Browsing by Author "John, Annie"

Now showing 1 - 9 of 9
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    A bioactive triphasic ceramic-coated hydroxyapatite promotes proliferation and osteogenic differentiation of human bone marrow stromal cells
    (JOURNAL OF BIOMEDICAL MATERIALS RESEARCH PART A, 2009)
    Hydroxyapatite (HA) ceramics are widely used as bone graft substitutes because of their biocompatibility and osteoconductivity. However, to enhance the success of therapeutic application, many efforts are undertaken to improve the bioactivity of HA. We have developed a triphasic, silica-containing ceramic-coated hydroxyapatite (HASi) and evaluated its performance as a scaffold for cell-based tissue engineering applications. Human bone marrow stromal cells (hBMSCs) were seeded on both HASi and HA scaffolds and cultured with and without osteogenic supplements for a period of 4 weeks. Cellular responses were determined in vitro in terms of cell adhesion, viability, proliferation, and osteogenic differentiation, where both materials exhibited excellent cytocompatibility. Nevertheless, an enhanced rate of cell proliferation and higher levels of both alkaline phosphatase expression and activity were observed for cells cultured on HASi with osteogenic supplements. These findings indicate that the bioactivity of HA endowed with a silica-containing coating has definitely influenced the cellular activity, projecting HASi as a suitable candidate material for bone regenerative therapy. (C) 2008 Wiley Periodicals, Inc. J Biomed Mater Res 90A: 533-542, 2009
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    A triphasic ceramic-coated porous hydroxyapatite for tissue engineering application
    (ACTA BIOMATERIALIA, 2008)
    Scaffolds which encourage the incorporation of a cell source for tissue engineering applications are critical determinants for clinical defects. Over the years, a number of biomaterials have emerged for cell support and growth, but only a few have demonstrated clinical efficacy. We therefore investigated an in-house-developed silica-based bioactive ceramic for its ability to support and sustain the growth of bone marrow-derived mesenchymal stem cells (BMSCs) in vitro. For this, MSCs aspirated from goat bone marrow were isolated and culture expanded on a novel triphasic ceramic composite coated hydroxyapatite (HASi) scaffold comprising hydroxyapatite, tricalcium phosphate and calcium silicate. The viability of cells that harbored on and within the material was ensured through fluorescence-activated cell sorting and confocal laser scanning microscope and for their anchorage sites by scanning electron microscopy. Interestingly, over the days in culture, cell-cell interactions gradually morphed into woven cell-sheets that spanned across the surface of the HASi, forming a canopy. To conclude, we have attempted to carry out the preliminary cytocompatibility studies of this novel ceramic to establish its appropriateness for bone tissue engineering application which is an important criterion in orthopaedic transplantation and regenerative surgery. (C) 2007 Acta Materialia Inc. Published by Elsevier Ltd. All rights reserved.
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    Adipogenesis on biphasic calcium phosphate using rat adipose-derived mesenchymal stem cells: In vitro and in vivo
    (JOURNAL OF BIOMEDICAL MATERIALS RESEARCH PART A, 2012)
    Developing adipose tissue-engineered construct to mend soft tissue defects arising from traumatic injury, tumor resections, and maxillofacial abnormalities is of prime importance in plastic and reconstructive surgical procedures. It is apparent that the clinical outcome of classic techniques like adipose tissue transplantation is unpredictable, with graft resorption, lack of vascularization, and impaired functionality. In this prospective, the concept of tissue engineering was adopted to fabricate a combination product with biphasic calcium phosphate (BCP) and rat adipose-derived mesenchymal stem cells (ASCs) toward the development of an adipose tissue construct. BCP, a combination of hydroxyapatite and a-tricalcium phosphate, was characterized for its physiochemical properties, and ASCs were characterized for their stemness. The cellceramic interactions were demonstrated in vitro, whereas adipogenesis was picturesquely depicted by Nile red-stained multilocular adipocyte-like cells. Subsequently, the three-dimensional cellceramic-engineered construct was implanted in the rat dorsal muscle for a period of 3 weeks to demonstrate the efficacy of the tissue construct in vivo. Interestingly, the histology of the postimplanted tissue construct revealed the distribution of chicken wire net-like fat cells within the vicinity of the construct. The efficacy of cell transplantation via the scaffold was traced using fluorescent in situ hybridization by labeling the Y chromosome. Thus, the ceramic-based construct may be a good option for reconstruction therapies. (C) 2012 Wiley Periodicals, Inc. J Biomed Mater Res Part A:, 2012.
