Assessment of inhalation exposure to microplastic particles when disposable masks are repeatedly used.

Wearing masks to prevent infectious diseases, especially during the COVID-19 pandemic, is common. However, concerns arise about inhalation exposure to microplastics (MPs) when disposable masks are improperly reused. In this study, we assessed whether disposable masks release inhalable MPs when reused in simulated wearing conditions. All experiments were conducted using a controlled test chamber setup with a constant inspiratory flow. Commercially available medical masks with a three-layer material, composition comprising polypropylene (PP in the outer and middle layers) and polyethylene (PE in the inner layer), were used as the test material. Brand-new masks with and without hand rubbing, as well as reused medical masks, were tested. Physical properties (number, size, and shape) and chemical composition (polymers) were identified using various analytical techniques such as fluorescence staining, fluorescence microscopy, and micro-Fourier Transform Infrared Spectroscopy (µFTIR). Scanning Electron Microscopy (SEM) was used to scrutinize the surface structure of reused masks across different layers, elucidating the mechanism behind the MP generation. The findings revealed that brand-new masks subjected to hand rubbing exhibited a higher cumulative count of MPs, averaging approximately 1.5 times more than those without hand rubbing. Fragments remained the predominant shape across all selected size classes among the released MPs from reused masks, primarily through a physical abrasion mechanism, accounting for >90 % of the total MPs. The numbers of PE particles were higher than PP particles, indicating that the inner layer of the mask contributed more inhalable MPs than the middle and outer layers combined. The released MPs from reused masks reached their peak after 8 h of wearing. This implies that regularly replacing masks serves as a preventive measure and mitigates associated health risks of inhalation exposure to MPs.

Hierarchically patterned polyurethane microgrooves featuring nanopillars or nanoholes for neurite elongation and alignment.

Surface micro- and nanostructures profoundly affect the functional performance of nerve regeneration implants by modulating neurite responses. However, few studies have investigated the impact of discrete nanostructures, such as nanopillars and nanoholes, and their combination with microgrooves on neurite outgrowth and alignment. Furthermore, numerous techniques have been developed for surface micro-/nanopatterning, but simple and low-cost approaches are quite limited. In this work, we show that nanopillars and nanoholes, and their combination with microgrooves, can be patterned on polyurethane (PU) films using a low-cost, reusable photoresist master mold prepared via nanosphere lens lithography and UV-LED photolithography, with specific "reinforcement" methods for overcoming the inherent drawbacks of using photoresist masters. We show that the PU nanopillars and nanoholes increase the neurite length of pheochromocytoma 12 (PC12) cells through unique growth cone interactions. Moreover, we demonstrate, for the first time, that hierarchically patterned nano-/microstructured PU films enhance both PC12 neurite elongation and alignment, showing the potential use of our proposed method for the micro-/nanopatterning of polymers for nerve tissue engineering.

3D-Cultured Adipose-Derived Stem Cell Spheres Using Calcium-Alginate Scaffolds for Osteoarthritis Treatment in a Mono-Iodoacetate-Induced Rat Model.

Osteoarthritis (OA) is a degenerative disease that causes pain, cartilage deformation, and joint inflammation. Mesenchymal stem cells (MSCs) are potential therapeutic agents for OA treatment. However, the 2D culture of MSCs could potentially affect their characteristics and functionality. In this study, calcium-alginate (Ca-Ag) scaffolds were prepared for human adipose-derived stem cell (hADSC) proliferation with a homemade functionally closed process bioreactor system; the feasibility of cultured hADSC spheres in heterologous stem cell therapy for OA treatment was then evaluated. hADSC spheres were collected from Ca-Ag scaffolds by removing calcium ions via ethylenediaminetetraacetic acid (EDTA) chelation. In this study, 2D-cultured individual hADSCs or hADSC spheres were evaluated for treatment efficacy in a monosodium iodoacetate (MIA)-induced OA rat model. The results of gait analysis and histological sectioning showed that hADSC spheres were more effective at relieving arthritis degeneration. The results of serological and blood element analyses of hADSC-treated rats indicated that the hADSC spheres were a safe treatment in vivo. This study demonstrates that hADSC spheres are a promising treatment for OA and can be applied to other stem cell therapies or regenerative medical treatments.

A Tumor Accelerator Based on Multicomponent Bone Scaffolds and Cancer Cell Homing.

