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Journal articles on the topic 'Ex vivo bone'

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Author: Grafiati

Published: 10 December 2022

Last updated: 28 January 2023

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1

Stirpe, Fiorenzo, Luigi Barbieri, Pier Luigi Tazzari, Angelo Dinota, and Marco Gobbi. "Ex vivo bone marrow purging with immunotoxins." European Journal of Haematology 43, S51 (April 24, 2009): 173–75. http://dx.doi.org/10.1111/j.1600-0609.1989.tb01512.x.

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2

Rutherford, R. B., B. Nussenbaum, and P. H. Krebsbach. "Bone morphogenetic protein 7 ex vivo gene therapy." Drug News & Perspectives 16, no. 1 (2003): 5. http://dx.doi.org/10.1358/dnp.2003.16.1.829301.

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3

FERNANDES, M. G., E. M. M. FONSECA, R. N. JORGE, M. VAZ, and M. I. DIAS. "THERMAL ANALYSIS IN DRILLING OF EX VIVO BOVINE BONES." Journal of Mechanics in Medicine and Biology 17, no. 05 (July 12, 2017): 1750082. http://dx.doi.org/10.1142/s0219519417500828.

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Bone drilling is a common procedure in Medicine, mainly in traumatology and orthopedic procedure for fractures fixation and in reconstructive surgery. The success of this surgical procedure is dependent on many factors, namely, on heat generation control during the bone drilling. The main concern in bone drilling is the mechanical and thermal damage of the bone induced by inappropriate parameters such as drill speed and feed-rate during the drilling. This study focuses on the temperature generated during drilling of cortical bone tissue (bovine origin) and solid rigid polyurethane foams with similar mechanical properties to the human bone tissue. Different parameters such as drill speed, feed-rate and hole depth were tested. All results showed that improvement of the drilling parameters and the drill temperatures can be estimated. It was concluded that when the drill speed and feed-rate were higher, the bone temperature increase was lower. The obtained results of temperature in the drilling process of polyurethane foam blocks or bovine bone were compared with a good agreement in between both.

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4

Cramer, E. E. A., K. Ito, and S. Hofmann. "Ex vivo Bone Models and Their Potential in Preclinical Evaluation." Current Osteoporosis Reports 19, no. 1 (January 11, 2021): 75–87. http://dx.doi.org/10.1007/s11914-020-00649-5.

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Abstract Purpose of Review Novel therapies for damaged and diseased bone are being developed in a preclinical testing process consisting of in vitro cell experiments followed by in vivo animal studies. The in vitro results are often not representative of the results observed in vivo. This could be caused by the complexity of the natural bone environment that is missing in vitro. Ex vivo bone explant cultures provide a model in which cells are preserved in their native three-dimensional environment. Herein, it is aimed to review the current status of bone explant culture models in relation to their potential in complementing the preclinical evaluation process with specific attention paid to the incorporation of mechanical loading within ex vivo culture systems. Recent Findings Bone explant cultures are often performed with physiologically less relevant bone, immature bone, and explants derived from rodents, which complicates translatability into clinical practice. Mature bone explants encounter difficulties with maintaining viability, especially in static culture. The integration of mechanical stimuli was able to extend the lifespan of explants and to induce new bone formation. Summary Bone explant cultures provide unique platforms for bone research and mechanical loading was demonstrated to be an important component in achieving osteogenesis ex vivo. However, more research is needed to establish a representative, reliable, and reproducible bone explant culture system that includes both components of bone remodeling, i.e., formation and resorption, in order to bridge the gap between in vitro and in vivo research in preclinical testing.

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5

Denis, I., G. Cournot, H. Lacroix, C. Colin, E. Zerath, and A. Pointillart. "In vivo bone metabolism and ex vivo bone marrow osteoprogenitors in vitamin D-deprived pigs." Bone 26, no. 5 (May 2000): 491–98. http://dx.doi.org/10.1016/s8756-3282(00)00257-x.

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6

Walker, Mary M., Molly E. Baumann, John H. Alexander, Britani N. Blackstone, Christopher B. Morgan, Thomas J. Scharschmidt, and Heather M. Powell. "Mechanical strain induces ex vivo expansion of periosteum." PLOS ONE 17, no. 12 (December 30, 2022): e0279519. http://dx.doi.org/10.1371/journal.pone.0279519.

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Segmental bone defects present complex clinical challenges. Nonunion, malunion, and infection are common sequalae of autogenous bone grafts, allografts, and synthetic bone implants due to poor incorporation with the patient’s bone. The current project explores the osteogenic properties of periosteum to facilitate graft incorporation. As tissue area is a natural limitation of autografting, mechanical strain was implemented to expand the periosteum. Freshly harvested, porcine periosteum was strained at 5 and 10% per day for 10 days with non-strained and free-floating samples serving as controls. Total tissue size, viability and histologic examination revealed that strain increased area to a maximum of 1.6-fold in the 10% daily strain. No change in tissue anatomy or viability via MTT or Ki67 staining and quantification was observed among groups. The osteogenic potential of the mechanical expanded periosteum was then examined in vivo. Human cancellous allografts were wrapped with 10% per day strained, fresh, free-floating, or no porcine periosteum and implanted subcutaneously into female, athymic mice. Tissue was collected at 8- and 16-weeks. Gene expression analysis revealed a significant increase in alkaline phosphatase and osteocalcin in the fresh periosteum group at 8-weeks post implantation compared to all other groups. Values among all groups were similar at week 16. Additionally, histological assessment with H&E and Masson-Goldner Trichrome staining showed that all periosteal groups outperformed the non-periosteal allograft, with fresh periosteum demonstrating the highest levels of new tissue mineralization at the periosteum-bone interface. Overall, mechanical expansion of the periosteum can provide increased area for segmental healing via autograft strategies, though further studies are needed to explore culture methodology to optimize osteogenic potential.

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7

Steck, R., C. Gatzka, E. Schneider, P. Niederer, and M. L. Tate. "Measurement of bone surface strains on the sheep metacarpus in vivo and ex vivo." Veterinary and Comparative Orthopaedics and Traumatology 16, no. 01 (2003): 38–43. http://dx.doi.org/10.1055/s-0038-1632754.

