Development of bioinks for 3D printing microporous, sintered calcium phosphate scaffolds.

Beta-tricalcium phosphate (ß-TCP)-based bioinks were developed to support direct-ink 3D printing-based manufacturing of macroporous scaffolds. Binding of the gelatin:ß-TCP ink compositions was optimized by adding carboxymethylcellulose (CMC) to maximize the ß-TCP content while maintaining printability. Post-sintering, the gelatin:ß-TCP:CMC inks resulted in uniform grain size, uniform shrinkage of the printed structure, and included microporosity within the ceramic. The mechanical properties of the inks improved with increasing ß-TCP content. The gelatin:ß-TCP:CMC ink (25:75 gelatin:ß-TCP and 3% CMC) optimized for mechanical strength was used to 3D print several architectures of macroporous scaffolds by varying the print nozzle tip diameter and pore spacing during the 3D printing process (compressive strength of 13.1 ± 2.51 MPa and elastic modulus of 696 ± 108 MPa was achieved). The sintered, macroporous ß-TCP scaffolds demonstrated both high porosity and pore size but retained mechanical strength and stiffness compared to macroporous, calcium phosphate ceramic scaffolds manufactured using alternative methods. The high interconnected porosity (45-60%) and fluid conductance (between 1.04 ×10-9 and 2.27 × 10-9 m4s/kg) of the ß-TCP scaffolds tested, and the ability to finely tune the architecture using 3D printing, resulted in the development of novel bioink formulations and made available a versatile manufacturing process with broad applicability in producing substrates suitable for biomedical applications.

Reactions: Antibacterial and bioactive dental restorative materials: Do they really work?

PURPOSE: The study and development of antibacterial materials for use in dental applications is growing with the development of novel materials and procedures. Examination of the effects of such antibacterial materials on oral pathogens as well as on stability and longevity of dental restorations is of paramount importance to the field. RESULTS: This review addressed the range of topics covered by the manuscripts presented at the Seoul symposium on antibacterial dental materials. CLINICAL SIGNIFICANCE: Based on the presented works, it seems that the emerging antibacterial and bioactive mate-rials can potentially benefit restorative dentistry; however, like many other subjects in clinical dentistry, good quality evidence on their effectiveness under clinical situations is yet to be accumulated.

Effect of silver nanoparticle geometry on methicillin susceptible and resistant Staphylococcus aureus, and osteoblast viability.

Orthopedic implant failure as a result of bacterial infection affects approximately 0.5-5% of patients. These infections are often caused by Staphylococcus aureus which is capable of attaching and subsequently forming a biofilm on the implant surface, making it difficult to eradicate with systemic antibiotics. Further, with the emergence of antibiotic-resistant bacteria, alternative treatments are necessary. Silver nanoparticles have received much attention for their broad spectrum antibacterial activity which has been reported to be both size and shape dependent. The purpose of this study was therefore to evaluate the effect of three different geometries on their effect on microbial susceptibility as well as evaluate their effect on bone cell viability. Silver nanoparticles of spherical, triangular and cuboid shapes were synthesized by chemical reduction methods. The susceptibility of S. aureus and methicillin-resistant S. aureus was evaluated a 24 h period and determined using a colorimetric assay. Further, the viability of human fetal osteoblast (hFOB) cells in the presence of the silver nanoparticles was evaluated over a period of 7 days by AlmarBlue fluorescence assay. hFOB morphology was also evaluated by light microscopy imaging. Results indicated that silver nanoparticle geometry did not have an effect on microbiota susceptibility or hFOB viability. However, high concentrations of silver nanoparticles (0.5 nM) conferred significant susceptibility towards the bacteria and significantly reduced hFOB viability. It was also found that the hFOBs exhibited an increasingly reduced viability to lower silver nanoparticle concentrations with an increase in exposure time.

Design of a paclitaxel prodrug conjugate for active targeting of an enzyme upregulated in breast cancer cells.

