Hybrid-Aligned Fibers of Electrospun Gelatin with Antibiotic and Polycaprolactone Composite Membranes as an In Vitro Drug Delivery System to Assess the Potential Repair Capacity of Damaged Cornea.

The cornea lacks the ability to repair itself and must rely on transplantation to repair damaged tissue. Therefore, creating alternative therapies using dressing membranes based on tissue engineering concepts to repair corneal damage before failure has become a major research goal. Themost outstanding features that are important in reconstructing a damaged cornea are the mechanical strength and transparency of the membrane, which are the most important standard considerations. In addition, preventing infection is an important issue, especially in corneal endothelial healing processes. The purpose of this study was to produce aligned fibers via electrospinning technology using gelatin (Gel) composite polycaprolactone (PCL) as an optimal transport and antibiotic release membrane. The aim of the composite membrane is to achieve good tenacity, transparency, antibacterial properties, and in vitro biocompatibility. Results showed that the Gel and PCL composite membranes with the same electrospinning flow rate had the best transparency. The Gel impregnated with gentamicin antibiotic in composite membranes subsequently exhibited transparency and enhanced mechanical properties provided by PCL and could sustainably release the antibiotic for 48 h, achieving good antibacterial effects without causing cytotoxicity. This newly developed membrane has the advantage of preventing epidermal tissue infection during clinical operations and is expected to be used in the reconstruction of damaged cornea in the future.

Synergistic effect of drug/antibiotic-impregnated micro/nanohybrid mesoporous bioactive glass/calcium phosphate composite bone cement on antibacterial and osteoconductive activities.

Calcium phosphate bone cements (CPC) can be used in minimally invasive surgery because of their injectability, and they can also be used to repair small and irregular bone defects. This study aimed to release the antibiotic gentamicin sulfate (Genta) to reduce tissue inflammation and prevent infection in the early stages of bone recovery. Subsequently, the sustained release of the bone-promoting drug ferulic acid (FA) mimicked the response of osteoprogenitor D1 cells interaction, thereby accelerating the healing process of the overall bone repair. Accordingly, the different particle properties of micro-nano hybrid mesoporous bioactive glass (MBG), namely, micro-sized MBG (mMBG) and nano-sized MBG (nMBG), were explored separately to generate different dose releases in MBG/CPC composite bone cement. Results show that nMBG had better sustained-release ability than mMBG when impregnated with the same dose. When 10 wt% of mMBG hybrid nMBG and composite CPC were used, the amount of MBG slightly shortened the working/setting time and lowered the strength but did not hinder the biocompatibility, injectability, anti-disintegration, and phase transformation of the composite bone cement. Furthermore, compared with 2.5wt%Genta@mMBG/7.5 wt% FA@nMBG/CPC, 5wt.%Genta@mMBG/5wt.%FA@nMBG/CPC exhibited better antibacterial activity, better compressive strength, stronger mineralization of osteoprogenitor cell, and similar 14-day slow-release trend of FA. The MBG/CPC composite bone cement developed can be used in clinical surgery to achieve the synergistic sustained release of antibacterial and osteoconductive activities.

Enhanced Cell Osteogenic Differentiation in Alendronate Acid and Flufenamic Acid Drug-Impregnated Nanoparticles of Mesoporous Bioactive Glass Composite Calcium Phosphate Bone Cement In Vitro.