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SummaryBone surface strains were measured on the dorsal ovine metacarpus during normal locomotion on a treadmill at different walking speeds to determine physiological strain levels. These measured strains were related to the strains measured in an ex vivo model of the sheep forelimb with two types of load application: loading by two Schanz-screws and loading via the radius. In vivo, the average surface strains were found to be dependent upon body weight as well as the walking speed. The orientation of the peak principal strain corresponded to the longitudinal axis of the bone. Ex vivo, loads applied via Schanz screws in the screw-loading model lead to strains on the dorsal metacarpus that corresponds to strains experienced in vivo during intermittent peak loads. Screw loading imparted primarily a bending load to the metacarpus, with the dorsal aspect in compression and the palmar aspect in tension. Loads, applied via the radius and the hoof in the radius-loading model, resulted in bone surface strains comparable to those measured during slow walking in vivo. In both ex vivo loading situations, peak strain orientation was parallel to the longitudinal axis of the sheep metacarpus. In conclusion, the results show that although the ex vivo loading models do not exactly replicate the load experienced in vivo, the magnitude and orientation of the principal strains on the dorsal metacarpus are within the range of strains occurring during normal physiological loading. These data validate the physiological significance of the ex vivo model and aid in understanding effects of mechanical loading on interstitial fluid flow and mass transport through bone.

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8

Buduo, Christian A. Di, Alessandra Balduini, and David L. Kaplan. "Translational approaches to functional platelet production ex vivo." Thrombosis and Haemostasis 115, no. 02 (March 2016): 250–56. http://dx.doi.org/10.1160/th15-07-0570.

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SummaryPlatelets, which are released by megakaryocytes, play key roles in haemostasis, angiogenesis, immunity, tissue regeneration and wound healing. The scarcity of clinical cures for life threatening platelet diseases is in a large part due to limited insight into the mechanisms that control the developmental process of megakaryocytes and the mechanisms that govern the production of platelets within the bone marrow. To overcome these limitations, functional human tissue models have been developed and studied to extrapolate ex vivo outcomes for new insight on bone marrow functions in vivo. There are many challenges that these models must overcome, from faithfully mimicking the physiological composition and functions of bone marrow, to the collection of the platelets generated and validation of their viability and function for human use. The overall goal is to identify innovative instruments to study mechanisms of platelet release, diseases related to platelet production and new therapeutic targets starting from human progenitor cells.

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9

Giulivi, Antonio, Mike Halpenny, Paul Birch, Lin Yang, and Lisa Martin. "Ex-Vivo Expansion of Megakaryocyte Progenitors." Blood 104, no. 11 (November 16, 2004): 2885. http://dx.doi.org/10.1182/blood.v104.11.2885.2885.

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Abstract Following Bone Marrow Transplant, thrombocytopenia and neutropenia always occur and patients require additional post transplant support in the form of platelet transfusions. Megakaryocytes (Mk), the precursors of platelets, are contained in hematopoietic progenitor cell products but their number is variable and relatively low. The infusion of ex vivo expanded Mk precursors could be beneficial by shortening the time to platelet engraftment and therefore reducing the amount of platelet transfusion support required by bone marrow transplant patients. The objective of this project was to investigate the expansion of Mk progenitors from peripheral blood stem cell (PBSC) harvests from patients with haematological malignancies. Briefly, CD34+ cells were isolated and cultured in serum free media supplemented with thrombopoietin (TPO) and Interleukin 1 (IL-1) then incubated at 37°C /5% CO2 for 8 – 12 days. Megakaryocyte progenitor analysis was accomplished using flow cytometry analysis (CD34+/41+, CD41+, CD61+) and Mk culture analysis (CFU-Mk) (Stem Cell Technologies). Mk progenitor expansion efficiency was determined as “fold expansion” of Mk progenitors produced over input levels. After 8 days of culture, a mean expansion of 46 fold (range 1.2 – 327.0, n=10) in megakaryocytic cells (CD61+) and a 15 fold expansion (1.2 – 41.7, n=10) in megakaryocyte progenitor cells (CD34+/41+) was observed. After 12 days, a 116 fold expansion (1.5 – 286, n=7) in megakaryocytic cells (CD61+) and a 19 fold expansion (2.4 – 40, n=7) in Mk progenitors (CD34+/CD41+) was observed. This study demonstrates that CD34+ cells can be used to effectively expand megakaryocytic cells using just two cytokines for an incubation period of 8 – 12 days. This data could be used to develop future protocols for use in clinical applications.

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10

Davies, H. M. S. "Ex vivo calibration and validation of in vivo equine bone strain measures." Equine Veterinary Journal 41, no. 3 (March 2009): 225–28. http://dx.doi.org/10.2746/042516409x396317.

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11

Infante, Teresa, Elena Cesario, Michele Gallo, Flavio Fazioli, Annarosaria De Chiara, Cristina Tutucci, Gaetano Apice, and Filomena de Nigris. "Ex Vivo Behaviour of Human Bone Tumor Endothelial Cells." Cancers 5, no. 4 (April 11, 2013): 404–17. http://dx.doi.org/10.3390/cancers5020404.

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12

Parrilla, Claudio, Wanda Lattanzi, Anna Rita Fetoni, Raffaella Marchese, and Gaetano Paludetti. "Ex Vivo Gene Therapy for Rat Mandibular Bone Regeneration." Otolaryngology–Head and Neck Surgery 143, no. 2_suppl (August 2010): P48. http://dx.doi.org/10.1016/j.otohns.2010.06.035.

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13

Panoskaltsis, Nicki, Athanasios Mantalaris, and J. H. David Wu. "Engineering a mimicry of bone marrow tissue ex vivo." Journal of Bioscience and Bioengineering 100, no. 1 (July 2005): 28–35. http://dx.doi.org/10.1263/jbb.100.28.

