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Shaohong Cheng, Patrick Aghajanian, Sheila Pourteymoor, Catrina Alarcon, Subburaman Mohan
Endochondral ossification plays an important role in the formation of the primary ossification centers (POCs) and secondary ossification centers (SOCs) of mammalian long bones. However, the molecular mechanisms that regulate POC and SOC formation are different. We recently demonstrated that Prolyl Hydroxylase Domain-containing Protein 2 (Phd2) is a key mediator of vitamin C effects on bone. We investigated the role of Phd2 on endochondral ossification of the epiphyses by conditionally deleting the Phd2 gene in osteoblasts and chondrocytes...
October 24, 2016: Scientific Reports
Chia-Feng Liu, William E Samsa, Guang Zhou, Véronique Lefebvre
A milestone in the evolutionary emergence of vertebrates was the invention of cartilage, a tissue that has key roles in modeling, protecting and complementing the bony skeleton. Cartilage is elaborated and maintained by chondrocytes. These cells derive from multipotent skeletal progenitors and they perform highly specialized functions as they proceed through sequential lineage commitment and differentiation steps. They form cartilage primordia, the primary skeleton of the embryo. They then transform these primordia either into cartilage growth plates, temporary drivers of skeletal elongation and endochondral ossification, or into permanent tissues, namely articular cartilage...
October 19, 2016: Seminars in Cell & Developmental Biology
Katie Bardsley, Agnieska Kwarciak, Christine Freeman, Ian Brook, Paul Hatton, Aileen Crawford
The regeneration of large bone defects remains clinically challenging. The aim of our study was to use a rat model to use nasal chondrocytes to engineer a hypertrophic cartilage tissue which could be remodelled into bone in vivo by endochondral ossification. Primary adult rat nasal chondrocytes were isolated from the nasal septum, the cell numbers expanded in monolayer culture and the cells cultured in vitro on polyglycolic acid scaffolds in chondrogenic medium for culture periods of 5-10 weeks. Hypertrophic differentiation was assessed by determining the temporal expression of key marker genes and proteins involved in hypertrophic cartilage formation...
October 11, 2016: Biomaterials
Anurati Saha, Rebecca Rolfe, Simon Carroll, Daniel J Kelly, Paula Murphy
Chondrogenesis in vivo is precisely controlled in time and space. The entire limb skeleton forms from cells at the core of the early limb bud that condense and undergo chondrogenic differentiation. Whether they form stable cartilage at the articular surface of the joint or transient cartilage that progresses to hypertrophy as endochondral bone, replacing the cartilage template of the skeletal rudiment, is spatially controlled over several days in the embryo. Here, we follow the differentiation of cells taken from the early limb bud (embryonic day 11...
October 22, 2016: Cell and Tissue Research
Fiona E Freeman, Laoise McNamara
Tissue engineering and regenerative medicine have significant potential to treat bone pathologies by exploiting the capacity for bone progenitors to grow and produce tissue constituents under specific biochemical and physical conditions. However, conventional tissue engineering approaches, which combine stem cells with biomaterial scaffolds, are limited as the constructs often degrade, due to a lack of vascularisation, and lack the mechanical integrity to fulfil loading bearing functions, and as such are not yet widely used for clinical treatment of large bone defects...
October 19, 2016: Tissue Engineering. Part B, Reviews
Kai Hu, Bjorn R Olsen
Vascular Endothelial Growth factor A (VEGF) is a critical regulator of vascular development, postnatal angiogenesis and homeostasis, and it is essential for bone development and repair. Blood vessels serve both as structural templates for bone formation and they provide essential cells, growth factors and minerals needed for synthesis and mineralization, as well as turnover, of the extracellular matrix in bone. Through its regulation of angiogenesis, VEGF contributes to coupling of osteogenesis to angiogenesis, and it directly controls the differentiation and function of osteoblasts and osteoclasts...
October 17, 2016: Developmental Dynamics: An Official Publication of the American Association of Anatomists
Tsai-Jung Lu, Fang-Yao Chiu, Hsiao-Ying Chiu, Ming-Chau Chang, Shih-Chieh Hung
Articular cartilage has a very limited capacity for self-repair, and mesenchymal stem cells (MSCs) have the potential to treat cartilage defects and osteoarthritis. However, in-depth mechanistic studies regarding their applications are required. Here, we demonstrated the use of chitosan film culture for promoting chondrogenic differentiation of MSCs. We found that MSCs formed spheres two days after seeding on dishes coated with chitosan. When MSCs were induced in a chondrogenic induction medium on chitosan films, the size of spheres continuously increased for up to 21 days...