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    Platelet-Rich Plasma and Fibrin Glue-Coated Bioactive Ceramics Enhance Growth and Differentiation of Goat Bone Marrow-Derived Stem Cells
    (TISSUE ENGINEERING PART A, 2009)
    New biotechnologies such as tissue engineering require functionally active cells within supportive matrices where the physical and chemical stimulus provided by the matrix is indispensable to determine the cellular behavior. This study has investigated the influence of platelet-rich plasma (PRP) and fibrin glue (FG) on the functional activity of goat bone marrow-derived mesenchymal stem cells (gBMSCs) that differentiated into the osteogenic lineage. To achieve this goal, PRP and FG were separately coated on bioactive ceramics like hydroxyapatite (HA) and silica-coated HA (HASi), on which gBMSCs were seeded and induced to differentiate into the osteogenic lineage for 28 days. The cells were then analyzed for viability (lactate dehydrogenase assay: acridine orange and ethidium bromide staining), morphology (scanning electron microscopy), proliferation (picogreen assay), cell cycle assay (propidium iodide staining), and differentiation (alkaline phosphatase [ALP] activity and real-time PCR analysis of ALP, osteocalcin, and osteopontin gene). It has been observed that PRP and FG have appreciably favored the viability, spreading, and proliferation of osteogenic-induced gBMSCs. The osteopontin and osteocalcin expression was significantly enhanced on PRP- and FG-coated HA and HASi, but PRP had effect on neither ALP expression nor ALP activity. The results of this study have depicted that FG-coated ceramics were better than PRP- coated and bare matrices. Among all, the excellent performance was shown by FG coated HASi, which may be attributed to the communal action of the stimulus emanated by Si in HASi and the temporary extracellular matrix provided by FG over HASi. Thus, we can conclude that PRP or FG in combination with bioactive ceramics could possibly enhance the functional activity of cells to a greater extent, promoting the hybrid composite as a promising candidate for bone tissue engineering applications.
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    Reconstruction of goat femur segmental defects using triphasic ceramic-coated hydroxyapatite in combination with autologous cells and platelet-rich plasma
    (ACTA BIOMATERIALIA, 2009)
    Segmental bone defects resulting from trauma or pathology represent a common and significant clinical problem. In this study, a triphasic ceramic (calcium silicate, hydroxyapatite and tricalcium phosphate)-coated hydroxyapatite (HASi) having the benefits of both HA (osteointegration, osteoconduction) and silica (degradation) was used as a bone substitute for the repair of segmental defect (2 cm) created in a goat femur model. Three experimental goat femur implant groups - (a) bare HASi, (b) osteogenic-induced goat bone marrow-derived mesenchymal stem cells cultured HASi (HASi + C) and (c) osteogenic-induced goat bone marrow-derived mesenchymal stem cells cultured HASi + platelet-rich plasma (HASi + CP) - were designed and efficacy performance in the healing of the defect was evaluated. In all the groups, the material united with host bone without any inflammation and an osseous callus formed around the implant. This reflects the osteoconductivity of HASi where the cells have migrated from the cut ends of host bone. The most observable difference between the groups appeared in the mid region of the defect. In bare HASi groups, numerous osteoblast-like cells could be seen together with a portion of material. However, in HASi + C and HASi + CP, about 60-70% of that area was occupied by woven bone, in line with material degradation. The interconnected porous nature (50-500 mu m), together with the chemical composition of the HASi, facilitated the degradation of HASi, thereby opening up void spaces for cellular ingrowth and bone regeneration. The combination of HASi with cells and PRP was an added advantage that could promote the expression of many osteoinductive proteins, leading to faster bone regeneration and material degradation. Based on these results, we conclude that bare HASi can aid in bone regeneration but, with the combination of cells and PRP, the sequence of healing events are much faster in large segmental bone defects in weight-bearing areas in goats. (c) 2009 Acta Materialia Inc. Published by Elsevier Ltd. All rights reserved.
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    Short-term studies using ceramic scaffolds in lapine model for osteochondral defect amelioration.
    (Biomedical materials (Bristol, England), 2012)
    This study was undertaken to glean preliminary information on the role of triphasic ceramic coated hydroxyapatite (HASi) and biphasic (alpha-tricalcium phosphate and hydroxyapatite based) calcium phosphate (BCP) for the development of osteochondral constructs. The proposed constructs were tested for performance in vitro with rabbit adipose-derived mesenchymal stem cells (RADMSCs) and further analysed in vivo in a lapine model for osteochondral defect amelioration. Desirable scaffolding architecture ensuring favourable conditions for cell attachment, nutrient exchange and neo-tissue organization was achieved by the synthesis of porous ceramic blocks and characterizations were carried out using x-ray diffraction and Fourier transform infrared spectroscopy. The cytocompatibility of the scaffold-cell combination product was evaluated using microscopy techniques that proved the scaffold to be non-cytotoxic and favourable for cell growth and proliferation. Short-term implantation studies were conducted with bare cylindrical HASi and BCP scaffolds, press fit deep into the bony bed of the median femoral condyles of the rabbit, which resulted in favourable specific in vivo response of de novo cartilage-like cells on the surface and sub-surface bony trabeculae. The generated pilot data will help to assess the severity of proposed procedures before embarking on scaled-up efforts.