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14

Zhou, Donghui, Khuchtumur Bum-Erdene, David Xu, Degang Liu, Doug Tompkins, Rania S. Sulaiman, Timothy W. Corson, John M. Chirgwin, and Samy O. Meroueh. "Small molecules inhibit ex vivo tumor growth in bone." Bioorganic & Medicinal Chemistry 26, no. 23-24 (December 2018): 6128–34. http://dx.doi.org/10.1016/j.bmc.2018.11.025.

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15

Bussard, Karen M., and Andrea M. Mastro. "Ex-vivo Analysis of the Bone Microenvironment in Bone Metastatic Breast Cancer." Journal of Mammary Gland Biology and Neoplasia 14, no. 4 (December 2009): 387–95. http://dx.doi.org/10.1007/s10911-009-9159-z.

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16

Matsugaki, Aira, Yumi Kimura, Ryota Watanabe, Fumihito Nakamura, Ryo Takehana, and Takayoshi Nakano. "Impaired Alignment of Bone Matrix Microstructure Associated with Disorganized Osteoblast Arrangement in Malignant Melanoma Metastasis." Biomolecules 11, no. 2 (January 20, 2021): 131. http://dx.doi.org/10.3390/biom11020131.

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Malignant melanoma favors spreading to bone, resulting in a weakened bone with a high fracture risk. Here, we revealed the disorganized alignment of apatite crystals in the bone matrix associated with the homing of cancer cells by developing an artificially controlled ex vivo melanoma bone metastasis model. The ex vivo metastasis model reflects the progressive melanoma cell activation in vivo, resulting in decreased bone mineral density and expression of MMP1-positive cells. Moreover, less organized intercellular connections were observed in the neighboring osteoblasts in metastasized bone, indicating the abnormal and randomized organization of bone matrix secreted by disconnected osteoblasts. Our study revealed that the deteriorated microstructure associated with disorganized osteoblast arrangement was a determinant of malignant melanoma-related bone dysfunction.

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17

Chandra, Sundeep, Patrizia Cristofori, Carlos Fonck, and Charles A. O’Neill. "Ex Vivo Gene Therapy: Graft-versus-host Disease (GVHD) in NSG™ (NOD.Cg-Prkdcscid Il2rgtm1Wjl/SzJ) Mice Transplanted with CD34+ Human Hematopoietic Stem Cells." Toxicologic Pathology 47, no. 5 (May 7, 2019): 656–60. http://dx.doi.org/10.1177/0192623319844484.

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A therapeutic option for monogenic disorders is gene therapy with ex vivo-transduced autologous hematopoietic stem cells (HSCs). Safety or efficacy studies of ex vivo-modified HSCs are conducted in humanized mouse models after ablation of the murine bone marrow and transfer of human CD34+ HSCs. Engrafted human CD34+ cells migrate to bone marrow and differentiate into various human hematopoietic lineages. A 12-week study was conducted in NSG™ mice to evaluate engraftment, differentiation, and safety of human CD34+ cells that were transduced ( ex vivo) with a proprietary lentiviral vector encoding a human gene (BMRN-1) or a mock (green fluorescent protein) vector. Several mice intravenously injected with naive CD34+ cells or transduced CD34+ cells had variable lymphohistiocytic inflammatory cell infiltrates and microgranulomas in the liver and lungs consistent with graft-versus-host disease (GVHD). Spleen, bone marrow, stomach, reproductive tract, but not the skin had similar inflammatory changes. Ex vivo viral transduction of CD34+ cells did not impact engraftment or predispose to xenogeneic GVHD.

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18

Pakhmurin, Denis, Viktoriya Pakhmurina, Alexander Kashin, Alexey Kulkov, Igor Khlusov, Evgeny Kostyuchenko, Ivan Sidorov, and Ilya Anisenya. "Compressive Strength Characteristics of Long Tubular Bones after Hyperthermal Ablation." Symmetry 14, no. 2 (February 2, 2022): 303. http://dx.doi.org/10.3390/sym14020303.

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Thermoablation is used in the treatment of tumorous bones. However, little is known about the influence such thermal treatment has on the mechanical properties of bone tissue. The purpose of this work was to study the influence of thermal treatment in a range of 60–100 °C (in increments of 10 °C) on the structural properties of pig femurs using an original approach that involved a periosteal arrangement of heating elements providing gradual dry heating of the bone from its periphery to its center. Heating of freshly extracted bone tissue segments was performed ex vivo using surface heaters of a Phoenix-2 local hyperthermia hardware system. Mechanical testing followed the single-axis compression scheme (traverse velocity of 1 mm/min). In the 60–90 °C range of heating, no attributes of reduced structural characteristics were found in the samples subjected to thermoablation in comparison to the control samples taken from symmetric areas of adjacent cylinders of healthy bones and not subjected to heat treatment. The following statistically significant changes were found as a result of compressing the samples to 100 °C after the heat treatment: reduced modulus of elasticity and increased elastic strain (strain-to-failure), mainly due to increases in plastic deformation. This finding may serve as evidence of a critical ex vivo change in the biomechanical behavior of bone tissues at such temperatures. Thus, ex vivo treatment of bone tissue in the thermal range of 60–90 °C may be used in studies of thermoablation efficiency against tumor involvement of bones.

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19

Ehnert, Sabrina, Helen Rinderknecht, Romina H. Aspera-Werz, Victor Häussling, and Andreas K. Nussler. "Use of in vitro bone models to screen for altered bone metabolism, osteopathies, and fracture healing: challenges of complex models." Archives of Toxicology 94, no. 12 (September 10, 2020): 3937–58. http://dx.doi.org/10.1007/s00204-020-02906-z.

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Abstract Approx. every third hospitalized patient in Europe suffers from musculoskeletal injuries or diseases. Up to 20% of these patients need costly surgical revisions after delayed or impaired fracture healing. Reasons for this are the severity of the trauma, individual factors, e.g, the patients’ age, individual lifestyle, chronic diseases, medication, and, over 70 diseases that negatively affect the bone quality. To investigate the various disease constellations and/or develop new treatment strategies, many in vivo, ex vivo, and in vitro models can be applied. Analyzing these various models more closely, it is obvious that many of them have limits and/or restrictions. Undoubtedly, in vivo models most completely represent the biological situation. Besides possible species-specific differences, ethical concerns may question the use of in vivo models especially for large screening approaches. Challenging whether ex vivo or in vitro bone models can be used as an adequate replacement for such screenings, we here summarize the advantages and challenges of frequently used ex vivo and in vitro bone models to study disturbed bone metabolism and fracture healing. Using own examples, we discuss the common challenge of cell-specific normalization of data obtained from more complex in vitro models as one example of the analytical limits which lower the full potential of these complex model systems.