This study aims to compare the anti-osteoporotic drugs alendronic acid (ALN) and flufenamic acid (FA) alone impregnate into nanoparticles of mesoporous bioactive glass (nMBG), which further composites calcium phosphate cement (CPC) and investigates their in vitro performance. The drug release, physicochemical properties, and biocompatibility of nMBG@CPC composite bone cement are tested, and the effect of the composites on improving the proliferation and differentiation efficiency of mouse precursor osteoblasts (D1 cells) is also investigated. Drug release shows that FA impregnates nMBG@CPC composite, a large amount of FA is released rapidly within 8 h, gradually reaching a stable release within 12 h, followed by a slow and sustained release within 14 days, and then reaches a plateau within 21 days. The release phenomenon confirms that the drug-impregnated nBMG@CPC composite bone cement effectively achieves slow drug delivery. The working time and setting time of each composite are within 4-10 min and 10-20 min, respectively, meeting the operational requirements of clinical applications. The addition of nMBG nanoparticles in the CPC matrix did not prevent the aggregation phenomenon under microstructural observation, thus resulting in a decrease in the strength of the nMBG@CPC composite. However, after 24 h of immersed reaction, the strength of each 5 wt.% nMBG impregnated with different concentrations of FA and ALN is still greater than 30 MPa, which is higher than the general trabecular bone strength. The drug-impregnated nMBG@CPC composites did not hinder the product formation and exhibit biocompatibility. Based on the proliferation and mineralization of D1 cells, the combination of nMBG with abundant FA and ALN in CPC is not conducive to the proliferation of D1 cells. However, when D1 cells are contact cultured for 21 days, alkaline phosphatase (ALP) enzyme activity shows higher ALP secretion from drug-impregnated nMBG@CPC composites than drug-free composites. Accordingly, this study confirms that nMBG can effectively impregnate the anti-osteoporosis drugs FA and ALN, and enhance the mineralization ability of osteoblasts. Furthermore, drug-impregnated nMBG applications can be used alone or in combination with CPC as a new option for osteoporotic bone-filling surgery.

In Vitro Characterizations of Post-Crosslinked Gelatin-Based Microspheres Modified by Phosphatidylcholine or Diammonium Phosphate as Antibiotic Delivery Vehicles.

Hydrogel-based microspheres prepared by emulsification have been widely used as drug carriers, but biocompatibility remains a challenging issue. In this study, gelatin was used as the water phase, paraffin oil was used as the oil phase, and Span 80 was used as the surfactant. Microspheres were prepared using a water-in-oil (W/O) emulsification. Diammonium phosphate (DAP) or phosphatidylcholine (PC) were further used to improve the biocompatibility of post-crosslinked gelatin microspheres. The biocompatibility of DAP-modified microspheres (0.5-10 wt.%) was better than that of PC (5 wt.%). The microspheres soaked in phosphate-buffered saline (PBS) lasted up to 26 days before fully degrading. Based on microscopic observation, the microspheres were all spherical and hollow inside. The particle size distribution ranged from 19 µm to 22 µm in diameter. The drug release analysis showed that the antibiotic gentamicin loaded on the microspheres was released in a large amount within 2 h of soaking in PBS. It was stabilized until the amount of microspheres integrated was significantly reduced after soaking for 16 days and then released again to form a two-stage drug release curve. In vitro experiments showed that DAP-modified microspheres at concentrations less than 5 wt.% had no cytotoxicity. Antibiotic-impregnated and DAP-modified microspheres had good antibacterial effects against Staphylococcus aureus and Escherichia coli, but these drug-impregnated groups hinder the biocompatibility of hydrogel microspheres. The developed drug carrier can be combined with other biomaterial matrices to form a composite for delivering drugs directly to the affected area in the future to achieve local therapeutic effects and improve the bioavailability of drugs.

Statin use in patients with type 2 diabetes has lower risk of hip fractures: A Taiwan national population-based study.

AIMS: Type 2 diabetes mellitus (T2DM) frequently co-exists with osteoporosis and dyslipidemia. Statins have been commonly used in the treatment of dyslipidemia. Recent studies have indicated a therapeutic role of statins in decreasing the risk of osteoporosis and fractures, but conflicting results have been reported. This study investigated the association between statin use and hip fracture (HFx) risk among T2DM patients. MATERIALS AND METHODS: A retrospective Taiwan population-based propensity-matched cohort study was performed using the Diabetes Mellitus Health Database from Taiwan National Health Insurance Research Database. Patients with newly diagnosed with T2DM between 2010 and 2014 were identified. Patients who previously used statins and had ever suffered HFx before the index date were excluded. HFx that occurred from 2010 to 2019 was collected to compute the cumulative rate of HFx. Hazard ratios (HRs) were calculated for the HFx risk according to the use or non-use of statins. To evaluate the dose-effect relationship of statins, sensitivity analyses were conducted. RESULTS: After propensity score matching for age and sex, 188,588 patients were identified as statin users and non-statin users. Statin use after T2DM diagnosis was associated with a decreased HFx risk with an adjusted HR (aHR) of 0.69 (P < 0.001). A dose-effect relationship was identified. The aHRs for developing HFx were 1.29, 0.67, and 0.36 for patients who used 28-174, 175-447, and >447 cumulative defined daily doses of statins, respectively (P < 0.001). CONCLUSIONS: Statin use in adults with T2DM showed a lower risk of HFx by demonstrating a dose-response relationship.

