Indirect Tissue Scaffold Fabrication via Fused Deposition Modeling and Biomimetic Mineralization

To alleviate material limitations of the additive manufacture of tissue scaffolds, researchers have looked to indirect fabrication approaches. The feature resolution of these processes is limited however, due to the viscous ceramic slurries that are typically employed. To alleviate these limitations, the authors look to an indirect fabrication process wherein a pattern, created using Fused Deposition Modeling, is biomimetically mineralized with an aqueous simulated body fluid, which forms a bonelike hydroxyapatite throughout the scaffold pattern. Mineralized patters are then heat treated to pyrolyze the pattern and sinter the minerals. With this process, scaffolds were created with wall thicknesses as small as 150 m and internal channel diameters of 280-340 m, an appropriate range for bone tissue engineering.Mechanical Engineerin

Impact of Secondary Reactive Species on the Apparent Decoupling of Poly(Ethylene Glycol) Diacrylate Hydrogel Average Mesh Size and Modulus

Poly(ethylene glycol) diacrylate (PEGDA) hydrogels are widely used in biotechnology due to their in situ crosslinking capacity and tunable physical properties. However, as with all single component hydrogels, the modulus of PEGDA networks cannot be tailored independently of mesh size. This interdependence places significant limitations on their use for defined, 3D cell-microenvironment studies and for certain controlled release applications. The incorporation of secondary reactive species (SRS) into PEGDA hydrogels has previously been shown to allow the identification of up to 6 PEGDA hydrogel formulations for which distinct moduli can be obtained at consistent average mesh size (or vice versa). However, the modulus and mesh size ranges which can be probed by these formulations are quite restricted. This work presents an in-depth study of SRS incorporation into PEGDA hydrogels, with the goal of expanding the space for which decoupled examination of modulus and mesh size effects is achievable. Towards this end, over 100 PEGDA hydrogels containing either N-vinyl pyrrolidone or star PEG-tetraacrylate as SRS were characterized. To our knowledge, this is the first study to demonstrate that SRS incorporation allows for the identification of a number of modulus ranges that can be probed at consistent average mesh size (or vice versa)

Identifying Specific Combinations of Matrix Properties that Promote Controlled and Sustained Release of a Hydrophobic Drug from Electrospun Meshes

Despite advances in the development of degradable polymers for drug delivery, effective translation of drug-loaded materials is often hindered due to a poor understanding of matrix property combinations that promote controlled and sustained release. In this study, we investigated the influence of dominant factors on the release of a hydrophobic glucocorticoid dexamethasone (DEX) from electrospun meshes. Polycaprolactone meshes released 98% of the drug within 24 h, while poly(l-lactide) meshes exhibited negligible release even after 28 days despite both polymers being slow-degrading. Differences in drug-polymer interactions and drug-polymer miscibility—but neither matrix degradation nor differences in bulk hydrophobicity—influenced DEX release from these semi-crystalline matrices. Poly(d,l-lactide-co-glycolide) 50:50 meshes possessing two different fiber diameters exhibited a sequential burst and sustained release, while poly(d,l-lactide-co-glycolide) 85:15 meshes cumulatively released 26% drug in a controlled manner. Although initial drug release from these matrices was driven by differences in matrix architecture and solid-state drug solubility, release toward the later stages was influenced by a combination of fiber swelling and matrix degradation as evidenced by gross and microstructural changes to the mesh network. We suggest that drug release from polymeric matrices can be better understood via investigation of critical matrix characteristics influencing release, as well as concomitant examination of drug-polymer interactions and miscibility. Our findings offer rational matrix design criteria to achieve controlled/extended drug release for promoting sustained biological responses

Dehydrated Human Amnion/Chorion Membrane Allograft Nerve Wrap Around the Prostatic Neurovascular Bundle Accelerates Early Return to Continence and Potency Following Robot-assisted Radical Prostatectomy: Propensity Score–matched Analysis

Abstract We present a propensity-matched analysis of patients undergoing placement of dehydrated human amnion/chorion membrane (dHACM) around the neurovascular bundle (NVB) during nerve-sparing (NS) robot-assisted laparoscopic prostatectomy (RARP). From March 2013 to July 2014, 58 patients who were preoperatively potent (Sexual Health Inventory for Men [SHIM] score >19) and continent (no pads) underwent full NS RARP. Postoperative outcomes were analyzed between propensity-matched graft and no-graft groups, including time to return to continence, potency, and biochemical recurrence. dHACM use was not associated with increased operative time or blood loss or negative oncologic outcomes ( p >0.500). Continence at 8 wk returned in 81.0% of the dHACM group and 74.1% of the no-dHACM group ( p =0.373). Mean time to continence was enhanced in group 1 patients (1.21 mo) versus (1.83 mo; p =0.033). Potency at 8 wk returned in 65.5% of the dHACM patients and 51.7% of the no-dHACM group ( p =0.132). Mean time to potency was enhanced in group 1, (1.34 mo), compared to group 2 (3.39 mo; p =0.007). Graft placement enhanced mean time to continence and potency. Postoperative SHIM scores were higher in the dHACM group at maximal follow-up (mean score 16.2 vs 9.1). dHACM allograft use appears to hasten the early return of continence and potency in patients following RARP

