Solaiman Tarafder

Calcium phosphate ceramic systems in growth factor and drug delivery for bone tissue engineering: A review

Calcium phosphates (CaPs) are the most widely used bone substitutes in bone tissue engineering due to their compositional similarities to bone mineral and excellent biocompatibility. In recent years, CaPs, especially hydroxyapatite and tricalcium phosphate, have attracted significant interest in simultaneous use as bone substitute and drug delivery vehicle, adding a new dimension to their application. CaPs are more biocompatible than many other ceramic and inorganic nanoparticles. Their biocompatibility and variable stoichiometry, thus surface charge density, functionality, and dissolution properties, make them suitable for both drug and growth factor delivery. CaP matrices and scaffolds have been reported to act as delivery vehicles for growth factors and drugs in bone tissue engineering. Local drug delivery in musculoskeletal disorder treatments can address some of the critical issues more effectively and efficiently than the systemic delivery. CaPs are used as coatings on metallic implants, CaP cements, and custom designed scaffolds to treat musculoskeletal disorders. This review highlights some of the current drug and growth factor delivery approaches and critical issues using CaP particles, coatings, cements, and scaffolds towards orthopedic and dental applications.

Microwave-sintered 3D printed tricalcium phosphate scaffolds for bone tissue engineering

This study reports the manufacturing process of 3D interconnected macroporous tricalcium phosphate (TCP) scaffolds with controlled internal architecture by direct 3D printing (3DP), and high mechanical strength obtained by microwave sintering. TCP scaffolds with 27%, 35% and 41% designed macroporosity with pore sizes of 500 μm, 750 μm and 1000 μm, respectively, were manufactured by direct 3DP. These scaffolds were then sintered at 1150 °C and 1250 °C in conventional electric muffle and microwave furnaces, respectively. Total open porosity between 42% and 63% was obtained in the sintered scaffolds due to the presence of intrinsic micropores along with designed pores. A significant increase in compressive strength between 46% and 69% was achieved by microwave compared to conventional sintering as a result of efficient densification. Maximum compressive strengths of 10.95 ± 1.28 MPa and 6.62 ± 0.67 MPa were achieved for scaffolds with 500 μm designed pores (~ 400 μm after sintering) sintered in microwave and conventional furnaces, respectively. An increase in cell density with a decrease in macropore size was observed during in vitro cell‐material interactions using human osteoblast cells. Histomorphological analysis revealed that the presence of both micro‐ and macropores facilitated osteoid‐like new bone formation when tested in femoral defects of Sprague–Dawley rats. Our results show that bioresorbable 3D‐printed TCP scaffolds have great potential in tissue engineering applications for bone tissue repair and regeneration. Copyright

Understanding of dopant-induced osteogenesis and angiogenesis in calcium phosphate ceramics

General trends in synthetic bone grafting materials are shifting towards approaches that can illicit osteoinductive properties. Pharmacologics and biologics have been used in combination with calcium phosphate (CaP) ceramics, however, they have recently become the target of scrutiny over safety. The importance of trace elements in natural bone health is well documented. Ions, for example, lithium, zinc, magnesium, manganese, silicon, strontium, etc., have been shown to increase osteogenesis and neovascularization. Incorporation of dopants (trace metal ions) into CaPs can provide a platform for safe and efficient delivery in clinical applications where increased bone healing is favorable. This review highlights the use of trace elements in CaP biomaterials, and offers an insight into the mechanisms of how metal ions can enhance both osteogenesis and angiogenesis.

Understanding the influence of MgO and SrO binary doping on the mechanical and biological properties of β-TCP ceramics

The objective of this study was to evaluate the influence of MgO and SrO doping on the mechanical and biological properties of beta-tricalcium phosphate (beta-TCP). beta-TCP was doped with two different binary compositions, 0.25 and 1.0wt.% SrO along with 1.0wt.% MgO. MgO and SrO doping increased the beta phase stability at a sintering temperature of 1250 degrees C and marginally decreased the compressive strength of beta-TCP. An in vitro cell-material interaction study, using human fetal osteoblast cells (hFOB), indicated that doped beta-TCP was non-toxic, and MgO/SrO dopants improved cell attachment and growth. beta-TCP implants doped with 1.0wt.% MgO and 1.0wt.% SrO showed good in vivo biocompatibility when tested in male Sprague-Dawley rats for 16 weeks. Histomorphology analysis indicated that MgO/SrO-doped beta-TCP promoted more osteogenesis than pure beta-TCP. In vivo osteocalcin and type I collagen assay also revealed faster bone formation in rats with doped beta-TCP implant compared to rats with pure beta-TCP implant. Low Ca(2+) concentration in the urine of rats with doped beta-TCP implant confirmed slower degradation of MgO/SrO-doped beta-TCP than pure beta-TCP.