The Role of Autophagy in Osteoarthritic Cartilage.

Osteoarthritis (OA) is one of the most common diseases leading to physical disability, with age being the main risk factor, and degeneration of articular cartilage is the main focus for the pathogenesis of OA. Autophagy is a crucial intracellular homeostasis system recycling flawed macromolecules and cellular organelles to sustain the metabolism of cells. Growing evidences have revealed that autophagy is chondroprotective by regulating apoptosis and repairing the function of damaged chondrocytes. Then, OA is related to autophagy depending on different stages and models. In this review, we discuss the character of autophagy in OA and the process of the autophagy pathway, which can be modulated by some drugs, key molecules and non-coding RNAs (microRNAs, long non-coding RNAs and circular RNAs). More in-depth investigations of autophagy are needed to find therapeutic targets or diagnostic biomarkers through in vitro and in vivo situations, making autophagy a more effective way for OA treatment in the future. The aim of this review is to introduce the concept of autophagy and make readers realize its impact on OA. The database we searched in is PubMed and we used the keywords listed below to find appropriate article resources.

Morphological Changes, Antibacterial Activity, and Cytotoxicity Characterization of Hydrothermally Synthesized Metal Ions-Incorporated Nanoapatites for Biomedical Application.

The objective of this study was to prepare hydroxyapatite (HA) with potential antibacterial activity against gram-negative and gram-positive bacteria by incorporating different atomic ratios of Cu2+ (0.1-1.0%), Mg2+ (1.0-7.0%), and Zn2+ (1.0-7.0%) to theoretically replace Ca2+ ions during the hydrothermal synthesis of grown precipitated HA nanorods. This study highlights the role of comparing different metal ions on synthetic nanoapatite in regulating the antibacterial properties and toxicity. The comparisons between infrared spectra and between diffractograms have confirmed that metal ions do not affect the formation of HA phases. The results show that after doped Cu2+, Mg2+, and Zn2+ ions replace Ca2+, the ionic radius is almost the same, but significantly smaller than that of the original Ca2+ ions, and the substitution effect causes the lattice distance to change, resulting in crystal structure distortion and reducing crystallinity. The reduction in the length of the nanopatites after the incorporation of Cu2+, Mg2+, and Zn2+ ions confirmed that the metal ions were mainly substituted during the growth of the rod-shape nanoapatite Ca2+ distributed along the longitudinal site. The antibacterial results show that nanoapatite containing Cu2+ (0.1%), Mg2+ (3%), and Zn2+ (5-7%) has obvious and higher antibacterial activity against gram-positive bacteria Staphylococcus aureus within 2 days. The antibacterial effect against the gram-negative bacillus Escherichia coli is not as pronounced as against Staphylococcus aureus. The antibacterial effect of Cu2+ substituted Ca2+ with an atomic ratio of 0.1~1.0% is even better than that of Mg2+- and Zn2+- doped with 1~7% groups. In terms of cytotoxicity, nanoapatites with Cu2+ (~0.2%) exhibit cytotoxicity, whereas Mg2+- (1-5%) and Zn2+- (~1%) doped nanoapatites are biocompatible at low concentrations but become cytotoxic as ionic concentration increases. The results show that the hydrothermally synthesized nanoapatite combined with Cu2+ (0.2%), Mg2+ (3%), and Zn2+ (3%) exhibits low toxicity and high antibacterial activity, which provides a good prospect for bypassing antibiotics for future biomedical applications.

In Vitro Evaluation of Calcium Phosphate Bone Cement Composite Hydrogel Beads of Cross-Linked Gelatin-Alginate with Gentamicin-Impregnated Porous Scaffold.