Bone tissue attracts cancer cell homing biologically, mechanically, or chemically. It is difficult and time consuming to identify their complex cross-talk using existed methods. In this study, a multi-component bone matrix was fabricated using gelatin, hydroxyapatite (HAp), and epidermal growth factor (EGF) as raw materials to investigate how "acellular" bone matrix affects cancer cell homing in bone. Then, EGF-responsive cancer cells were cultured with the scaffold in a dynamical bioreactor. For different culture periods, the effects of HAp, gelatin, and EGF on the cell adhesion, proliferation, 3D growth, and migration of cancer were evaluated. The results indicated that a small amount of calcium ion released from the scaffolds accelerated cancer MDA-MB-231 adhesion on the surface of inner pores. Moreover, degradable gelatin key caused cancer cell growth on the scaffold surface to turn into a 3D aggregation. Despite this, the formation of cancer spheroids was slow, and required 14 days of dynamic culture. Thankfully, EGF promoted cancer cell adhesion, proliferation, and migration, and cancer spheroids were observed only after 3-day culture. We concluded that the combination of the multiple components in this scaffold allows cancer cells to meet multiple requirements of cancer dynamic progression.

Measurement of Low Concentration of Micro-Plastics by Detection of Bioaffinity-Induced Particle Retention Using Surface Plasmon Resonance Biosensors.

The issue of micro-plastics is becoming more and more important due to their ubiquity and the harm they cause to the human body. Therefore, evaluating the biological-physical interaction of micro-plastics with health cells has become the focus of many research efforts. This study focuses on the movement mode and low concentration detection development for micro-plastics in surface plasmon resonance (SPR). Firstly, 20-micrometer micro-plastics were prepared by grinding and filtering, and the movement mode was explored; then, the characteristics were investigated by SPR. Chromatographic analysis showed that the surface charge of micro-plastics dominated the elution time, and estrogen receptors (ERs) played a supporting role. A difference of micro-plastics in SPR sensorgram was observed, inferring the micro-plastics' movement in rolling mode on the ERs. Characteristics analysis indicated that the low particle number of micro-plastics on SPR showed a linear relationship with the response unit (RU). When ERs were immobilized on the biosensor, the force of the binding of micro-plastics to ERs under an ultra-low background was equivalent to the dissociation rate constant shown as follows: PS (0.05 nM) > PVC (0.09 nM) > PE (0.14 nM). The ELISA-like magnetic beads experiment verified the specificity between ERs and micro-plastics. Therefore, by using the SPR technique, a biological-derived over-occupation of PS was found via higher binding force with ERs and longer retention time. In the future, there will be considerable potential for micro-plastics issues, such as identification in natural samples, biomarking, real-time detection in specific environments/regions and human health subject.

A Cyclic BMP-2 Peptide Upregulates BMP-2 Protein-Induced Cell Signaling in Myogenic Cells.

In the current study, we designed four cyclic peptide analogues by incorporating two cysteine residues in a BMP-2 linear knuckle epitope in such a way that the active region of the peptide could be either inside or outside the cyclic ring. Bone morphogenetic protein receptor BMPRII was immobilized on the chip surface, and the interaction of the linear and cyclic peptide analogues was studied using surface plasmon resonance (SPR). From the affinity data, the peptides with an active region inside the cyclic ring had a higher binding affinity in comparison to the other peptides. To confirm that our affinity data are in line in vitro, we studied the expression levels of RUNX2 (runt-related transcription factor) and conducted an osteogenic marker alkaline phosphatase (ALP) assay and staining. Based on the affinity data and the in vitro experiments, peptide P-05 could be a suitable candidate for osteogenesis, with higher binding affinity and increased RUNX2 and ALP expression in comparison to the linear peptides.

An MRI-Guided Ring High-Intensity Focused Ultrasound System for Noninvasive Breast Ablation.

High-intensity focused ultrasound (HIFU) has been used for noninvasive treatment of breast tumors, but the present magnetic resonance imaging (MRI)-guided HIFU (MRI-HIFU) systems encounter skin burn. In this study, a novel MRI-HIFU breast ablation system was developed to improve the above problem. The system consisted of the ring HIFU phased-array transducer, a commercial power amplifier, the mechanical positioner, and the graphical user interface control software. MRI thermometry was also established to monitor the temperature in the HIFU-treated tissue. Ablation of pork and the in vivo rabbit leg were carried out to validate the developed system. Results of fat-surrounding pork ablation showed that the ring HIFU system reached a safe margin of 3 mm without fat burn. Moreover, precision of the positioner moving the HIFU focal zone was within 6% error under MRI circumstances. The representative MRI temperature images show that the peak temperatures among the five ablations ranged between 66 °C and 91 °C, and their thermal doses were over 10000. The system could also ablate the biceps femoris of a rabbit without skin burn to form a lesion of 2.5 mm beneath the skin. With the HIFU dose of 315 W/10 s, the MRI temperature map revealed that the maximum temperature and the thermal dose were 60 °C and 3380, respectively. The MRI-guided ring HIFU system can ablate the target tissue near subcutaneous fat without fat burn. The system prototype is a promising tool for clinical implementation.