Breast cancer is the second most common cause of cancer-related deaths in women. Chemotherapy is an important treatment modality, and paclitaxel (PTX) is often the first-line therapy for its metastatic form. The two most notable limitations related to PTX-based treatment are the poor hydrophilicity of the drug and the systemic toxicity due to the drug's nonspecific and indiscriminate distribution among the tissues. The present work describes an approach to counter both challenges by designing a conjugate of PTX with a hydrophilic macromolecule that is coupled through a biocleavable linker, thereby allowing for active targeting to an enzyme significantly upregulated in cancer cells. The resultant strategy would allow for the release of the active ingredient preferentially at the site of action in related cancer cells and spare normal tissue. Thus, PTX was conjugated to the hydrophilic poly(amdioamine) [PAMAM] dendrimer through the cathepsin B-cleavable tetrapeptide Gly-Phe-Leu-Gly. The PTX prodrug conjugate (PGD) was compared to unbound PTX through in vitro evaluations against breast cancer cells and normal kidney cells as well as through in vivo evaluations using xenograft mice models. As compared to PTX, PGD demonstrated a higher cytotoxicity specific to cell lines with moderate-to-high cathepsin B activity; cells with comparatively lower cathepsin B activity demonstrated an inverse of this relationship. Regression analysis between the magnitude of PGD-induced cytotoxic increase over PTX and cathepsin B expression showed a strong, statistically significant correlation (r(2) = 0.652, p < 0.05). The PGD conjugate also demonstrated a markedly higher tumor reduction as compared to PTX treatment alone in MDA-MB-231 tumor xenograft models, with PGD-treated tumor volumes being 48% and 34% smaller than PTX-treated volumes at weeks 2 and 3 after treatment initiation.

Electrospun composite-coated endotracheal tubes with controlled siRNA and drug delivery to lubricate and minimize upper airway injury.

Endotracheal Tubes (ETTs) maintain and secure a patent airway; however, prolonged intubation often results in unintended injury to the mucosal epithelium and inflammatory sequelae which complicate recovery. ETT design and materials used have yet to adapt to address intubation associated complications. In this study, a composite coating of electrospun polycaprolactone (PCL) fibers embedded in a four-arm polyethylene glycol acrylate matrix (4APEGA) is developed to transform the ETT from a mechanical device to a dual-purpose device capable of delivering multiple therapeutics while preserving coating integrity. Further, the composite coating system (PCL-4APEGA) is capable of sustained delivery of dexamethasone from the PCL phase and small interfering RNA (siRNA) containing polyplexes from the 4APEGA phase. The siRNA is released rapidly and targets smad3 for immediate reduction in pro-fibrotic transforming growth factor-beta 1 (TGFϐ1) signaling in the upper airway mucosa as well as suppressing long-term sequelae in inflammation from prolonged intubation. A bioreactor was used to study mucosal adhesion to the composite PCL-4APEGA coated ETTs and investigate continued mucus secretory function in ex vivo epithelial samples. The addition of the 4APEGA coating and siRNA delivery to the dexamethasone delivery was then evaluated in a swine model of intubation injury and observed to restore mechanical function of the vocal folds and maintain epithelial thickness when observed over 14 days of intubation. This study demonstrated that increase in surface lubrication paired with surface stiffness reduction significantly decreased fibrotic behavior while reducing epithelial adhesion and abrasion.

Aspirin/amoxicillin loaded chitosan microparticles and polydopamine modified titanium implants to combat infections and promote osteogenesis.

It is known that titanium (Ti) implant surfaces exhibit poor antibacterial properties and osteogenesis. In this study, chitosan particles loaded with aspirin, amoxicillin or aspirin + amoxicillin were synthesized and coated onto implant surfaces. In addition to analysing the surface characteristics of the modified Ti surfaces, the effects of the modified Ti surfaces on the adhesion and viability of rat bone marrow-derived stem cells (rBMSCs) were evaluated. The metabolic activities of Staphylococcus aureus (S. aureus) and Escherichia coli (E. coli) biofilms on the modified Ti surfaces were also measured in vitro. Moreover, S. aureus was tested for its antibacterial effect by coating it in vivo. Using water as the droplet medium, the contact angles of the modified Ti surfaces increased from 44.12 ± 1.75° to 58.37 ± 4.15°. In comparison to those of the other groups tested, significant increases in rBMSC adhesion and proliferation were observed in the presence of aspirin + amoxicillin-loaded microspheres, whereas a significant reduction in the metabolic level of biofilms was observed in the presence of aspirin + amoxicillin-loaded microspheres both in vitro and in vivo. Aspirin and amoxicillin could be used in combination to coat implant surfaces to mitigate bacterial activities and promote osteogenesis.