October 12, 2016: Cell Transplantation
Garyfallia Papaioannou, Fatemeh Mirzamohammadi, Tatsuya Kobayashi
During endochondral bone development, osteoblasts are continuously differentiated from locally residing progenitor cells. However, the regulation of such endogenous osteoprogenitor cells is still poorly understood mainly due to the difficulty in identifying such cells in vivo. In this paper, we genetically labeled different cell populations of the osteoblast linage using stage-specific, tamoxifen-inducible Cre transgenic mice to investigate their responses to a proliferative stimulus. We have found that overactivation of Kras signaling in type II collagen-positive, immature osteoprogenitor cells, but not in mature osteoblasts, substantially increases the number of their descendant stromal cells and mature osteoblasts, and subsequently increases bone mass...
October 13, 2016: Cell Death & Disease
Gabrielle Martel, Charles-André Couture, Guillaume Gilbert, Stéphane Bancelin, Hélène Richard, Thomas Moser, Sabrina Kiss, François Légaré, Sheila Laverty
Osteochondrosis is an ischemic chondronecrosis of epiphyseal growth cartilage that results in focal failure of endochondral ossification and osteochondritis dissecans at specific sites in the epiphyses of humans and animals, including horses. The upstream events leading to the focal ischemia remain unknown. The epiphyseal growth cartilage matrix is composed of proteoglycan and collagen macromolecules and encases its vascular tree in canals. The matrix undergoes major dynamic changes in early life that could weaken it biomechanically and predispose it to focal trauma and vascular failure...
October 2016: Journal of Orthopaedic Research: Official Publication of the Orthopaedic Research Society
Takako Hattori, Shinsuke Itoh, Masaharu Takigawa
Recent progress in gene-editing technology has provided a strong impact for improved our understanding of molecular functions in living organisms. Here we describe our method to generate transgene-overexpressing mouse models, which method involves the use of tissue-specific promoters for analyzing a certain molecule (s) in special tissues. The protocol described in this chapter uses the Col2a1 promoter-enhancer, which is known for driving specific and strong transgene expression in cartilage and is based on several of our studies showing a positive role of the connective tissue growth factor (CCN2) in cartilage-bone development and maintenance of articular cartilage...
2017: Methods in Molecular Biology
Tsuyoshi Shimo, Masaharu Takigawa
Angiogenesis, the process of generating new blood vessels from an existing vasculature, is essential in normal developmental processes such as endochondral ossification and in numerous kinds of pathogenesis including tumor growth. A part from the action of angiogenic factor or antiangiogenic factor, it is still unknown at which stage of the angiogenic cascade these agents affect angiogenesis. Here, we describe methods for the use of connective tissue growth factor (CTGF/CCN2) and CCN2 neutralizing antibody in the currently used principal angiogenesis assays, including those in vitro ones for the proliferation, migration, adhesion, and tube formation of endothelial cells and in vivo assays such as those utilizing type I collagen implantation and the chick chorioallantoic membrane (CAM)...
2017: Methods in Molecular Biology
Connie Y Chang, Daniel I Rosenthal, Deborah M Mitchell, Atsuhiko Handa, Susan V Kattapuram, Ambrose J Huang
Metabolic bone diseases are a diverse group of diseases that result in abnormalities of (a) bone mass, (b) structure mineral homeostasis, (c) bone turnover, or (d) growth. Osteoporosis, the most common metabolic bone disease, results in generalized loss of bone mass and deterioration in the bone microarchitecture. Impaired chondrocyte development and failure to mineralize growth plate cartilage in rickets lead to widened growth plates and frayed metaphyses at sites of greatest growth. Osteomalacia is the result of impaired mineralization of newly formed osteoid, which leads to characteristic Looser zones...
October 2016: Radiographics: a Review Publication of the Radiological Society of North America, Inc
Murong You, Juehua Jing, Dasheng Tian, Jun Qian, Guangrong Yu
Dioscin has been shown to play important roles in suppression of osteoclast maturation. It is proposed as a potential natural product for the treatment of osteoclast-related diseases. We hypothesized in this study that treatment of dioscin on bone marrow mesenchymal stem cells (BMSCs) could increase the osteo-chondrogenic differentiation of BMSCs and promote endochondral ossification of BMSCs in bone fracture environment. BMSCs were extracted from femur and tibia of male C57b mice. Stemness of BMSCs was studied by performing proliferation assay and multilineage differentiation...