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Nakajima, Keiji, Yusuke Komiyama, Hironori Hojo, Shinsuke Ohba, Fumiko Yano, Naoko Nishikawa, Hiroyuki Aburatani, Tsuyoshi Takato, and Ung-il Chung. "Enhancement of bone formation ex vivo and in vivo by a helioxanthin-derivative." Biochemical and Biophysical Research Communications 395, no. 4 (May 2010): 502–8. http://dx.doi.org/10.1016/j.bbrc.2010.04.041.

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21

Kuznetsov, Sergei A., Mahesh H. Mankani, and Pamela Gehron Robey. "EFFECT OF SERUM ON HUMAN BONE MARROW STROMAL CELLS: EX VIVO EXPANSION AND IN VIVO BONE FORMATION." Transplantation 70, no. 12 (December 2000): 1780–87. http://dx.doi.org/10.1097/00007890-200012270-00018.

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22

Klüter, Tim, Rywan Hassan, Alexander Rasch, Hendrik Naujokat, Fanlu Wang, Peter Behrendt, Sebastian Lippross, et al. "An Ex Vivo Bone Defect Model to Evaluate Bone Substitutes and Associated Bone Regeneration Processes." Tissue Engineering Part C: Methods 26, no. 1 (January 1, 2020): 56–65. http://dx.doi.org/10.1089/ten.tec.2019.0274.

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23

Jakubikova, Jana, Danka Cholujova, Richard W. Groen, Jungnam Joo, Sun-Young Kong, Teru Hideshima, Rikio Suzuki, et al. "Mimicking Myeloma Niche Ex Vivo." Blood 124, no. 21 (December 6, 2014): 2076. http://dx.doi.org/10.1182/blood.v124.21.2076.2076.

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Abstract Introduction: Recent studies have elucidated the importance of using 3-dimensional rather than 2-dimensional models in order to create an experimental system recapitulating the specialized properties of the bone marrow microenvironment. Since the neoplastic bone marrow (BM) milieu plays important roles in multiple myeloma (MM) pathogenesis, novel models to study the MM cell in its neoplastic microenvironment are needed. Methods: To mimic the neoplastic BM microenvironment of MM patients, we have established a special hydrogel-based 3-dimensional (3-D) model by ex-vivo culturing MM patient-derived mesenchymal stem cells (MM-MSCs), the predominant cellular component of the marrow niche, which promotes greater mineralization and differentiation than a 2-dimensional (2-D) system. Results: To characterize MM-MSCs in different stages of MM, we utilized an 11 multi-color flow cytometry panel. The percentage of MSCs (CD73+CD90+CD105+lin-CD45-CD34-HLA-DR-) population in BM aspirate samples of 50 MM patients (MGUS, smoldering MM, newly diagnosed MM, and relapsed or relapsed/refractory MM) was evaluated, and correlated with the distribution of (CD38+ CD138+) plasma cells. MSCs were less frequent (10x) than plasma cells, and increased with disease progression to relapsed/refractory MM. We seeded MM-MSCs (N=34) which had been expanded by adhesion methods in 2-D versus 3-D models in order to create an ex-vivo MM niche-like structure. In the hydrogel-based 3-D model, MM-MSCs formed compact clusters with active fibrous connections and meshwork-like structures at day 3 to 7. Moreover, calcium mineralization of clusters was observed, associated with the capacity for differentiation towards the osteoblastogenic or adipogenic lineage when cultured with differentiation media. Furthermore, the production of osteopontin (OPN) and angiopoietin-2 (Ang-2) was significantly higher in 3-D vs. 2-D MM-MSCs, assessed by multiplex luminex technology. Phenotypic profiling of 3-D MM-MSCs clusters revealed high expression of CD73+CD90+CD105+ and lack of expression of CD45, CD34 and HLA-DR, as in to 2-D MM-MSCs. MSC-specific markers including CD166 and HLA-ABC did not reveal any significant changes in 3-D vs. 2-D MM-MSCs; however, 3-D MM-MSCs had significantly decreased expression of CD271 and CD146 compared to 2-D cultures. We also observed significantly higher expression of extracellular matrix (ECM) molecules including fibronectin, laminin, collagen I, and collagen IV (p<0.001) in 3-D vs. 2-D MM-MSCs. Similarly, activation of integrins including VLA-2, VLA-4 and VLA-5 on the MSCs surface was also increased in 3-D MM-MSCs, as determined by confocal microscopy and flow cytometry analysis. Importantly, MM-MSCs cultured in 3-D vs. 2-D model have higher expression of N-cadherin and CXCL12 and decreased expression of nestin, reflecting the MM BM niche. Gene expression analyses of 3-D MM-MSCs revealed upregulation of BMP-2, MGP, PTGIS, COL14A1 and other genes and down-regulation of DKK1, ADAM9, OPCML genes and others compared to 2-D MM-MSCs. We also measured significantly higher production of IL-6 (p=0.002), IL-8, MCP-1(MCAF), RANTES, VEGF and HGF (p<0.001) in 3-D vs. 2-D MM-MSCs, by multiplex luminex analysis. Next, we co-cultured tumor cells from MM patients (12 MM patients) with either autologous or allogeneic MM-MSCs in 3-D vs. 2-D model. Plasma (CD38/CD138+) cells in 3-D co-culture were increased in 8/12 MM patients and equivalent to 2-D in 4/12 patients. By co-culturing MM cell lines (OPM1, RPMI-S, OCIMY5 and KMS11) labeled with CFSE fluorescent dye with various MSCs, we evaluated expression of side population (SP) cells, identified by Hoechst staining, and gating on CFSE positive MM cells, as low Hoechst stained cells. Our results showed that the SP fraction was significantly lower in 3-D compared to 2-D in co-cultures of various MM-MSCs with all 4 MM cell lines. Finally, we validated drug resistance to melphalan, bortezomib, lenalidomide, and carfilzomib in 3-D co-cultures of CFSE labeled primary tumor cells with various MM-MSCs. Conclusions: This 3-D co-culture system closely mimics the myeloma BM niche, and therefore may be useful to identify and validate novel targeted therapies. Disclosures No relevant conflicts of interest to declare.