Spatial Recombinant Human Bone Morphogenetic Protein 2 Delivery from Hydroxyapatite Scaffolds Sustains Bone Regeneration in Rabbit Radius.

Regenerating large bone defects requires a multifaceted approach combining optimal scaffold designs with appropriate growth factor delivery. Supraphysiological doses of recombinant human bone morphogenetic protein 2 (rhBMP2), typically used for the regeneration of large bone defects clinically in conjunction with an acellular collagen sponge (ACS), have resulted in many complications. In this study, we develop a hydroxyapatite/collagen I (HA/Col) scaffold to improve the mechanical properties of the HA scaffolds, while maintaining open connected porosity. Varying rhBMP2 dosages were then delivered from a collagenous periosteal membrane and paired with HA or HA/Col scaffolds to treat critical-sized (15 mm) diaphyseal radial defect in New Zealand white rabbits. The groups examined were ACS +76 µg rhBMP2 (clinically used INFUSE dosage), HA +76 µg rhBMP2, HA +15 µg rhBMP2, HA/Col +15 µg rhBMP2, and HA/Col +15 µg rhBMP2 + bone marrow-derived stromal cells (bMSCs). After 8 weeks of implantation, all regenerated bones were evaluated using microcomputed tomography, histology, histomorphometry, and torsional testing. It was observed that the bone volume regenerated in the HA/Col +15 µg rhBMP2 group was significantly higher than that in the groups with 76 µg rhBMP2. The same scaffold and growth factor combination resulted in the highest bone mineral density of the regenerated bone, and the most bone apposition on the scaffold surface. Both the HA and HA/Col scaffolds paired with 15 µg rhBMP2 had sustained ingrowth of the mineralization front after 2 weeks compared to the groups with 76 µg rhBMP2, which had far greater mineralization in the first 2 weeks after implantation. Complete bridging of the defect site and no significant difference in torsional strength, stiffness, or angle at failure were observed across all groups. No benefit of additional bMSC seeding was observed on any of the quantified metrics, while bone-implant apposition was reduced in the cell-seeded group. This study demonstrated that the controlled spatial delivery of rhBMP2 at the periosteum at significantly lower doses can be used as a strategy to improve bone regeneration around space maintaining scaffolds. Tweet Inside-out or outside-in: growth factors delivered from the outside of porous mineral-collagen scaffolds, maintain strength and regrow bone better in a rabbit study. Twitter handle for senior author (@Guda_Lab) and sponsoring institution (@UTSA) Impact Statement This study provides insights on bone regeneration in the presence of spatially controlled delivery of recombinant human bone morphogenetic protein 2 (rhBMP2) from porous hydroxyapatite scaffolds coated with collagen I films. Using critical-sized defects created in the radial diaphysis of skeletally mature New Zealand White rabbits, microcomputed tomography and histomorphometry indicated significantly higher bone regeneration, bone mineral density, and bone-implant contact, as well as sustained regeneration over longer durations with lower dosage of rhBMP2 delivered periosteally.

In vivo performance of bilayer hydroxyapatite scaffolds for bone tissue regeneration in the rabbit radius.