A study of Drynaria fortunei in modulation of BMP­2 signalling by bone tissue engineering

Background/aim: Drynaria fortunei (Gusuibu; GSB) is a popular traditional Chinese medicine used for bone repair. An increasing number of studies have reported that GSB induces osteogenic differentiation in bone marrow mesenchymal stem cells (BMSCs). These results provide insight into the application of GSB for bone tissue engineering techniques used to repair large bone defects. However, few studies have described the molecular mechanisms of GSB. Materials and methods: In the present study, the effects of GSB and naringin, a marker compound, on the binding of BMP-2 to BMPR and BMP-2-derived signal transduction were investigated using surface plasmon resonance (SPR) and coculturing with BMPR- expressed cell line, C2C12, respectively. Furthermore, naringin was also used to prepare naringin contained scaffolds for bone tissue engineering. The physical and chemical properties of these scaffolds were analysed using scanning electron microscopy (SEM) and highperformance liquid chromatography (HPLC). These scaffolds were cocultured with rabbit BMSCs in vitro and implanted into rabbit calvarial defects for bone repair assessment. Results: The results showed that GSB and naringin affect the binding of BMP and BMPR in SPR experiments. GSB is a subtle BMP modulator that simultaneously inhibits the binding of BMP-2 to BMPR-1A and enhances its binding to BMPR-1B. In contrast, naringin inhibited BMP-2 binding to BMPR-1A. In vitro studies involving the phosphorylation of signals downstream of BMPR and Smad showed that GSB and naringin affected stem cell differentiation by inhibiting BMPR-1A signalling. When using GSB for bone tissue engineering, naringin exhibited a higher capacity for slow and gradual release from the scaffold, which promotes bone formation via osteoinduction. Moreover, control and naringin scaffolds were implanted into rabbit calvarial defects for 4 weeks, and naringin enhanced bone regeneration in vivo significantly. Conclusions: GSB and its marker compound (naringin) could inhibit the binding of BMP-2 and BMPR-1A to control cell differentiation by blocked BMPR-1A signalling and enhanced BMPR-1B signalling. GSB and naringin could be good natural BMP regulators for bone tissue engineering.

Enhancement of Neurite Outgrowth by Warming Biomaterial Ultrasound Treatment.

Ultrasound is a method for enhancing neurite outgrowth because of its thermal effect. In order to reach the working temperature to enhance neurite outgrowth, long-time treatment by ultrasound is necessary, while acknowledging that the treatment poses a high risk of damaging nerve cells. To overcome this problem, we developed a method that shortens the ultrasonic treatment time with a warming biomaterial. In this study, we used Fe3O4 nanoparticle-embedded polycaprolactone (PCL) as a sonosensitized biomaterial, which has an excellent heating rate due to its high acoustic attenuation. With this material, the ultrasonic treatment time for enhancing neurite outgrowth could be effectively shortened. Ultrasonic treatment could also increase neuronal function combined with the warming biomaterial, with more promoter neuronal function than only ultrasound. Moreover, the risk of overexposure can be avoided by the use of the warming biomaterial by reducing the ultrasonic treatment time, providing better effectiveness.

Development of an MRI-Compatible High-Intensity Focused Ultrasound Phased Array Transducer Dedicated for Breast Tumor Treatment.

High-intensity focused ultrasound (HIFU) under magnetic resonance imaging (MRI) guidance can achieve a noninvasive and precise ablation of the solid tumor. In the study, an MRI-compatible 1-MHz 16-channel ring-shaped transducer was developed to minimize the burn risk of breast skin and perform volumetric ablation for short treatment time. The measured electroacoustic conversion efficiency of the transducer was 50.90% ± 5. The transducer could produce a point and a quasi-hollow-cylinder lesion in a thermal-sensitive phantom or an ex vivo pork by tuning the phase of each element. It may achieve volumetric ablation of 1.5 cm3 when the point lesion is located inside the hollow lesion. Ex vivo ablation experiments showed that the transducer could cause a coagulative necrosis in the pork from the surrounded subcutaneous fat by 5 mm without fat damage. The temperature and region of the pork ablation were quantified by MRI technique. There was no MRI interference from HIFU and vice versa while both systems operated concurrently.

