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The process of new bone formation following trauma requires the temporal recruitment of cells to the site, including mesenchymal stem cells, preosteoblasts, and osteoblasts, the latter of which deposit minerals. Hence, bone repair, a process that is assessed by the extent of mineralization within the defect, can take months before it is possible to determine if a treatment is successful. Here, a fluorescently tagged Osterix, an early key gene in the bone formation cascade, is used as a predictive measure of bone formation. Using a calvarial defect model in mice, the ability to noninvasively track the Osterix transcription factor in an Osterix-mCherry mouse model is evaluated as a measure for bone formation following treatment with recombinant human Bone-Morphogenetic-Protein 2 (rhBMP-2). Two distinct delivery materials are utilized, an injectable nanocomposite hydrogel and a collagen sponge, that afford distinct release kinetics and it is found that cherry-fluorescent protein can be detected as early as 2 weeks following treatment. Osterix intensity correlates with subsequent bone formation and hence can serve as a rapid screening tool for osteogenic drugs or for the evaluation and optimization of delivery platforms., (© 2019 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.)

Hydrogels are an attractive class of biomaterials for minimally invasive local drug delivery given their injectability, tunability, high water content, and biocompatibility. Broad applicability though is challenged: relatively modest mechanical properties restrict use to soft tissues, while flow properties necessary for injectability limit implantation to dried, enclosed tissues to minimize material migration during gelation. To address these dual concerns, we designed an injectable nanocomposite hydrogel based on dextran aldehyde and a poly(amido amine) dendrimer doped with phyllosilicate nanoplatelet fillers. Balance of components allows for exfoliation of nanoplatelets, significantly changing macromer solution flow, facilitating injection and manipulation in a wide variety of implantation contexts while enhancing compressive modulus of hydrogels at low loading. Importantly, rheological and mechanical effects were dependent on aspect ratio, with high aspect ratio nanoplatelets having much stronger effects on mechanics and low aspect ratio nanoplatelets having stronger effects on rheology, enabling nearly independent control of rheological and mechanical properties. Nanoplatelets enhanced hydrogel properties at a filler loading substantially lower than that of comparably sized nanoparticles. We present a model to explain the role that aspect ratio plays in control of rheology and mechanics in nanoplatelet-containing hydrogels, with lessons for further nanocomposite hydrogel development. This low-cost biocompatible material may be useful as a drug delivery platform in challenging implantation environments.

Background: ALPPS (Associating Liver Partition and Portal vein ligation for Staged hepatectomy) is a new surgical approach for the treatment of liver tumors. It is indicated in cases where the future liver remnant is not sufficient to maintain postoperative liver function. We report a totally laparoscopic ALPPS with selective hepatic artery clamping. Pneumoperitoneum itself results in up to 53% of portal vein flow and selective hepatic artery clamping can reduce blood loss while maintaining hepatocellular function. Therefore, the combination of both techniques may result in effective control of bleeding with no damage in the liver function that may have direct impact in the result of ALPPS procedure., Methods: A 65-year-old man with colorectal liver metastases in all liver segments, except liver segment 1 (S1), were evaluated as unresectable. He underwent chemotherapy with objective response and multidisciplinary board decided for ALPPS procedure. First stage was performed entirely by laparoscopy and consisted of enucleation of metastases from segments 2 and 3, ligation of the right portal vein and liver splitting under selective common hepatic artery clamping. The second stage was done 3 weeks later and consisted of laparoscopic right trisectionectomy by laparoscopy., Results: Operative time was 250 and 200 minutes, respectively. Estimated blood loss was 150 and 100 mL. There was no need for transfusion or hospitalization in intensive care. He was discharged on the 3rd and 5th postoperative day, respectively. Recovery was uneventful after both stages and patient did not present any sign of liver failure. Elevation of liver enzymes was minimal. Computerized tomography (CT) scan before second stage showed a liver hypertrophy of 53%, sFLR was 0.37 before second stage, or 33% of the total liver volume. CT scan shows no residual liver disease and optimum liver regeneration. Patient is well with no evidence of the disease 11 months after the procedure., Conclusions: Totally laparoscopic ALPPS is a feasible and safe approach for selected patients with liver tumors. The hypertrophy of the remaining liver was adequate and sequential procedures were performed without morbidity and no mortality. Selective hepatic artery clamping seems to be an interesting solution to decrease intraoperative blood loss without the harsh effect of Pringle maneuver., Competing Interests: The authors have no conflicts of interest to disclose.