Calcium phosphate bone cement (CPC) is in the form of a paste, and its special advantage is that it can repair small and complex bone defects. In the case of open wounds, tissue debridement is necessary before tissue repair and the subsequent control of wound infection; therefore, CPC composite hydrogel beads containing antibiotics provide an excellent option to fill bone defects and deliver antibiotics locally for a long period. In this study, CPC was composited with the millimeter-sized spherical beads of cross-linked gelatin-alginate hydrogels at the different ratios of 0 (control), 12.5, 25, and 50 vol.%. The hydrogel was impregnated with gentamicin and characterized before compositing with CPC. The physicochemical properties, gentamicin release, antibacterial activity, biocompatibility, and mineralization of the CPC/hydrogel composites were characterized. The compressive strength of the CPC/hydrogel composites gradually decreased as the hydrogel content increased, and the compressive strength of composites containing gentamicin had the largest decrease. The working time and setting time of each group can be adjusted to 8 and 16 min, respectively, using a hardening solution to make the composite suitable for clinical use. The release of gentamicin before the hydrogel beads was composited with CPC varied greatly with immersion time. However, a stable controlled release effect was obtained in the CPC/gentamicin-impregnated hydrogel composite. The 50 vol.% hydrogel/CPC composite had the best antibacterial effect and no cytotoxicity but had reduced cell mineralization. Therefore, the optimal hydrogel beads content can be 25 vol.% to obtain a CPC/gentamicin-impregnated hydrogel composite with adequate strength, antibacterial activity, and bio-reactivity. This CPC/hydrogel containing gentamicin is expected to be used in clinical surgery in the future to accelerate bone regeneration and prevent prosthesis infection after surgery.

In Vitro Evaluation of a Composite Gelatin-Hyaluronic Acid-Alginate Porous Scaffold with Different Pore Distributions for Cartilage Regeneration.

Although considerable achievements have been made in the field of regenerative medicine, since self-repair is not an advanced ability of articular cartilage, the regeneration of osteochondral defects is still a challenging problem in musculoskeletal diseases. Cartilage regeneration aims to design a scaffold with appropriate pore structure and biological and mechanical properties for the growth of chondrocytes. In this study, porous scaffolds made of gelatin, hyaluronic acid, alginate, and sucrose in different proportions of 2 g (SL2) and 4 g (SL4) were used as porogens in a leaching process. Sucrose with particle size ranges of 88-177 µm (Hµ) and 44-74 µm (SHµ) was added to the colloid, and the individually cross-linked hydrogel scaffolds with controllable pore size for chondrocyte culture were named Hµ-SL2, Hµ-SL4, SHµ-SL2 and SHµ-SL4. The perforation, porosity, mechanical strength, biocompatibility, and proliferation characteristics of the hydrogel scaffold and its influence on chondrocyte differentiation are discussed. Results show that the addition of porogen increases the porosity of the hydrogel scaffold. Conversely, when porogens with the same particle size are added, the pore size decreases as the amount of porogen increases. The perforation effect of the hydrogel scaffolds formed by the porogen is better at 88-177 µm compared with that at 44-74 µm. Cytotoxicity analysis showed that all the prepared hydrogel scaffolds were non-cytotoxic, indicating that no cross-linking agent residues that could cause cytotoxicity were found. In the proliferation and differentiation of the chondrocytes, the SHµ-SL4 hydrogel scaffold with the highest porosity and strength did not achieve the best performance. However, due to the compromise between perforation pores, pore sizes, and strength, as well as considering cell proliferation and differentiation, Hµ-SL4 scaffold provided a more suitable environment for the chondrocytes than other groups; therefore, it can provide the best chondrocyte growth environment for this study. The development of hydrogels with customized pore properties for defective cartilage is expected to meet the requirements of the ultimate clinical application.

Suramin attenuates intervertebral disc degeneration by inhibiting NF-κB signalling pathway.