A Coaxial Dual-element Focused Ultrasound Probe for Guidance of Epidural Catheterization: An Experimental Study.

Ultrasound guidance for epidural block has improved clinical blind-trial problems but the design of present ultrasonic probes poses operating difficulty of ultrasound-guided catheterization, increasing the failure rate. The purpose of this study was to develop a novel ultrasonic probe to avoid needle contact with vertebral bone during epidural catheterization. The probe has a central circular passage for needle insertion. Two focused annular transducers are deployed around the passage for on-axis guidance. A 17-gauge insulated Tuohy needle containing the self-developed fiber-optic-modified stylet was inserted into the back of the anesthetized pig, in the lumbar region under the guidance of our ultrasonic probe. The inner transducer of the probe detected the shallow echo signals of the peak-peak amplitude of 2.8 V over L3 at the depth of 2.4 cm, and the amplitude was decreased to 0.8 V directly over the L3 to L4 interspace. The outer transducer could detect the echoes from the deeper bone at the depth of 4.5 cm, which did not appear for the inner transducer. The operator tilted the probe slightly in left-right and cranial-caudal directions until the echoes at the depth of 4.5 cm disappeared, and the epidural needle was inserted through the central passage of the probe. The needle was advanced and stopped when the epidural space was identified by optical technique. The needle passed without bone contact. Designs of the hollow probe for needle pass and dual transducers with different focal lengths for detection of shallow and deep vertebrae may benefit operation, bone/nonbone identification, and cost.

Cardiac fibrosis in mouse expressing DsRed tetramers involves chronic autophagy and proteasome degradation insufficiency.

Proteinopathy in the heart which often manifests excessive misfolded/aggregated proteins in cardiac myocytes can result in severe fibrosis and heart failure. Here we developed a mouse model, which transgenically express tetrameric DsRed, a red fluorescent protein (RFP), in an attempt to mimic the pathological mechanisms ofcardiac fibrosis. Whilst DsRed is expressed and forms aggregation in most mouse organs, certain pathological defects are specifically recapitulated in cardiac muscle cells including mitochondria damages, aggresome-like residual bodies, excessive ubiquitinated proteins, and the induction of autophagy. The proteinopathy and cellular injuries caused by DsRed aggregates may be due to impaired or overburdened ubiquitin-proteasome system and autophagy-lysosome systems. We further identified that DsRed can be ubiquitinated and associated with MuRF1, a muscle-specific E3 ligase. Concomitantly, an activation of NF-κB signaling and a strong TIMP1 induction were noted, suggesting that RFP-induced fibrosis was augmented by a skewed balance between TIMP1 and MMPs. Taken together, our study highlights the molecular consequences of uncontrolled protein aggregation leading to congestive heart failure, and provides novel insights into fibrosis formation that can be exploited for improved therapy.

A potent inhibition of oxidative stress induced gene expression in neural cells by sustained ferulic acid release from chitosan based hydrogel.

Traumatic brain injury (TBI) is an extremely cataclysmic neurological disorder and the inhibition of oxidative stress following TBI could effectively protect the brain from further impairments. An injectable thermosensitive chitosan/gelatin/ß-Glycerol phosphate (C/G/GP) hydrogel for the controlled release of the phenolic antioxidant ferulic acid (FA) to inhibit the neurological oxidative stress was demonstrated. The C/G/GP hydrogel ensures an excellent clinical expediency with a gelation temperature of 32.6°C and gelation time of 75.58s. In-vitro cytotoxicity assays of C/G/GP hydrogel and FA have revealed an excellent biocompatibility with the Neuro-2a cells. 500µM of FA was considered to be an effective concentration to reduce the oxidative stress in Neuro-2a cells. TUNEL staining images evidenced that the H2O2 induced DNA fragmentation was comprehensively controlled after FA treatment. The mRNA gene expression profiles markedly authenticate the neuroprotectivity of FA by down-regulating ROS, inflammatory and apoptosis related markers. The outcomes of this study suggest that, C/G/GP hydrogel carrying ferulic acid could effectively protect further secondary traumatic brain injury associated impairments.

Injectable and thermoresponsive self-assembled nanocomposite hydrogel for long-term anticancer drug delivery.

