Developing a novel resorptive hydroxyapatite-based bone substitute for over-critical size defect reconstruction: physicochemical and biological characterization and proof of concept in segmental rabbit’s ulna reconstruction

2020 ◽  
Vol 65 (4) ◽  
pp. 491-505
Author(s):  
Milutin Micic ◽  
Djordje Antonijevic ◽  
Sanja Milutinovic-Smiljanic ◽  
Dijana Trisic ◽  
Bozana Colovic ◽  
...  

AbstractThe aim of this study was to develop novel hydroxyapatite (HAP)-based bioactive bone replacement materials for segmental osteotomy reconstruction. Customized three-dimensional (3D) bone construct was manufactured from nanohydroxyapatite (nHAP) with poly(lactide-co-glycolide) (PLGA) coating using 3D models derived from the computed tomography (CT) scanning of the rabbit’s ulna and gradient 3D printing of the bone substitute mimicking the anatomical shape of the natural bone defect. Engineered construct revealed adequate micro-architectural design for successful bone regeneration having a total porosity of 64% and an average pore size of 256 μm. Radiography and micro-CT analysis depicted new bone apposition through the whole length of the reconstructed ulna with a small area of non-resorbed construct in the central area of defect. Histological analysis revealed new bone formation with both endochondral and endesmal type of ossification. Immunohistochemistry analysis depicted the presence of bone formation indicators – bone morphogenetic protein (BMP), osteocalcin (OCN) and osteopontin (OPN) within newly formed bone. Manufactured personalized construct acts as a “smart” responsive biomaterial capable of modulating the functionality and potential for the personalized bone reconstruction on a clinically relevant length scale.

2017 ◽  
Vol 54 (3) ◽  
pp. 181-201
Author(s):  
Rebecca Johnson ◽  
Mark Longman ◽  
Brian Ruskin

