Caiyun Mu

Dendrimerlike Mesoporous Silica Nanoparticles as pH-Responsive Nanocontainers for Targeted Drug Delivery and Bioimaging

In this work, we employed dendrimerlike mesoporous silica nanoparticles with hierarchical pores (HPSNs) to fabricate drug delivery system bioimaging and targeted tumor therapy in vivo. N,N-phenylenebis(salicylideneimine)dicarboxylic acid (Salphdc) was used both as the gatekeeper of HPSNs via pH-responsive coordination bonds between -COOH of Salphdc and In(3+) ions and as a fluorescence imaging agent. Folic acid was then conjugated to Salphdc as the targeting unit. The results revealed that the system could deliver model drug DOX to the tumor site with high efficiency and then cause cell apoptosis and tumor growth inhibition. Moreover, the conjugated Salphdc was proved to be a promising fluorescence probe for tracing distribution of the system in vivo. The study affords a potential nanoconainer for cancer therapy and biological imaging.

Alendronate-loaded hydroxyapatite-TiO2 nanotubes for improved bone formation in osteoporotic rabbits

Early mechanical fixation between an implant and native bone is critically important for successful orthopedic implantation, especially for hosts suffering osteoporosis with reduced bone mass. To endow a titanium-based implant with a desirable local anti-osteoporosis property for enhancing its early osseointegration, alendronate-loaded hydroxyapatite-TiO2 nanotube (TNT-HA-Aln) substrates were fabricated and systematically characterized in this study. The results of Aln/Ca2+ release and Ca2+ concentration in an osteoclast medium verified that the release of Aln was significantly accelerated along with the acidity rise caused by osteoclast differentiation. Other in vitro tests, such as CCK-8, alkaline phosphatase (ALP), mineralization, gene expression (Runx2, Osterix, ALP, Col I, OPN, OC, OPG and RANKL), protein production (OPG and RANKL) and tartrate-resistant acid phosphatase (TRAP), proved that TNT-HA-Aln substrates have great potential for improving osteoblast proliferation/differentiation and inhibiting osteoclast differentiation. Moreover, in vivo tests, such as the push-out test, micro-CT and H&E staining proved that TNT-HA-Aln implants could efficiently improve local osseointegration after implantation for 3 months. The study provides an alternative to exploiting drug-device combinations to enhance early osseointegration in osteoporosis.

A pH-responsive nanocontainer based on hydrazone-bearing hollow silica nanoparticles for targeted tumor therapy

A pH-responsive drug delivery system based on hollow mesoporous silica nanoparticles (HMSNs) was fabricated for targeted tumor therapy by using hydrazone bonds as pH-sensitive linkers and hyaluronic acid (HA) molecules as both blocking and targeting agents. HMSNs were synthesized with good dispersion and dimensions of around 88 nm. Detailed material characterisation suggested that the drug delivery system was successfully constructed. It displayed a fast pH stimulus response for controlled drug release in vitro. Besides, systematic biological investigations revealed that the drug delivery system had good biocompatibility, which could effectively target tumor cells and deliver therapeutic cargo to induce tumor cell apoptosis in vitro and suppression of tumor growth in vivo. This study reports a promising drug delivery system for potential clinical application against tumor therapy.

Influence of strontium ions incorporated into nanosheet-pore topographical titanium substrates on osteogenic differentiation of mesenchymal stem cells in vitro and on osseointegration in vivo

Biophysical cues or biochemical cues were proved to efficiently regulate the fate of mesenchymal stem cells (MSCs), but their synergistic effects on the biological functions of MSCs remain to be further investigated. In this study, titanium (Ti) substrates were fabricated with distinct sub-micrometer nanosheet-pore topography via a vapor alkaline treatment method. Strontium (Sr) ions were then incorporated into the Ti substrates via ion exchange. Apart from the influence of biophysical cues from topography, MSCs were simultaneously affected by the biochemical cues from the continuously released Sr ions. The MSCs grown onto Ti substrates with Sr incorporated in them displayed higher (p < 0.05 or p < 0.01) cellular functions than those of pure Ti substrates, including proliferation, the genes and proteins expressions of osteogenic markers and mineralization potential when comparing them with the results of those MSCs grown onto pure Ti substrates. Furthermore, the in vivo investigations demonstrated that the Sr incorporated Ti implants promoted new bone formation. All the results indicated that the incorporated Sr ions and the nanosheet-pore topography of the Ti substrates synergistically enhanced the osteogenic differentiation of MSCs in vitro and osseointegration in vivo. This study advances the understanding of the synergistic influence of biophysical cues and biochemical cues on MSC osteogenic differentiation.

