment of Osteochondral defects, repairing the Tissues to their original state remains an unmet need. Such limitations have thus prompted the development of various constructs for Osteochondral Tissue regeneration. An in vivo model that is both clinically relevant and economically practical is necessary to evaluate the efficacy of different Tissue engineered constructs. In this article, we present a full thickness rabbit femoral condyle defect model and describe the analytical techniques to assess the regeneration of Osteochondral Tissue.
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click functionalized Tissue specific hydrogels for Osteochondral Tissue engineering
Journal of Biomedical Materials Research Part A, 2020Co-Authors: Jason L Guo, Yu Seon Kim, Brandon T Smith, Virginia Y Xie, Emma Watson, Gang Bao, Antonios G MikosAbstract:Osteochondral repair requires the induction of both articular cartilage and subchondral bone development, necessitating the presentation of multiple Tissue-specific cues for these highly distinct Tissues. To provide a singular hydrogel system for the repair of either Tissue type, we have developed biofunctionalized, mesenchymal stem cell-laden hydrogels that can present in situ biochemical cues for either chondrogenesis or osteogenesis by simple click modification of a crosslinker, poly(glycolic acid)-poly(ethylene glycol)-poly(glycolic acid)-di(but-2-yne-1,4-dithiol) (PdBT). After modifying PdBT with either cartilage-specific biomolecules (N-cadherin peptide, chondroitin sulfate) or bone-specific biomolecules (bone marrow homing peptide 1, glycine-histidine-lysine peptide), the biofunctionalized, PdBT-crosslinked hydrogels can selectively promote the desired bone- or cartilage-like matrix synthesis and Tissue-specific gene expression, with effects dependent on both biomolecule selection and concentration. Our findings establish the versatility of this click functionalized hydrogel system as well as its ability to promote in vitro development of Osteochondral Tissue phenotypes.
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fabrication and mechanical characterization of 3d printed vertical uniform and gradient scaffolds for bone and Osteochondral Tissue engineering
Acta Biomaterialia, 2019Co-Authors: Sean M Bittner, Brandon T Smith, Luis Diazgomez, Carrigan D Hudgins, Anthony J Melchiorri, David Scott, J Fisher, Antonios G MikosAbstract:Abstract Recent developments in 3D printing (3DP) research have led to a variety of scaffold designs and techniques for Osteochondral Tissue engineering; however, the simultaneous incorporation of multiple types of gradients within the same construct remains a challenge. Herein, we describe the fabrication and mechanical characterization of porous poly(e-caprolactone) (PCL) and PCL-hydroxyapatite (HA) scaffolds with incorporated vertical porosity and ceramic content gradients via a multimaterial extrusion 3DP system. Scaffolds of 0 wt% HA (PCL), 15 wt% HA (HA15), or 30 wt% HA (HA30) were fabricated with uniform composition and porosity (using 0.2 mm, 0.5 mm, or 0.9 mm on-center fiber spacing), uniform composition and gradient porosity, and gradient composition (PCL-HA15-HA30) and porosity. Micro-CT imaging and porosity analysis demonstrated the ability to incorporate both vertical porosity and pore size gradients and a ceramic gradient, which collectively recapitulate gradients found in native Osteochondral Tissues. Uniaxial compression testing demonstrated an inverse relationship between porosity, ϕ, and compressive modulus, E, and yield stress, σy, for uniform porosity scaffolds, however, no differences were observed as a result of ceramic incorporation. All scaffolds demonstrated compressive moduli within the appropriate range for trabecular bone, with average moduli between 86 ± 14–220 ± 26 MPa. Uniform porosity and pore size scaffolds for all ceramic levels had compressive moduli between 205 ± 37–220 ± 26 MPa, 112 ± 13–118 ± 23 MPa, and 86 ± 14–97 ± 8 MPa respectively for porosities ranging between 14 ± 4–20 ± 6%, 36 ± 3–43 ± 4%, and 54 ± 2–57 ± 2%, with the moduli and yield stresses of low porosity scaffolds being significantly greater (p 0.05) to those of the highest porosity uniform scaffolds (porosity gradient scaffolds 98 ± 23–107 ± 6 MPa, and 102 ± 7 MPa for dual composition/porosity gradient scaffolds), indicating that these properties are more heavily influenced by the weakest section of the gradient. The compression data for uniform scaffolds were also readily modeled, yielding scaling laws of the form E ∼ (1 − ϕ)1.27 and σy ∼ (1 − ϕ)1.37, which demonstrated that the compressive properties evaluated in this study were well-aligned with expectations from previous literature and were readily modeled with good fidelity independent of polymer scaffold geometry and ceramic content. All uniform scaffolds were similarly deformed and recovered despite different porosities, while the large-pore sections of porosity gradient scaffolds were significantly more deformed than all other groups, indicating that porosity may not be an independent factor in determining strain recovery. Moving forward, the technique described here will serve as the template for more complex multimaterial constructs with bioactive cues that better match native Tissue physiology and promote Tissue regeneration. Statement of significance This manuscript describes the fabrication and mechanical characterization of “dual” porosity/ceramic content gradient scaffolds produced via a multimaterial extrusion 3D printing system for Osteochondral Tissue engineering. Such scaffolds are designed to better address the simultaneous gradients in architecture and mineralization found in native Osteochondral Tissue. The results of this study demonstrate that this technique may serve as a template for future advances in 3D printing technology that may better address the inherent complexity in such heterogeneous Tissues.