The Three Forks Formation, which is about 230 ft thick along the southern Nesson Anticline (McKenzie County, ND), has four “benches” with distinct petrographic and petrophysical characteristics that impact reservoir quality. These relatively clean benches are separated by slightly more illitic (higher gamma-ray) intervals that range in thickness from 10 to 20 ft. Here we compare pore sizes observed in scanning electron microscope (SEM) images of the benches to the total porosity calculated from binned precession decay times from a suite of 13 nuclear magnetic resonance (NMR) logs in the study area as well as the logarithmic mean of the relaxation decay time (T2 Log Mean) from these NMR logs. The results show that the NMR log is a valid tool for quantifying pore sizes and pore size distributions in the Three Forks Formation and that the T2 Log Mean can be correlated to a range of pore sizes within each bench of the Three Forks Formation. The first (shallowest) bench of the Three Forks is about 35 ft thick and consists of tan to green silty and shaly laminated dolomite mudstones. It has good reservoir characteristics in part because it was affected by organic acids and received the highest oil charge from the overlying lower Bakken black shale source rocks. The 13 NMR logs from the study area show that it has an average of 7.5% total porosity (compared to 8% measured core porosity), and ranges from 5% to 10%. SEM study shows that both intercrystalline pores and secondary moldic pores formed by selective partial dissolution of some grains are present. The intercrystalline pores are typically triangular and occur between euhedral dolomite rhombs that range in size from 10 to 20 microns. The dolomite crystals have distinct iron-rich (ferroan) rims. Many of the intercrystalline pores are partly filled with fibrous authigenic illite, but overall pore size typically ranges from 1 to 5 microns. As expected, the first bench has the highest oil saturations in the Three Forks Formation, averaging 50% with a range from 30% to 70%. The second bench is also about 35 ft thick and consists of silty and shaly dolomite mudstones and rip-up clast breccias with euhedral dolomite crystals that range in size from 10 to 25 microns. Its color is quite variable, ranging from green to tan to red. The reservoir quality of the second bench data set appears to change based on proximity to the Nesson anticline. In the wells off the southeast flank of the Nesson anticline, the water saturation averages 75%, ranging from 64% to 91%. On the crest of the Nesson anticline, the water saturation averages 55%, ranging from 40% to 70%. NMR porosity is consistent across the entire area of interest - averaging 7.3% and ranging from 5% to 9%. Porosity observed from samples collected on the southeast flank of the Nesson Anticline is mainly as intercrystalline pores that have been extensively filled with chlorite clay platelets. In the water saturated southeastern Nesson Anticline, this bench contains few or no secondary pores and the iron-rich rims on the dolomite crystals are less developed than those in the first bench. The chlorite platelets in the intercrystalline pores reduce average pore size to 500 to 800 nanometers. The third bench is about 55 ft thick and is the most calcareous of the Three Forks benches with 20 to 40% calcite and a proportionate reduction in dolomite content near its top. It is also quite silty and shaly with a distinct reddish color. Its dolomite crystals are 20 to 50 microns in size and partly abraded and dissolved. Ferroan dolomite rims are absent. This interval averages 7.1% porosity and ranges from 5% to 9%, but the pores average just 200 nanometers in size and occur mainly as microinterparticle pores between illite flakes in intracrystalline pores in the dolomite crystals. This interval has little or no oil saturation on the southern Nesson Anticline. Unlike other porosity tools, the NMR tool is a lithology independent measurement. The alignment of hydrogen nuclei to the applied magnetic field and the subsequent return to incoherence are described by two decay time constants, longitudinal relaxation time (T1) and transverse relaxation time (T2). T2 is essentially the rate at which hydrogen nuclei lose alignment to the external magnetic field. The logarithmic mean of T2 (T2 Log Mean) has been correlated to pore-size distribution. In this study, we show that the assumption that T2 Log Mean can be used as a proxy for pore-size distribution changes is valid in the Three Forks Formation. While the NMR total porosity from T2 remains relatively consistent in the three benches of the Three Forks, there are significant changes in the T2 Log Mean from bench to bench. There is a positive correlation between changes in T2 Log Mean and average pore size measured on SEM samples. Study of a “type” well, QEP’s Ernie 7-2-11 BHD (Sec. 11, T149N, R95W, McKenzie County), shows that the 1- to 5-micron pores in the first bench have a T2 Log Mean relaxation time of 10.2 msec, whereas the 500- to 800-nanometer pores in the chlorite-filled intercrystalline pores in the second bench have a T2 Log Mean of 4.96 msec. This compares with a T2 Log Mean of 2.86 msec in 3rd bench where pores average just 200 nanometers in size. These data suggest that the NMR log is a useful tool for quantifying average pore size in the various benches of the Three Forks Formation.


2014 ◽  
Vol 1004-1005 ◽  
pp. 670-674 ◽  
Author(s):  
Dian Quan Dong ◽  
Hong Bo Dai ◽  
Jian Guo Zheng

Polystyrene microspheres with 120nm diameter were synthesized by emulsion polymerization and three-dimensionally ordered colloidal crystal templates were obtained by centrifugal sedimentation.Three dimensionally ordered nanopore (3DON) manganese oxide lithium ion-sieve was prepared after filtration, two heated roasting and acid modified by using precursor solution filling the colloidal crystal templates, which was prepared by Lithium salt, manganese salt and citric acid. SEM, XRD, and saturated exchange capacity test were used to characterize the roasting condition, appearance, structure, and ion exchange performance of the oxide. The results showed that, the optimum roasting condition of preparing lithium ion-sieve precursors were found as follows: heating rate at 2°C/min, 300 °C roasting 4h and 800 °C roasting 8h, The 3DON Li4Mn5O12lithium ion sieve precursor has the shape of three-dimensional cross-linked connected into the network structure. Li4Mn5O12was regularly arranged and the hole wall was integrity,average pore size of approximately 90nm.3DON Li4Mn5O12 showed good stability for acid and the retrofit of lithium ion-sieve showed a high selectivity for Li+. The saturated exchange capacity of Li+is 51.98mgLi+/g.