A Rare Case of Renal Gastrinoma

We present a rare case of renal gastrinoma. To the best of our knowledge, only one case of renal gastrinoma has been reported in the literature so far. An African American male was diagnosed with Zollinger Ellison syndrome at the age of 15 years, when he underwent surgery for peritonitis secondary to duodenal ulcer perforation. Further evaluation was deferred and proton pump inhibitors were prescribed. Later evaluation showed a left renal mass. Serum gastrin levels were 4,307 pg/ml. A CAT scan of the abdomen showed 4- x 4-cm heterogeneous solid mass in the interpolar region of the left kidney with central hypodensity. Somatostatin scintigraphy confirmed a receptor-positive mass in the same location. Nephrectomy was done and the tumor was diagnosed on histopathological examination as a gastrinoma. At 6-month follow-up, gastrin levels were 72 pg/ml. After a follow-up of 6 years, the patient has no recurrent symptoms

Collagen-Mimetic Hydrogels Promote Human Endothelial Cell Adhesion, Migration and Phenotypic Maturation

This work evaluates the response of human aortic endothelial cells (HAECs) to thromboresistant collagen-mimetic hydrogel coatings toward improving the biocompatibility of existing off-the-shelf small-caliber vascular grafts. Specifically, bioactive hydrogels-previously shown to support α1/α2 integrin-mediated cell adhesion but to resist platelet activation-were fabricated by combining poly(ethylene glycol) (PEG) with a 120 kDa, triple-helical collagen-mimetic protein (Scl2-2) containing the GFPGER adhesion sequence. Analysis of HAECs seeded onto the resulting PEG-Scl2-2 hydrogels demonstrated that HAEC adhesion increased with increasing Scl2-2 concentration, while HAEC migration rate decreased over this same concentration range. In addition, evaluation of HAEC phenotype at confluence indicated significant differences in the gene expression of NOS3, thrombomodulin, and E-selectin on the PEG-Scl2-2 hydrogels relative to PEG-collagen controls. At the protein level, however, only NOS3 was significantly different between the PEG-Scl2-2 and PEG-collagen surfaces. Furthermore, PECAM-1 and VE-cadherin expression on PEG-Scl2-2 hydrogels versus PEG-collagen controls could not be distinguished at either the gene or protein level. Cumulatively, these data indicate the PEG-Scl2-2 hydrogels warrant further investigation as off-the-shelf graft coatings. In future studies, the Scl2-2 protein can potentially be modified to include additional extracellular matrix or cytokine binding sites to further improve endothelial cell responses

PERIOPERATIVE AND ONCOLOGICAL OUTCOMES IN HIGH RISK ELDERLY PATIENTS

Aim of the study ontroversy continues to exist concerning the treatment of choice for D\u2019Amico high risk elderly patients. The purpose of this study is to compare the perioperative and oncologic outcomes of robotic assisted radical prostatectomy (RARP) in D\u2019Amico high risk, propensity score-matched elderly and younger cohorts. Materials and methods From January 2008 through August 2012, 3818 patients underwent RARP at our institution by a single surgeon (VP). Retrospective analysis of prospectively collected data from our Institutional Review Board approved registry identified 80 D\u2019Amico high risk patients, 70 years of age and over. A propensity scorematch analysis was conducted using multivariable analysis to compare elderly patients (age 70 and over) to those under 70. The final two study cohorts \u2013 D\u2019Amico high risk elderly patients (n=80) and D\u2019Amico high risk younger patients (n=80) constituted the clinical material for this comparative study of perioperative and oncologic outcomes. Results Preoperative clinical characteristics were similar for the two matched groups. The operative time, transfusion rate and intra-operative complications were similar for the two groups.The mean estimated blood loss was significantly greater in younger patients (156.1 \ub1 84.2 mLvs 113.6 \ub1 67.7; p=0.002). No significant differences were observed in laterality, ease of nerve sparing or surgeon subjectively assessed anastomosis and pathological outcomes between the groups. No significant differences were found in postoperative complication rates, overall pain scores, length of stay or duration of indwelling catheterization. At follow-up, freedom from biochemical recurrence (BCR) in elderly patients was 85.0% vs. 83.8% in younger patients. The mean time to BCR in elderly patients was 15.0 months (range, 2.3 to 38.8) and 14.5 months (range, 5.2 to 35.1) in younger patients. Discussion Conclusions This study clearly demonstrates that RARP can be performed in D\u2019Amico high risk elderly patients without increasing perioperative morbidity and with oncologic outcomes comparable to high risk younger patients. RARP in elderly patients presenting with localized prostate cancer should be considered a viable treatment alternative based on the individual\u2019s life expectancy