Microwave-processed nanocrystalline hydroxyapatite: Simultaneous enhancement of mechanical and biological properties

Despite the excellent bioactivity of hydroxyapatite (HA) ceramics, poor mechanical strength has limited the applications of these materials primarily to coatings and other non-load-bearing areas as bone grafts. Using synthesized HA nanopowder, dense compacts with grain sizes in the nanometer to micrometer range were processed via microwave sintering between 1000 and 1150 degrees C for 20 min. Here we demonstrate that the mechanical properties, such as compressive strength, hardness and indentation fracture toughness, of HA compacts increased with a decrease in grain size. HA with 168 +/- 86 nm grain size showed the highest compressive strength of 395 +/- 42 MPa, hardness of 8.4+/-0.4 GPa and indentation fracture toughness of 1.9 +/- 0.2 MPa m(1/2). To study the in vitro biological properties, HA compacts with grain size between 168 nm and 1.16 microm were assessed for in vitro bone cell-material interactions with human osteoblast cell line. Vinculin protein expression for cell attachment and bone cell proliferation using MTT assay showed that surfaces with finer grains provided better bone cell-material interactions than coarse-grained samples. Our results indicate simultaneous improvements in mechanical and biological properties in microwave sintered HA compacts with nanoscale grain size.

Understanding in vivo response and mechanical property variation in MgO, SrO and SiO2 doped β-TCP

The aim of this work is to evaluate the influence of MgO, SrO and SiO₂ doping on mechanical and biological properties of β-tricalcium phosphate (β-TCP) to achieve controlled resorption kinetics of β-TCP system for maxillofacial and spinal fusion application. We prepared dense TCP compacts of four different compositions, i) pure β-TCP, ii) β-TCP with 1.0wt.% MgO+1.0wt.% SrO, iii) β-TCP with 1.0wt.% SrO+0.5wt.% SiO₂, and iv) β-TCP with 1.0wt.% MgO+1.0wt.% SrO+0.5wt.% SiO₂, by uniaxial pressing and sintering at 1250°C. β phase stability is observed at 1250°C sintering temperature due to MgO doping in β-TCP. In vitro mineralization in simulated body fluid (SBF) for 16 weeks shows excellent apatite growth on undoped and doped samples. Strength degradation of TCP samples in SBF is significantly influenced by both dopant chemistry and amount of dopant. Compressive strengths for all samples show degradation in SBF over the 16 week time period with varying degradation kinetics. MgO/SrO/SiO₂ doped sample shows no strength loss, while undoped TCP shows the maximum strength loss from 419 ± 28 MPa to 158 ± 28 MPa over the 16 week study. In case of MgO/SrO doped TCP, strength loss is slow and gradual. TCP doped with 1.0wt.% MgO and 1.0wt.% SrO shows excellent in vivo biocompatibility when tested in male Sprague-Dawley rats for 16 weeks. Histomorphology analysis reveals that MgO/SrO doped TCP promoted osteogenesis by excellent early stage bone remodeling as compared to undoped TCP.

3D printed tricalcium phosphate bone tissue engineering scaffolds: effect of SrO and MgO doping on in vivo osteogenesis in a rat distal femoral defect model

The presence of interconnected macro pores is important in tissue engineering scaffolds for guided tissue regeneration. This study reports in vivo biological performance of interconnected macro porous tricalcium phosphate (TCP) scaffolds due to the addition of SrO and MgO as dopants in TCP. We have used direct three dimensional printing (3DP) technology for scaffold fabrication followed by microwave sintering. Mechanical strength was evaluated for scaffolds with 500 μm, 750 μm, and 1000 μm interconnected designed pore sizes. Maximum compressive strength of 12.01 ± 1.56 MPa was achieved for Sr–Mg doped scaffold with 500 μm interconnected designed pore size. In vivo biological performance of the microwave sintered pure TCP and Sr–Mg doped TCP scaffolds was assessed by implanting 350 μm designed interconnected macro porous scaffolds in rat distal femoral defect. Sintered pore size of these 3D printed scaffolds were 311 ± 5.9 μm and 245 ± 7.5 μm for pure and SrO–MgO doped TCP scaffolds, respectively. These 3D printed scaffolds possessed multiscale porosity, i.e., 3D interconnected designed macro pores along with intrinsic micro pores. Histomorphology and histomorphometric analysis revealed a significant increase in osteoid like new bone formation, and accelerated mineralization inside SrO and MgO doped 3D printed TCP scaffolds as compared to pure TCP scaffolds. An increase in osteocalcin and type I collagen level was also observed in rat blood serum with SrO and MgO doped TCP scaffolds compared to pure TCP scaffolds. Our results show that these 3D printed SrO and MgO doped TCP scaffolds with multiscale porosity contributed to early healing through accelerated osteogenesis.