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    Tissue regeneration and repair of goat segmental femur defect with bioactive triphasic ceramic-coated hydroxyapatite scaffold
    (JOURNAL OF BIOMEDICAL MATERIALS RESEARCH PART A, 2009)
    Bone tissue engineering which is a developing and challenging field of science, is expected to enhance the regeneration and repair of bone lost from injury or disease and ultimately to gain its aesthetic contour. The objective of this study was to fabricate a tissue-engineered construct in vitro using a triphasic ceramic-coated hydroxypatite (HASi) in combination with stem cells and to investigate its potential in healing segmental defect in goat model. To accomplish this attempt, mesenchymal stern cells isolated from goat bone marrow were seeded onto HASi scaffolds and induced to differentiate into the osteogenic lineage in vitro. Scanning electron microscopy and light microscopy revealed adhesion and spread-out cells, which eventually formed a cell-sheet like canopy over the scaffold. Cells migrated and distributed themselves within the internal voids of the porous ceramic. Concurrently, the neoosteogenesis of the tissue-engineered construct was validated in vivo in comparison with bare HASi (without cells) in goat femoral diaphyseal segmental defect (2 cm) at 4 months postimplantation through radiography, computed tomography, histology, histomorphometry, scanning electron microscopy and inductively coupled plasma spectrometry. Good osteointegration and osteoconduction was observed in bare and tissue-engineered HASi. The performance of tissue-engineered HASi was better and faster which was evident by the lamellar bone organization of newly formed bone throughout the defect together with the degradation of the material. On the contrary with bare HASi, immature woven bony bridges still intermingled with scattered small remnants of the material was observed in the mid region of the defect at 4 months. Encouraging results from this preclinical study has proved the capability of the tissue-engineered HASi as a promising candidate for the reconstruction of similar bony defects in humans. (C) 2008 Wiley Periodicals, Inc. J Biomed Mater Res 91A: 855-865, 2009
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    Treatment of Goat Femur Segmental Defects with Silica-Coated Hydroxyapatite-One-Year Follow-Up
    (TISSUE ENGINEERING PART A, 2010)
    Segmental bone defects caused by tumor resections, trauma, and skeletal abnormalities such as osteomyelitis remain a major problem in orthopedics because of the lack of predictability in attaining functional bone after the treatment. The objective of this study was to propose an indigenous porous biodegradable triphasic ceramic (calcium silicate, tricalcium phosphate, and hydroxyapatite [HA])-coated HA (core) (HASi) for the repair of such segmental defects. With respect to the synthesis of HASi, HA blocks were prepared by wet precipitation, dipped in silica sol (sol gel method), sintered at 1200 degrees C, polished in the form of hollow cylinder (2 cm long with an outer and inner diameter of 2 cm and 7 mm, respectively), and implanted into a 2-cm segmental defect created in the goat femur diaphysis. This study prolonged for 12 uneventful months and thereafter neo-osteogenesis in par with material degradation was analyzed through radiography, histology, histomorphometry, scanning electron microscope (SEM)-energy dispersive spectrum, micro-computed tomography, and inductively coupled plasma spectrometry. HASi proved to be osteoconductive, osteointegrative, and degradative in nature, without the intervention of fibrous tissue formation at the defect site. Histologically, the newly formed bone reorganized, mineralized, and attained the appearance and contour of the original femoral diaphysis in 1 year. The interconnected porous structure with silica composition aided progressive bone regeneration and repair in par with degradation of the material. Thus the study proposed the possibility of using HASi as a suitable material in clinical orthopedic reconstructive surgery, which remains a formidable challenge.
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    Triphasic ceramic coated hydroxyapatite as a niche for goat stem cell-derived osteoblasts for bone regeneration and repair.
    (Journal of materials science. Materials in medicine, 2009)
    Current treatment strategies for the repair or replacement of bone use synthetic implants with stem cells and their progeny--a new approach to address unmet medical needs. This study has evaluated the effect of a silica-coated bioactive ceramic, namely HASi in comparison to hydroxyapatite (HA) on the adhesion, proliferation and osteogenic differentiation of goat bone marrow-derived mesenchymal stem cells in vitro in a prolonged culture of 28 days. The cellular activities were significantly enhanced on HASi signifying the role of silica to stimulate osteoblast cells. The fabrication of such a 'cell-ceramic construct using autologous MSCs' is aimed for the transplantation to a large bone defect site in the goat femur model which still remains a formidable challenge in Orthopedic surgery.