AIMS: Interleukin (IL)-1ß is one of the major pathogenic regulators during the pathological development of intervertebral disc degeneration (IDD). However, effective treatment options for IDD are limited. Suramin is used to treat African sleeping sickness. This study aimed to investigate the pharmacological effects of suramin on mitigating IDD and to characterize the underlying mechanism. METHODS: Porcine nucleus pulposus (NP) cells were treated with vehicle, 10 ng/ml IL-1ß, 10 µM suramin, or 10 µM suramin plus IL-1ß. The expression levels of catabolic and anabolic proteins, proinflammatory cytokines, mitogen-activated protein kinase (MAPK), and nuclear factor (NF)-κB-related signalling molecules were assessed by Western blotting, quantitative real-time polymerase chain reaction (qRT-PCR), and immunofluorescence analysis. Flow cytometry was applied to detect apoptotic cells. The ex vivo effects of suramin were examined using IDD organ culture and differentiation was analyzed by Safranin O-Fast green and Alcian blue staining. RESULTS: Suramin inhibited IL-1ß-induced apoptosis, downregulated matrix metalloproteinase (MMP)-3, MMP-13, a disintegrin and metalloproteinase with thrombospondin motifs (ADAMTS)-4, and ADAMTS-5, and upregulated collagen 2A (Col2a1) and aggrecan in IL-1ß-treated NP cells. IL-1ß-induced inflammation, assessed by IL-1ß, IL-8, and tumour necrosis factor α (TNF-α) upregulation, was alleviated by suramin treatment. Suramin suppressed IL-1ß-mediated proteoglycan depletion and the induction of MMP-3, ADAMTS-4, and pro-inflammatory gene expression in ex vivo experiments. CONCLUSION: Suramin administration represents a novel and effectively therapeutic approach, which could potentially alleviate IDD by reducing extracellular matrix (ECM) deposition and inhibiting apoptosis and inflammatory responses in the NP cells. Cite this article: Bone Joint Res 2021;10(8):498-513.

Evaluation of the Grafting Efficacy of Active Biomolecules of Phosphatidylcholine and Type I Collagen on Polyether Ether Ketone: In Vitro and In Vivo.

Biomolecule grafting on polyether ether ketone (PEEK) was used to improve cell affinity caused by surface inertness. This study demonstrated the sequence-polished (P) and sulfonated (SA) PEEK modification to make a 3D structure, active biomolecule graftings through PEEK silylation (SA/SI) and then processed with phosphatidylcholine (with silylation of SA/SI/PC; without SA/PC) and type I collagen (COL I, with silylation of SA/SI/C; without SA/C). Different modified PEEKs were implanted for 4, 8, and 12 weeks for histology. Sulfonated PEEK of SA showed the surface roughness was significantly increased; after the silylation of SA/SI, the hydrophilic nature was remarkably improved. The biomolecules were effectively grafted through silylation, and the cells showed improved attachment after 1 h. Furthermore, the SA/SI/PC group showed good in vitro mineralization. The new bone tissues were integrated into the 3D porous structures of SA/SI/PC and SA/SI/C in vivo making PEEK a potential alternative to metals in orthopedic implants.

Strength and Biocompatibility of Heparin-Based Calcium Phosphate Cement Grafted with Ferulic Acid.

The biomimetic synthesis of carbonated apatites by biomolecule-based templates is a promising way for broadening apatite applications in bone tissue regeneration. In this work, heparin was used as an organic template to prepare uniform carbonate-based apatite nanorods (CHA) and graft ferulic acid (F-CHA) for enhanced bone mineralization. Next, by combining calcium phosphate cement (CPC) with different F-CHA/CPC ratios, a new type of injectable bone cement combined with F-CHA bioactive apatite was developed (CPC + F-CHA). The physicochemical properties, biocompatibility, and mineralization potential of the CPC + F-CHA composites were determined in vitro. The experimental results confirmed the preparation of highly biocompatible CHA and the compatibility of F-CHA with CPC. Although CPC + F-CHA composites with F-CHA (2.5 wt%, 5 wt%, and 10 wt%) showed a significant reduction in compressive strength (CS), compositing CPC with 10 wt% F-CHA yielded a CS suitable for orthopedic repair (CS still larger than 30 MPa). Spectroscopic and phase analyses revealed that the phase of the hydrothermally synthesized CHA product was not modified by the heparin template. Injection and disintegration tests indicated that the CPC + F-CHA composites have good biocompatibility even at 10 wt% F-CHA. D1 osteoprogenitor cells were cultured with the composites for 7 days in vitro, and the CPC + 10%F-CHA group demonstrated significantly promoted cell mineralization compared with other groups. Given these results, the use of over 10% F-CHA in CPC composites should be avoided if the latter is to be applied to load-bearing areas. A stress-shielding device may also be recommended to stabilize these areas. These newly developed biocompatible CPC + F-CHA have great potential as osteoconductive bone fillers for bone tissue engineering.