Topography of calcium phosphate ceramics regulates primary cilia length and TGF receptor recruitment associated with osteogenesis

The surface topography of synthetic biomaterials is known to play a role in material-driven osteogenesis. Recent studies show that TGFβ signalling also initiates osteogenic differentiation. TGFβ signalling requires the recruitment of TGFβ receptors (TGFβR) to the primary cilia. In this study, we hypothesize that the surface topography of calcium phosphate ceramics regulates stem cell morphology, primary cilia structure and TGFβR recruitment to the cilium associated with osteogenic differentiation. We developed a 2D system using two types of tricalcium phosphate (TCP) ceramic discs with identical chemistry. One sample had a surface topography at micron-scale (TCP-B, with a bigger surface structure dimension) whilst the other had a surface topography at submicron scale (TCP-S, with a smaller surface structure dimension). In the absence of osteogenic differentiation factors, human bone marrow stromal cells (hBMSCs) were more spread on TCP-S than on TCP-B with alterations in actin organization and increased primary cilia prevalence and length. The cilia elongation on TCP-S was similar to that observed on glass in the presence of osteogenic media and was followed by recruitment of transforming growth factor-β RII (p-TGFβ RII) to the cilia axoneme. This was associated with enhanced osteogenic differentiation of hBMSCs on TCP-S, as shown by alkaline phosphatase activity and gene expression for key osteogenic markers in the absence of additional osteogenic growth factors. Similarly, in vivo after a 12-week intramuscular implantation in dogs, TCP-S induced bone formation while TCP-B did not. It is most likely that the surface topography of calcium phosphate ceramics regulates primary cilia length and ciliary recruitment of p-TGFβ RII associated with osteogenesis and bone formation. This bioengineering control of osteogenesis via primary cilia modulation may represent a new type of biomaterial-based ciliotherapy for orthopedic, dental and maxillofacial surgery applications. Statement of Significance The surface topography of synthetic biomaterials plays important roles in material-driven osteogenesis. The data presented herein have shown that the surface topography of calcium phosphate ceramics regulates mesenchymal stromal cells (e.g., human bone marrow mesenchymal stromal cells, hBMSCs) with respect to morphology, primary cilia structure and TGFβR recruitment to the cilium associated with osteogenic differentiation in vitro. Together with bone formation in vivo, our results suggested a new type of biomaterial-based ciliotherapy for orthopedic, dental and maxillofacial surgery by the bioengineering control of osteogenesis via primary cilia modulation

Discovery of synergistic material-topography combinations to achieve immunomodulatory osteoinductive biomaterials using a novel in vitro screening method: The ChemoTopoChip

Human mesenchymal stem cells (hMSCs) are widely represented in regenerative medicine clinical strategies due to their compatibility with autologous implantation. Effective bone regeneration involves crosstalk between macrophages and hMSCs, with macrophages playing a key role in the recruitment and differentiation of hMSCs. However, engineered biomaterials able to simultaneously direct hMSC fate and modulate macrophage phenotype have not yet been identified. A novel combinatorial chemistry-topography screening platform, the ChemoTopoChip, is used here to identify materials suitable for bone regeneration by screening 1008 combinations in each experiment for human immortalized mesenchymal stem cell (hiMSCs) and human macrophage response. The osteoinduction achieved in hiMSCs cultured on the “hit” materials in basal media is comparable to that seen when cells are cultured in osteogenic media, illustrating that these materials offer a materials-induced alternative to osteo-inductive supplements in bone-regeneration. Some of these same chemistry-microtopography combinations also exhibit immunomodulatory stimuli, polarizing macrophages towards a pro-healing phenotype. Maximum control of cell response is achieved when both chemistry and topography are recruited to instruct the required cell phenotype, combining synergistically. The large combinatorial library allows us for the first time to probe the relative cell-instructive roles of microtopography and material chemistry which we find to provide similar ranges of cell modulation for both cues. Machine learning is used to generate structure-activity relationships that identify key chemical and topographical features enhancing the response of both cell types, providing a basis for a better understanding of cell response to micro topographically patterned polymers