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Osteochondral Tissue regeneration using a bilayered composite hydrogel with modulating dual growth factor release kinetics in a rabbit model
Journal of Controlled Release, 2013Co-Authors: Kyobum Kim, John A Jansen, Mark E Wong, Antonios G Mikos, Johnny Lam, Patrick P Spicer, Aline Lueckgen, Yasuhiko Tabata, Kurtis F KasperAbstract:Biodegradable oligo(poly(ethylene glycol) fumarate) (OPF) composite hydrogels have been investigated for the delivery of growth factors (GFs) with the aid of gelatin microparticles (GMPs) and stem cell populations for Osteochondral Tissue regeneration. In this study, a bilayered OPF composite hydrogel that mimics the distinctive hierarchical structure of native Osteochondral Tissue was utilized to investigate the effect of transforming growth factor-β3 (TGF-β3) with varying release kinetics and/or insulin-like growth factor-1 (IGF-1) on Osteochondral Tissue regeneration in a rabbit full-thickness Osteochondral defect model. The four groups investigated included (i) a blank control (no GFs), (ii) GMP-loaded IGF-1 alone, (iii) GMP-loaded IGF-1 and gel-loaded TGF-β3, and (iv) GMP-loaded IGF-1 and GMP-loaded TGF-β3 in OPF composite hydrogels. The results of an in vitro release study demonstrated that TGF-β3 release kinetics could be modulated by the GF incorporation method. At 12 weeks post-implantation, the quality of Tissue repair in both chondral and subchondral layers was analyzed based on quantitative histological scoring. All groups incorporating GFs resulted in a significant improvement in cartilage morphology compared to the control. Single delivery of IGF-1 showed higher scores in subchondral bone morphology as well as chondrocyte and glycosaminoglycan amount in adjacent cartilage Tissue when compared to a dual delivery of IGF-1 and TGF-β3, independent of the TGF-β3 release kinetics. The results suggest that although the dual delivery of TGF-β3 and IGF-1 may not synergistically enhance the quality of engineered Tissue, the delivery of IGF-1 alone from bilayered composite hydrogels positively affects Osteochondral Tissue repair and holds promise for Osteochondral Tissue engineering applications.
Thiago Domingues Stocco - One of the best experts on this subject based on the ideXlab platform.
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cell viability of porous poly d l lactic acid vertically aligned carbon nanotubes nanohydroxyapatite scaffolds for Osteochondral Tissue engineering
Materials, 2019Co-Authors: Thiago Domingues Stocco, Eliane Antonioli, Conceicao De Maria Vaz Elias, Bruno V M Rodrigues, Idalia A W B Siqueira, Mario Ferretti, Fernanda Roberta Marciano, Anderson Oliveira LoboAbstract:Treatment of articular cartilage lesions remains an important challenge. Frequently the bone located below the cartilage is also damaged, resulting in defects known as Osteochondral lesions. Tissue engineering has emerged as a potential approach to treat cartilage and Osteochondral defects. The principal challenge of Osteochondral Tissue engineering is to create a scaffold with potential to regenerate both cartilage and the subchondral bone involved, considering the intrinsic properties of each Tissue. Recent nanocomposites based on the incorporation of nanoscale fillers into polymer matrix have shown promising results for the treatment of Osteochondral defects. In this present study, it was performed using the recently developed methodologies (electrodeposition and immersion in simulated body fluid) to obtain porous superhydrophilic poly(d,l-lactic acid)/vertically aligned carbon nanotubes/nanohydroxyapatite (PDLLA/VACNT-O:nHAp) nanocomposite scaffolds, to analyze cell behavior and gene expression of chondrocytes, and then assess the applicability of this nanobiomaterial for Osteochondral regenerative medicine. The results demonstrate that PDLLA/VACNT-O:nHAp nanocomposite supports chondrocytes adhesion and decreases type I Collagen mRNA expression. Therefore, these findings suggest the possibility of novel nanobiomaterial as a scaffold for Osteochondral Tissue engineering applications.