The purpose of this study is to develop an injectable thermoresponsive hydrogel system that can undergo sol-gel phase transition by the stimulation of body temperature with improved mechanical stability and biocompatibility as a controlled drug delivery carrier for cancer therapy. Hexamethylene diisocyanate (HDI) was introduced into Pluronic F127 as a chain extender to improve the mechanical stability. HDI-Pluronic F127 copolymer was then incorporated with hyaluronic acid to develop a thermoresponsive nanocomposite hydrogel system. The physiochemical properties were characterized. The anticancer drug release profile and effect to inhibit tumor cells growth were analyzed in vitro and in vivo. The results showed that HDI-Pluronic F127/hyaluronic acid thermoresponsive hydrogel could undergo sol-gel transition as temperature increased to 37 °C. The nanocomposite polymer can spontaneously self-assemble into micellar structure with size of 100-200 nm. The release of doxorubicin (DOX) from HDI-PF127/HA composite hydrogel was a zero-order profile and maintained sustained release for over 28 days. The viability of tumor cells and size of tumor significantly decreased with incubation time, indicating the potential to have a therapeutic effect for cancer therapy. The injectable thermoresponsive nanocomposite hydrogel system was biocompatible and degradable and had the slow controlled release property for anticancer drugs with potential applications in the field of drug delivery.

Autologous bone marrow stromal cells loaded onto porous gelatin scaffolds containing Drynaria fortunei extract for bone repair.

GGT-GSB composite was prepared by mixing a biodegradable GGT composite containing genipin-crosslinked gelatin and ß-tricalcium phosphate with Gu-Sui-Bu extract (GSB) (Drynaria fortunei (Kunze) J. Sm.), a traditional Chinese medicine. Then, porous GGT and GGT-GSB scaffolds were fabricated using a salt-leaching method. The GGT and GGT-GSB scaffolds thus obtained had a macroporous structure and high porosity. Rabbit bone marrow stromal cells (BMSCs) were seeded onto GGT and GGT-GSB scaffolds. The biological response of rabbit calvarial bone to these scaffolds was considered to evaluate the potential of the scaffolds for use in bone tissue engineering. After 8 weeks of implantation, each scaffold induced new bone formation at a cranial bone defect, as was verified by X-ray microradiography. The BMSC-seeded GGT-GSB scaffolds induced more new bone formation than the BMSC-seeded GGT and acellular scaffolds. These observations suggest that an autologous BMSCs-seeded porous GGT-GSB scaffold can be adopted in bone engineering in vivo and has great potential for regenerating defective bone tissue.

Preparation and characterization of dexamethasone-immobilized chitosan scaffold.

Chitosan film (CSF), 2-dimensional scaffold, was modified with dexamethasone (DEX) in the present work via amino-alcohol forming reaction between amino group of chitosan and carbonyl group of DEX. Successful immobilization was identified by ATR-FTIR spectroscopy and, the immobilized amounts were determined by weighting methods (WM) and integrated area analysis method (AM) of HPLC. For experiments of cell culture of osteogenic differentiation, our results showed that DEX immobilization has more efficiency than the other group (such as both groups of CSF and CSF with free DEX, CSF/fDEX), which was demonstrated as indicated by cell image analysis and ALP activity assay. The results show, as AD-MSCs cultured on DEX-CSF, its shape of cell transformed to polygonal or cubical. The ALP activity assay indicates that of DEX-CSF increase about 3.5 folds than that of CSF's. The results suggest that immobilization of DEX can make chitosan scaffolds to induce differentiation of AD-MSCs toward osteoblastic lineage for bone tissue engineering application.

Induction of the mitochondria apoptosis pathway by phytohemagglutinin erythroagglutinating in human lung cancer cells.

BACKGROUND: Deregulation of apoptosis will influence the balance of cell proliferation and cell death, resulting in various fatal diseases that can include cancer. In prior research reports related to cancer therapy, phytohemagglutinin, a lectin extracted from red kidney beans, demonstrated the ability to inhibit the growth of human cancer cells. However, one of its isoforms, erythroagglutinating, has yet to be evaluated on its anticancer effects. METHODS: PHA-E was used to induce apoptosis of A-549 lung cancer cells and the possible signal transduction pathway was elucidated, as measured by the MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay, G6PD release assay, flow cytometry, and Western blot analysis. RESULTS: PHA-E treatment caused a dose-dependent increase of cell growth inhibition and cytotoxicity on A-549 cells. In annexin V/propidium iodide [i.e., PI] and TUNEL (terminal deoxynucleotidyl transferase dUTP nick-end labeling)/PI assay, we found that the rate of apoptotic cells was raised as the concentration of PHA-E increased. Treatment of A-549 cells with PHA-E resulted in enhancing the release of cytochrome c, which thus activated an increase in caspase 9 and caspase 3, the upregulation of Bax and Bad, the downregulation of Bcl-2 and phosphorylated Bad, and finally the inhibition of the epidermal growth factor receptor and its downstream signal pathway PI3K/Akt and MEK/ERK. CONCLUSIONS: PHA-E can induce growth inhibition and cytotoxicity of lung cancer cells, which is mediated through an activation of the mitochondria apoptosis pathway. These results suggest that PHA-E can be developed into a new therapeutic treatment that can be applied as an effective anti-lung cancer drug in the near future.

