Current protocols for differentiation of stem cells make use of multiple treatments of soluble signals and/or matrix factors and result typically in partial differentiation to mature cells with under-or overexpression of adult tissue-specific genes. We developed a strategy for rapid and efficient differentiation of stem cells using substrata of biomatrix scaffolds, tissue-specific extracts enriched in extracellular matrix, and associated growth factors and cytokines, in combination with a serum-free, hormonally defined medium (HDM) tailored for the adult cell type of interest. Biomatrix scaffolds were prepared by a novel, four-step perfusion decellularization protocol using conditions designed to keep all collagen types insoluble. The scaffolds maintained native histology, patent vasculatures, and 1% of the tissue's proteins but >95% of its collagens, most of the tissue's collagen-associated matrix components, and physiological levels of matrix-bound growth factors and cytokines. Collagens increased from almost undetectable levels to >15% of the scaffold's proteins with the remainder including laminins, fibronectins, elastin, nidogen/entactin, proteoglycans, and matrix-bound cytokines and growth factors in patterns that correlate with histology. Human hepatic stem cells (hHpSCs), seeded onto liver biomatrix scaffolds and in an HDM tailored for adult liver cells, lost stem cell markers and differentiated to mature, functional parenchymal cells in 1 week, remaining viable and with stable mature cell phenotypes for more than 8 weeks. Conclusion: Biomatrix scaffolds can be used for biological and pharmaceutical studies of lineage-restricted stem cells, for maintenance of mature cells, and, in the future, for implantable, vascularized engineered tissues or organs. (HEPATOLOGY 2011;53:293-305) T he ongoing revolution in stem cell research has made possible the identification and isolation of stem cell populations including those from fetal and postnatal tissues.1 The potential of human hepatic stem cells (hHpSCs) and other stem/progenitors for pharmaceutical research, cell-based therapies,

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Several studies have indicated differences in bond strength of dental materials to crown and root dentin. To investigate the potential differences in matrix properties between these locations, we analyzed upper root and crown dentin in human third molars for ultimate tensile strength and collagen biochemistry. In both locations, tensile strength tested perpendicular to the direction of dentinal tubules (undemineralized crown = 140.4 +/- 48.6/root = 95.9 +/- 26.1; demineralized crown = 16.6 +/- 6.3/root = 29.0 +/- 12.4) was greater than that tested parallel to the tubular direction (undemineralized crown = 73.1 +/- 21.2/root = 63.2 +/- 22.6; demineralized crown = 9.0 +/- 3.9/root = 16.2 +/- 8.0). The demineralized specimens showed significantly greater tensile strength in root than in crown. Although the collagen content was comparable in both locations, two major collagen cross-links, dehydrodihydroxylysinonorleucine/its ketoamine and pyridinoline, were significantly higher in the root (by ~ 30 and ~ 55%, respectively) when compared with those in the crown. These results indicate that the profile of collagen cross-linking varies as a function of anatomical location in dentin and that the difference may partly explain the site-specific tensile strength.

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©Operative Dentistry, 2006, 31-6, 677-681 PNR Pereira • MF Nunes PA Miguez • EJ Swift Jr Clinical RelevanceThe 1-step self-etching adhesive had significantly higher mean bond strength to normal dentin than to caries-affected dentin. Although Single Bond had a similar tendency, bonds to normal dentin and caries-affected dentin were not significantly different. SUMMARYThis study was designed to evaluate the bond strengths of a 1-step self-etching system and a 2-step "etch and rinse" adhesive system to cariesaffected dentin and normal dentin. In addition, the micromorphology of the adhesive interfaces was analyzed using scanning electron microscopy (SEM). Extracted human molars with occlusal caries that had been stored frozen were ground in order to expose the caries-affected dentin and surrounding normal dentin. The teeth were then bonded using either Adper Prompt LPop or Single Bond (3M ESPE) and restored with Filtek Z250 (3M ESPE). After storage in water for 24 hours at 37°C, the teeth were sectioned, prepared for microtensile bond strength test and tested in tension at a crosshead speed of 1-mm/minute. After debonding of the interfaces, microhardness of the dentin underlying the interface of all specimens was measured. The thickness of the hybrid layers was observed under SEM. The results of this study indicate that the bond strength of Adper Prompt L-Pop adhesive was significantly higher to normal dentin than to caries-affected dentin (p<0.05) and that the bond strength of Single Bond to both normal and caries-affected dentin was not significantly different (p>0.05). Additionally, the thickness of the hybrid layers produced by both adhesive systems was thicker for caries-affected dentin.

Purpose: To evaluate the effects of dentin collagen modifications induced by various cross-linkers on the stability of collagen matrix and the inhibition of root caries. Materials and Methods: The following cross-linkers were tested: 5% glutaraldehyde (GA), 0.5% proanthocyanidin (PA), 0.625% genipin (GE). In the first experiment, cross-linker-treated demineralized human root dentin was digested with bacterial collagenase, centrifuged, and the supernatants were subjected to amino acid analysis to determine collagen content. The residues were analyzed by SDS-PAGE and hydroxyproline analysis. In the second experiment, bovine root surfaces were conditioned with phosphoric acid, treated with the cross-linkers, incubated with Streptococcus mutans and Lactobacillus acidophilus for 1 week and the root caries inhibition was evaluated with confocal microscopy. Lastly, the ability of the bacteria to colonize the root surface was evaluated. In this experiment slabs of bovine root were treated with the cross-linkers and incubated in a suspension of S. mutans and L. acidophilus. The slabs were washed, resuspended in water, glucose was added, and the pH measured. Results: While all collagen was digested with collagenase in the control groups, only a small proportion was solubilized in the GA-, PA-, and GE-treated groups. The root caries was significantly inhibited by treatment with PA or GA. Drops in pH in the cross-linker-treated groups were essentially the same as in the untreated group. Conclusion: Naturally occurring cross-linkers, especially PA, could be used to modify root dentin collagen to efficiently stabilize collagen and to increase its resistance against caries.

Structured Abstract Objective The pore size of the scaffold is a critical factor in repairing large bone defect. Here, we investigated the potential of bone regeneration using novel nanocomposite polydopamine‐laced hydroxyapatite collagen calcium silicate (HCCS‐PDA) scaffolds with two different pore sizes, 250 and 500 μm. Samples/Setting A total of 12 male Sprague‐Dawley rats were implanted with HCCS‐PDA scaffold with pore size of either 250 or 500 μm into surgically created critical‐sized defect (CSD). Methods HCCS‐PDA scaffolds were fabricated using mould printing technique. The effect of pore size on mechanical strength of the scaffolds was assessed by compression testing. After seeding with rat mesenchymal stem cells (rMSCs), the scaffolds were implanted, and new bone formation was evaluated using microCT and histomorphometric analysis after 8 weeks. Results MicroCT and histology analysis demonstrated restricted peripheral new bone formation in either dural or periosteal side and limited new bone formation in the 250 μm pore scaffold. Conversely, the 500‐μm pore scaffold showed more penetration of new bone into the scaffold and greater bone regeneration in the rat CSD. Conclusion Based on our results, which demonstrated improved new bone formation in 500 μm pores scaffold, we can conclude that effective scaffold pore size that induces osteointegration and bone regeneration is around 500 μm for HCCS‐PDA nanocomposite scaffold.