Co-Culture of Mesenchymal Stem Cells and Ligamentocytes on Triphasic Embroidered Poly(L-lactide-co-ε-caprolactone) and Polylactic Acid Scaffolds for Anterior Cruciate Ligament Enthesis Tissue Engineering.

Successful anterior cruciate ligament (ACL) reconstructions strive for a firm bone-ligament integration. With the aim to establish an enthesis-like construct, embroidered functionalized scaffolds were colonized with spheroids of osteogenically differentiated human mesenchymal stem cells (hMSCs) and lapine (l) ACL fibroblasts in this study. These triphasic poly(L-lactide-co-ε-caprolactone) and polylactic acid (P(LA-CL)/PLA) scaffolds with a bone-, a fibrocartilage transition- and a ligament zone were colonized with spheroids directly after assembly (DC) or with 14-day pre-cultured lACL fibroblast and 14-day osteogenically differentiated hMSCs spheroids (=longer pre-cultivation, LC). The scaffolds with co-cultures were cultured for 14 days. Cell vitality, DNA and sulfated glycosaminoglycan (sGAG) contents were determined. The relative gene expressions of collagen types I and X, Mohawk, Tenascin C and runt-related protein (RUNX) 2 were analyzed. Compared to the lACL spheroids, those with hMSCs adhered more rapidly. Vimentin and collagen type I immunoreactivity were mainly detected in the hMSCs colonizing the bone zone. The DNA content was higher in the DC than in LC whereas the sGAG content was higher in LC. The gene expression of ECM components and transcription factors depended on cell type and pre-culturing condition. Zonal colonization of triphasic scaffolds using spheroids is possible, offering a novel approach for enthesis tissue engineering.

Venous injection of a triphasic calcium-based implant in a sheep model of pulmonary embolism demonstrates minimal acute systemic effects.

PURPOSE: Implant leakage is the most common complication of vertebral augmentation. Alternative injectable materials must demonstrate intravascular safety comparable to or better than polymethyl methacrylate (PMMA). This study assessed the systemic effects of a triphasic calcium-based implant or PMMA injected directly into the femoral vein in a large animal model designed to mimic severe intravascular implant leakage. METHODS: Six skeletally mature female sheep were randomly assigned (n = 3) to either the PMMA or the triphasic implant (AGN1, composition: calcium sulfate, ß-tricalcium phosphate, brushite) treatment group. Femoral veins of each sheep were directly injected with 0.5 mL of implant material to mimic leakage volumes reported during PMMA vertebroplasty. To compare acute systemic effects of the materials, cardiovascular parameters, laboratory coagulation markers, and calcium and sulfate serum levels were monitored for 60 min after implant injection. Thrombotic and embolic events were evaluated by radiologic imaging, necropsy, and histopathology. RESULTS: Heart rate, systemic arterial blood pressure, arterial oxygenation, arterial carbon dioxide content, and coagulation markers remained within physiological range after either AGN1 or PMMA injection. No blood flow interruption in the larger pulmonary vessels was observed in either group. Lung histopathology revealed that the severity of thrombotic changes after AGN1 injection was minimal to slight, while changes after PMMA injection were minimal to massive. CONCLUSION: Acute systemic effects of intravascular AGN1 appeared to be comparable to or less than that of intravascular PMMA. Furthermore, in this preliminary study, the severity and incidence of pulmonary histological changes were lower for AGN1 compared to PMMA.

Local osteo-enhancement of osteoporotic vertebra with a triphasic bone implant material increases strength-a biomechanical study.

PURPOSE: The aim of this study was to assess the biomechanical properties of intact vertebra augmented using a local osteo-enhancement procedure to inject a triphasic calcium sulfate/calcium phosphate implant material. METHODS: Twenty-one fresh frozen human cadaver vertebra (Th11-L2) were randomized into three groups: treatment, sham, and control (n = 7 each). Treatment included vertebral body access, saline lavage to displace soft tissue and marrow elements, and injection of the implant material to fill approximately 20% of the vertebral body by volume. The sham group included all treatment steps, but without injection of the implant material. The control group consisted of untreated intact osteoporotic vertebra. Load at failure and displacement at failure for each of the three groups were measured in axial compression loading. RESULTS: The mean failure load of treated vertebra (4118 N) was significantly higher than either control (2841 N) or sham (2186 N) vertebra (p < 0.05 for: treatment vs. control, treatment vs. sham). Treated vertebra (1.11 mm) showed a significantly higher mean displacement at failure than sham vertebra (0.80 mm) (p < 0.05 for: treatment vs. sham). In the control group, the mean displacement at failure was 0.99 mm. CONCLUSIONS: This biomechanical study shows that a local osteo-enhancement procedure using a triphasic implant material significantly increases the load at failure and displacement at failure in cadaveric osteoporotic vertebra.

