NIR-Responsive Methotrexate-Modified Iron Selenide Nanorods for Synergistic Magnetic Hyperthermic, Photothermal, and Chemodynamic Therapy.

Breast cancer is a malignant tumor with a high mortality rate among women. Therefore, it is necessary to develop novel therapies to effectively treat this disease. In this study, iron selenide nanorods (FeSe2 NRs) were designed for use in magnetic hyperthermic, photothermal, and chemodynamic therapy (MHT/PTT/CDT) for breast cancer. To illustrate their efficacy, FeSe2 NRs were modified with the chemotherapeutic agent methotrexate (MTX). MTX-modified FeSe2 (FeSe2-MTX) exhibited excellent controlled drug release properties. Fe2+ released from FeSe2 NRs induced the release of •OH from H2O2 via a Fenton/Fenton-like reaction, enhancing the efficacy of CDT. Under alternating magnetic field (AMF) stimulation and 808 nm laser irradiation, FeSe2-MTX exerted potent hyperthermic and photothermal effects by suppressing tumor growth in a breast cancer nude mouse model. In addition, FeSe2 NRs can be used for magnetic resonance imaging in vivo by incorporating their superparamagnetic characteristics into a single nanomaterial. Overall, we presented a novel technique for the precise delivery of functional nanosystems to tumors that can enhance the efficacy of breast cancer treatment.

Inorganic nanoparticles for photothermal treatment of cancer.

In recent years, inorganic nanoparticles (NPs) have attracted increasing attention as potential theranostic agents in the field of oncology. Photothermal therapy (PTT) is a minimally invasive technique that uses nanoparticles to produce heat from light to kill cancer cells. PTT requires two essential elements: a photothermal agent (PTA) and near-infrared (NIR) radiation. The role of PTAs is to absorb NIR, which subsequently triggers hyperthermia within cancer cells. By raising the temperature in the tumor microenvironment (TME), PTT causes damage to the cancer cells. Nanoparticles (NPs) are instrumental in PTT given that they facilitate the passive and active targeting of the PTA to the TME, making them crucial for the effectiveness of the treatment. In addition, specific targeting can be achieved through their enhanced permeation and retention effect. Thus, owing to their significant advantages, such as altering the morphology and surface characteristics of nanocarriers comprised of PTA, NPs have been exploited to facilitate tumor regression significantly. This review highlights the properties of PTAs, the mechanism of PTT, and the results obtained from the improved curative efficacy of PTT by utilizing NPs platforms.

Polyelectrolyte multilayer composite coating on 316 L stainless steel for controlled release of dual growth factors accelerating restoration of bone defects.

A composite coating of polyelectrolyte multilayers (PEMs) consisting of collagen, a chitosan barrier, and poly-γ-glutamic acid was fabricated using a spin coating technique to investigate and overcome the limited osseointegration capacity of 316 L stainless steel (316 L SS). To further enhance the biocompatibility, bone morphogenetic protein 2 (BMP-2) and basic fibroblast growth factor-2 (FGF-2) were loaded separately as dual growth factors, allowing for progressive drug release following the natural process of bone regeneration. The first burst release of FGF-2 triggered the proliferation of surrounding cells, and the subsequent release of BMP-2 stimulated their differentiation. The microstructure, surface potential, hardness, reduced Young's modulus, and wettability were assessed using scanning electron microscopy, nanoindentation, and water contact angle. The formation of apatite layers after immersion in simulated body fluid confirmed the bioactivity of this PEM. PEMs loaded with BMP-2 and FGF-2 showed a long sustained release of growth factors for up to 48 days. The biological properties were studied in vitro with rat bone mesenchymal stem cells (rBMSCs) and in vivo using a rat critical-sized calvarial defect model. PEMs loaded with growth factors further stimulated the proliferation and osteogenic differentiation of rBMSCs and the histology results indicated that new bone tissues could directly grow onto the PEMs. These findings suggest that PEM composite coating possesses significant potential for surface modification and long-term drug release of metallic implants to assist with bone restoration.

