Synergistic effect between bioactive glass foam and a perfusion bioreactor on osteogenic differentiation of human adipose stem cells
ABSTRACT Tissue engineering is a multidisciplinary science which combines a structural scaffold and cells to form a construct able to promote regeneration of injured tissue. Bioactive glass foam produced by sol-gel is an osteoinductive material with a network of interconnected macropores necessary for cell colonization. The use of human adipose-derived stem cell (hASC) presents advantages as the potential for a large number of cells, rapid expansion in vitro and the capability of differentiating into osteoblasts. The use of a bioreactor in three-dimensional cell culture enables greater efficiency for cell nutrition and application of mechanical forces, important modulators of bone physiology. The hASC seeded in a bioactive glass scaffold and cultured in osteogenic Leibovitz L-15 medium in a bioreactor with a flow rate of 0.1 mL/min demonstrated a significant increase in cell proliferation and viability and ALP activity peak after 14 days. The immunofluorescence assay revealed an expression of osteopontin, osteocalcin and type I collagen from 7 to 21 days after culture. The cells changed from a spindle shape to a cuboidal morphology characteristic of osteoblasts. The PCR assay confirmed that osteopontin, osteocalcin and ALP genes were expressed. These results indicate that hASCs differentiated into an osteogenic phenotype when cultured in bioactive glass scaffold, osteogenic Leibovitz L-15 medium and a perfusion bioreactor. Therefore, these results highlight the synergism between a bioactive glass scaffold and the effect of perfusion on cells and indicate the differentiation into an osteogenic phenotype.
- SourceAvailable from: Deepak Kalaskar
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- "ADSCs can undergo osteogenic differentiation in vitro by exposure to a combination of ascorbate, β-glycerophosphate, various bone morphogenetic proteins (BMPs), dexamethasone, and/or vitamin D3, confirming bone formation using Alizarin Red or Von Kossa staining, usually over a two-week period . Several studies have shown that ADSCs will express multiple markers for osteogenesis in these conditions including cbfa-1, alkaline phosphatase, osteopontin, osteocalcin and collagen I [46, 47]. "
ABSTRACT: Craniofacial malformations, have devastating psychosocial implications for many adults and children and causes huge socioeconomic burden. Currently craniofacial defects require soft tissue transfer, bone grafting techniques or difficult procedures such as microvascular free flaps. Such tissues are often limited in quantity, their harvest causes secondary large donor site defects and they lack the capability to fully restore previous form and function. Stem cell technology is being utilised for various tissue and organs of the body and consequently surgeons are eager to transfer these principles for craniofacial surgery. Adipose derived stem cells (ADSCs) are an exciting stem cell source for craniofacial surgeons due to their easy and painless isolation, relatively large abundance and familiarity with the harvesting procedure. ADSCs also have multiple desirable properties including adipogenic, osteogenic and chondrogenic potential, enhancement of angiogenesis and immunodulatory function. Due to these advantageous characteristics, ASDCs have been explored to repair craniofacial bone, soft tissue and cartilage. The desirable characteristics of ADSCs for craniofacial surgical applications will be explained. We report the experimental and clinical studies that have explored the use of ADSCs for bone, cartilage and soft tissue craniofacial defects. We conclude by establishing the key questions that are preventing the clinical application of ADSCs for craniofacial surgery.Stem Cell Reviews and Reports 06/2014; 10(5). DOI:10.1007/s12015-014-9522-3 · 2.77 Impact Factor
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- "Also, in the study performed by Declercq and colleagues , After 6 weeks of dynamic culture, scaffolds were highly colonized and the osteogenic gene expression was higher compared to static cultures. Recently, Silva and colleagues  demonstrated that ASCs differentiate towards the osteogenic phenotype when cultured in a bioactive glass scaffold, with the osteogenic Leibovitz L-15 medium and a perfusion bioreactor, as indicated both the significant increase in cell proliferation and viability, the increased ALP activity, and the expression of osteospecific protein (i.e., osteocalcin and osteopontin) 2-to-3 weeks after culture. Furthermore, a coculture model of human osteoblast and endothelial lineage cells has been established by seeding and culturing cells freshly isolated from the SVF of AT within porous 3D ceramic scaffolds . "
ABSTRACT: Adipose tissue represents a hot topic in regenerative medicine because of the tissue source abundance, the relatively easy retrieval, and the inherent biological properties of mesenchymal stem cells residing in its stroma. Adipose-derived mesenchymal stem cells (ASCs) are indeed multipotent somatic stem cells exhibiting growth kinetics and plasticity, proved to induce efficient tissue regeneration in several biomedical applications. A defined consensus for their isolation, classification, and characterization has been very recently achieved. In particular, bone tissue reconstruction and regeneration based on ASCs has emerged as a promising approach to restore structure and function of bone compromised by injury or disease. ASCs have been used in combination with osteoinductive biomaterial and/or osteogenic molecules, in either static or dynamic culture systems, to improve bone regeneration in several animal models. To date, few clinical trials on ASC-based bone reconstruction have been concluded and proved effective. The aim of this review is to dissect the state of the art on ASC use in bone regenerative applications in the attempt to provide a comprehensive coverage of the topics, from the basic laboratory to recent clinical applications.11/2013; 2013(416391):416391. DOI:10.1155/2013/416391
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ABSTRACT: This review analyzes the literature of bone grafts and introduces tissue engineering as a strategy in this field of orthopedic surgery. We evaluated articles concerning bone grafts; analyzed characteristics, advantages, and limitations of the grafts; and provided explanations about bone-tissue engineering technologies. Many bone grafting materials are available to enhance bone healing and regeneration, from bone autografts to graft substitutes; they can be used alone or in combination. Autografts are the gold standard for this purpose, since they provide osteogenic cells, osteoinductive growth factors, and an osteoconductive scaffold, all essential for new bone growth. Autografts carry the limitations of morbidity at the harvesting site and limited availability. Allografts and xenografts carry the risk of disease transmission and rejection. Tissue engineering is a new and developing option that had been introduced to reduce limitations of bone grafts and improve the healing processes of the bone fractures and defects. The combined use of scaffolds, healing promoting factors, together with gene therapy, and, more recently, three-dimensional printing of tissue-engineered constructs may open new insights in the near future.Journal of Orthopaedic Surgery and Research 03/2014; 9(1):18. DOI:10.1186/1749-799X-9-18 · 1.39 Impact Factor