The objective of this study was to investigate the in vivo biomechanical performance of bone defects implanted with novel bilayer hydroxyapatite (HAp) scaffolds that mimic the cortical and cancellous organization of bone. The scaffolds maintained architectural continuity in a rabbit radius segmental defect model and were compared to an untreated defect group (negative control) and autologous bone grafts (positive control). Micro-CT evaluations indicated total bone and scaffold volume in the experimental group was significantly greater than the defect group but lesser than the autologous bone graft treatment. The flexural toughness of the scaffold and the autograft groups was significantly greater than the flexural toughness of the defect group. Interestingly, the absolute density of the bone mineral as well as calcium to phosphorus (Ca/P) ratio in that mineral for the scaffold and autograft contralateral bones was significantly higher than those for the defect contralaterals suggesting that the scaffolds contributed to calcium homeostasis. It was concluded from this study that new bone regenerated in the bilayer HAp scaffolds was comparable to the empty defects and while the HAp scaffolds provided significant increase in modulus when compared to empty defect and their flexural toughness was comparable to autografts after 8 weeks of implantation.

Electrically Stimulated Tunable Drug Delivery From Polypyrrole-Coated Polyvinylidene Fluoride.

Electrical stimulus-responsive drug delivery from conducting polymers such as polypyrrole (PPy) has been limited by lack of versatile polymerization techniques and limitations in drug-loading strategies. In the present study, we report an in-situ chemical polymerization technique for incorporation of biotin, as the doping agent, to establish electrosensitive drug release from PPy-coated substrates. Aligned electrospun polyvinylidene fluoride (PVDF) fibers were used as a substrate for the PPy-coating and basic fibroblast growth factor and nerve growth factor were the model growth factors demonstrated for potential applications in musculoskeletal tissue regeneration. It was observed that 18-h of continuous polymerization produced an optimal coating of PPy on the surface of the PVDF electrospun fibers with significantly increased hydrophilicity and no substantial changes observed in fiber orientation or individual fiber thickness. This PPy-PVDF system was used as the platform for loading the aforementioned growth factors, using streptavidin as the drug-complex carrier. The release profile of incorporated biotinylated growth factors exhibited electrosensitive release behavior while the PPy-PVDF complex proved stable for a period of 14 days and suitable as a stimulus responsive drug delivery depot. Critically, the growth factors retained bioactivity after release. In conclusion, the present study established a systematic methodology to prepare PPy coated systems with electrosensitive drug release capabilities which can potentially be used to encourage targeted tissue regeneration and other biomedical applications.

Ti-9Mn ß-type alloy exhibits better osteogenicity than Ti-15Mn alloy in vitro.

Ti-9Mn and Ti-15Mn were prepared using an arc furnace in order to understand their osteogenic behavior as a biomedical implant. Ti-9Mn surface showed a significantly lower contact angle value (41%) as compared with the Ti-15Mn surface. The higher Ra and lower hydrophilicity values of Ti-9Mn alloy as compared with Ti-15Mn alloy indicates that Ti-9Mn can have better osteoconductive properties. ALP activity of the osteoblast cells on the Ti-9Mn alloy was elevated by 45% on day 7 and 20% on day 14 as compared to the Ti-15Mn alloy that reflects faster induction of osteoblast phenotypes of MG63 cells. Filopodia and lamellipodia structures were spread more on the Ti-9Mn specimens as compared to the Ti-15Mn alloy. Cell viability on Ti-9Mn alloy increased by 25% and 32%, respectively after 7 and 14 days of culture as compared to Ti-15Mn alloy. On day 14 of culture, the relative expression of RUNX2, COL1, and OC on Ti-9Mn alloy were elevated by 35%, 21%, and 30% respectively than the Ti-15Mn alloy. Ti-9Mn alloy also exhibited an inductive effect on the cell proliferation, and upregulation in the expression of ALP, RUNX2, and OC that is, the genes related to osteoblastic differentiation. Hence, the present in vitro results suggest that Ti-9Mn can be a preferred implant material than the Ti-15Mn alloy.

Development of a Bioinspired, Self-Adhering, and Drug-Eluting Laryngotracheal Patch.