Reconstruction of calvarial defect using a tricalcium phosphate-oligomeric proanthocyanidins cross-linked gelatin composite.

A biodegradable GTP composite which was composed of oligomeric proanthocyanidins (OPCs) cross-linked gelatin mixed with tricalcium phosphate was developed as a bone substitute. The subcutaneous implantation in rats was examined to determine the in vivo degradation and biocompatibility of the GTP composites with various cross-linking densities. Experimental results indicated that the rate of in vivo degradation was markedly attenuated as the concentration of OPCs increased above 5.0wt%. Furthermore, this study examined the biological response of rabbit calvarial bone to GTP composite to evaluate its potential for use as an osteoconductive bone substitute. Bone defects (10mm in diameter) in New Zealand white rabbits were filled with the GTP composite. The de-protenized bovine cancellous bone matrix was employed as the control material. The results of radiographic analyses demonstrated obviously greater new bone ingrowth in the GTP composite than in the de-proteinized bovine bone at the same implantation time. Progressive replacement of the GTP composite by new bone proceeded by a combination of osteoconduction and biodegradation. The biodegradable GTP composite thus has great potential for improving bone repair.

Blocking effect of an immuno-suppressive agent, cynarin, on CD28 of T-cell receptor.

PURPOSE: Cynarin, a potential immunosuppressant that blocks the interaction between the CD28 of T-cell receptor and CD80 of antigen presenting cells, was found in Echinacea purpurea by a new pharmaceutical screening method: After Flowing Through Immobilized Receptor (AFTIR; Dong et al., J Med Chem, 49: 1845-1854, 2006). This Echinacea component is the first small molecule that is able to specifically block "signal 2" of T-cell activation. METHODS: In this study, we used the AFTIR method to further confirm that cynarin effectively blocked the binding between CD80 of B-cells and CD28 of T-cells, and provide details of its mechanism of action. RESULTS: The experimental results showed that cynarin blocked about 87% of the CD28-dependent "signal 2" pathway of T-cell activation under the condition of one to one ratio of T-cell and B-cell in vitro. Theoretical structure modeling showed that cynarin binds to the "G-pocket" of CD28 (Evans et al., Nat Immunol, 6:271-279, 2005), and thus interrupts the site of interaction between CD28 and CD80. CONCLUSIONS: These results confirm both that AFTIR is a promising method for screening selective active compounds from herbal medicine and that cynarin has great potential as an immuno-suppressive agent.

A novel bone substitute composite composed of tricalcium phosphate, gelatin and drynaria fortunei herbal extract.

The Chinese herb, Gu-Sui-Bu (GSB) (Drynaria fortunei J. Sm.) has been anecdotally reported to enhance bone healing. We had previously confirmed in vitro the efficacy and safety of GSB in bone healing, and showed that it influenced both osteoblast and osteoclast activity. For clinically useful application of these bone regenerative effects, a satisfactory delivery system for GSB is required. In this study, we determined the optimal concentration of GSB for regenerative activity in rat bone cells via MTT, alkaline phosphatase (ALP), nodule formation and TRAP assays, and designed and tested a GSB-rich bone composite material. The composite was fabricated by mixing a biodegradable GGT composite, containing genipin cross-linked gelatin and tricalcium phosphate, with the predetermined concentration of GSB (GGT-GSB). Neonatal rat calvarial culture and animal implantation were employed to evaluate and compare in vitro and in vivo the potential of GGT-GSB and GGT in regeneration of defective bone tissue. The most effective concentration of GSB was 100 mug/mL, which significantly increased osteoblast numbers, intracellular ALP levels and nodule numbers, without influencing osteoclast activity. In vitro and in vivo tests also showed that GGT-GSB accelerated bone regeneration compared to GGT. GGT-GSB thus has great potential for improved bone repair.