Polymers ◽  
2019 ◽  
Vol 11 (3) ◽  
pp. 457 ◽  
Author(s):  
Rodrigo Urruela-Barrios ◽  
Erick Ramírez-Cedillo ◽  
A. Díaz de León ◽  
Alejandro Alvarez ◽  
Wendy Ortega-Lara

Three-dimensional (3D) printing technologies have become an attractive manufacturing process to fabricate scaffolds in tissue engineering. Recent research has focused on the fabrication of alginate complex shaped structures that closely mimic biological organs or tissues. Alginates can be effectively manufactured into porous three-dimensional networks for tissue engineering applications. However, the structure, mechanical properties, and shape fidelity of 3D-printed alginate hydrogels used for preparing tissue-engineered scaffolds is difficult to control. In this work, the use of alginate/gelatin hydrogels reinforced with TiO2 and β-tricalcium phosphate was studied to tailor the mechanical properties of 3D-printed hydrogels. The hydrogels reinforced with TiO2 and β-TCP showed enhanced mechanical properties up to 20 MPa of elastic modulus. Furthermore, the pores of the crosslinked printed structures were measured with an average pore size of 200 μm. Additionally, it was found that as more layers of the design were printed, there was an increase of the line width of the bottom layers due to its viscous deformation. Shrinkage of the design when the hydrogel is crosslinked and freeze dried was also measured and found to be up to 27% from the printed design. Overall, the proposed approach enabled fabrication of 3D-printed alginate scaffolds with adequate physical properties for tissue engineering applications.


2007 ◽  
Vol 330-332 ◽  
pp. 967-970 ◽  
Author(s):  
B. Otsuki ◽  
Mitsuru Takemoto ◽  
Shunsuke Fujibayashi ◽  
Masashi Neo ◽  
Tadashi Kokubo ◽  
...  

A porous structure comprises pores and pore throats with a complex three-dimensional network structure, and many investigators have described the relationship between average pore size and the amount of bone ingrowth. However, the influence of network structure or pore throats for tissue ingrowth has rarely been discussed. Bioactive porous titanium implants with 48% porosity were analyzed using specific algorithms for three-dimensional analysis of interconnectivity based on a micro focus X-ray computed tomography system. In vivo histological analysis was performed using the very same implants implanted into the femoral condyles of male rabbits for 6 weeks. This matching study revealed that more poorly differentiated pores tended to have narrow pore throats, especially in their shorter routes to the outside. Data obtained suggest that this sort of novel analysis is useful for evaluating bone and tissue ingrowth into porous biomaterials.


2020 ◽  
Vol 10 (2) ◽  
pp. 20190079 ◽  
Author(s):  
Yassen Abbas ◽  
Lucia G. Brunel ◽  
Michael S. Hollinshead ◽  
Ridma C. Fernando ◽  
Lucy Gardner ◽  
...  

The endometrium is the secretory lining of the uterus that undergoes dynamic changes throughout the menstrual cycle in preparation for implantation and a pregnancy. Recently, endometrial organoids (EO) were established to study the glandular epithelium. We have built upon this advance and developed a multi-cellular model containing both endometrial stromal and epithelial cells. We use porous collagen scaffolds produced with controlled lyophilization to direct cellular organization, integrating organoids with primary isolates of stromal cells. The internal pore structure of the scaffold was optimized for stromal cell culture in a systematic study, finding an optimal average pore size of 101 µm. EO seeded organize to form a luminal-like epithelial layer, on the surface of the scaffold. The cells polarize with their apical surface carrying microvilli and cilia that face the pore cavities and their basal surface attaching to the scaffold with the formation of extracellular matrix proteins. Both cell types are hormone responsive on the scaffold, with hormone stimulation resulting in epithelial differentiation and stromal decidualization.