OBJECTIVES/HYPOTHESIS: Novel laryngotracheal wound coverage devices are limited by complex anatomy, smooth surfaces, and dynamic pressure changes and airflow during breathing. We hypothesize that a bioinspired mucoadhesive patch mimicking how geckos climb smooth surfaces will permit sutureless wound coverage and also allow drug delivery. STUDY DESIGN: ex-vivo. METHODS: Polycaprolactone (PCL) fibers were electrospun onto a substrate and polyethylene glycol (PEG) - acrylate flocks in varying densities were deposited to create a composite patch. Sample topography was assessed with laser profilometry, material stiffness with biaxial mechanical testing, and mucoadhesive testing determined cohesive material failure on porcine tracheal tissue. Degradation rate was measured over 21 days in vitro along with dexamethasone drug release profiles. Material handleability was evaluated via suture retention and in cadaveric larynges. RESULTS: Increased flocking density was inversely related to cohesive failure in mucoadhesive testing, with a flocking density of PCL-PEG-2XFLK increasing failure strength to 6880 ± 1810 Pa compared to 3028 ± 791 in PCL-PEG-4XFLK density and 1182 ± 262 in PCL-PEG-6XFLK density. The PCL-PEG-2XFLK specimens had a higher failure strength than PCL alone (1404 ± 545 Pa) or PCL-PEG (2732 ± 840). Flocking progressively reduced composite stiffness from 1347 ± 15 to 763 ± 21 N/m. Degradation increased from 12% at 7 days to 16% after 10 days and 20% after 21 days. Cumulative dexamethasone release at 0.4 mg/cm2 concentration was maintained over 21 days. Optimized PCL-PEG-2XFLK density flocked patches were easy to maneuver endoscopically in laryngeal evaluation. CONCLUSIONS: This novel, sutureless, patch is a mucoadhesive platform suitable to laryngeal and tracheal anatomy with drug delivery capability. LEVEL OF EVIDENCE: NA Laryngoscope, 131:1958-1966, 2021.

Regeneration enhanced in critical-sized bone defects using bone-specific extracellular matrix protein.

Extracellular matrix (ECM) products have the potential to improve cellular attachment and promote tissue-specific development by mimicking the native cellular niche. In this study, the therapeutic efficacy of an ECM substratum produced by bone marrow stem cells (BM-MSCs) to promote bone regeneration in vitro and in vivo were evaluated. Fluorescence-activated cell sorting analysis and phenotypic expression were employed to characterize the in vitro BM-MSC response to bone marrow specific ECM (BM-ECM). BM-ECM encouraged cell proliferation and stemness maintenance. The efficacy of BM-ECM as an adjuvant in promoting bone regeneration was evaluated in an orthotopic, segmental critical-sized bone defect in the rat femur over 8 weeks. The groups evaluated were either untreated (negative control); packed with calcium phosphate granules or granules+BM-ECM free protein and stabilized by collagenous membrane. Bone regeneration in vivo was analyzed using microcomputed tomography and histology. in vivo results demonstrated improvements in mineralization, osteogenesis, and tissue infiltration (114 ± 15% increase) in the BM-ECM complex group from 4 to 8 weeks compared to mineral granules only (45 ± 21% increase). Histological observations suggested direct apposition of early bone after 4 weeks and mineral consolidation after 8 weeks implantation for the group supplemented with BM-ECM. Significant osteoid formation and greater functional bone formation (polar moment of inertia was 71 ± 0.2 mm4 with BM-ECM supplementation compared to 48 ± 0.2 mm4 in untreated defects) validated in vivo indicated support of osteoconductivity and increased defect site cellularity. In conclusion, these results suggest that BM-ECM free protein is potentially a therapeutic supplement for stemness maintenance and sustaining osteogenesis.

Silver (Ag) doped magnesium phosphate microplatelets as next-generation antibacterial orthopedic biomaterials.