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Guidi, Novella, Gina Marka, Vadim Sakk, Yi Zheng, Maria Carolina Florian, and Hartmut Geiger. "An Aged Bone Marrow Niche Restrains Rejuvenated Hematopoietic Stem Cells." Stem Cells 39, no. 8 (April 13, 2021): 1101–6. http://dx.doi.org/10.1002/stem.3372.

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Abstract Aging-associated leukemia and aging-associated immune remodeling are in part caused by aging of hematopoietic stem cells (HSCs). An increase in the activity of the small RhoGTPase cell division control protein 42 (Cdc42) within HSCs causes aging of HSCs. Old HSCs, treated ex vivo with a specific inhibitor of Cdc42 activity termed CASIN, stay rejuvenated upon transplantation into young recipients. We determined in this study the influence of an aged niche on the function of ex vivo rejuvenated old HSCs, as the relative contribution of HSCs intrinsic mechanisms vs extrinsic mechanisms (niche) for aging of HSCs still remain unknown. Our results show that an aged niche restrains the function of ex vivo rejuvenated HSCs, which is at least in part linked to a low level of the cytokine osteopontin found in aged niches. The data imply that sustainable rejuvenation of the function of aged HSCs in vivo will need to address the influence of an aged niche on rejuvenated HSCs.

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Anesi, Alexandre, Mattia Di Bartolomeo, Arrigo Pellacani, Marzia Ferretti, Francesco Cavani, Roberta Salvatori, Riccardo Nocini, Carla Palumbo, and Luigi Chiarini. "Bone Healing Evaluation Following Different Osteotomic Techniques in Animal Models: A Suitable Method for Clinical Insights." Applied Sciences 10, no. 20 (October 14, 2020): 7165. http://dx.doi.org/10.3390/app10207165.

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Osteotomy is a common step in oncological, reconstructive, and trauma surgery. Drilling and elevated temperature during osteotomy produce thermal osteonecrosis. Heat and associated mechanical damage during osteotomy can impair bone healing, with consequent failure of fracture fixation or dental implants. Several ex vivo studies on animal bone were recently focused on heating production during osteotomy with conventional drill and piezoelectric devices, particularly in endosseous dental implant sites. The current literature on bone drilling and osteotomic surface analysis is here reviewed and the dynamics of bone healing after osteotomy with traditional and piezoelectric devices are discussed. Moreover, the methodologies involved in the experimental osteotomy and clinical studies are compared, focusing on ex vivo and in vivo findings.

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Delguste, C., G. Perona, P. Lebecque, F. Duboeuf, O. Lepage, W. Martin-Rosset, and M. Donabedian. "Third metacarpal bone mineral density assessment in the standing horse by dual X-ray absorptiometry." Veterinary and Comparative Orthopaedics and Traumatology 18, no. 01 (2005): 26–30. http://dx.doi.org/10.1055/s-0038-1632924.

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SummaryBone mineral density (BMD) is correlated to mechanical properties of bone. In the horse, dual energy X-ray absorptiometry (DXA) has yet only been performed ex-vivo, but a new portable DXA device would be ideal for in-vivo BMD measurement. We explored field suitability, precision and accuracy of this device for in-vivo third metacarpal density assessment. Precision was analysed by calculating measurement variation under repeated measurement tests with (reproducibility) and without (repeatability) limb repositioning. Repeatability and reproducibility were tested ex-vivo, at the same time that intra- and inter-operator reproducibility were assessed in-vivo. In order to test accuracy, bone mineral content (BMC) of several bone samples determined by DXA and ashing were compared. Repeatability was 1.47% and reproducibility 1.69% ex-vivo. In-vivo reproducibility varied between 2.91 and 4.06% for intraoperator test and between 3.13 and 5.53% for interoperator test. BMC measured by DXA and ash weight were highly correlated (R2>0.99). In conclusion, under described conditions this DXA device is usable, accurate and precise. Its sensitiveness reaches 8.23% in an individual longitudinal monitoring. Using the third metacarpal bone as an example, we have shown that this device is suitable for experimental or clinical monitoring.

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Dana M., Marzocco, Lee Sean, Kurtz Kenneth S, Fawad Javed, Delgado-Ruiz Rafael, and Romanos Georgios E. "Temperature changes in bone using an air scaler Ex Vivo." STOMATOLOGY EDU JOURNAL 7, no. 4 (2020): 252–58. http://dx.doi.org/10.25241/stomaeduj.2020.7(4).art.3.

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Introduction Frictional forces induced by osteotomy devices may induce an unwanted temperature increase in bone. This experimental study aimed to evaluate temperature changes produced in dense bone by three different osteotomies produced by an air scaler device. Methodology Under the same parameters, forty-five linear osteotomies were prepared on the cortical layer of fresh porcine ribs resembling dense bone with three different air scaler insert tips: sagittal saw (Tip A), diamond ball (Tip B) and square chisel (Tip C). The length of the osteotomies was standardized to 10 mm in length. The depths of cuts ranged from 0.5 mm to 2.0 mm. The future osteotomy areas were marked with a graphite pen, and thermocouple microprobes were placed 1 mm lateral at both sides of the marks. The maximum temperature, differential temperature, and time for cut completion were recorded. Analysis of Variance and Kruskal Wallis test were used for the group comparisons. Results Tip A induced the highest of the maximum temperature recordings (Tip A: 48.0 o C). Tip B and C produced comparable maximum temperatures (Tip B: 43.6 o C and Tip C: 44.0 o C). Total mean temperature change increased more for Tip B (4.13) and less in Tip C (0.2). Timing of cuts ranged from 30 seconds to 5 minutes (2.30 ± 1.76 min). Overall average temperature change was less than 100 o C within one minute. Conclusion Osseous site preparation can be achieved with the Air scaler and different air scaler inserts without inducing significant critical thermal changes in bone.