Study on proanthocyanidins crosslinked collagen membrane for guided bone tissue regeneration.

The goal of this study is to understand the ability of a newly developed barrier membrane to enhance bone tissue regeneration. Here in this study we present the in vitro characterization of the barrier membrane made from type I collagen and crosslinked by oligomeric proanthocyanidins (OPCs). The effects of the membrane (P-C film) on cell cycle, proliferation, alkaline phosphatase activity, and mineralization were evaluated using the human osteoblast cell line MG-63, while the barrier ability was examined using MG-63 cells, as well as the human skin fibroblast cell line WS-1. The pore size is one of the factors that plays a key role in tissue regeneration, therefore, we evaluated the pore size of the membrane using a capillary flow porometer. Our results showed that the mean pore size of the P-C film was approximately 7-9 µm, the size known to inhibit cell migration across the membrane. The P-C film also demonstrated excellent cell viability and good biocompatibility, since the cell number increased with time, with MG-63 cells proliferating faster on the P-C film than in the cell culture flask. Furthermore, the P-C film promoted osteoblast differentiation, resulting in higher alkaline phosphatase activity and mineralization. Therefore, our results suggest that this P-C film has a great potential to be used in guided bone regeneration during periodontal regeneration and bone tissue engineering.

Hyperthermia-Induced Controlled Local Anesthesia Administration Using Gelatin-Coated Iron-Gold Alloy Nanoparticles.

The lack of optimal methods employing nanoparticles to administer local anesthesia often results in posing severe risks such as non-biocompatibility, in vivo cytotoxicity, and drug overdose to patients. Here, we employed magnetic field-induced hyperthermia to achieve localized anesthesia. We synthesized iron-gold alloy nanoparticles (FeAu Nps), conjugated an anesthetic drug, Lidocaine, and coated the product with gelatin to increase the biocompatibility, resulting in a FeAu@Gelatin-Lidocaine nano-complex formation. The biocompatibility of this drug-nanoparticle conjugate was evaluated in vitro, and its ability to trigger local anesthesia was also evaluated in vivo. Upon exposure to high-frequency induction waves (HFIW), 7.2 ± 2.8 nm sized superparamagnetic nanoparticles generated heat, which dissociated the gelatin coating, thereby triggering Lidocaine release. MTT assay revealed that 82% of cells were viable at 5 mg/mL concentration of Lidocaine, indicating that no significant cytotoxicity was induced. In vivo experiments revealed that unless stimulated with HFIW, Lidocaine was not released from the FeAu@Gelatin-Lidocaine complex. In a proof-of-concept experiment, an intramuscular injection of FeAu@Gelatin-Lidocaine complex was administered to the rat posterior leg, which upon HFIW stimulation triggered an anesthetic effect to the injected muscle. Based on our findings, the FeAu@Gelatin-Lidocaine complex can deliver hyperthermia-induced controlled anesthetic drug release and serve as an ideal candidate for site-specific anesthesia administration.

Ultra-highly sensitive organophosphorus biosensor based on chitosan/tin disulfide and British housefly acetylcholinesterase.

Pesticides have been extensively applied worldwide to protect crops from worms and insects; however, the continuous use of pesticides affects ecosystems, agricultural product safety, nontarget organisms, and human health. In this paper, we report a highly sensitive biosensor for the determination of pesticides based on tin sulfide (SnS2) and chitosan (CHIT) nanocomposites decorated with a unique British housefly acetylcholinesterase (AChE). The hydrothermally synthesized nano-SnS2 mixed with chitosan solution (CHIT-SnS2) was drop-casted onto a glassy carbon electrode (GCE). Subsequently, the British housefly AChE was immobilized on the CHIT/SnS2-coated GCE that was then employed for pesticide detection. The developed biosensor showed an ultra-high sensitivity and wide linear detection range from 0.02 nM to 20000 nM with a detection limit of 0.02 nM for the detection of chlorpyrifos as the model pesticide. Furthermore, the AChE/CHIT-SnS2/GCE exhibited acceptable storage stability, good reproducibility, and selectivity.