Polycaprolactone-Coated 3D Printed Tricalcium Phosphate Scaffolds for Bone Tissue Engineering: In Vitro Alendronate Release Behavior and Local Delivery Effect on In Vivo Osteogenesis

The aim of this work was to evaluate the effect of in vitro alendronate (AD) release behavior through polycaprolactone (PCL) coating on in vivo bone formation using PCL-coated 3D printed interconnected porous tricalcium phosphate (TCP) scaffolds. Higher AD and Ca2+ ion release was observed at lower pH (5.0) than that at higher pH (7.4). AD and Ca2+ release, surface morphology, and phase analysis after release indicated a matrix degradation dominated AD release caused by TCP dissolution. PCL coating showed its effectiveness for controlled and sustained AD release. Six different scaffold compositions, namely, (i) TCP (bare TCP), (ii) TCP + AD (AD-coated TCP), (iii) TCP + PCL (PCL-coated TCP), (iv) TCP + PCL + AD, (v) TCP + AD + PCL, and (vi) TCP + AD + PCL + AD were tested in the distal femoral defect of Sprague–Dawley rats for 6 and 10 weeks. An excellent bone formation inside the micro and macro pores of the scaffolds was observed from histomorphology. Histomorphometric analysis revealed maximum new bone formation in TCP + AD + PCL scaffolds after 6 weeks. No adverse effect of PCL on bioactivity of TCP and in vivo bone formation was observed. All scaffolds with AD showed higher bone formation and reduced TRAP (tartrate resistant acid phosphatase) positive cells activity compared to bare TCP and TCP coated with only PCL. Bare TCP scaffolds showed the highest TRAP positive cells activity followed by TCP + PCL scaffolds, whereas TCP + AD scaffolds showed the lowest TRAP activity. A higher TRAP positive cells activity was observed in TCP + AD + PCL compared to TCP + AD scaffolds after 6 weeks. Our results show that in vivo local AD delivery from PCL-coated 3DP TCP scaffolds could further induce increased early bone formation.

Electrically Polarized Biphasic Calcium Phosphates: Adsorption and Release of Bovine Serum Albumin

In this study, we applied electrical polarization technique to increase adsorption and control protein release from biphasic calcium phosphate (BCP). Three different biphasic calcium phosphate (BCP) composites, with hydroxyapatite (HAp) and β-tricalcium phosphate (β-TCP), were processed and electrically polarized. Our study showed that stored charge was increased in the composites with the increase in HAp percentage. Adsorption of bovine serum albumin (BSA), as a model protein, on the poled as well as unpoled surfaces of the composites was studied. The highest amount of BSA adsorption was obtained on positively poled surfaces of each composite. Adsorption isotherm study suggested a multilayer adsorption of BSA on the BCP composites. The effect of electrical polarization on BSA release kinetics from positively charged BCP surfaces was studied. A gradual increase in percent BSA release from positively charged BCP surfaces with decreasing stored charge was observed. Our study showed that the BCP based composites have the potential to be used as a drug or growth factor delivery vehicle.

SrO- and MgO-doped microwave sintered 3D printed tricalcium phosphate scaffolds: mechanical properties and in vivo osteogenesis in a rabbit model.

The presence of interconnected macro pores allows guided tissue regeneration in tissue engineering scaffolds. However, highly porous scaffolds suffer from having poor mechanical strength. Previously, we showed that microwave sintering could successfully be used to improve mechanical strength of macro porous tricalcium phosphate (TCP) scaffolds. This study reports the presence of SrO and MgO as dopants in TCP scaffolds improves mechanical and in vivo biological performance. We have used direct three dimensional printing (3DP) technology for scaffold fabrication. These 3DP scaffolds possessed multiscale porosity, that is, 3D interconnected designed macro pores along with intrinsic micro pores. A significant increase in mechanical strength, between 37 and 41%, was achieved due to SrO and MgO doping in TCP as compared with pure TCP. Maximum compressive strengths of 9.38 ± 1.86 MPa and 12.01 ± 1.56 MPa were achieved by conventional and microwave sintering, respectively, for SrO-MgO-doped 3DP scaffolds with 500 μm designed pores. Histomorphological and histomorphometric analysis revealed a significantly higher osteoid, bone and haversian canal formation induced by the presence of SrO and MgO dopants in 3DP TCP as compared with pure TCP scaffolds when tested in rabbit femoral condyle defect model. Increased osteon and thus enhanced network of blood vessel formation, and osteocalcin expression were observed in the doped TCP scaffolds. Our results show that these 3DP SrO-MgO-doped TCP scaffolds have the potential for early wound healing through accelerated osteogenesis and vasculogenesis.