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Arellano, Danna L., Andrea Verdugo-Meza, Florian Drescher, Felipe Olvera-Rodriguez, Patricia Juarez, and Pierrick Fournier. "T cell suppression by the bone microenvironment increases bone metastases from breast cancer in mice." Journal of Immunology 202, no. 1_Supplement (May 1, 2019): 138.10. http://dx.doi.org/10.4049/jimmunol.202.supp.138.10.

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Abstract Bone metastases are a frequent complication of breast cancer and cannot be cured. Immunotherapy could be used for their treatment. However, its efficiency could be limited by the ability of T cells to increase bone resorption that supports cancer cell proliferation and bone metastases. To characterize the effect of T cells on bone metastases, we used a syngeneic mouse model with 4T1 breast cancer cells. When inoculated in normal mice, 4T1 caused more osteolysis as measured on x-rays, compared to SCID and T cell-depleted mice (253% and 61% more, respectively). Histology confirmed that normal mice have an increased tumor burden and number of osteoclasts at the tumor/bone interface compared to SCID mice. T cells isolated from bone metastases suppressed osteoclast formation ex vivo, in contrast with in vivo data. However, this effect was due to the ex vivo activation of T cells, consistent with decreased levels of Rankl and increased expression of anti-osteoclastic Ifng and Il4 mRNA in activated T cells. In vivo, Ifng was not detected, and levels of pro-osteoclastic Rankl and Tnfa were higher in T cells from bone metastases, suggesting they are pro-osteoclastic, unlike activated T cells. Culturing bone metastasis T cells ex vivo with ConA or anti-CD3/CD28, failed to increase the expression of the activation marker CD69, confirming the presence of suppressive factors in this microenvironment. Accordingly, there was an increase of MDSCs, including monocytic MDSCs that are PD-L1+ (87%) in 4T1 bone metastases. They could suppress the activation of T cells that are PD-1+ (71%) in bone metastases. These results suggest that unactivated T cells increase bone metastases and their activation by immunotherapy could be used for the treatment of bone metastases.

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Rowley, SD, RJ Jones, S. Piantadosi, HG Braine, OM Colvin, J. Davis, R. Saral, S. Sharkis, J. Wingard, and AM Yeager. "Efficacy of ex vivo purging for autologous bone marrow transplantation in the treatment of acute nonlymphoblastic leukemia." Blood 74, no. 1 (July 1, 1989): 501–6. http://dx.doi.org/10.1182/blood.v74.1.501.501.

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Abstract We used an in vitro measure of drug activity to predict the efficacy of ex vivo purging of leukemic cells from autologous bone marrow grafts. We previously found that the myeloid progenitor cell (CFU-GM) content of the marrow grafts after ex vivo purging with 4- hydroperoxycyclophosphamide (4-HC) correlates with time to hematologic recovery after autologous bone marrow transplantation in patients with acute nonlymphoblastic leukemia. We observed that variable red blood cell concentration of the bone marrow incubation mixture results in differential cytotoxic activity of 4-HC. The CFU-GM content of the graft after the ex vivo treatment is a measure of this 4-HC activity. We analyzed the disease-free survival of 45 patients with acute nonlymphoblastic leukemia undergoing autologous bone marrow transplantation with 4-HC purged grafts. Patients who relapsed after transplantation had 4.2 +/- 1.1% of graft CFU-GM surviving the ex vivo purge, compared with 1.1 +/- 0.4% for patients who achieved a sustained remission (P = .06). Twenty-three patients with a CFU-GM content after 4-HC purging of greater than 1% of the pretreatment value had an actuarial disease-free survival of 12%, compared to 36% for 22 patients with a less than or equal to 1% CFU-GM content after purging (P = .006). Therefore, percent CFU-GM survival as a measure of 4-HC cytotoxicity identified a group of patients with insufficient purging. Although no randomized clinical trials have documented the need for ex vivo purging, our results suggest that effective bone marrow purging is important for the optimal application of autologous transplantation in the treatment of acute nonlymphoblastic leukemia.

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Rowley, SD, RJ Jones, S. Piantadosi, HG Braine, OM Colvin, J. Davis, R. Saral, S. Sharkis, J. Wingard, and AM Yeager. "Efficacy of ex vivo purging for autologous bone marrow transplantation in the treatment of acute nonlymphoblastic leukemia." Blood 74, no. 1 (July 1, 1989): 501–6. http://dx.doi.org/10.1182/blood.v74.1.501.bloodjournal741501.

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We used an in vitro measure of drug activity to predict the efficacy of ex vivo purging of leukemic cells from autologous bone marrow grafts. We previously found that the myeloid progenitor cell (CFU-GM) content of the marrow grafts after ex vivo purging with 4- hydroperoxycyclophosphamide (4-HC) correlates with time to hematologic recovery after autologous bone marrow transplantation in patients with acute nonlymphoblastic leukemia. We observed that variable red blood cell concentration of the bone marrow incubation mixture results in differential cytotoxic activity of 4-HC. The CFU-GM content of the graft after the ex vivo treatment is a measure of this 4-HC activity. We analyzed the disease-free survival of 45 patients with acute nonlymphoblastic leukemia undergoing autologous bone marrow transplantation with 4-HC purged grafts. Patients who relapsed after transplantation had 4.2 +/- 1.1% of graft CFU-GM surviving the ex vivo purge, compared with 1.1 +/- 0.4% for patients who achieved a sustained remission (P = .06). Twenty-three patients with a CFU-GM content after 4-HC purging of greater than 1% of the pretreatment value had an actuarial disease-free survival of 12%, compared to 36% for 22 patients with a less than or equal to 1% CFU-GM content after purging (P = .006). Therefore, percent CFU-GM survival as a measure of 4-HC cytotoxicity identified a group of patients with insufficient purging. Although no randomized clinical trials have documented the need for ex vivo purging, our results suggest that effective bone marrow purging is important for the optimal application of autologous transplantation in the treatment of acute nonlymphoblastic leukemia.