Chemical cross-linking on gelatin-hyaluronan loaded with hinokitiol for the preparation of guided tissue regeneration hydrogel membranes with antibacterial and biocompatible properties.

The mechanical properties and structural stability of hydrogels and their performance in antidegradation can be enhanced by cross-linking them with N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide hydrochloride (EDC). However, residual EDC compromises the biocompatibility of cross-linked hydrogels and the formability of un-cross-linked hydrogels. In this study, a facile process for preparing hydrogel regenerative membranes exerting antibacterial effects and containing gelatin/hyaluronic acid (G/HA) through solution casting was proposed. The membranes were cross-linked with EDC (G/HA-Ec-0H) and impregnated with two concentrations of the antibacterial agent of hinokitiol (G/HA-Ec-2H and G/HA-Ec-4H). Amide bonds formed, and the rate of active amino acid fixation was higher than 90%, which was directly proportional to the degree of cross-linking. The G/HA-Ec-2H and G/HA-Ec-4H groups with hinokitiol showed good antibacterial properties. The rate of hydrogel degradation decreased, and the integrity of sample morphology was maintained at more than 80% for over 3 days in the immersion. Then, the hydrogel structures relaxed and disintegrated through a rapid degradation reaction within 24 h. The biocompatibility results showed that low concentrations of hinokitiol did not affect cell viability. Moreover, hydrogel membranes after 14 days of cell incubation showed good cell adhesion and proliferation. In summary, the membrane biostability of the cross-linked gelatin/hyaluronan hydrogels was enhanced by EDC at a biocompatible concentration, and the functionalized group of G/HA-Ec-2H shows potential as a biodegradable material for biocompatible tissue-guarded regeneration membranes with antibacterial properties.

Impact of type 2 diabetes on postoperative outcome after hip fracture: nationwide population-based study in Taiwan.

The impact of diabetes mellitus (DM) on hip fracture (HFx) is still controversial. We used nationwide population-based data in Taiwan to observe postoperative outcomes of HFx in patients with type 2 diabetes mellitus (T2DM) and found that the impact of T2DM may be related to medication of blood glucose control. OBJECTIVE: Published studies evaluating diabetic patients with HFx have shown controversial outcomes. We assessed the impact of T2DM on postoperative outcomes after HFx in elderly patients using the nationwide population database in Taiwan. RESEARCH DESIGN AND METHODS: We used data from the National Health Research Institute in Taiwan to recruit patients who had undergone operations for HFx between 2000 and 2009. The recruited patients with T2DM were divided into the oral antidiabetic drug (OAD) cohort and the insulin cohort according to the use or non-use of insulin. Patients without DM were propensity score matched in a 1:1 ratio by four variables. We used the χ2 test, linear regression and Cox proportional hazards model to assess variables, including length of hospital stay, medical cost, complications, early readmission, and 1-year mortality. RESULTS: We identified 5490 subjects in total. The insulin cohort exhibited prolonged hospital stay (11.8 days), higher medical costs, more complications within 30 and 90 after hip surgery, earlier readmission, and higher 1 year mortality rate (25.8%) than the OAD and non-DM cohorts. The OAD cohort had longer hospital stay (10.1 days) and higher readmission rate but fewer complications and mortality rates (14.9%) than the non-DM cohort. CONCLUSIONS: After matching confounding factors, the T2DM with OAD control groups were not associated with higher complication or mortality rates but were associated with higher readmission rates. However, diabetic patients with insulin control have poor outcome. The impact of T2DM on the postoperative outcomes of patients with HFx may be related to blood glucose control medication.