This article reports for the first time our successful result in the synthesis of antibacterial single-phase newberyite (NB, MgHPO4 .3H2 O), an important magnesium phosphate (MgP) bioceramic. The prime novelty lies in the fact that we explore novel MgPs as next-generation orthopedic biomaterials as opposed to conventional calcium phosphates (CaP). While NB has already shown great promise, unlike its competitor struvite (ST, MgNH4 PO4 .6H2 O), NB is not intrinsically antibacterial. Given the havoc created by surgical site infections (SSI) in orthopedics, it would be worthwhile to explore if antibacterial NB can be synthesized cost-effectively. To accomplish that central goal, we used silver ion (Ag+ ) containing precursor solutions and exposed those to microwave irradiation. This action resulted in the rapid synthesis of NB microplatelets. Besides, three other specific objectives are addressed. First, Ag-doping was optimized to preserve the single-phase nature for sustained dopant release. Second, Ag+ release kinetics against common infection causing bacterial strains was analyzed. Finally, we inspected for any harmful effect of Ag-doped NB on MC3T3 preosteoblasts. Interestingly, the single-phase nature of NB microplatelets can be retained until 2 wt % Ag-doping and they exhibit good antibacterial and cytocompatible properties. Even though 3 wt % Ag-doped compositions (composites) were 100% antibacterial; they were cytotoxic.

Hard and soft tissue evaluation of titanium dental implants and abutments with nanotubes in canines.

BACKGROUND: Little is known regarding the interaction of dental implant surface nanotubes and oral soft and hard tissues. The purpose of this study was to evaluate both histologically and radiographically the qualitative and quantitative effects of dental implant surface nanotubes on hard and soft tissue in a canine model. METHODS: Three subgroups consisting of a combination of test and control implants and abutments (Group A: control implant/control abutment, Group B: control implant/test abutment: Group C: test implant/test abutment) were placed in edentulous mandibles of six large-breed canines. Implants and abutments were placed on one side at baseline, and on the opposite side of the mandible at week 10; sacrifice occurred at week 12. Quantitative and qualitative analyses were used to measure newly formed hard and soft tissues histologically and radiographically. RESULTS: The mean radiographic change in marginal bone level from weeks 0 to 12 between implant groups was not statistically significant (P > 0.05). Mean soft tissue contact (junctional epithelium + connective tissue) for Groups A, B, and C were 2.29, 2.33, and 2.31 mm, respectively, with no statistically significant difference (P > 0.05) between the groups. All connective tissue fibers were oriented parallel to the abutment regardless of surface treatment. CONCLUSIONS: The findings of this study suggest that healing of hard and soft tissues around implants and abutments is similar when comparing grit-blasted surfaces to machined, turned surfaces with nanotubes. Both resulted in similar soft tissue contact values, as well as connective tissue fiber orientation.

In vivo hydroxyapatite scaffold performance in infected bone defects.

Critically sized bone defects are often compounded by infectious complications. The standard of care consists of bone autografts with systemic antibiotics. These injuries and treatments lead to donor site morbidity, antibiotic resistant strains of bacteria, and often end stage amputation. This study proposes an alternative to the autograft using a porous, hydroxyapatite (HA) scaffold evaluated with and without infection and antibiotics. Twenty-four New Zealand white rabbits received either our HA scaffold or a pulverized autograft (PBA) within a surgically created critical-sized defect in the femur. The two grafts were evaluated in either septic or aseptic defects and with or without antibiotic treatment. The HA scaffolds were characterized with micro computed tomography. Post-euthanasia, micro computed tomography, histology, and white blood cells component analysis were completed. The HA had significantly greater (p < .001) mineralization to total volume than the PBA groups with 27.56% and 14.88%, respectively, and the septic HA groups were significantly greater than the aseptic groups both with and without antibiotics (p = .016). The bone quality denoted by bone mineral density was also significantly greater (p < .001) in the HA groups (67.01 ± 0.38 mgHA/cm3 ) than the PBA groups (64.66 ± 0.85 mgHA/cm3 ). The HA scaffold is a viable alternative to the bone autograft in defects with and without infection as shown by the quality and quantity of bone.

Scaffold Architecture and Matrix Strain Modulate Mesenchymal Cell and Microvascular Growth and Development in a Time Dependent Manner.