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Watanabe, Ryota, Aira Matsugaki, Takuya Ishimoto, Ryosuke Ozasa, Takuya Matsumoto, and Takayoshi Nakano. "A Novel Ex Vivo Bone Culture Model for Regulation of Collagen/Apatite Preferential Orientation by Mechanical Loading." International Journal of Molecular Sciences 23, no. 13 (July 4, 2022): 7423. http://dx.doi.org/10.3390/ijms23137423.

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The anisotropic microstructure of bone, composed of collagen fibers and biological apatite crystallites, is an important determinant of its mechanical properties. Recent studies have revealed that the preferential orientation of collagen/apatite composites is closely related to the direction and magnitude of in vivo principal stress. However, the mechanism of alteration in the collagen/apatite microstructure to adapt to the mechanical environment remains unclear. In this study, we established a novel ex vivo bone culture system using embryonic mouse femurs, which enabled artificial control of the mechanical environment. The mineralized femur length significantly increased following cultivation; uniaxial mechanical loading promoted chondrocyte hypertrophy in the growth plates of embryonic mouse femurs. Compressive mechanical loading using the ex vivo bone culture system induced a higher anisotropic microstructure than that observed in the unloaded femur. Osteocytes in the anisotropic bone microstructure were elongated and aligned along the long axis of the femur, which corresponded to the principal loading direction. The ex vivo uniaxial mechanical loading successfully induced the formation of an oriented collagen/apatite microstructure via osteocyte mechano-sensation in a manner quite similar to the in vivo environment.

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Abrams, Kraig, Scott S. Graves, Maura H. Parker, and Rainer Storb. "CD94 Ex Vivo Cultures in a Bone Marrow Transplantation Setting." Transplantation Direct 6, no. 12 (November 16, 2020): e632. http://dx.doi.org/10.1097/txd.0000000000001082.

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Wroblewski, B. Mike, Max Esser, and David W. Srigley. "Release of gentamicin from bone cement an ex-vivo study." Acta Orthopaedica Scandinavica 57, no. 5 (January 1986): 413–14. http://dx.doi.org/10.3109/17453678609014759.

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Smith, Emma L., Matthew Locke, Rachel J. Waddington, and Alastair J. Sloan. "An Ex Vivo Rodent Mandible Culture Model for Bone Repair." Tissue Engineering Part C: Methods 16, no. 6 (December 2010): 1287–96. http://dx.doi.org/10.1089/ten.tec.2009.0698.

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Sloan, A. J., S. Y. Taylor, E. L. Smith, J. L. Roberts, L. Chen, X. Q. Wei, and R. J. Waddington. "A Novel Ex vivo Culture Model for Inflammatory Bone Destruction." Journal of Dental Research 92, no. 8 (July 15, 2013): 728–34. http://dx.doi.org/10.1177/0022034513495240.

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36

Ammi, Azzdine Y., Douglas T. Mast, I‐Hua Huang, Todd A. Abruzzo, Constantin C. Coussios, George J. Shaw, and Christy K. Holland. "Characterization of Ultrasound Propagation Through Ex‐vivo Human Temporal Bone." Journal of the Acoustical Society of America 123, no. 5 (May 2008): 3632. http://dx.doi.org/10.1121/1.2934870.

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Ammi, Azzdine Y., T. Douglas Mast, I. Hua Huang, Todd A. Abruzzo, Constantin-C. Coussios, George J. Shaw, and Christy K. Holland. "Characterization of Ultrasound Propagation Through Ex-vivo Human Temporal Bone." Ultrasound in Medicine & Biology 34, no. 10 (October 2008): 1578–89. http://dx.doi.org/10.1016/j.ultrasmedbio.2008.02.012.

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38

Pantawane, Mangesh V., Yee-Hsien Ho, William B. Robertson, Riaz J. K. Khan, Daniel P. Fick, and Narendra B. Dahotre. "Thermal Assessment of Ex Vivo Laser Ablation of Cortical Bone." ACS Biomaterials Science & Engineering 6, no. 4 (February 4, 2020): 2415–26. http://dx.doi.org/10.1021/acsbiomaterials.9b01559.

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39

Hervé, P., and J. Y. Cahn. "Ex vivo and conditioning chemotherapy for autologous bone marrow transplantation." Baillière's Clinical Haematology 4, no. 1 (January 1991): 223–46. http://dx.doi.org/10.1016/s0950-3536(05)80292-6.

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40

Bell, A. J. "Ex vivo manipulation of bone marrow grafts with monoclonal antibodies." Transfusion Science 10, no. 1 (January 1989): 39–50. http://dx.doi.org/10.1016/0955-3886(89)90007-6.

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41

Nahirnyak, Volodymyr M., and Christy K. Holland. "Ultrasound hyperthermia in clotted blood and ex vivo cranial bone." Journal of the Acoustical Society of America 119, no. 5 (May 2006): 3377. http://dx.doi.org/10.1121/1.4808927.

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42

Riglet, L., C. Confavreux, P. Chaudier, J. B. Pialat, F. Bermond, M. Gardegaront, H. Follet, and D. Mitton. "Ex vivo experiments on femurs to assess metastatic bone strength." Computer Methods in Biomechanics and Biomedical Engineering 23, sup1 (October 19, 2020): S260—S261. http://dx.doi.org/10.1080/10255842.2020.1815312.

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43

Paprocka, M., A. Wiedlocha, and Cz Radzikowski. "Comparative studies on mouse leukemic bone marrow purging ex vivo." European Journal of Cancer and Clinical Oncology 27 (January 1991): S55. http://dx.doi.org/10.1016/0277-5379(91)91386-w.

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Tsiridis, Eleftherios, Neelam Gurav, Guy Bailey, Rod Sambrook, and Lucy Di Silvio. "A novel ex vivo culture system for studying bone repair." Injury 37, no. 3 (September 2006): S10—S17. http://dx.doi.org/10.1016/j.injury.2006.08.019.