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Cell Viability of Porous Poly(d,l-lactic acid)/Vertically Aligned Carbon Nanotubes/Nanohydroxyapatite Scaffolds for Osteochondral Tissue Engineering
'MDPI AG', 2019Co-Authors: Thiago Domingues Stocco, Eliane Antonioli, Conceicao De Maria Vaz Elias, Bruno V M Rodrigues, Mario Ferretti, Fernanda Roberta Marciano, Idália Aparecida Waltrick De Brito Siqueira, Anderson Oliveira LoboAbstract:Treatment of articular cartilage lesions remains an important challenge. Frequently the bone located below the cartilage is also damaged, resulting in defects known as Osteochondral lesions. Tissue engineering has emerged as a potential approach to treat cartilage and Osteochondral defects. The principal challenge of Osteochondral Tissue engineering is to create a scaffold with potential to regenerate both cartilage and the subchondral bone involved, considering the intrinsic properties of each Tissue. Recent nanocomposites based on the incorporation of nanoscale fillers into polymer matrix have shown promising results for the treatment of Osteochondral defects. In this present study, it was performed using the recently developed methodologies (electrodeposition and immersion in simulated body fluid) to obtain porous superhydrophilic poly(d,l-lactic acid)/vertically aligned carbon nanotubes/nanohydroxyapatite (PDLLA/VACNT-O:nHAp) nanocomposite scaffolds, to analyze cell behavior and gene expression of chondrocytes, and then assess the applicability of this nanobiomaterial for Osteochondral regenerative medicine. The results demonstrate that PDLLA/VACNT-O:nHAp nanocomposite supports chondrocytes adhesion and decreases type I Collagen mRNA expression. Therefore, these findings suggest the possibility of novel nanobiomaterial as a scaffold for Osteochondral Tissue engineering applications
Conceicao De Maria Vaz Elias - One of the best experts on this subject based on the ideXlab platform.
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cell viability of porous poly d l lactic acid vertically aligned carbon nanotubes nanohydroxyapatite scaffolds for Osteochondral Tissue engineering
Materials, 2019Co-Authors: Thiago Domingues Stocco, Eliane Antonioli, Conceicao De Maria Vaz Elias, Bruno V M Rodrigues, Idalia A W B Siqueira, Mario Ferretti, Fernanda Roberta Marciano, Anderson Oliveira LoboAbstract:Treatment of articular cartilage lesions remains an important challenge. Frequently the bone located below the cartilage is also damaged, resulting in defects known as Osteochondral lesions. Tissue engineering has emerged as a potential approach to treat cartilage and Osteochondral defects. The principal challenge of Osteochondral Tissue engineering is to create a scaffold with potential to regenerate both cartilage and the subchondral bone involved, considering the intrinsic properties of each Tissue. Recent nanocomposites based on the incorporation of nanoscale fillers into polymer matrix have shown promising results for the treatment of Osteochondral defects. In this present study, it was performed using the recently developed methodologies (electrodeposition and immersion in simulated body fluid) to obtain porous superhydrophilic poly(d,l-lactic acid)/vertically aligned carbon nanotubes/nanohydroxyapatite (PDLLA/VACNT-O:nHAp) nanocomposite scaffolds, to analyze cell behavior and gene expression of chondrocytes, and then assess the applicability of this nanobiomaterial for Osteochondral regenerative medicine. The results demonstrate that PDLLA/VACNT-O:nHAp nanocomposite supports chondrocytes adhesion and decreases type I Collagen mRNA expression. Therefore, these findings suggest the possibility of novel nanobiomaterial as a scaffold for Osteochondral Tissue engineering applications.
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Cell Viability of Porous Poly(d,l-lactic acid)/Vertically Aligned Carbon Nanotubes/Nanohydroxyapatite Scaffolds for Osteochondral Tissue Engineering
'MDPI AG', 2019Co-Authors: Thiago Domingues Stocco, Eliane Antonioli, Conceicao De Maria Vaz Elias, Bruno V M Rodrigues, Mario Ferretti, Fernanda Roberta Marciano, Idália Aparecida Waltrick De Brito Siqueira, Anderson Oliveira LoboAbstract:Treatment of articular cartilage lesions remains an important challenge. Frequently the bone located below the cartilage is also damaged, resulting in defects known as Osteochondral lesions. Tissue engineering has emerged as a potential approach to treat cartilage and Osteochondral defects. The principal challenge of Osteochondral Tissue engineering is to create a scaffold with potential to regenerate both cartilage and the subchondral bone involved, considering the intrinsic properties of each Tissue. Recent nanocomposites based on the incorporation of nanoscale fillers into polymer matrix have shown promising results for the treatment of Osteochondral defects. In this present study, it was performed using the recently developed methodologies (electrodeposition and immersion in simulated body fluid) to obtain porous superhydrophilic poly(d,l-lactic acid)/vertically aligned carbon nanotubes/nanohydroxyapatite (PDLLA/VACNT-O:nHAp) nanocomposite scaffolds, to analyze cell behavior and gene expression of chondrocytes, and then assess the applicability of this nanobiomaterial for Osteochondral regenerative medicine. The results demonstrate that PDLLA/VACNT-O:nHAp nanocomposite supports chondrocytes adhesion and decreases type I Collagen mRNA expression. Therefore, these findings suggest the possibility of novel nanobiomaterial as a scaffold for Osteochondral Tissue engineering applications