BACKGROUND: Volumetric tissue-engineered constructs are limited in development due to the dependence on well-formed vascular networks. Scaffold pore size and the mechanical properties of the matrix dictates cell attachment, proliferation and successive tissue morphogenesis. We hypothesize scaffold pore architecture also controls stromal-vessel interactions during morphogenesis. METHODS: The interaction between mesenchymal stem cells (MSCs) seeded on hydroxyapatite scaffolds of 450, 340, and 250 µm pores and microvascular fragments (MVFs) seeded within 20 mg/mL fibrin hydrogels that were cast into the cell-seeded scaffolds, was assessed in vitro over 21 days and compared to the fibrin hydrogels without scaffold but containing both MSCs and MVFs. mRNA sequencing was performed across all groups and a computational mechanics model was developed to validate architecture effects on predicting vascularization driven by stiffer matrix behavior at scaffold surfaces compared to the pore interior. RESULTS: Lectin staining of decalcified scaffolds showed continued vessel growth, branching and network formation at 14 days. The fibrin gel provides no resistance to spread-out capillary networks formation, with greater vessel loops within the 450 µm pores and vessels bridging across 250 µm pores. Vessel growth in the scaffolds was observed to be stimulated by hypoxia and successive angiogenic signaling. Fibrin gels showed linear fold increase in VEGF expression and no change in BMP2. Within scaffolds, there was multiple fold increase in VEGF between days 7 and 14 and early multiple fold increases in BMP2 between days 3 and 7, relative to fibrin. There was evidence of yap/taz based hippo signaling and mechanotransduction in the scaffold groups. The vessel growth models determined by computational modeling matched the trends observed experimentally. CONCLUSION: The differing nature of hypoxia signaling between scaffold systems and mechano-transduction sensing matrix mechanics were primarily responsible for differences in osteogenic cell and microvessel growth. The computational model implicated scaffold architecture in dictating branching morphology and strain in the hydrogel within pores in dictating vessel lengths.

Immediate Dental Implant Stabilization in a Canine Model Using a Novel Mineral-Organic Adhesive: 4-Month Results.

PURPOSE: This study evaluated a novel injectable, self-setting, osteoconductive, resorbable adhesive that provides immediate implant stabilization. MATERIALS AND METHODS: Twenty-six large canines had the mandibular second through fourth premolars and the first molar removed bilaterally. After 3 months, oversized osteotomies were prepared with only the apical 2 mm of the implant engaging native bone. One site had a novel resorbable, self-setting, mineral-organic adhesive (TN-SM) placed around the implant, a second site received bone graft, and a third site received only blood clot. Removal torque, standardized radiography, and histology were used to evaluate implant stability and tissue contact after 24 hours, 10 days, and 4 months. RESULTS: Mean removal torque values after 24 hours were 1.4, 1.3, and 22.2 Ncm for the control, bone graft, and mineral-organic adhesive, respectively. After 10 days, these values were 5.7, 6.2, and 45.7 Ncm and at 4 months increased to 88.7, 77.8, and 104.7 Ncm, respectively. Clinical, radiographic, and histologic evaluations showed a lack of inflammatory reaction. Control defects were initially radiolucent in the coronal area; grafted sites revealed particles in the gap, with both conditions gradually filling with bone over time. At 10 days, histologic evaluation demonstrated excellent biocompatibility and intimate contact of mineral-organic adhesive to both the implant and bone, providing an osseointegration-like bond; control sites revealed no bone contact in the defect area, while the bone-grafted sites revealed unattached graft particles. At 4 months, much of the mineral-organic adhesive was replaced with bone; the control and grafted sites had some bone fill, and many of the defects demonstrated no bone-to-implant contact and were filled with soft tissue or isolated graft particles. CONCLUSION: The mineral-organic adhesive provides immediate (osseointegration-like) and continued implant stabilization over 4 months in sites lacking primary stability. Experimental sites demonstrated maintenance of crestal bone levels adjacent to the mineral-organic adhesive and soft tissue exclusion without the use of membranes in this canine model. These results demonstrate that this novel mineral-organic adhesive can enable implant osseointegration in a site where insufficient native bone exists to allow immediate implant placement.

Histomorphometric analysis of delayed implantation after horizontal distraction osteogenesis of the mandible in dogs.