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45

Burgner, J., M. Müller, J. Raczkowsky, and H. Wörn. "Ex vivo accuracy evaluation for robot assisted laser bone ablation." International Journal of Medical Robotics and Computer Assisted Surgery 6, no. 4 (November 11, 2010): 489–500. http://dx.doi.org/10.1002/rcs.366.

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46

Abubakar, Adamu Abdul, Sahar Mohammed Ibrahim, Ahmed Khalaf Ali, Kareem Obayes Handool, Mohammad Shuaib Khan, Mohamed Noordin Mustapha, Tengku Azmi Ibrahim, Ubedullah Kaka, and Loqman Mohamad Yusof. "Postnatal ex vivo rat model for longitudinal bone growth investigations." Animal Models and Experimental Medicine 2, no. 1 (February 20, 2019): 34–43. http://dx.doi.org/10.1002/ame2.12051.

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47

Rutherford, R. B., P. Racenis, S. Fatherazi, and K. Izutsu. "Bone Formation by BMP-7-transduced Human Gingival Keratinocytes." Journal of Dental Research 82, no. 4 (April 2003): 293–97. http://dx.doi.org/10.1177/154405910308200410.

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BMPs are a family of pleiotropic signaling molecules involved at various stages in the formation of bones and teeth. In addition, recombinant BMP can induce bone and dentin regeneration when applied directly to adult tissues. We have shown that fibroblasts transduced ex vivo by BMP cDNA delivered by recombinant adenoviruses induce bone formation and convert to osteoblasts upon implantation in vivo. To determine if this osteogenic capacity was limited to fibroblasts, we found that BMP-7-transduced human oral keratinocyte cells (HOKC) also formed ectopic bone. The ossicles formed by the BMP-7-transduced HOKC were smaller and more dense than those formed by BMP-7-transduced human gingival fibroblasts (HGF). Implanted HOKC were localized adjacent to the developing bone by immunocytochemical detection of keratin expression. However, no bone-like tissue formed when HOKC were implanted into diffusion chambers in vivo. We conclude that BMP-transduced HOKC secrete BMP and form bone in vivo but, unlike BMP-transduced HGF, do not transdifferentiate to osteoblasts.

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48

Laranga, Roberta, Serena Duchi, Toni Ibrahim, Ania Naila Guerrieri, Davide Maria Donati, and Enrico Lucarelli. "Trends in Bone Metastasis Modeling." Cancers 12, no. 8 (August 17, 2020): 2315. http://dx.doi.org/10.3390/cancers12082315.

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Bone is one of the most common sites for cancer metastasis. Bone tissue is composed by different kinds of cells that coexist in a coordinated balance. Due to the complexity of bone, it is impossible to capture the intricate interactions between cells under either physiological or pathological conditions. Hence, a variety of in vivo and in vitro approaches have been developed. Various models of tumor–bone diseases are routinely used to provide valuable information on the relationship between metastatic cancer cells and the bone tissue. Ideally, when modeling the metastasis of human cancers to bone, models would replicate the intra-tumor heterogeneity, as well as the genetic and phenotypic changes that occur with human cancers; such models would be scalable and reproducible to allow high-throughput investigation. Despite the continuous progress, there is still a lack of solid, amenable, and affordable models that are able to fully recapitulate the biological processes happening in vivo, permitting a correct interpretation of results. In the last decades, researchers have demonstrated that three-dimensional (3D) methods could be an innovative approach that lies between bi-dimensional (2D) models and animal models. Scientific evidence supports that the tumor microenvironment can be better reproduced in a 3D system than a 2D cell culture, and the 3D systems can be scaled up for drug screening in the same way as the 2D systems thanks to the current technologies developed. However, 3D models cannot completely recapitulate the inter- and intra-tumor heterogeneity found in patients. In contrast, ex vivo cultures of fragments of bone preserve key cell–cell and cell–matrix interactions and allow the study of bone cells in their natural 3D environment. Moreover, ex vivo bone organ cultures could be a better model to resemble the human pathogenic metastasis condition and useful tools to predict in vivo response to therapies. The aim of our review is to provide an overview of the current trends in bone metastasis modeling. By showing the existing in vitro and ex vivo systems, we aspire to contribute to broaden the knowledge on bone metastasis models and make these tools more appealing for further translational studies.

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Shih, Yuru Vernon, and Shyni Varghese. "Tissue engineered bone mimetics to study bone disorders ex vivo: Role of bioinspired materials." Biomaterials 198 (April 2019): 107–21. http://dx.doi.org/10.1016/j.biomaterials.2018.06.005.

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Pelled, Gadi, Dmitriy Sheyn, Wafa Tawackoli, Deuk Soo Jun, Youngdo Koh, Susan Su, Doron Cohn Yakubovich, et al. "BMP6-Engineered MSCs Induce Vertebral Bone Repair in a Pig Model: A Pilot Study." Stem Cells International 2016 (2016): 1–8. http://dx.doi.org/10.1155/2016/6530624.

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Osteoporotic patients, incapacitated due to vertebral compression fractures (VCF), suffer grave financial and clinical burden. Current clinical treatments focus on symptoms’ management but do not combat the issue at the source. In this pilot study, allogeneic, porcine mesenchymal stem cells, overexpressing the BMP6 gene (MSC-BMP6), were suspended in fibrin gel and implanted into a vertebral defect to investigate their effect on bone regeneration in a clinically relevant, large animal pig model. To check the effect of the BMP6-modified cells on bone regeneration, a fibrin gel only construct was used for comparison. Bone healing was evaluated in vivo at 6 and 12 weeks and ex vivo at 6 months. In vivo CT showed bone regeneration within 6 weeks of implantation in the MSC-BMP6 group while only minor bone formation was seen in the defect site of the control group. After 6 months, ex vivo analysis demonstrated enhanced bone regeneration in the BMP6-MSC group, as compared to control. This preclinical study presents an innovative, potentially minimally invasive, technique that can be used to induce bone regeneration using allogeneic gene modified MSCs and therefore revolutionize current treatment of challenging conditions, such as osteoporosis-related VCFs.

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