There is an age-associated reduction in the bone healing activity of bone morphogenetic protein-2 (BMP-2) that is currently addressed by administering higher doses of BMP-2 in elderly patients. The unwanted medical complications from high dose BMP-2 motivated this investigation to determine whether the addition of a low dose of fibroblast growth factor 2 (FGF-2) could enhance the ability of a lower dose of BMP-2 to heal calvarial bone defects in old mice (18-20 months old). FGF-2 (5 ng) and BMP-2 (2 μg) were administered by a controlled release two-phase biomaterial scaffold placed into the bone defect. FGF-2 released more rapidly and completely in vitro than BMP-2 (40% vs 2%). In vivo, both BMP-2 and FGF-2+BMP-2 groups formed more new bone in calvarial defects than scaffold alone (p < 0.001) or FGF-2 only groups (p < 0.01). The overall total volume of new bone was not statistically increased by the addition of FGF-2 to BMP-2 as measured by microCT, but the pattern of bone deposition was different. In old mice, but not young, there was enhanced bony fill in the central bone defect area when the BMP-2 was supplemented with FGF-2. Histological analysis of the center of the defect revealed an increased bone volume (%BV/TV (p = 0.004)) from the addition of FGF-2. These studies suggest that combining a low dose of FGF-2 with a low dose of BMP-2 has the potential to increase bone healing in old mice relative to BMP-2 alone., (Copyright © 2015 Elsevier Inc. All rights reserved.)

The in vivo osteogenesis potential of mesenchymal-like cells derived from human embryonic stem cells (hESC-MCs) was evaluated in vivo by implantation on collagen/hydroxyapatite scaffolds into calvarial defects in immunodeficient mice. This study is novel because no osteogenic or chondrogenic differentiation protocols were applied to the cells prior to implantation. After 6 weeks, X-ray, microCT, and histological analysis showed that the hESC-MCs had consistently formed a highly vascularized new bone that bridged the bone defect and seamlessly integrated with host bone. The implanted hESC-MCs differentiated in situ to functional hypertrophic chondrocytes, osteoblasts, and osteocytes forming new bone tissue via an endochondral ossification pathway. Evidence for the direct participation of the human cells in bone morphogenesis was verified by two separate assays: with Alu and by human mitochondrial antigen positive staining in conjunction with co-localized expression of human bone sialoprotein in histologically verified regions of new bone. The large volume of new bone in a calvarial defect and the direct participation of the hESC-MCs far exceeds that of previous studies and that of the control adult hMSCs. This study represents a key step forward for bone tissue engineering because of the large volume, vascularity, and reproducibility of new bone formation and the discovery that it is advantageous to not over-commit these progenitor cells to a particular lineage prior to implantation. The hESC-MCs were able to recapitulate the mesenchymal developmental pathway and were able to repair the bone defect semi-autonomously without preimplantation differentiation to osteo- or chondroprogenitors.

