Polycaprolactone- and polycaprolactone/ceramic-based 3D-bioplotted porous scaffolds for bone regeneration: A comparative study.

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.

Trypsin as enhancement in cyclical tracheal decellularization: Morphological and biophysical characterization.

There are different types of tracheal disorders (e.g. cancer, stenosis and fractures). These can cause respiratory failure and lead to death of patients. Several attempts have been made for trachea replacement in order to restore the airway, including anastomosis and implants made from synthetic or natural materials. Tracheal allotransplantation has shown high rejection rates, and decellularization has emerged as a possible solution. Decellularization involves the removal of antigens from cells in the organ or tissue, leaving a matrix that can be used as 3D cell-scaffold. Although this process has been used for tracheal replacement, it usually takes at least two months and time is critical for patients with tracheal disorders. Therefore, there is necessary to develop a tracheal replacement process, which is not only effective, but also quick to prepare. The aim of this research was to develop a faster trachea decellularization protocol using Trypsin enzyme and Ethylenediaminetetraacetic acid (EDTA) as decellularization agents. Three protocols of cyclic trachea decellularization (Protocols A, B, and C) were compared. Following Protocol A (previously described in the literature), 15 consecutive cycles were performed over 32 days. Protocol B (a variation of Protocol A) ­ EDTA being added ­ with 15 consecutive cycles performed over 60 days. Finally, Protocol C, with the addition of Trypsin as a decellularization agent, 5 consecutive cycles being performed over 10 days. For the three protocols, hematoxylin­eosin (H&E) staining and DNA residual content quantification were performed to establish the effectiveness of the decellularization process. Scanning Electron Microscopy (SEM) was used to observe the changes in porosity and microarrays. To evaluate the structural matrices integrity, Thermogravimetric Analysis (TGA) and biomechanical test were used. None of the protocols showed significant alteration or degradation in the components of the extracellular matrix (ECM). However, in Protocol C, more cellular components were removed in less time, making it the most efficient process. In addition, the cell tracking and viability was evaluated with chondrocytes seeding on the scaffold obtained by Protocol C, which showed an adequate cell scaffolding ability of this matrix.

Thermoluminescent characteristics of synthetic hydroxyapatite (SHAp).

This paper presents the experimental results of the thermoluminescent (TL) characteristics of synthetic hydroxyapatite (SHAp) obtained by the sol-gel method. For preparation of the SHAp powders, phosphorus pentoxide (P2O5) and calcium nitrate tetrahydrated (Ca(NO3)2-4H2O) were used. The powders obtained were submitted at different temperatures. The structural and morphological characterization were carried out using X-ray diffraction (XRD) and scanning electron microscopy techniques. TL glow curve exhibited two peaks centered at around 200 °C and 300 °C. TL response of SHAp as a function of gamma absorbed dose was linear over a wide dose range. Fading of the storage information in the samples irradiated was also studied. The experimental results show that the synthetic hydroxyapatite obtained by the sol-gel method may have used in gamma radiation dosimetry applications.

Characterization of Bone Cements Prepared with Either Hydroxyapatite, α-TCP or Bovine Bone / Caracterización de cementos óseos preparados con hidroxiapatita, α-TCP o hueso bovino

In this work, we report the preparation of bone cements by using methyl methacrylate (MMA) as a base monomer and either hydroxyapatite (HA), alpha tricalcium phosphate (α-TCP) or bovine bone particles as bioactive fillers. In general, it was observed that curing times increased by the addition of any of these fillers (from 4 to 6.7 min). Maximum temperatures decrease slightly by the addition of 20 wt.% of either α-TCP or bovine bone (80.3°C and 73.2°C respectively) but it did not change by the addition of HA (84.3°C) with respect to PMMA only bone cement used as control. Residual monomer content was lower than 4% in the bioactive bone cements. By using α-TCP or bovine bone compressive strength increased with respect to the unfilled bone cement but it was reduced when HA was used. However, all these formulations fulfill the 70 MPa required for bone cement use. Flexural strength was increased by using either a-TCP o bovine bone but the addition of HA decreased this properties compared to the base bone cement. However, the minimum flexural strength (50 MPa) was fulfilled only in those experimental formulations containing low amounts of α-TCP. The minimum tensile strength (30 MPa) was satisfied by all formulations but it was always lower than the exhibited by the unfilled bone cement.

Este trabajo reporta la preparación de cementos óseos utilizando metacrilato de metilo (MMA) como monómero base y rellenos bioactivos tales como hidroxiapatita (HA), fosfato tricálcico alfa (α-TCP) o hueso bovino. En general, los tiempos de curado aumentaron con la inclusión de estos refuerzos (de 4 hasta 6.7 min). La temperatura máxima alcanzada durante la polimerización del cemento disminuyó ligeramente al adicionar 20% de α-TCP o hueso bovino (80.3°C y 73.2°C respectivamente) y se mantuvo sin cambio en las formulaciones con HA (84.3°C) con respecto al control de solo PMMA. El contenido de monómero residual en los cementos bioactivos fue menor al 4%. La presencia de α-TCP o hueso bovino aumentó la resistencia a la compresión del cemento base y la adición de HA la disminuyó, cumpliendo en todos los casos con la resistencia mínima a la compresión (70 MPa) sugerida para su uso como cemento óseo. La adición de α-TCP o hueso bovino aumentó la resistencia a la flexión del cemento base pero la adición de HA la redujo aunque el requerimiento mínimo de resistencia a la flexión (50 MPa) fue cumplido solamente al usar concentraciones bajas de α-TCP. La resistencia tensil mínima (30 MPa) fue satisfecha por todas las formulaciones aunque siempre fue menor que la exhibida por el cemento base.