Engineering Triphasic Nanocomposite Coatings on Pretreated Mg Substrates for Biomedical Applications.

Biodegradable polymer-based nanocomposite coatings provide multiple advantages to modulate the corrosion resistance and cytocompatibility of magnesium (Mg) alloys for biomedical applications. Biodegradable poly(glycerol sebacate) (PGS) is a promising candidate used for medical implant applications. In this study, we synthesized a new PGS nanocomposite system consisting of hydroxyapatite (HA) and magnesium oxide (MgO) nanoparticles and developed a spray coating process to produce the PGS nanocomposite layer on pretreated Mg substrates, which improved the coating adhesion at the interface and their cytocompatibility with bone marrow derived mesenchymal stem cells (BMSCs). Prior to the spray coating process of polymer-based nanocomposites, the Mg substrates were pretreated in alkaline solutions to enhance the interfacial adhesion strength of the polymer-based nanocomposite coatings. The addition of HA and MgO nanoparticles (nHA and nMgO) to the PGS matrix, as well as the alkaline pretreatment of the Mg substrates, significantly enhanced the interfacial adhesion strength when compared with the PGS coating on the nontreated Mg control. The average BMSC adhesion densities were higher on the PGS/nHA/nMgO coated Mg than the noncoated Mg controls under direct contact conditions. Moreover, the addition of nHA and nMgO to the PGS matrix and coating the nanocomposite onto Mg substrates increased the average BMSC adhesion density when compared with the PGS/nHA/nMgO coated titanium (Ti) and PGS coated Mg controls under direct contact. Therefore, the spray coating process of PGS/nHA/nMgO nanocomposites on Mg substrates or other biodegradable metal substrates could provide a promising surface treatment strategy for biodegradable implant applications.

Polymers with intrinsic microporosity (PIMs) for targeted CO2 reduction to ethylene.

CO2 reduction offers an attractive alternative green synthetic route for ethylene, especially where CO2 could be sourced from industrial exhausts and in combination with green power sources. However, practical applications are currently limited due to the unfortunately low selectivity of cathode materials towards ethylene. This work uses polymers with intrinsic microporosity (PIMs) to improve the performance of copper gas diffusion electrodes for CO2 reduction to ethylene. We report an improved selectivity and activity towards ethylene with the addition of a thin PIMs layer, which is seen as improved Faradaic efficiency, increased stability and a shift in the reduction to lower overpotential. This improvement is highly dependent on the thickness of the added polymer layer, with too thick a layer having a detrimental impact on the hydrophobicity of the gas diffusion layer. With a compromise in loading, PIMs can be used to enhance the activity and selectivity of catalysts for targeted CO2 reduction to ethylene.

Composite Alloplastic Biomaterial vs. Autologous Platelet-Rich Fibrin in Ridge Preservation.

AIM: The aim of this study was to examine the clinical and histological differences of using a combination of alloplastic beta triphasic calcium phosphate (ß-TCP) and a cross-linked collagen membrane versus autologous platelet-rich fibrin (PRF-L) in ridge preservation after dental extraction. MATERIAL AND METHODS: Fifty-one patients were included in this observational case-series study. Dental extractions were performed, after which 25 patients were grafted with ß-TCP and 26 with PRF-L. After four months of healing, clinical, radiological, histomorphometric and histological evaluations were performed. RESULTS: A significantly higher percentage of mineralized tissue was observed in samples from the PRF-L grafted areas. Cellularity was higher in PRF-L grafted areas (osteocytes in newly formed bone per mm² = 123.25 (5.12) vs. 84.02 (26.53) for PRF-L and ß-TCP, respectively, p = 0.01). However, sockets grafted with PRF-L showed a higher reduction in the bucco-lingual dimension after four months of healing (2.19 (0.80) vs. 1.16 (0.55) mm, p < 0.001), as well as a higher alteration in the final position of the mid muco-gingival junction (1.73 (1.34) vs. 0.88 (0.88) mm, p < 0.01). CONCLUSION: PRF-L concentrate accelerates wound healing in post-extraction sockets in terms of new mineralized tissue component. However, the use of ß-TCP biomaterial appears to be superior to maintain bucco-lingual volume and the final position of the muco-gingival junction.