(-)-Epigallocatechin-3-gallate (EGCG) enhances healing of femoral bone defect.

BACKGROUND: Previously, we found that (-)-epigallocatechin-3-gallate (EGCG) enhanced osteogenic differentiation of murine bone marrow mesenchymal stem cells by increasing the mRNA expression of osteogenesis-related genes, alkaline phosphatase activity and eventually mineralization. We further found EGCG supplementation preserved bone mass and microarchitecture in female rats during estrogen deficiency in the proximal tibia and lumbar spine at least in part by increasing bone morphogenetic protein-2 (BMP2). BMP2 can enhance de novo bone formation. PURPOSE: In this study, we evaluate the effect of local EGCG application in de novo bone formation in bone defect healing. METHODS: Twenty-four rats aged 4 months were weight-matched and randomly allocated to 2 groups: defect control with vehicle treatment (control) and defect with 10 µM EGCG treatment (EGCG). Daily vehicle and EGCG were applied locally by percutaneous local injection 2 days after defect creation for 2 weeks. Four weeks after treatment, animals were sacrificed for micro-computed tomography (µ-CT) and biomechanical analysis. RESULTS: Local EGCG at femoral defect can enhance de novo bone formation by increasing bone volume and subsequently improve mechanical properties including max load, break point, stiffness, area under the max load curve, area under the break point curve and ultimate stress. CONCLUSIONS: Local EGCG may enhance bone defect healing via at least partly by the de novo bone formation of BMP-2.

Modulating the release of proteins from a loaded carrier of alginate/gelatin porous spheres immersed in different solutions.

BACKGROUND: A biodegradable porous particle for the controlled biofactor delivery which assembly of pores in scaffolds can improve the permeation and diffusion of drugs or growth factors. OBJECTIVE: Porous-spheres in millimeter scale were prepared by mixing sodium alginate and gelatin interpenetrating networks with cross-linkers; interconnected open pores were fabricated through solvent casting and particulate leaching. METHODS: Morphological characteristics, degradation, and bovine serum albumin (BSA) release rates of the porous-spheres immersed in three different solutions, namely, deionized distilled water, simulated body fluid (SBF), and phosphate-buffered saline (PBS), were detected. RESULTS: Porous-spheres with a large amount of gelatin exhibited an increase in water absorption rates without affecting scaffold strength and no cytotoxicity was elicited. Highly interconnected pores with a diameter of 100-200 µm were uniformly distributed in scaffolds. The weight loss in PBS was faster than that in other solutions; the highest release rate of BSA in SBF was observed for 2 h. The release rates also exhibited linear patterns from 2 h to 24 h in all of the groups. CONCLUSIONS: After 1 d of immersion in solutions, BSA release rates in scaffolds logarithmically decreased for 14 d. The degradation of porous-spheres also showed an inverse pattern.

An Innovative Anchoring Technique for Anterior Transfer of the Tibialis Posterior Tendon.

Favorable results have been reported for tibialis posterior tendon transfers, which can effectively restore the dorsiflexion of the ankle and normal heel-to-toe gait. However, the commonly used methods for anchoring the transplanted tendon have some drawbacks. Therefore, we developed a new tendon-anchoring method to improve fixation of the transferred tendon and reduce the related complications. The new method entails tying the anchoring suture to the navicular bone instead of the button on the plantar foot to avoid wound complications. It requires no additional skin incisions or special equipment. We retrospectively evaluated 24 feet of 19 pediatric patients (13 [68.4%] females and 6 [31.6%] males) who had undergone anterior transfer of the tibialis posterior tendon with our new method from 2000 to 2013. All patients were clinically followed up. At the final follow-up visit, they were evaluated while standing and walking, and the range of motion of the foot was evaluated. The mean age at surgery was 7.8 (range 2 to 16) years. At the longest follow-up point, all the patients exhibited improved gait, except for 1 patient who required a secondary procedure. All the transferred tibialis posterior tendons could be palpated with certainty during active dorsiflexion or withdrawal of the foot. No tendon displacements, wound infections, or postoperative complications were observed. Fixation of a transferred tibialis posterior tendon by tying the suture to the navicular bone is simple and reliable. This technique can efficiently prevent the plantar ulcers that can develop with the traditional pull-out button method and provides a solution when appropriate-size bioabsorbable interference screws are unavailable.