Study on strontium doped tricalcium silicate synthesized through sol-gel process.

We successfully synthesized a strontium-doped tricalcium silicate (SrxCa3-xSiO5, Sr = 0 to 2 mol%) bone cement using the sol-gel process. The material properties including crystallinity, setting time, mechanical strength, and hydration products were characterized. Release of ions and pH values of simulated body fluid soaked with the bone cement were measured. In vitro biocompatibility of different concentrations of the material was evaluated by the viability of L929 cells. The setting times of as-prepared slurries were all <70 min. Doping with 0.5 mol% Sr reduced the final setting time by 20 min. After 14 days curing, 0.25 mol% Sr-doped SrxCa3-xSiO5 possessed the highest compressive strength of 45 MPa among all the Sr-doped groups with no statistical difference to Ca3SiO5. The bioactivity of the materials was confirmed with the formation of an apatite layer on the surface of the materials after immersion in simulated body fluid. In addition, the proliferation of L929 cells exposed to 1 mol% Sr was significantly promoted as compared to no Sr doping. SrxCa3-xSiO5 is a novel and advanced material that has the potential to serve as a bone cement in bone restoration with appropriate mechanical strength and favorable biocompatibility.

Salivary Electrochemical Cortisol Biosensor Based on Tin Disulfide Nanoflakes.

Cortisol, a steroid hormone, is secreted by the hypothalamic-pituitary-adrenal system. It is a well-known biomarker of psychological stress and is hence known as the "stress hormone." If cortisol overexpression is prolonged and repeated, dysfunction in the regulation of cortisol eventually occurs. Therefore, a rapid point-of-care assay to detect cortisol is needed. Salivary cortisol electrochemical analysis is a non-invasive method that is potentially useful in enabling rapid measurement of cortisol levels. In this study, multilayer films containing two-dimensional tin disulfide nanoflakes, cortisol antibody (C-Mab), and bovine serum albumin (BSA) were prepared on glassy carbon electrodes (GCE) as BSA/C-Mab/SnS2/GCE, and characterized using electrochemical impedance spectroscopy and cyclic voltammetry. Electrochemical responses of the biosensor as a function of cortisol concentrations were determined using cyclic voltammetry and differential pulse voltammetry. This cortisol biosensor exhibited a detection range from 100 pM to 100 µM, a detection limit of 100 pM, and a sensitivity of 0.0103 mA/Mcm2 (R2 = 0.9979). Finally, cortisol concentrations in authentic saliva samples obtained using the developed electrochemical system correlated well with results obtained using enzyme-linked immunosorbent assays. This biosensor was successfully prepared and used for the electrochemical detection of salivary cortisol over physiological ranges, based on the specificity of antibody-antigen interactions.

Gamma-poly glutamate/gelatin composite hydrogels crosslinked by proanthocyanidins for wound healing.

A novel multifunctional poly(γ-glutamic acid) (γ-PGA)/gelatin hydrogel has been developed and used as a wound dressing. An ideal wound dressing should effectively provide a moist environment, absorb wound exudates and protect the wound from foreign microbes. Water soluble γ-PGA salts of sodium and calcium forms were chosen for their good biocompatibility, biodegradability and water absorption capacity. Oligomeric proanthocyanidins (OPCs), naturally occurring plant metabolites and potent antioxidants, were investigated as a non-toxic crosslinking agent in this study. The effects of hydrogels on the degree of crosslinking, swelling, in vitro degradation, mechanical properties and radical scavenging activity were systemically evaluated. A cell viability assay demonstrated that these OPCs crosslinked γ-PGA/gelatin (PGO) hydrogels were not cytotoxic to L929 fibroblasts. Dermal irritation and skin sensitization tests were examined using a guinea pig model; the hydrogels were considered to be neither allergic nor a dermal sensitizer in guinea pigs. Lastly, an in vivo wound healing model in rats was used to study the effects of the hydrogels on wound healing for 21 days. PGO hydrogels formed by both Na and Ca salts could accelerate wound contraction and re-epithelialization, in which Na-PGO hydrogel was significantly better than the untreated control group. The findings suggest that PGO hydrogels are promising wound dressing materials for the treatment for wound healing.

