Determination of hair cortisol in horses: comparison of immunoassay vs LC-HRMS/MS.

The measure of hair cortisol concentration (HCC) is becoming an emerging approach to monitor mid-/long-term stress in animals, so it is more and more important to develop accurate and reliable methods. In the light of this, the aim of the present study was to compare mane HCCs of 47 horses with different managements, by means of an immunoassay (ELISA) and liquid chromatography coupled to hybrid high-resolution mass spectrometry (LC-HRMS/MS). After the washing step, the ground hair was extracted with methanol. The extract was evaporated and redissolved in two different aqueous solutions, depending on the detection technique. The methods were validated according to EMA guideline for bioanalytical method validation, in the range 2-50 pg mg-1 (ELISA) and 1-100 pg mg-1 (LC-HRMS/MS). Satisfactory quantitative performances were obtained for both of the approaches, but this latter demonstrated better precision. The detected concentrations in real samples were encompassing the range 1.3-8.8 pg mg-1 and 2.0-17.9 pg mg-1 by means of LC-HRMS/MS and ELISA, respectively. Overall, HCCs measured with ELISA technique were 1.6 times higher. The overestimation of immunoassay results might be caused by cross-reactivity phenomena of laboratory reagents and other structurally similar hormones present in the mane.

Investigating the cecal microbiota in broiler poultry farms and its potential relationships with animal welfare.

The present study assessed the modulation of cecal microbiota and correlations with Campylobacter colonization and animal welfare status. For these purposes, we conducted a cross sectional study of the cecal microbiota from 187 broilers reared in 13 batches from 10 poultry farms by performing 16S rRNA sequencing (regions V3-4). The welfare of each batch was assessed using a simplified Welfare Quality® protocol, scoring higher in organic batches, compared to both antibiotic-free and conventional batches. The bioinformatics analyses were conducted in QIIME 2 and a linear discriminant analysis determined the association between microbiota and animals with different Campylobacter carriage status and welfare levels. In the microbiota from the subjects negative for Campylobacter or with high welfare scores, Bacteroidetes was the predominant phylum with the genus Megamonas significantly increased in abundance. A greater abundance of Parabacteroides, Phascolarctobacterium, Helicobacter in poultry negative for Campylobacter was also found at the genus level. Animals with the lowest welfare scores showed an increased abundance of Proteobacteria. The results suggested a different microbial composition and diversity in the analyzed groups.

A New Highly Hydrophilic Electrochemical Implant Titanium Surface: A Histological and Biomechanical In Vivo Study.

PURPOSE: The aim was to compare the osseointegration degree and secondary implant stability between implants with different surface treatments. MATERIALS AND METHODS: A novel electrochemical treatment was applied to modify the sandblasted and acid-etched surface (SLA) to obtain the new hydrophilic Feeling (FEL) surface presenting a highly soluble and homogenous film made of calcium and phosphorus nanocrystals. Twenty 3.8 × 10-mm dynamix implants (Cortex) were inserted in sheep iliac crests. Sheep were killed after 2 months. Bone-to-implant contact percentage (%BIC) and biomechanical parameters, such as implant stability quotient (ISQ) and value of actual micromotion (VAM), were evaluated for each implants. RESULTS: No implant failures were observed. Implants of test group showed %BIC value 30% higher in respect with control group (P = 0.001). No statistical differences were detected between the 2 groups in VAM and ISQ values. CONCLUSION: Both surface treatments were highly osteoconductive because they were able to significantly increase the bone density onto implant surface in respect with that in which they were inserted (D4 bone density). The hydrophilic FEL surface demonstrated an increase of about 216% in BIC in respect with host bone density and an additional 30% more in respect with SLA surface. Faster osseointegration process is desirable in case of early implant loading protocol.

Translational value of sheep as animal model to study sinus augmentation.

Sinus augmentation is a routine surgical procedure in dentistry. At present, various animal models are available for the research purpose on this topic. In particular, for the first time, we have performed a morphological study on sheep sinus, using cone beam computed tomography (CBCT), to precisely define the anatomy of the ovine sinus. Then, we compared the sheep and human sinus morphological parameters, in order to uniform the research approach to the sinus augmentation procedures and to standardize this experimental model. Six fresh heads of adult female sheep were studied with CBCT and histologic examination to determine the dimensions and the organization of the ovine maxillary sinus. The comparison of the dimensional values between man and sheep shows evident differences between the two species; CBCT offers detailed information for studying normal maxillary sinus. Human and sheep maxillary sinus show anatomical differences that must be taken into account in experimental procedures.

Effect of temperature on the dental implant osseointegration development in low-density bone: an in vivo histological evaluation.

