Browsing by Author "Varma, Harikrishna"
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Item 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, 2009Item 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.Item Laser surface modification of titanium substrate for pulsed laser deposition of highly adherent hydroxyapatite(JOURNAL OF MATERIALS SCIENCE-MATERIALS IN MEDICINE, 2011)Biomedical implant devices made out of titanium and its alloys are benefited by a modified surface or a bioactive coating to enhance bone bonding ability and to function effectively in vivo for the intended period of time. In this respect hydroxyapatite coating developed through pulsed laser deposition is a promising approach. Since the success of the bioactive ceramic coated implant depends mainly on the substrate-coating strength; an attempt has been made to produce micro patterned surface structure on titanium substrate for adherent hydroxyapatite coating. A pulsed Nd-YAG laser beam (355 nm) with 10 Hz repetition rate was used for surface treatment of titanium as well as hydroxyapatite deposition. The unfocussed laser beam was used to modify the substrate surface with 500-18,000 laser pulses while keeping the polished substrate in water. Hydroxyapatite deposition was done in a vacuum deposition chamber at 400A degrees C with the focused laser beam under 1 x 10(-3) mbar oxygen pressure. Deposits were analyzed to understand the physico-chemical, morphological and mechanical characteristics. The obtained substrate and coating surface morphology indicates that laser treatment method can provide controlled micro-topography. Scratch test analysis and microindentation hardness values of coating on laser treated substrate indicate higher mechanical adhesion with respect to coatings on untreated substrates.Item Preparation and analysis of chemically gradient functional bioceramic coating formed by pulsed laser deposition(JOURNAL OF MATERIALS SCIENCE-MATERIALS IN MEDICINE, 2012)Bioactive ceramic coatings based on calcium phosphates yield better functionality in the human body for a variety of metallic implant devices including orthopaedic and dental prostheses. In the present study chemically and hence functionally gradient bioceramic coating was obtained by pulsed laser deposition method. Calcium phosphate bioactive ceramic coatings based on hydroxyapatite (HA) and tricalcium phosphate (TCP) were deposited over titanium substrate to produce gradation in physico-chemical characteristics and in vitro dissolution behaviour. Sintered targets of HA and alpha-TCP were deposited in a multi target laser deposition system. The obtained deposits were characterized by X-ray diffraction, fourier transform infrared spectroscopy, scanning electron microscopy and energy dispersive X-ray analysis. Inductively coupled plasma spectroscopy was used to estimate the in vitro dissolution behaviour of coatings. The variation in mechanical property of the gradient layer was evaluated through scratch test and micro-indentation hardness. The bioactivity was examined in vitro with respect to the ability of HA layer to form on the surface as a result of contact with simulated body fluid. It could be inferred that chemically gradient functional bioceramic coating can be produced by laser deposition of multiple sintered targets with variable chemical composition.Item Pulsed laser deposition of hydroxyapatite on titanium substrate with titania interlayer(JOURNAL OF MATERIALS SCIENCE-MATERIALS IN MEDICINE, 2011)Pulsed laser deposition (PLD) has been used to deposit hydroxyapatite (HA) ceramic over titanium substrate with an interlayer of titania. PLD has been identified as a potential candidate for bioceramic coatings over metallic substrates to be used as orthopedic and dental implants because of better process control and preservation of phase identity of the coating component. However, direct deposition of hydroxyapatite on titanium at elevated temperature results in the formation of natural oxide layer along with some perovskites like calcium titanate at the interface. This leads to easy debonding of ceramic layer from the metal and thereby affecting the adhesion strength. In the present study, adherent and stable HA coating over Ti6Al4V was achieved with the help of an interlayer of titania. The interlayer was made to a submicron level and HA was deposited consecutively to a thickness of around one micron by exposing to laser ablation at a substrate temperature of 400A degrees C. The deposited phase was identified to be phase pure HA by X-ray diffraction, scanning electron microscopy, energy dispersive X-ray analysis, and inductively coupled plasma spectrometry. The mechanical behavior of coating evaluated by scratch test indicates that the adhesion strength of HA coating was improved with the presence of titania interlayer.Item 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.