RESUMO
One of the critical challenges that scaffolding faces in the organ and tissue regeneration field lies in mimicking the structure, and the chemical and biological properties of natural tissue. A high-level control over the architecture, mechanical properties and composition of the materials in contact with cells is essential to overcome such challenge. Therefore, definition of the method, materials and parameters for the production of scaffolds during the fabrication stage is critical. With the recent emergence of rapid prototyping (RP), it is now possible to create three-dimensional (3D) scaffolds with the essential characteristics for the proliferation and regeneration of tissues, such as porosity, mechanical strength, pore size and pore interconnectivity, and biocompatibility. In this study, we employed 3D bioplotting, a RP technology, to fabricate scaffolds made from (i) pure polycaprolactone (PCL) and (ii) a composite based on PCL and ceramic micro-powder. The ceramics used for the composite were bovine bone filling Nukbone® (NKB), and hydroxyapatite (HA) with 5%, 10% or 20% wt. CONTENT: The scaffolds were fabricated in a cellular lattice structure (i.e. meshing mode) using a 0/90° lay down pattern with a continuous contour filament in order to achieve interconnected porous reticular structures. We varied the temperature, as well as injection speed and pressure during the bioplotting process to achieve scaffolds with pore size ranging between 200 and 400µm and adequate mechanical stability. The resulting scaffolds had an average pore size of 323µm and an average porosity of 32%. Characterization through ATR-FTIR revealed the presence of the characteristic bands of hydroxyapatite in the PCL matrix, and presented an increase of the intensity of the phosphate and carbonyl bands as the ceramic content increased. The bioplotted 3D scaffolds have a Young's modulus (E) in the range between 0.121 and 0.171GPa, which is compatible with the modulus of natural bone. PCL/NKB scaffolds, particularly 10NKBP (10% NKB wt.) exhibited the highest proliferation optical density, demonstrating an evident osteoconductive effect when cultured in Dulbecco's Modified Eagle Medium (DMEM). Scanning electron microscopy (SEM) confirmed osteoblast anchorage to all composite scaffolds, but a low adhesion to the all-PCL scaffold, as well as cell proliferation. The results from this study demonstrate the potential of PCL/NKB 3D bioplotted scaffolds as viable platforms to enable osseous tissue formation, which can be used in several tissue engineering applications, including improvement of bone tissue regeneration.
Assuntos
Cerâmica , Animais , Regeneração Óssea , Bovinos , Durapatita , Poliésteres , Porosidade , Engenharia Tecidual , Alicerces TeciduaisRESUMO
Resumen: Los andamios fibrilares han recibido un enorme interés como futuros biomateriales con potencial aplicación en el campo de la biomedicina regenerativa. En este sentido, hemos optimizado los parámetros para la síntesis de diferentes concentraciones (6, 7, y 10 %) de andamios de ácido poli-láctico (PLA) por la técnica de hilado por propulsión de gas (AJS). Dichos andamios fueron caracterizados por Microscopía Electrónica de Barrido (SEM) y por espectrometría Infrarroja con Transformada de Fourier (FTIR). Nuestros resultados mostraron que los andamios son fibrilares con diámetros en escalas nanométricas. Asimismo; se estudió la biocompatibilidad celular in vitro al realizar ensayos de adhesión, proliferación y de interacción célula-material al cultivar células troncales mesenquimales derivadas de médula ósea. Nuestros datos indican que las membranas fibrilares de PLA aumentan la respuesta celular, no son citotóxicas al compararse con las películas delgadas de PLA. Por lo tanto; el método de síntesis propuesto tiene potencial para la fabricación de membranas hiladas con una facilidad de procesamiento y podría ser un prometedor biomaterial económico con futuras aplicaciones en la regeneración de tejidos.
Abstract: Fiber scaffolds have received increasing interest as promising biomaterials for potential application in the field of tissue regeneration. In this sense, we optimized the parameters for the synthesis of different concentrations (6, 7, and 10 %) of poly-lactic acid (PLA) scaffolds by air jet spinning technology (AJS). The PLA scaffolds were characterized by Scanning Electron Microscopy (SEM) and Fourier Transform Infrared Spectroscopy (FTIR) analysis. Our results by SEM micrographs showed that scaffolds have a fibrilar morphology with nanoscale diameter of fibers. Biocompatibility assay was observed through an in vitro experiment based on cell attachment, MTT and cell-material interaction assay when culturing bone marrow-derived mesenchymal stem cells onto the PLA spun membrane scaffolds. Our data indicate that fiber membrane of PLA scaffold increase the cellular response, are not cytotoxic when compared to thin films of PLA. Thus; the proposed synthesis method has potential for easy processing of spun fibrilar scaffolds with good biocompatibility and could be a promising economical biomaterial with future potential applications in tissue regeneration.
RESUMO
Little is known about the molecular mechanisms that regulate the cementogenesis process, because specific cementum markers are not yet available. To investigate whether a cementoblastoma-conditioned medium-derived protein (CP) could be useful as a cementum biological marker, we studied its expression and distribution in human periodontal tissues, human periodontal ligament, alveolar bone, and cementoblastoma-derived cells. In human periodontal tissues, immunoreactivity to anti-CP was observed throughout the cementoid phase of acellular and cellular cementum, cementoblasts, cementocytes, cells located in the endosteal spaces of human alveolar bone, and in cells in the periodontal ligament located near the blood vessels. Immunopurified CP promoted cell attachment on human periodontal ligament, alveolar bone-derived cells, and gingival fibroblasts. A monoclonal antibody against bovine cementum attachment protein (CAP) cross-reacted with CP. These findings indicate that CP identifies potential cementoblast progenitor cells, is immunologically related to CAP species, and serves as a biological marker for cementum.