PURPOSE: Insufficient alveolar width is known to impede successful implantation and is a major obstacle to successful oral reconstruction using intraosseous implants. The aim of this study was to evaluate the osseointegration at implantation after consolidation in distracted narrow alveolar bone. MATERIALS: Three adult mongrel dogs weighing 9 to 10 kg were studied. The lower premolars were extracted and horizontal distraction was performed using a distraction device after 8 weeks. Eight weeks after consolidation period, SLA surface implants were installed. The dogs were killed after 4 weeks implantation. RESULTS: The preoperative and postoperative buccolingual widths of alveolar ridge were 2.7 ± 2.1 mm and 4.8 ± 2.5 mm, respectively. Direct bone contact was achieved, with no significant difference in new bone formation observed between implants placed in the distracted and undistracted bone after 4 weeks. CONCLUSIONS: It was concluded from this study that horizontal distraction is a useful technique in augmenting a narrow alveolar ridge necessary for implant placement.

Hepatocyte-like cells derived from human amniotic epithelial, bone marrow, and adipose stromal cells display enhanced functionality when cultured on decellularized liver substrate.

Transplantation of primary hepatocytes has been used in treatments for various liver pathologies and end-stage liver disease. However, shortage of donor tissue and the inability of hepatocyte proliferation in vitro have lead to alternative methods such as stem cell-derived hepatocyte-like cells (HLCs). Mesenchymal stromal/stem cells, and amniotic epithelial cells were isolated from human bone marrow (BM-MSCs), lipoaspirates (ASCs), and amniotic tissue (AECs) respectively. All cells were differentiated into HLCs on plates coated with Type I collagen or Porcine Liver Extracellular Matrix (PLECM-AA) matrix. Flow cytometry of BM-MSCs and ASCs, and AECs showed high expression of MSC-specific and embryonic stem cell markers respectively. All cell types differentiated into osteocytes, chondrocytes, and adipocytes. All cell type-derived HLCs presented the typical cuboidal primary hepatocyte morphology on PLECM-AA and fewer vacuoles (AECs) compared to HLCs cultured on type I collagen. Gene analysis of all cell type-derived HLCs cultured on PLECM-AA revealed higher upregulation of genes involved in drug transportation and metabolism compared to HLCs cultured on type I collagen. Although, HLCs cultured on PLECM-AA displayed some hepatocyte-related function and bioactivity, overall gene expression was lower compared to that of primary hepatocytes suggesting that caution should be taken when considering using HLCs to replace total hepatocyte functionality.

The effect of titanium surface roughening on protein absorption, cell attachment, and cell spreading.

PURPOSE: The purpose of this study was to compare properties of roughened and polished titanium with respect to their ability to attach to cells and bind to protein as well as their cell spreading behavior. MATERIALS AND METHODS: Three different titanium surface treatments were compared for their ability to support cell attachment and spreading: sandblasted and acid-etched, resorbable blast media, and machine-polished titanium. The surface of the materials was characterized for surface roughness, surface energy, and surface chemistry. Osteoblast-like MG-63 cells were tested for in vitro attachment and spreading in the presence of serum proteins. Cell attachment was assessed by direct counting, dye binding, and microculture titanium assays. Cell spreading was determined by measuring area/cell in phalloidin-AlexaFluor 488 stained cells. Absorption of bovine serum albumin was determined by assay. RESULTS: Scanning electron micrography and x-ray diffractometry confirmed increased surface roughness of the roughened materials. All 3 materials had similar albumin binding kinetics. Three different methods confirmed that roughened surfaces enhance early cell attachment to titanium in the presence of serum. Cells spread better on smoother machined surfaces than on the roughened surfaces. CONCLUSION: Roughened titanium surfaces exhibited better early cell attachment than smooth surfaces in the presence of serum. The cells attached to roughened titanium were less spread than those attached to machined titanium. Although albumin binding was not different for roughened surfaces, it is possible that roughened surfaces preferentially bound to serum adhesive proteins to promote early cell attachment.