Preparation and in vivo investigation of oligomeric proanthocyanidins cross-linked collagen serving as synthesized tissue regeneration membrane.

In this study, type I collagen membranes were prepared using oligomeric proanthocyanidins (OPCs) as the cross-linking agent. The fabricated materials were evaluated to be applied as guided tissue regeneration membranes for periodontal defects. The mechanical strength of the cross-linked collagen membranes, namely OPCs-Col films, using different concentrations of OPCs ranged from 30 to 60 kPa. The cross-linked collagen membranes had better thermal stability than non-cross-linked one and could effectively resist the decomposition in collagenase solution as long as fifty days. The results of material characterization showed that 10% OPCs-Col film was ideal for our purpose. In vitro study using L929 and MG-63 cells revealed that 10% OPCs-Col film had great biocompatibility while OPC was demonstrated to be not cytotoxic as glutaraldehyde and genipin but even promote L929 cells. The material was further studied for in vivo studies with two models, subcutaneous and cranium defects in rat. The subcutaneous test showed that the regeneration membrane degraded till one month and the inflammatory response also reduced with implantation time. When implanted into the cranium defect, no lesions of the brain were caused and new bone tissue was observed inside the material. The results of in vivo studies showed that the synthesized membrane was helpful for tissue regeneration with long degradation time. The tissue regeneration membranes can barrier the rapid growing soft tissue, in order to save the capacity for the growth of neo bone.

Iron-gold alloy nanoparticles serve as a cornerstone in hyperthermia-mediated controlled drug release for cancer therapy.

INTRODUCTION: The efficacy of a chemotherapy drug in cancer therapy is highly determined by the ability to control the rate and extent of its release in vivo. However, the lack of techniques to accurately control drug release drastically limits the potency of a chemotherapy drug. MATERIALS AND METHODS: Here, we present a novel strategy to precisely monitor drug release under magnetic stimulation. Methotrexate (MTX), an anticancer drug, was covalently functionalized onto iron-gold alloy magnetic nanoparticles (Fe-Au alloy nanoparticles or NFAs) through 2-aminoethanethiol grafting and the ability of this drug-nanoparticle conjugate (NFA-MTX) in limiting HepG2 (liver carcinoma) cell growth was studied. Well-dispersed NFAs were prepared through pyrolysis. RESULTS AND DISCUSSION: Transmission electron microscopy revealed the average nanoparticle size to be 7.22±2.6 nm, while X-ray diffraction showed distinct 2θ peaks for iron and gold, confirming the presence of iron and gold nanoparticles. Inductively coupled plasma mass spectrometry revealed that the amount of NFA-MTX conjugate ingested by HepG2 cancer cells was 1.5 times higher than that ingested by L929 fibroblasts, thereby proving a higher selective ingestion by cancer cells compared to normal cells. Fourier-transform infrared spectroscopy revealed the breakage of Au-S bonds by the heat generated under magnetic field stimulation to release MTX from the NFA-MTX conjugate, triggering a 95% decrease in cellular viability at 100 µg/mL. CONCLUSION: The ability of NFA-MTX to dissociate under the influence of an applied magnetic field provides a new strategy to induce cancer cell death via hyperthermia. Applications in drug delivery, drug development, and cancer research are expected.

Composite Polyelectrolyte Multilayer and Mesoporous Bioactive Glass Nanoparticle Coating on 316L Stainless Steel for Controlled Antibiotic Release and Biocompatibility.