Nanosheet-pore topographical titanium substrates: a biophysical regulator of the fate of mesenchymal stem cells

Recent reports have demonstrated that nano- or micro-scale topography could enhance the cellular functions of stem cells. In this study, a sub-micrometer topography composed of nanosheet-pore structures was fabricated on the pure titanium surface by a simple vapor alkaline-treatment method to understand more profoundly sub-micrometer topography mediated stem cell behaviors. The topography was characterized by scanning electron microscopy, atomic force microscopy, X-ray photoelectron spectroscopy, X-ray diffraction and contact angle measurements, respectively. It specifically mediated cellular functions of rat bone marrow-derived mesenchymal stem cells (MSCs) on cellular and molecular levels under either normal medium or osteoinductive medium conditions. The experimental results indicated that the topography dramatically promoted the adhesion of MSCs grown on the surface, but the shape, morphology and spreading of cells were not significantly affected. In addition, the study demonstrated that the formation of focal adhesion complexes (FAs) were highly dependent on the topography, which in turn affects the subsequent biological functions of MSCs, especially accelerating osteogenic differentiation of MSCs under different conditions. Overall, the sub-micrometer topographical titanium substrate was an excellent biophysical regulator of the fate of mesenchymal stem cells, specifically inducing their differentiation into osteoblasts.

Regulation of local bone remodeling mediated by hybrid multilayer coating embedded with hyaluronan-alendronate/BMP-2 nanoparticles on Ti6Al7Nb implants

Osteoporosis, a common bone disease, has been identified as a major obstacle for successful implantation. Therefore, the promotion of early mechanical fixation between implants and the surrounding bone can strongly increase the success rate of orthopedic operation in osteoporosis patients. In this study, functional hyaluronan-alendronate/BMP-2 (HA-Aln/BMP-2) nanoparticles were embedded into the Gel/Chi multilayers on Ti6Al7Nb surfaces (namely Ti6Al7Nb/LBL/NP) to endow the Ti6Al7Nb-based implant with local anti-osteoporosis properties. The release test showed that the loaded BMP-2 only slowly released along with the degradation of multilayers, and no burst release emerged at the early stage. In vitro cell experiments, including cell morphology, viability, alkaline phosphatase (ALP) activity, mineralization capacity and tartrate-resistant acid phosphatase (TRAP), demonstrated that the prepared Ti6Al7Nb/LBL/NP implants not only improved the proliferation and differentiation of osteoblasts but also inhibited the maturity of osteoclasts. Moreover, the in vivo tests of the push-out test, micro-CT and histological stains further verified that the Ti6Al7Nb/LBL/NP implant was more beneficial to promoting the local osseointegration between the natural bone and the implant when compared to those of the control groups after implantation for 3 months in osteoporotic rabbits. The study demonstrated a flexible method for effectively enhancing the early osseointegration between the implant and the native osteoporotic bone.

Strontium folic acid derivative functionalized titanium surfaces for enhanced osteogenic differentiation of mesenchymal stem cells in vitro and bone formation in vivo

The introduction of the bioactive strontium (Sr) element has become an attractive method in the design of bio-functional layers on titanium surfaces. However, there are still no effective solutions to some of the associated problems including the toxicity of free Sr2+ ions and the rapid and irreversible loss of the strontium element from the bio-functional layers. In this study, we successfully fabricated a bioactive layer on Ti substrates with a strontium folic acid derivative (FASr). About 3.11 at% Sr was incorporated into the Ti surface. The characterization results showed that FASr was stable over a long period of time and minimal free Sr2+ ions were detected in simulated body fluid (SBF). In the in vitro experiment, the FASr could significantly promote the cell adhesion, proliferation and osteogenic differentiation of mesenchymal stem cells (MSCs) over a short period. Furthermore, it could dramatically accelerate the bone formation around the implant. In vivo, a total of 30 7-week old male Sprague Dawley (SD) rats were applied for implantation tests. The results showed that this positive stimulatory effect became more evident in the later stages of the in vivo observation. This study provides an effective strategy for designing and optimizing Ti-based implants.

Simultaneous delivery of BMP-2 factor and anti-osteoporotic drugs using hyaluronan-assembled nanocomposite for synergistic regulation on the behaviors of osteoblasts and osteoclasts in vitro