2021 ◽  
Author(s):  
Dominik Göldner ◽  
Fotios Alexandros Karakostis ◽  
Armando Falcucci

This protocol presents the first detailed step-by-step pipeline for the 3D scanning and post processing of large batches of lithic artefacts using a micro-computed tomography (micro-CT) scanner (i.e., a Phoenix v-tome-x S model by General Electronics MCC, Boston MA) and an Artec Space Spider scanner (Artec Inc., Luxembourg). This protocol was used to scan and analyze ca. 700 lithic artefacts from the Protoaurignacian layers at Fumane Cave in north-eastern Italy (Falcucci et al., in preparation). For this study several costly scanners and proprietary software packages were employed. Although it is not easy to find a low-budget alternative for the scanners, it is possible to use free and open-source software programs, such as 3D-Slicer (https://www.slicer.org/) or MorphoDig (https://morphomuseum.com/morphodig), to process CT data as well as MeshLab (Cignoni et al. 2008) to interact with the 3D models in general. However, if alternative software is used, the steps and their order described in this protocol might diverge significantly. A cost-effective alternative to create 3D models is digital photogrammetry using commercial cameras and freely available software like Meshroom (https://alicevision.org). Although photogrammetry is an affordable technique to create accurate 3D models of objects, this method might not be useful when scanning large batches of artefacts, as it will require a lot of computation time and processing capacity. Likewise, it could be difficult to generate accurate 3D models of very small and/or detailed tool shapes using 3D surface scanners because stone tools are often much smaller than the recommended minimum field of view. Similarly, the resolution of conventional medical CT scanners might not be sufficient to capture minor details of stone tools, such as the outline or dorsal scars. Thus, high-resolution micro-CT technology is the only reliable way to accurately capture the overall morphology of small stone tools. This protocol aims at providing the first detailed procedure dedicated to the scanning of small lithic implements for further three-dimensional analysis. Note that some of the steps must be repeated at different working stages throughout this protocol. In cases where a task must be done in the exact same way as described in a previous step, a reference to that step is provided. When slight changes were made, the step was modified and reported entirely. This protocol contains a few red and green colours (e.g., arrows or within-program colours) which might be perceived differently by people with dyschromatopsia. However, the display of these colours has been kept to a minimum. We recommend the reader to go over the entire protocol carefully, even if only some specific parts are required. A few points are in fact interdependent, and some of them must be applied simultaneously. Content: Part 1 – Styrofoam preparation Part 2 – Micro-CT scanning Part 3 – 3D model extraction of CT scanned stone artifacts using Avizo Part 4 – Cropping extracted surface model to separate Face A and B in Artec Studio Part 5 – Cropping Face A to separate the lines in Artec Studio Part 6 – Cropping each stone artefact from the lines in Artec Studio Part 7 – Virtually control measurements in MeshLab Part 8 – Artec scanning of larger artifacts Part 9 – Export meshes as non-binary ply models for successive analysis in geomorph Three-dimensional example (in ply format) of the effectivity of the StyroStone Protocol: You can download an example of one Styrofoam line in 3D obtained using our protocol to appreciate the result that can be achieved. We have selected a line where objects are characterized by different metric and morphological attributes. Notice the retouching well visible in the last five smaller artifacts (counting from the left when artifact are oriented with the dorsal face in front of the observer and the butt down), as well as the platforms and bulbs of all artifacts. For more information and examples, feel free to contact us!


2013 ◽  
Vol 774-776 ◽  
pp. 949-953
Author(s):  
Ming Hua Huang ◽  
Hui Dong ◽  
Di Ru Xu ◽  
Duan Cheng Wang ◽  
Yong Shun Cui ◽  
...  

KGM and Collagen II were selected as the main raw materials and ammonia served as the crosslinking agent to prepare the porous KGM / COLII composite cartilage scaffolds by blending method and freeze-drying method. The porosity, average pore size, compressive strength and water absorption were measured on the basis of the related standard. The scaffolds were characterized by SEM and XRD. The results show that the optimal program of preparing composite cartilage scaffolds is KGM (2g), COLII (1g), freeze temperature (-20 ° C) and ammonia (0.1 ml). The optimal cartilage scaffolds are porous three-dimensional network structures which the porosity is more than 90%; the average pore size is about 200μm; the compressive strength is about 0.75Mpa and the water absorption reaches up to 892%.