Self-reinforced composites (SRCs) are materials where both the matrix and fiber-reinforcing phase are made up of the same polymer. Improved bonding can be achieved with self-reinforced composites compared to traditional dual-polymer, fiber-reinforced composites owing to the identical chemistry of the components in SRCs. Bonding between the fiber and matrix phase is an important factor in applications where mechanical stability is required, such as in the field of bone repair. In this study, we prepared bioabsorbable poly(L-lactic acid)/hydroxyapatite (PLLA/HA) self-reinforced composites via a three-step process that includes surface etching of the fiber, the deposition of the HA coating onto the PLLA fibers through immersion in simulated body fluid (SBF), and hot compaction molding. Although coated with a layer of HA, self-reinforced composites were successfully generated by hot compaction. The effects of compaction time (15 and 30 min), compaction temperature (140, 150, 155, 160, 165, and 170 °C), and HA wt% (0, 5, 10, and 15 wt%) on flexural mechanical properties were studied. Mechanical test results indicated that in unfilled (no HA) PLLA SRCs, compaction time and temperature increased the flexural modulus of the composites tested. Based on the results obtained for unfilled composites, a single compaction time and temperature condition of 15 min and 170 °C were selected to study the effect of HA loading on the composite mechanical properties. HA was successfully loaded onto the fibers at 0, 5, 10, and 15 wt% before hot compaction and was found to significantly increase flexural modulus (P=0.0001). Modulus values ranged from 8.3 GPa±0.5 (0 wt% HA) to 9.7 GPa±0.6 (15 wt% HA). Microscopy results suggest that the HA in these composites forms a nodular-like structure along the fibers, which allows polymer-polymer contact yet prevents longitudinal shear. The procedure used successfully generated composites with flexural moduli near the lower range of bone that may have a possible clinical use for load-bearing bone-fixation devices., (Copyright © 2012 Elsevier Ltd. All rights reserved.)

These studies provide evidence for the ability of a commercially available, defined, hyaluronan-gelatin hydrogel, HyStem-C™, to maintain both mouse embryonic stem cells (mESCs) and human induced pluripotent stem cells (hiPSCs) in culture while retaining their growth and pluripotent characteristics. Growth curve and doubling time analysis show that mESCs and hiPSCs grow at similar rates on HyStem-C™ hydrogels and mouse embryonic fibroblasts and Matrigel™, respectively. Immunocytochemistry, flow cytometry, gene expression and karyotyping reveal that both human and murine pluripotent cells retain a high level of pluripotency on the hydrogels after multiple passages. The addition of fibronectin to HyStem-C™ enabled the attachment of hiPSCs in a xeno-free, fully defined medium., (Copyright © 2012 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.)

Currently, the bone-repair biomaterials market is dominated by high modulus metals and their alloys. The problem of stress-shielding, which results from elastic modulus mismatch between these metallic materials and natural bone, has stimulated increasing research into the development of polymer-ceramic composite materials that can more closely match the modulus of bone. In this study, we prepared poly(L: -lactic acid)/hydroxyapatite/poly(epsilon-caprolactone) (PLLA/HA/PCL) composites via a four-step process, which includes surface etching of the fiber, the deposition of the HA coating onto the PLLA fibers through immersion in simulated body fluid (SBF), PCL coating through a dip-coating process, and hot compression molding. The initial HA-coated PLLA fiber had a homogeneous and continuous coating with a gradient structure. The effects of HA: PCL ratio and molding temperature on flexural mechanical properties were studied and both were shown to be important to mechanical properties. Mechanical results showed that at low molding temperatures and up to an HA: PCL volume ratio of 1, the flexural strain decreased while the flexural modulus and strength increased. At higher mold temperatures with a lower viscosity of the PCL a HA: PCL ratio of 1.6 gave similar properties. The process successfully produced composites with flexural moduli near the lower range of bone. Such composites may have clinical use for load bearing bone fixation.

We have designed monolayers with weak intermolecular interactions for use as placeholders in intelligent self- and directed-assembly. We have shown that these 1-adamantanethiolate monolayers are labile with respect to displacement by exposing them to dilute solutions of alkanethiols. These self-assembled monolayers (SAMs) of 1-adamantanethiol on Au{111} were probed using ambient scanning tunneling microscopy (STM), and their assembled order was determined. Solution deposition of the molecules results in a highly ordered hexagonally close-packed molecular lattice with a measured nearest neighbor distance of 6.9 +/- 0.4 A. The SAMs exhibit several rotational domains, but lack the protruding domain boundaries typical of alkanethiolate SAMs, and are similarly stable at room temperature. Co-deposition of alkanethiol and 1-adamantanethiol from solution results in alkanethiolate SAMs, except when using extremely low alkanethiol to 1-adamantanethiol concentration ratios. Facile displacement of low interaction strength SAMs can be exploited to enhance patterning using soft nanolithography.