2016: American Journal of Translational Research
Adam O'Reilly, Daniel John Kelly
Developing successful tissue engineering strategies requires an understanding of how cells within an implanted scaffold interacts with the host environment. The objective of this study was to use a computational mechanobiological model to explore how the design of a cell laden scaffold influences the spatial formation of cartilage and bone within an osteochondral defect. Tissue differentiation was predicted using a previously developed model in which cell fate depends on the local oxygen tension and the mechanical environment within a damaged joint...
October 6, 2016: Tissue Engineering. Part A
Bin Zhou, Deheng Chen, Huazi Xu, Xiaolei Zhang
Chondrocyte plays a critical role in endochondral ossification and cartilage repair by maintaining the cartilaginous matrix. Statins have been widely used to lower the cholesterol level in patients with cardiovascular disorders. Previous research has demonstrated potential role of statins in chondrocyte proliferation. This study addresses the proliferation-regulatory effect of lovastatin in rabbit chondrocytes as well as the underlying signaling mechanisms, thereby exploring its potential application in chondrocyte-related disorders, such as cartilage damage and osteoarthritis...
October 3, 2016: In Vitro Cellular & Developmental Biology. Animal
Soonchul Lee, Jia Shen, Hsin Chuan Pan, Swati Shrestha, Greg Asatrian, Alan Nguyen, Carolyn Meyers, Vi Nguyen, Min Lee, Chia Soo, Kang Ting, Aaron W James
Hedgehog (Hh) signaling positively regulates both endochondral and intramembranous ossification. Use of small molecules for tissue engineering applications pose several advantages. Here, we examined whether use of an acellular scaffold treated with the small molecule Smoothened agonist (SAG) could aid in critical size mouse calvarial defect repair. First, we verified the pro-osteogenic effect of SAG in vitro, using primary neonatal mouse calvarial cells (NMCCs). Next, a 4 mm non-healing defect was created in the mid-parietal bone of ten week old CD-1 mice...
October 5, 2016: Tissue Engineering. Part A
Lei Shi, Guodong Cai, Jiangang Shi, Yongfei Guo, Dechun Chen, Deyu Chen, Haisong Yang
Ossification of the posterior longitudinal ligament (OPLL) involves ectopic calcification of the spinal ligament preferentially at the cervical spine. OPLL is associated with different diseases and occurs by endochondral ossification, which is associated with the activity of different transcription factors. However, the pathogenesis of OPLL remains unclear. Here, we investigated the role of osterix (Osx), a transcription factor that functions downstream of Runx2 and is an important regulator of osteogenesis, in the process of OPLL in a dexamethasone (Dex)-induced model of spinal ligament ossification...
September 29, 2016: Experimental Cell Research
Esperanza Macarena López-Pliego, Miguel Ángel Giráldez-Sánchez, Juan Mora-Macías, Esther Reina-Romo, Jaime Domínguez
INTRODUCTION: Bone transport (BT) for segmentary bone defects is a well-known technique as it enables correction with new bone formation, which is similar to the previous bone. Despite the high number of experimental studies of distraction osteogenesis in bone lengthening, the types of ossification and histological changes that occur in the regenerate of the bone transport process remain controversial. OBJECTIVE: The aim of this study is to provide the complete evolution of tissues and the types of ossification in the regenerate during the different phases of bone formation after BT until the end of the remodelling period...
September 2016: Injury
Holly E Weiss-Bilka, Megan E McGann, Matthew J Meagher, Ryan K Roeder, Diane R Wagner
Key aspects of native endochondral bone development and fracture healing can be mimicked in mesenchymal stem cells (MSCs) through standard in vitro chondrogenic induction. Exploiting this phenomenon has recently emerged as an attractive technique to engineer bone tissue, however relatively little is known about the best conditions for doing so. The objective of this study was to compare the bone forming capacity and angiogenic induction of hypertrophic cell constructs containing human adipose-derived stem cells (hASCs) primed for chondrogenesis in two different culture systems: high-density pellets and alginate bead hydrogels...
September 30, 2016: Journal of Tissue Engineering and Regenerative Medicine
Anna Neve, Nicola Maruotti, Addolorata Corrado, Francesco Paolo Cantatore
Despite intensive research in spondyloarthritis pathogenesis, some important questions still remain unanswered, particularly concerning enthesis new bone formation. Several evidences suggest that it prevalently occurs by endochondral ossification, however it remains to identify factors that can induce and influence its initiation and progression. Recent progress, achieved in animal models and in vitro and genetic associations studies, has led us to hypothesize that several systemic factors (adipokines and gut hormones) and local factors (BMP and Wnt signalling) as well as angiogenesis and mechanical stress are involved...
September 29, 2016: Annals of Medicine
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