Plasma electrolytic oxidation (PEO) of Ti-15Mo alloys conducted in electrolytes containing Ca and P compounds can be an efficient process with which to obtain bioactive coatings. This paper reports on the influence of the nature of the electrolyte, its concentration, and PEO process parameters on the properties of anodized layers on Ti-15Mo. A wide range of Ca- and P-containing alkaline and acidic solutions was employed to incorporate Ca and P ions into the anodized layer. The efficiency of the incorporation was evaluated by the Ca/P ratio in the coating as compared to that in the electrolyte. It was found that alkaline solutions are not suitable electrolytes for the formation of good quality, uniform PEO coatings. Only acidic electrolytes are appropriate for obtaining well-adherent homogeneous layers on Ti-15Mo. However, the maximum Ca/P ratios reached in the coatings were rather low (close to 1). The variation of electrical signal (negative-to-positive current ratio, frequency) and time of electrolysis do not result in a substantial change of this value. The processing time, however, did influence the coating thickness. Despite their low Ca/P ratio, the anodized layers demonstrate good biological activity, comparable to pure microrough titanium.

Angiogenesis and bone regeneration are closely interconnected processes. Whereas type-H blood vessels are abundantly found in the osteogenic zones during endochondral long bone development, their presence in flat bones’ development involving intramembranous mechanisms remains unclear. Here, we hypothesized that type-H-like capillaries that highly express CD31 and Endomucin (EMCN), may be present at sites of intramembranous bone development and participate in the control of osteogenesis. A rabbit model of calvarial bone augmentation was used in which bone growth was controlled over time (2–4 weeks) using a particulate bone scaffold. The model allowed the visualization of the entire spectrum of stages throughout bone growth in the same sample, i.e., active ossification, osteogenic activity, and controlled inflammation. Using systematic mRNA hybridization, the formation of capillaries subpopulations (CD31–EMCN staining) over time was studied and correlated with the presence of osteogenic precursors (Osterix staining). Type-H-like capillaries strongly expressing CD31 and EMCN were identified and described. Their presence increased gradually from the regenerative zone up to the osteogenic zone, at 2 and 4 weeks. Type-H-like capillaries may thus represent the initial vascular support encountered in flat bones’ development and which organize osteogenic niches.

This study evaluates the influence of a gelatin sponge on adipose-derived stromal cells (ASC). Transcriptomic data revealed that, compared to ASC in a monolayer, a cross-linked porcine gelatin sponge strongly influences the transcriptome of ASC. Wound healing genes were massively regulated, notably with the inflammatory and angiogenic factors. Proteomics on conditioned media showed that gelatin also acted as a concentrator and reservoir of the regenerative ASC secretome. This secretome promoted fibroblast survival and epithelialization, and significantly increased the migration and tubular assembly of endothelial cells within fibronectin. ASC in gelatin on a chick chorioallantoic membrane were more connected to vessels than an empty sponge, confirming an increased angiogenesis in vivo. No tumor formation was observed in immunodeficient nude mice to which an ASC gelatin sponge was transplanted subcutaneously. Finally, ASC in a gelatin sponge prepared from outbred rats accelerated closure and re-vascularization of ischemic wounds in the footpads of rats. In conclusion, we provide here preclinical evidence that a cross-linked porcine gelatin sponge is an optimal carrier to concentrate and increase the regenerative activity of ASC, notably angiogenic. This formulation of ASC represents an optimal, convenient and clinically compliant option for the delivery of ASC on ischemic wounds.

Deproteinized bovine bone mineral particles embedded in collagen (DBBM-C) are widely used for bone regenerations with excellent, albeit sometimes variable clinical outcomes. Clinicians usually prepare DBBM-C by mixing with blood. Replacing blood by saline represents an alternative. We investigated if saline treatment could improve DBBM-C i. handling in vitro and ii. biological performances in a rabbit calvarial model. In vitro, DBBM-C blocks soaked in saline or blood were submitted to compression tests. In vivo, four poly ether ether ketone (PEEK)cylinders were placed on 16 rabbit skulls, filled with DBBM-C soaked in blood or saline for 2–4–8–12 weeks before histomorphometry. DBBM-C blocks were fully hydrated after 30 s in saline when 120 s in blood could not hydrate blocks core. Stiffness gradually decreased 2.5-fold after blood soaking whereas a six-fold decrease was measured after 30 s in saline. In vivo, saline treatment allowed 50% more bone regeneration during the first month when compared to blood soaking. This difference was then no longer visible. New bone morphology and maturity were equivalent in both conditions. DBBM-C saline-soaking facilitated its handling and accelerated bone regeneration of highly qualitative tissues when compared to blood treatment. Saline pretreatment thus may increase the clinical predictability of bone augmentation procedures.

Analysis of marginal adaptation of dental adhesive interfaces using scanning electron microscopy has proven to be a powerful nondestructive method to evaluate the quality of adhesion. This methodology is, however, time-consuming and needs expensive equipment. The purpose of this study was to evaluate the possibility and efficiency of using a digital optical microscope (DOM) to perform marginal analysis and to compare it with the scanning electron microscope (SEM) analysis. Fifteen defect-free molars were selected for this study. Class V cavities were prepared and restored with resin composite, and epoxy replicas were obtained from silicone impressions of the restored teeth. Custom-made image analysis software was then used to measure the percentage of the noncontinuous margins (NCM) of each sample. To compare the DOM to the gold standard, SEM, each sample was analyzed 10 times using the DOM and three times using the SEM, by the same experienced operator. The repeatability coefficient and concordance were evaluated, and a Bland and Altman analysis was used for comparison of the two methods of measurements. To validate the DOM analysis method, an ANOVA approach (Gage R R) was used. Repeatability and reproducibility, which are two components of precision to validate the DOM analysis system, were calculated. For this, the same restorations were analyzed by two additional operators three times with the DOM. The duration of each step of the analysis using both methods was also recorded as a secondary outcome. Regarding the repeatability of each method, the Friedman test showed no statistically significant difference within the repetitions of measurements by SEM and DOM (p = 0.523 and p = 0.123, respectively). Moreover, the Bland-Altman analysis revealed a bias of 0.86 and concluded no statistically significant difference between the two methods, DOM and SEM. ANOVA evaluated DOM measurement system variation including the variances of repeatability and reproducibility that represented, respectively, 0.3% and 4% of the variance components. Reproducibility or inter-operator variability represented the principal source of variability with a statistically significant difference (p = 0.024). The time required for analysis with SEM was almost double that of DOM. The use of digital optical microscopy appears to be a valid alternative to the SEM for the analysis of marginal adaptation of dental adhesive interfaces. Further studies to evaluate the effect of training of operators in digital optical microscopy could reveal higher accuracy for this method and inter-operator agreement when experience is gained.