To treat the osteoporosis and regulate the biological behaviors of both osteoblasts and osteoclasts, we prepared a natural polysaccharide-derived nanocomposite, containing alendronate-grafted hyaluronate (HA-Aln) and bone morphogenetic protein 2 (BMP-2) and investigated its synergistic regulation on the behaviors of osteoblasts and osteoclasts in vitro. The HA-Aln/BMP-2 nanocomposite was fabricated through the electrostatic interactions between the HA-Aln molecule and BMP-2 molecule. Here, BMP-2 was used to improve the osteoblast-mediated bone formation. Alendronate (Aln), a targeting ligand to bone matrix, was used to inhibit the osteoclast-mediated bone resorption. In vitro results showed that HA-Aln/BMP-2 nanocomposite could effectively maintain the bioactivity of loaded drugs. The osteoblasts that treated with the HA-Aln/BMP-2 nanocomposite presented a higher level of cell motility, alkaline phosphatase (ALP) activity, mineralization capacity, and osteoblast-related gene expressions (runt-related transcription factor 2, osterix, ALP, collagen type I, osteocalcin, and osteopontin), as compared to that of control group. Besides, the RAW264.7 cells that were treated with HA-Aln/BMP-2 nanocomposite showed a lower level of osteoclastic differentiation. Overall, the HA-Aln/BMP-2 nanocomposite exhibits promising potential as an efficient carrier for co-delivery of anti-osteoporotic drug and growth factors to promote osteoblastic differentiation and bone formation while suppressing osteoclastic activity.

Sustained raloxifene release from hyaluronan-alendronate-functionalized titanium nanotube arrays capable of enhancing osseointegration in osteoporotic rabbits

To enhance the localized bone remodeling at titanium-based implants under osteoporotic conditions, TiO2 nanotube arrays (TNT) were used as nanoreserviors for raloxifene (Ral) and then covered with the hybrid multilayered coating of chitosan and alendronate grafted hyaluronic acid (HA-Aln) via a spin-assisted layer-by-layer technique. The fabrication of this system (TNT/Ral/LBL-Aln) was characterized by field emission scanning electron microscopy (SEM), atomic force microscope (AFM) and X-ray photoelectron spectroscopy (XPS), respectively. The release test showed that the composited multilayers onto Ral-loaded TiO2 nanotube substrate (TNT/Ral) could prevent the burst release of Ral from TiO2 nanotube arrays and maintain stable Ral concentration at the implant site even after 192h. The TNT/Ral/LBL-Aln system demonstrated higher alkaline phosphatase (ALP) activity, mineralization capability in osteoblasts as well as lower tartrate-resistant acid phosphatase (TRAP) activity in osteoclasts compared to both bare TiO2 nanotube and TNT/Ral substrate, respectively. Moreover, the in vivo tests of micro-CT, histological staining and push-out testing showed that TNT/Ral/LBL-Aln implant could efficiently enhance the formation of new bone around the implant and promote bone binding in osteoporotic rabbits. The study indicated the potential application of TNT/Ral/LBL-Aln system for bone remodeling under osteoporotic condition.

Multilayered coating of titanium implants promotes coupled osteogenesis and angiogenesis in vitro and in vivo

We used surface-modified titanium (Ti) substrates with a multilayered structure composed of chitosan-catechol (Chi-C), gelatin (Gel) and hydroxyapatite (HA) nanofibers, which were previously shown to improve osteogenesis, as a platform to investigate the interaction of osteogenesis and angiogenesis during bone healing. Combined techniques of Transwell co-culture, wound healing assay, enzyme linked immunosorbent assay (ELISA), quantitative real-time polymerase chain reaction (qRT-PCR), western blotting and immunohistochemical staining were used to evaluate adhesion, morphology and migration of adipose-derived mesenchymal stem cells (Ad-MSCs) and human umbilical vein endothelial cells (HUVECs) grown on different Ti substrates. We investigated the effect of substrates on the osteogenic differentiation of Ad-MSCs and reciprocal paracrine effects of Ad-MSCs on HUVECs or vice versa. The multilayered Ti substrates directly regulated the cellular functions of Ad-MSCs and angiogenic HUVECs and mediated communication between them by enhancing paracrine effects via cell-matrix interactions in vitro. The in vivo results showed that the change of microenvironment induced by surface-modified Ti implants promoted the adhesion, recruitment and proliferation of MSCs and facilitated coupled osteogenesis and angiogenesis in bone healing. The study proved that multilayer-film-coated Ti substrates positively mediated cellular biological function in vitro and improved bone healing in vivo. STATEMENT OF SIGNIFICANCE Recent studies have revealed that osteogenesis and angiogenesis are coupled, and that communication between osteoblasts and endothelial cells is essential for bone healing and remodeling processes; however, these conclusions only result from in vitro studies or in vivo studies using transgenic murine models. Relatively little is known about the communication between osteoblasts and endothelial cells in peri-implants during bone healing processes. Our results revealed the cellular/molecular mechanism of how multilayered Ti substrates mediate reciprocal paracrine effects between adipose-derived mesenchymal stem cells and human umbilical vein endothelial cells; moreover, the interactions between the cell-matrix and peri-implant was proven in vivo with enhanced bone healing. This study contributes to our understanding of the fundamental mechanisms of angiogenesis and osteogenesis that affect peri-implantation, and thus, provides new insights into the design of future high-quality orthopedic implants.