PeerJ ◽  
2020 ◽  
Vol 8 ◽  
pp. e9690 ◽  
Author(s):  
Sérgio Ferreira-Cardoso ◽  
Pierre-Henri Fabre ◽  
Benoit de Thoisy ◽  
Frédéric Delsuc ◽  
Lionel Hautier

Background Ecological adaptations of mammals are reflected in the morphological diversity of their feeding apparatus, which includes differences in tooth crown morphologies, variation in snout size, or changes in muscles of the feeding apparatus. The adaptability of their feeding apparatus allowed them to optimize resource exploitation in a wide range of habitats. The combination of computer-assisted X-ray microtomography (µ-CT) with contrast-enhancing staining protocols has bolstered the reconstruction of three-dimensional (3D) models of muscles. This new approach allows for accurate descriptions of muscular anatomy, as well as the quick measurement of muscle volumes and fiber orientation. Ant- and termite-eating (myrmecophagy) represents a case of extreme feeding specialization, which is usually accompanied by tooth reduction or complete tooth loss, snout elongation, acquisition of a long vermiform tongue, and loss of the zygomatic arch. Many of these traits evolved independently in distantly-related mammalian lineages. Previous reports on South American anteaters (Vermilingua) have shown major changes in the masticatory, intermandibular, and lingual muscular apparatus. These changes have been related to a functional shift in the role of upper and lower jaws in the evolutionary context of their complete loss of teeth and masticatory ability. Methods We used an iodine staining solution (I2KI) to perform contrast-enhanced µ-CT scanning on heads of the pygmy (Cyclopes didactylus), collared (Tamandua tetradactyla) and giant (Myrmecophaga tridactyla) anteaters. We reconstructed the musculature of the feeding apparatus of the three extant anteater genera using 3D reconstructions complemented with classical dissections of the specimens. We performed a description of the musculature of the feeding apparatus in the two morphologically divergent vermilinguan families (Myrmecophagidae and Cyclopedidae) and compared it to the association of morphological features found in other myrmecophagous placentals. Results We found that pygmy anteaters (Cyclopes) present a relatively larger and architecturally complex temporal musculature than that of collared (Tamandua) and giant (Myrmecophaga) anteaters, but shows a reduced masseter musculature, including the loss of the deep masseter. The loss of this muscle concurs with the loss of the jugal bone in Cyclopedidae. We show that anteaters, pangolins, and aardvarks present distinct anatomies despite morphological and ecological convergences.


2021 ◽  
Vol 11 (24) ◽  
pp. 12163
Author(s):  
Sujuan Pan ◽  
Changqing Wang ◽  
Yibo Wang ◽  
Rongmin Wang

In this work, feather keratin was extracted from the waste feather of chicken via alkyd pretreatment and reduction method, the extraction rate is above 85%. The molecular weight and aggregation morphology of feather keratin in an aqueous environment were characterized by 18-angle laser light scattering gel permeation chromatography and field emission transmission electron microscopy. The relationship between the structure and properties of feather keratin is discussed. The 1-(3-dimethylaminopropyl) -3-ethylcarbondiimide hydrochloride and N-hydroxysuccinimide were used as activation system and cross-linkage. The gallic acid was used as modification reagent and was bonded to feather keratin chains; meanwhile, feather keratin chains were cross-linked through covalent bonds obtained the novel adsorbent (named as GA-FK gel). The GA-FK gel was investigated by IR, SEM, TGA, XRD, and BET methods. The results indicated that GA molecules successfully bonded to feather keratin chains and cross-linked between feather keratin chains. The GA-FK gel was found to have a three-dimensional network structure with abundant mesopores. Its pore size range is 1.8~90 nm; average pore size is 19.6 nm. Its specific surface area is 7.17 m2·g−1. In addition, GA-FK gel was applied to remove Fe(III) in water. The maximum adsorption capacity was 319.0 mg·g−1. The adsorption process of GA-FK gel to Fe(III) presents a typical two-stage pattern accompanied with swelling. The adsorption kinetics of GA-FK gel to Fe(III) follows the quasi-second-order model, the adsorption isotherm follows the Freundlich model. Therefore, the adsorption mechanism is non-specific adsorption.


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