Transparent hydroxyapatite (HAP) was prepared by sintering gel-cast powder compacts at 1000°C for 2 h; the resultant HAP material was studied using X-ray diffractometry, transmission electron microscopy, scanning electron microscopy, and microhardness measurement. Nanoscale HAP crystallites were prepared using a precipitation method that involved calcium nitrate and ammonium dihydrogen orthophosphate solutions; the preparation was conducted at a temperature of 0°C. The precipitate was gel-cast and sintered at 1000°C in the form of a transparent ceramic that had a uniform grain size of 250 m. The maximum Vickers microhardness obtained for a sample sintered at 1000°C was 6.57 GPa. The sintering behavior of gel-cast samples prepared from high-temperature-precipitated HAP was compared with that of material prepared at 0°C.
Scaffolds to aid in repair, replacement, or regeneration of bony tissues have been developed using a wide spectra of materials. Under clinical conditions, assessment of healing and implant placement is guided radiographically. In this context, strontium's role in osteostimulation and its relevance in radio-opacity are known. Therefore to aid in assessment and to ensure tissue regeneration, a bone mimetic porous strontium calcium phosphate (SrCaPO(4) ) was synthesized in-house, which was non-cytotoxic (ISO 10993 (Part V) and subsequently characterized for its crystallinity, functional groups, and 3D porous topography. Furthermore, to assess the feasibility of the bioactive ceramic scaffolds in bone repair, SrCaPO(4) and hydroxyapatite (HA-Control) scaffolds were implanted in the segmental ulna bone critical-sized defect (1.5 cm) of New Zealand White Rabbits (leporine model-Oryctolagus cuniculus) for a period of 4 and 12 weeks, respectively. Healing of the defects was uneventful without any inflammation or infection. Radio-opacity of SrCaPO(4) within the defect site enabled easy assessment of implant placement and osteointegration. Again, histological evaluation coupled with micro-CT and histomorphometrical analysis indicated that SrCaPO(4) favored significant de novo bone formation in par with material degradation at 4 and 12 weeks post-implantation compared to HA at 4 and 12 weeks. Investigations on this radio-opaque SrCaPO(4) established its role in the repair of critical-sized segmental defects, proposing it as a suitable bone substitute for clinical reconstructive surgery with easy radiographic evaluation.
The development of bone replacement materials is an important objective in the field of orthopaedic surgery. Due to the drawbacks of treating bone defects with autografts, synthetic bone graft materials have become optional. So in this work, a bone tissue engineering approach with radiopaque bioactive strontium incorporated calcium phosphate was proposed for the preliminary cytocompatibility studies for bone substitutes. Accumulating evidence indicates that strontium containing biomaterials promote enhanced bone repair and radiopacity for easy imaging. Hence, strontium calcium phosphate (SrCaPO4) and hydroxyapatite scaffolds have been investigated for its ability to support and sustain the growth of rabbit adipose-derived mesenchymal stem cells (RADMSCs) in vitro. They were characterized via Micro-CT for pore size distribution. Cells used were isolated from New Zealand White rabbit adipose tissue, characterized by FACS and via differentiation into the osteogenic lineage by alkaline phosphatase, Masson's trichome, Alizarin Red and von Kossa staining on day 28. Material-cell interaction was observed by SEM imaging of cell morphology on contact with material. Live-Dead analysis was done by confocal laser scanning microscopy and cell cluster analysis via μCT. The in vitro biodegradation, elution and nucleation of apatite formation of the material was evaluated using simulated body fluid and phosphate buffered saline in static regime up to 28 days at 37 °C. These results demonstrated that SrCaPO4 is a good candidate for bone tissue engineering applications and with osteogenically-induced RADMSCs, they may serve as potential implants for the repair of critical-sized bone defects.
Calcium sulfate cement (CSC) has emerged as a potential bone filler material mainly because of the possibility of incorporating therapeutic agents. Delivery of the cement through a needle or cannula will make it more useful in clinical applications. However, it was not possible to make CSC injectable because of the inherent lack of viscosity. The present work demonstrates the design development of a viscous and fully-injectable CSC by incorporating hydrogen orthophosphate ions, which does not hamper the biocompatibility of the material. The effect of addition of hydrogen orthophosphate on the rheological properties of the CSC paste was studied using a custom made capillary rheometer. The physicochemical changes associated with cement setting process were examined using X-ray diffraction and Fourier transform infrared spectroscopy and the thermal changes were measured through isothermal differential scanning calorimetry. Micromorphological features of different compositions were observed in environmental scanning electron microscopy and the presence of phosphate ions was identified with energy dispersive X-ray spectroscopic analysis and inductively coupled plasma-optical emission spectroscopy. The results indicated that HPO4 (2-) ions have profound effects on the rheological properties and setting of the CSC paste. Significant finding is that the HPO4 (2-) ions are getting substituted in the calcium sulfate dihydrate crystals during setting. The variations of setting time and compressive strength of the cement with the additive concentration were investigated. An optimum concentration of 2.5 % w/w gave a fully-injectable cement with clinically relevant setting time (below 20 min) and compressive strength (12 MPa). It was possible to inject the optimised cement paste from a syringe through an 18-gauge needle with thumb pressure. This cement will be useful both as bone filler and as a local drug delivery medium and it allows minimally invasive bone defect management.
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 α-tricalcium phosphate, was characterized for its physiochemical properties, and ASCs were characterized for their stemness. The cell-ceramic interactions were demonstrated in vitro, whereas adipogenesis was picturesquely depicted by Nile red-stained multilocular adipocyte-like cells. Subsequently, the three-dimensional cell-ceramic-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.
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