Bacterial infection in wounds or implants can cause osteomyelitis, eventually leading to orthopedic implant failure. In this study, polyelectrolyte multilayer (PEM) coating comprising collagen as the cationic layer, chitosan as the barrier layer and γ-poly-glutamic acid as the anionic layer were fabricated onto a 316L stainless steel substrate by spin coating technique. Tetracycline-loaded 57S mesoporous bioactive glass nanoparticles (57S MBG, SiO2:CaO:P2O5 = 57:33:10 by wt%) were introduced into the γ-poly-glutamic acid layers. Herein, 57S MBG nanoparticles were successfully incorporated into the PEMs with a total thickness of ∼53 µm on 316L stainless steel (SS-PEMs-57S), which exhibited good hydrophilicity with a contact angle of 18.71°. The hardness of SS-PEMs-57S was 0.66 GPa while the Young's modulus was 11.5 GPa; these values are similar to those for the cortical bone. The surface roughness of MBG nanoparticle-incorporated PEMs increased from 231 to 384 nm. Controlled release of tetracycline loaded in MBG nanoparticles resulted in sustained antibacterial effect for up to 7 days, with higher release efficacy at low pH, which may be induced by inflammation or infection. Tetracycline loaded in SS-PEMs-57S showed good bacterial inhibition and maintained good cell viability in rat bone marrow mesenchymal stem cells (BMSCs) in the MTT assay. Moreover, SS-PEMs-57S also promoted mineralization of BMSCs. Therefore, this surface modification technology has great potential for endowing orthopedic implants with antibacterial and osteoconductive properties.

FTIR characterization and release of bovine serum albumin from bioactive glasses.

BACKGROUND: Bioactive glass has attracted substantial interest in orthopedics, but it has been less explored as a drug carrier. This study investigated the bovine serum albumin (BSA) release from bioactive 13-93B0 and 13-93B3 glasses. METHODS: Glass disks (13-93B0 and 13-93B3; n = 5) were loaded with 4 mg of BSA and coated under different chitosan-coating conditions. The amount of BSA released in phosphate-buffered saline (PBS) was evaluated, and a degradation study was performed to find out the weight loss and pH of PBS. Secondary structures of BSA on 13-93B0 were characterized by Fourier transform infrared (FTIR) spectroscopy. RESULTS: One hundred percent protein release occurred by 24 hours for all 13-93B3 groups. However, chitosan coating delayed 100% release up to 72 hours in 13-93B0 groups. The 13-93B3 glass showed higher degradation rates than 13-93B0 regardless of chitosan-coating status. Multilayer and sandwich chitosan coatings further delayed BSA release from 13-93B0. FTIR analysis revealed that α-helical structure was the highest among all groups and significantly higher in the 2% sandwich chitosan coating group (32.0% ± 2.1%), compared with uncoated and 4% chitosan groups. CONCLUSIONS: Chitosan coating can delay the burst release of BSA from 13-93B0 glass and be a potential coating on bioactive glass for drug delivery purposes.

Angiogenesis Assays for the Evaluation of Angiogenic Properties of Orthopaedic Biomaterials - A General Review.

Vascularization is an essential process in bone formation, remodeling and regeneration during both bone development and fracture repair. Vascularization remains a big challenge directly leading to the final success of newly regenerated bone. In this review, the advantages and disadvantages of different angiogenesis assays and bone defect models are described in details for investigating revascularization of materials of interest. Unlike conventional angiogenesis study with growth factors or pharmaceutical molecules performed in two-dimension, special considerations are taken into account whether these assays can be translated for testing three-dimensional implantable devices. Over the years, accurate and quantifiable in vitro, ex vivo and in vivo assays have been extensively demonstrated to be useful in examining how new blood vessels grow. These methods can contribute to the fundamental understanding of angiogenic properties of the materials, but a bone defect model is still pivotal in order to understand the cascade actions of angiogenesis along with bone formation. Finally, angiogenesis and osteogenesis are both complex processes interacting with each other, the choice of which assay to be performed should adequately address the clinical relevance and reflect the sequence of responses of revascularization of the test materials.

Fabrication of Poly-l-lactic Acid/Dicalcium Phosphate Dihydrate Composite Scaffolds with High Mechanical Strength-Implications for Bone Tissue Engineering.