Osteoregenerative capacities of dicalcium phosphate-rich calcium phosphate bone cement.

Calcium phosphate cement (CPC) is a widely used bone substitute. However, CPC application is limited by poor bioresorption, which is attributed to apatite, the stable product. This study aims to systematically survey the biological performance of dicalcium phosphate (DCP)-rich CPC. DCP-rich CPC exhibited a twofold, surface-modified DCP anhydrous (DCPA)-to-tetracalcium phosphate (TTCP) molar ratio, whereas conventional CPC (c-CPC) showed a onefold, surface unmodified DCPA-to-TTCP molar ratio. Cell adhesion, morphology, viability, and alkaline phosphatase (ALP) activity in the two CPCs were examined with bone cell progenitor D1 cultured in vitro. Microcomputed tomography and histological observation were conducted after CPC implantation in vivo to analyze the residual implant ratio and new bone formation rate. D1 cells cultured on DCP-rich CPC surfaces exhibited higher cell viability, ALP activity, and ALP quantity than c-CPC. Histological evaluation indicated that DCP-rich CPC showed lesser residual implant and higher new bone formation rate than c-CPC. Therefore, DCP-rich CPC can improve bioresorption. The newly developed DCP-rich CPC exhibited potential therapeutic applications for bone reconstruction.

Biphasic products of dicalcium phosphate-rich cement with injectability and nondispersibility.

In this study, a calcium phosphate cement was developed using tetracalcium phosphate and surface-modified dicalcium phosphate anhydrous (DCPA). This developed injectable bone graft substitute can be molded to the shape of the bone cavity and set in situ through the piping system that has an adequate mechanical strength, non-dispersibility, and biocompatibility. The materials were based on the modified DCPA compositions of calcium phosphate cement (CPC), where the phase ratio of the surface-modified DCPA is higher than that of the conventional CPC for forming dicalcium phosphate (DCP)-rich cement. The composition and morphology of several calcium phosphate cement specimens during setting were analyzed via X-ray diffractometry and transmission electron microscopy coupled with an energy dispersive spectroscopy system. The compressive strength of DCP-rich CPCs was greater than 30MPa after 24h of immersion in vitro. The reaction of the CPCs produced steady final biphasic products of DCPs with apatite. The composites of calcium phosphate cements derived from tetracalcium phosphate mixed with surface-modified DCPA exhibited excellent mechanical properties, injectability, and interlocking forces between particles, and they also featured nondispersive behavior when immersed in a physiological solution.

Bicondylar tibial plateau fracture treated by open reduction and fixation with unilateral locked plating.

The management of bicondylar tibial plateau fractures is challenging. A lateral locking plate offers an alternative method to traditional dual plating to avoid further stripping of soft tissue. Nevertheless, the rate of malreduction and fixation loss remains high. From 2007 to 2009, we performed open reduction and fixation with unilateral locked plating to directly reduce the fracture in 15 patients with bicondylar plateau fracture. The average follow-up duration was 16.2 months (range: 12-30 months), and the average age of the patients was 43 years (range: 19-64 years). All fractures were Orthopaedic Trauma Association type 41-C. Postoperative radiographic alignment was evaluated immediately and at 2-4 weeks, 8-12 weeks, 5-7 months, and 11-13 months. Both Oxford knee score and Hospital for Special Surgery knee score were used to evaluate functional outcomes. The average duration within which union was achieved was 4.8 months (range: 2-10 months). One patient incurred wound dehiscence; however, there was no case of deep infection. Malreduction occurred in one patient (6.7%) while fixation loss occurred in three patients (20%) with subsidence of the posteromedial fragment and varus malalignment. Despite the malreduction rate being lower in our study than in previous studies involving unilateral locked plating, a high rate of fixation loss was recorded. Per our limited experience, we believe that unilateral locked plating may have limitations in patients with selective patterns of bicondylar tibial plateau fractures.