OBJECTIVES: To make an in vivo evaluation of the effects of 2 different bone temperatures, on the development of implant osseointegration, in low-density bone. MATERIALS AND METHODS: Fifteen implant osteotomic sites were prepared in the iliac crests of sheep. Before the implant insertion, 5 sites were heated to 50°C for 1 minute, 5 sites to 60°C for 1 minute, and 5 sites were not overheated. Fifteen titanium dental implants (Cortex, Israel) were inserted. After a healing period of 2 months, the histomorphometric parameters calculated for each implant were the Bone-Implant Contact percentage (%BIC) and the infrabony pocket depth. Unpaired t test was applied to find statistical differences between groups. RESULTS: No implants failed. Statistical significant differences in %BIC and periimplant bone loss were found between the 60°C group and control group. No significant differences were found between the 50°C group and control group, although bone suffering signs were present. CONCLUSION: An osteotomic site overheating up to 60°C for 1 minute in low-density bone, before implant insertion, did not lead to implant failure, but it induced significant crestal bone loss during healing and lower %BIC.

Effect of 50 to 60°C heating on osseointegration of dental implants in dense bone: an in vivo histological study.

OBJECTIVES: To evaluate, in vivo, the effects of bone temperatures increased up to 60°C introduced before implant insertion on titanium implant osseointegration. MATERIALS AND METHODS: Twenty-four acid etched and sandblasted implants (Cortex Dental Implants) were inserted in the inferior edge of sheep mandibles. Osteotomic sites were randomly divided into 3 groups before inserting the implant. In test 1 group and in test 2 group, implant sites were overheated, respectively, up to 50°C for 1 minute and 60°C for 1 minute, with an electronic controlled probe of 3 mm in diameter and 10 mm in length. Osteotomic sites in control group were not overheated. Implants were inserted according to standard procedures. After 2 months healing, % bone implant contact (%BIC) and infrabony pockets' depth were measured. Unpaired t test was applied to find statistical differences between groups. RESULTS: No implant failure occurred. No statistical significant difference in %BIC was found among groups. Histological analysis showed that mean infrabony pockets were statistically deeper in 60°C group than in other groups. CONCLUSIONS: Bone temperature up to 60°C for 1 minute does not seem to significantly impair titanium dental implant osseointegration. Bone damage signs evident in the 60°C group suggest that careful drilling procedure with sufficient irrigation is necessary to avoid periimplant infrabony pockets' formation. More in vivo evaluations are needed to identify what is the value of bone temperature increase for irreversible inhibition of implant osseointegration.

Role of amniotic fluid mesenchymal cells engineered on MgHA/collagen-based scaffold allotransplanted on an experimental animal study of sinus augmentation.

OBJECTIVES: The present research has been performed to evaluate whether a commercial magnesium-enriched hydroxyapatite (MgHA)/collagen-based scaffold engineered with ovine amniotic fluid mesenchymal cells (oAFMC) could improve bone regeneration process in vivo. MATERIALS AND METHODS: Bilateral sinus augmentation was performed on eight adult sheep in order to compare the tissue regeneration process at 45 and 90 days after implantation of the oAFMC-engineered scaffold (Test Group) or of the scaffold alone (Ctr Group). The process of tissue remodeling was analyzed through histological, immunohistochemical, and morphometric analyses by calculating the proliferation index (PI) of oAFMC loaded on the scaffold, the total vascular area (VA), and vascular endothelial growth factor (VEGF) expression levels within the grafted area. RESULTS: MgHA/collagen-based scaffold showed high biocompatibility preserving the survival of oAFMC for 90 days in grafted sinuses. The use of oAFMC increased bone deposition and stimulated a more rapid angiogenic reaction, thus probably supporting the higher cell PI recorded in cell-treated sinuses. A significantly higher VEGF expression (Test vs. Ctr Group; p = 0.0004) and a larger total VA (p = 0.0006) were detected in the Test Group at 45 days after surgery. The PI was significantly higher (p = 0.027) at 45 days and became significantly lower at 90 days (p = 0.0007) in the Test Group sinuses, while the PI recorded in the Ctr Group continued to increase resulting to a significantly higher PI at day 90 (CTR day 45 vs. CTR day 90; p = 0.022). CONCLUSIONS: The osteoinductive effect of a biomimetic commercial scaffold may be significantly improved by the presence of oAFMC. CLINICAL RELEVANCE: The amniotic fluid mesenchymal cell (AFMC) may represent a novel, largely and easily accessible source of mesenchymal stem cells to develop cell-based therapy for maxillofacial surgery.