Scaffolds were fabricated from poly-l-lactic acid (PLLA)/dicalcium phosphate dihydrate (DCPD) composite by indirect casting. Sodium citrate and PLLA were used to improve the mechanical properties of the DCPD scaffolds. The resulting PLLA/DCPD composite scaffold had increased diametral tensile strength and fracture energy when compared to DCPD only scaffolds (1.05 vs. 2.70 MPa and 2.53 vs. 12.67 N-mm, respectively). Sodium citrate alone accelerated the degradation rate by 1.5 times independent of PLLA. Cytocompatibility of all samples were evaluated using proliferation and differentiation parameters of dog-bone marrow stromal cells (dog-BMSCs). The results showed that viable dog-BMSCs attached well on both DCPD and PLLA/DCPD composite surfaces. In both DCPD and PLLA/DCPD conditioned medium, dog-BMSCs proliferated well and expressed alkaline phosphatase (ALP) activity indicating cell differentiation. These findings indicate that incorporating both sodium citrate and PLLA could effectively improve mechanical strength and biocompatibility without increasing the degradation time of calcium phosphate cement scaffolds for bone tissue engineering purposes.

Phytomolecule icaritin incorporated PLGA/TCP scaffold for steroid-associated osteonecrosis: Proof-of-concept for prevention of hip joint collapse in bipedal emus and mechanistic study in quadrupedal rabbits.

Steroid-associated osteonecrosis (SAON) may lead to joint collapse and subsequent joint replacement. Poly lactic-co-glycolic acid/tricalcium phosphate (P/T) scaffold providing sustained release of icaritin (a metabolite of Epimedium-derived flavonoids) was investigated as a bone defect filler after surgical core-decompression (CD) to prevent femoral head collapse in a bipedal SAON animal model using emu (a large flightless bird). The underlying mechanism on SAON was evaluated using a well-established quadrupedal rabbit model. Fifteen emus were established with SAON, and CD was performed along the femoral neck for the efficacy study. In this CD bone defect, a P/T scaffold with icaritin (P/T/I group) or without icaritin (P/T group) was implanted while no scaffold implantation was used as a control. For the mechanistic study in rabbits, the effects of icaritin and composite scaffolds on bone mesenchymal stem cells (BMSCs) recruitment, osteogenesis, and anti-adipogenesis were evaluated. Our efficacy study showed that P/T/I group had the significantly lowest incidence of femoral head collapse, better preserved cartilage and mechanical properties supported by more new bone formation within the bone tunnel. For the mechanistic study, our in vitro tests suggested that icaritin enhanced the expression of osteogenesis related genes COL1α, osteocalcin, RUNX2, and BMP-2 while inhibited adipogenesis related genes C/EBP-ß, PPAR-γ, and aP2 of rabbit BMSCs. Both P/T and P/T/I scaffolds were demonstrated to recruit BMSCs both in vitro and in vivo but a higher expression of migration related gene VCAM1 was only found in P/T/I group in vitro. In conclusion, both efficacy and mechanistic studies show the potential of a bioactive composite porous P/T scaffold incorporating icaritin to enhance bone defect repair after surgical CD and prevent femoral head collapse in a bipedal SAON emu model.

The effect of diluted triple and double antibiotic pastes on dental pulp stem cells and established Enterococcus faecalis biofilm.

OBJECTIVES: To investigate the effect of various dilutions of antibiotic medicaments used in endodontic regeneration on the survival of human dental pulp stem cells (DPSCs) and to determine their antibacterial effect against established Enterococcus faecalis biofilm. MATERIALS AND METHODS: The cytotoxic and antibacterial effects of different triple (TAP) and double antibiotic paste (DAP) dilutions (0.125, 0.25, 0.5, 1, and 10 mg/ml) were tested against Enterococcus faecalis established biofilm and DPSC. Established bacterial biofilm were exposed to antibiotic dilutions for 3 days. Then, biofilms were collected, spiral plated, and the numbers of bacterial colony forming units (CFU/ml) were determined. For the cytotoxic effect, lactate dehydrogenase activity assays (LDH) and cell viability assays (WST-1) were used to measure the percentage of DPSC cytotoxicity after 3-day treatment with the same antibiotic dilutions. A general linear mixed model was used for statistical analyses (α = 0.05). RESULTS: All antibiotic dilutions significantly decreased the bacterial CFU/ml. For WST-1 assays, all antibiotic dilutions except 0.125 mg/ml significantly reduced the viability of DPSC. For LDH assays, the three lowest tested concentrations of DAP (0.5, 0.25, 0.125 mg/ml) and the two lowest concentrations of TAP (0.25 and 0.125 mg/ml) were non-toxic to DPSC. CONCLUSIONS: All tested dilutions had an antibacterial effect against E. faecalis. However, 0.125 mg/ml of DAP and TAP showed a significant antibacterial effect with no cytotoxic effects on DPSCs. CLINICAL RELEVANCE: Using appropriate antibiotic concentrations of intracanal medicament during endodontic regeneration procedures is critical to disinfect root canal and decrease the adverse effects on stem cells.