Immobilisation of solvatochromic dyes on transparent cellulose films; an improved methodology for the study of homeopathic potencies

Highly diluted and succussed solutions (homeopathic potencies) have been shown to interact with a wide range of solvatochromic dyes based on changes in their UV-visible spectra. Studies so far have involved free dyes in solution, but there is a pressing need to find ways to investigate the potency-dye interaction using isolated dye molecules in order to ask more searching physico-chemical questions regarding the fundamental nature of potencies. Aims and Methods:The aims of the present study have been tolook for ways to covalently immobilizesolvatochromic dyes onto transparent cellulose films and hence be in a position to investigate dye-potency interactions without the complication of dye-dye interactions, including dye aggregation, which can occur with free dyes in solution. Results: to date a total of nine different dyes have been immobilised on cellulose films using epoxide activation of hydroxyl groups on the cellulose surface. Using this methodology studies have begun looking at the time course of potency action on one of these immobilised dyes, Brooker's merocyanine. Results show that the interaction ofArsenicum10M with this dye consists of three phases ­an initial growth phase, a sustained plateau of interaction and a final decline phase lasting several days.Conclusions: amethodology has been developed that successfully immobilises solvatochromic dyes onto transparent cellulose film. These films can then be used in a spectrophotometer to study at a much more detailed level how potencies interact with dyes compared with using free dyes in solution. Results indicate that the information gained in this way provides new insights regarding the fundamental nature of potencies. Specifically, studies using immobilised Brooker's merocyanine with Arsenicum10M reveal that the lifetime of the potency is much longer than expected and that its action consists of three distinct phases, suggesting a resonant interaction with the dye.

Pulsed laser deposition and in vitro characteristics of triphasic - HASi composition on titanium.

Pulsed laser deposition was used to deposit bioactive triphasic glass-ceramic composition (HASi) over titanium substrate using dense HASi target. Bioactive glass compositions are considered the most useful synthetic materials for immediate bone attachment because of its bioresorption, osteoconduction and osteointegration characteristics under in vivo conditions. The disadvantage of its brittleness associated with bioactive glass-ceramics has prompted its coating over metallic implants for the combination of duo mechanical and bioactive properties. The hard HASi target was able to undergo laser ablation under ambient gas pressure without bulk erosion of the target. Laser deposition was found to be efficient in depositing triphasic composition for immediate bone integration. The target and deposits were analyzed for the phase, composition and microstructural characteristics by means of X-ray diffraction, Fourier transform infrared spectroscopy, energy-dispersive X-ray analysis and scanning electron microscopy. Simultaneously, the adherent nature and mechanical behaviour of deposits were confirmed by scratch test and micro-indentation methods. Further, the in vitro dissolution and bioactivity were assessed by soaking in simulated body fluid followed by elemental analysis using inductively coupled plasma spectroscopy. The deposits were found to be cell-friendly, which was indicated by the phenomenology of stem cells under in vitro conditions.

Evaluation of oriented electrospun fibers for periosteal flap regeneration in biomimetic triphasic osteochondral implant.

Osteochondral defects represent a serious clinical problem. Although the cell-scaffold complexes have been reported to be effective for repairing osteochondral defects, a periosteal flap is frequently needed to arrest leakage of the implanted cells into the defect and to contribute to the secretion of cytokines to stimulate cartilage repair. The electrospun mesh mimicking the function of the flap assists tissue regeneration by preventing cell leakage and merits favorable outcomes in the cartilaginous region. In this study, an oriented poly(ε-caprolactone) (PCL) fibrous membrane (OEM) was fabricated by electrospinning as a periosteal scaffold and then freeze-dried with a collagen type I and hyaluronic acid cartilage scaffold (CH) and finally, freeze-dried with a tricalcium phosphate (TCP) bone substratum. Scanning electron microscopic images show obvious microstructure formation of the trilayered scaffolds, and electrospun fibrous membranes have an oriented fibrous network structure for the periosteal phase. Also shown are opened and interconnected pores with well designed three-dimensional structure, able to be bound in the CH (chondral phase) and TCP (osseous phase) scaffolds. In vitro results showed that the OEM can promote the orientation of bone marrow mesenchymal stem cell (BMSCs) and BMSCs can penetrate into the CH and TCP. After successfully combining the BMSCs, the tissue-engineered cartilage which contained the OEM and TCP complex was successfully used to regenerate the osteochondral defects in the rabbit model with greatly improved repair effects.