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.

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.

Osteogenic activity of the mixture of chitosan and particulate dentin.

To evaluate the healing effect of a mixture of chitosan and particulate dentin, a 8-mm-diameter critical size defect was created in the calvarium of 75 rats. The rats were divided into four experimental groups and a control group (no treatment). The defects in the experimental groups were grafted either with pig particulate dentin (group 1), a mixture of particulate dentin and plaster of Paris (group 2), particulate dentin and chitosan (group 3), or chitosan only (group 4). Rats in each group were sacrificed 2, 4, and 8 weeks after implantation. All experimental groups showed more new bone formation when compared to the control group. Additionally, all groups exhibited more bone growth at 8 weeks than at 4 weeks after implantation. It was concluded from this study that defects treated with particulate dentin powder-chitosan mixture may mediate an excellent effect on the formation of new bones.

Effect of chemically modified titanium surfaces on protein adsorption and osteoblast precursor cell behavior.

PURPOSE: To investigate the effects of different chemically modified titanium surfaces on protein adsorption and the osteoblastic differentiation of human embryonic palatal mesenchymal (HEPM) cells. MATERIALS AND METHODS: Three different surfaces were evaluated. The first, a machined surface (Ti-M), was considered a control. The second surface was acid etched (Ti-AE). The third surface was prepared by exposing the Ti-AE samples to sodium hydroxide (NaOH) solution (Ti-AAE). The surface characteristics of chemically modified titanium were investigated by means of scanning electron microscopy (SEM), Fourier transform infrared spectroscopy (FTIR), and profilometry. To evaluate the production of biomarkers, commercial kits were utilized. RESULTS: Surface composition and morphology affected the kinetics of protein adsorption. Ti-AE surfaces manifested a greater affinity for fibronectin adsorption compared to Ti-M or Ti-AAE surfaces. It was observed that Ti-AE and Ti-AAE surfaces promoted significantly greater cell attachment compared to Ti-M surfaces. Statistically significant differences were also observed in the expression of alkaline phosphatase (ALP) activity, osteocalcin, and osteopontin on all 3 titanium surfaces. ALP activity and osteocalcin production up to day 12 suggested that differentiation of the cells into osteoblasts had occurred and that cells were expressing a bone-forming phenotype. CONCLUSIONS: It was thus concluded from this study that surface morphology and composition play a critical role in enhancing HEPM cell proliferation and differentiation into osteoblast cells.

Properties and cyclic fatigue of glass infiltrated tape cast alumina cores produced using a water-based solvent.

OBJECTIVE: The purpose of this study was to investigate the properties of tape cast alumina composite produced using a water-based solvent and its possible clinical use as an all ceramic crown system in a fixed partial denture. Durability of the system will be measured by fatigue test to simulate the masticating conditions of the oral cavity. METHODS: The optimal weight ratio of water-based alumina tape was determined by tensile strength, shrinkage ratio and durability. The coefficient of thermal expansion, fracture toughness, biaxial flexural strength and flexural strength after fatigue test of a composite produced from alumina tape at optimal weight ratios were determined and compared to In-Ceram alumina core (control). RESULTS: The weight ratio of alumina/(alumina+binder+plasticizer) of 0.84 and binder/(binder+plasticizer) of 0.5 was observed to be the optimal composition for achieving excellent composite properties. Coefficient of thermal expansion of the sintered alumina tape was observed to be 7.3x10(-6)/degrees C, and this value was increased to 7.5x10(-6)/degrees C after infiltrating the sintered tape with glass. The fracture toughness and biaxial flexural strength of glass infiltrated alumina tape was observed to be 4.6 MPa m(1/2) and 498 MPa, respectively. After cyclic loading for 10(2)-10(6) cycles, no significant change in the biaxial flexural strength was observed between the glass infiltrated alumina core and the In-Ceram alumina core (p>0.05). SIGNIFICANCE: The observed properties provide evidence that the water-based tape cast alumina-glass composite is suitable for clinical use as an all ceramic crown system in a fixed partial denture.

Protein adsorption and osteoblast precursor cell attachment to hydroxyapatite of different crystallinities.

PURPOSE: The effect of hydroxyapatite (HA) crystallinity on protein adsorption and osteoblast precursor cell attachment to HA was investigated. MATERIALS AND METHODS: Different weight ratios of 100% crystalline HA and 100% amorphous calcium phosphate powders were mixed and pressed into disks (0.5 g) of different crystallinities--either 0% (HAO), 30% (HA30), 50% (HA50), 70% (HA70), or 100% (HA100). RESULTS: X-ray diffraction indicated differences in HA crystallinities. In addition, dissolution of the HA was dependent on its crystallinity, with an increase in phosphorus dissolution as the degree of crystallinity was decreased. No significant difference in albumin adsorption and initial osteoblast precursor cell attachment was observed in the range of HA0 to HA70 surfaces. However, a significantly lower albumin adsorption and initial osteoblast precursor cell attachment were observed on HA100. DISCUSSION: It was suggested that changes in ionic interactions as a result of a change in crystallinity affect the amount of calcium ion ligands readily available to electrostatically bind to proteins. CONCLUSION: It was thus concluded from this study that HA crystallinity affects the amount of albumin adsorbed and initial osteoblast attachment.

Surface characteristics and structure of anodic oxide films containing Ca and P on a titanium implant material.

An anodic oxide film that formed on titanium with a mixture of beta-glycerophosphate sodium (beta-GP) and calcium acetate was investigated. The anodic oxide had interconnected pores (ca. 1-2 microm in diameter) and intermediate roughness (0.60-1.50 microm). In addition, it contained a mixture of amorphous, anatase, and rutile oxides. With an increase in the anodizing voltage and/or concentration of calcium incorporated into the oxide, the degree of oxide crystallinity increased. However, with an increase in the concentration of beta-GP, the degree of oxide crystallinity decreased. It was concluded that the surface roughness, oxide crystallinity, and surface composition of the anodic oxide were dependent on the voltage, current density, and concentration of the electrolyte. It was also concluded that the anodized surface could be optimized for maximum osseointegration.