The effects of 3D bioactive glass scaffolds and BMP-2 on bone formation in rat femoral critical size defects and adjacent bones.

Reconstruction of critical size defects in the load-bearing area has long been a challenge in orthopaedics. In the past, we have demonstrated the feasibility of using a biodegradable load-sharing scaffold fabricated from poly(propylene fumarate)/tricalcium phosphate (PPF/TCP) loaded with bone morphogenetic protein-2 (BMP-2) to successfully induce healing in those defects. However, there is limited osteoconduction observed with the PPF/TCP scaffold itself. For this reason, 13-93 bioactive glass scaffolds with local BMP-2 delivery were investigated in this study for inducing segmental defect repairs in a load-bearing region. Furthermore, a recent review on BMP-2 revealed greater risks in radiculitis, ectopic bone formation, osteolysis and poor global outcome in association with the use of BMP-2 for spinal fusion. We also evaluated the potential side effects of locally delivered BMP-2 on the structures of adjacent bones. Therefore, cylindrical 13-93 glass scaffolds were fabricated by indirect selective laser sintering with side holes on the cylinder filled with dicalcium phosphate dehydrate as a BMP-2 carrier. The scaffolds were implanted into critical size defects created in rat femurs with and without 10 µg of BMP-2. The x-ray and micro-CT results showed that a bridging callus was found as soon as three weeks and progressed gradually in the BMP group while minimal bone formation was observed in the control group. Degradation of the scaffolds was noted in both groups. Stiffness, peak load and energy to break of the BMP group were all higher than the control group. There was no statistical difference in bone mineral density, bone area and bone mineral content in the tibiae and contralateral femurs of the control and BMP groups. In conclusion, a 13-93 bioactive glass scaffold with local BMP-2 delivery has been demonstrated for its potential application in treating large bone defects.

Development of a cell-derived matrix: effects of epidermal growth factor in chemically defined culture.

Extracellular matrices without animal components and with high mechanical strength are needed for the development of the next generation of viable skin replacements. The goal of this study was to determine the optimal concentration of epidermal growth factor (EGF) to maximize the strength and collagen content of cell-derived matrix (CDM) produced by fibroblasts in vitro in serum-free media. Scaffold-free CDM samples were produced by human dermal fibroblasts in the presence of 0-50 ng/mL EGF in chemically defined media. After 21 days of culture, a membrane inflation system was used to measure the biaxial tensile strength, failure stretch ratio, and thickness of each treatment group. The fibroblasts treated with 5 ng/mL EGF produced the thickest matrix (270 microm). A thinner (130 microm) matrix, produced when the fibroblasts were treated with 0.5 ng/mL, had an ultimate tensile strength (895 kPa), greater than two times that of the other treatment groups. The fibroblasts treated with 0.5 ng/mL also had the highest collagen density (23.5 mg/cm(3)). Fibroblasts stimulated with the lowest (0.05 ng/mL) and highest (50 ng/mL) concentrations of EGF produced significantly weaker matrices and lower collagen densities. There was no significant correlation between UTS and collagen density suggesting that mechanisms other than density contribute to the strength of the matrix. Taken together, these data indicate that the optimal EGF concentration depends upon the relative importance of matrix strength and volume in a given application and that 0.5-5.0 ng/mL EGF promotes production of a robust extracellular matrix in only 3 weeks.