Tetracalcium Phosphate Biocement Hardened with a Mixture of Phytic Acid-Phytase in the Healing Process of Osteochondral Defects in Sheep.

Hyaline articular cartilage has unique physiological, biological, and biomechanical properties with very limited self-healing ability, which makes the process of cartilage regeneration extremely difficult. Therefore, research is currently focused on finding new and potentially better treatment options. The main objective of this in vivo study was to evaluate a novel biocement CX consisting of tetracalcium phosphate-monetit biocement hardened with a phytic acid-phytase mixture for the regeneration of osteochondral defects in sheep. The results were compared with tetracalcium phosphate-monetit biocement with classic fast-setting cement systems and untreated defects. After 6 months, the animals were sacrificed, and the samples were evaluated using macroscopic and histologic methods as well as X-ray, CT, and MR-imaging techniques. In contrast to the formation of fibrous or fibrocartilaginous tissue on the untreated side, treatment with biocements resulted in the formation of tissue with a dominant hyaline cartilage structure, although fine fibres were present (p < 0.001). There were no signs of pathomorphological changes or inflammation. Continuous formation of subchondral bone and hyaline cartilage layers was present even though residual biocement was observed in the trabecular bone. We consider biocement CX to be highly biocompatible and suitable for the treatment of osteochondral defects.

Comparative Analysis of Methods for Somatic Cell Counting in Cow's Milk and Relationship between Somatic Cell Count and Occurrence of Intramammary Bacteria.

The aim of this study was to compare three on-farm commercial methods for the indirect detection of subclinical mastitis in dairy cows: the California mastitis test (CMT), the Porta side somatic cell count milk test (Porta SCC), and the DeLaval cell counter (DCC), with the Fossomatic cell count (FSCC), and to evaluate the relationship between the determined somatic cell count SCC and the occurrence of intramammary pathogens in the milk of dairy cows. A total of 284 sensory unchanged mixed milk samples, collected during the milking on a dairy farm, were analyzed in this study for somatic cell counts by the mentioned on-farm tests. Quarter milk samples (n = 583) from all the selected cows were cultured. The agreement, sensitivity, and specificity of the three indirect commercial diagnostic tests (the CMT, the Porta SCC, and the DeLaval cell counter) were calculated, and the FSCC was used as the gold standard. The results were analyzed statistically using the Pearson correlation test and the paired t-test. The CMT matched with the FSCC in 83.1% of the samples, with the Porta SCC in 80.6%, and with the DCC in 80.3% of the samples. The sensitivity and specificity reached 81.0% and 92.9% for the CMT, 79.4% and 90.7% for the Porta SCC, and 75.8% and 97.5% for the DCC, respectively. The correlation between the FSCC and the Porta SCC was 0.86 (p < 0.0001), and between the FSCC and the DCC, it was 0.92 (p < 0.0001). The differences between them were insignificant. Bacteria were detected in 130 (22.3%) quarter milk samples. The most prevalent bacteria were Enterococcus spp. (36.2%), followed by E. coli (20%), coagulase-negative staphylococci (13.1%), A. viridans (9.2%), Streptococcus spp. (9.2%), Proteus spp. (6.2%), and S. intermedius (3.9%). Contagious isolates (S. aureus) were detected in 3 quarter milk samples (2.3%). The agreement between the individual tests and the microbiological culture was as follows: 69.2% for the CMT; 73.7% for the Porta SCC; 71.6% for the DCC; and 76.5% for the FSCC. Higher SCCs were detected in the milk samples contaminated with bacteria than in the healthy milk (p < 0.001). No significance was found between the presence of individual species of intramammary pathogens and the different levels of SCCs. Based on the results, bacteria are the predominant cause of subclinical mastitis. The increased SCC of some milk samples with no presence of bacteria meant that the increase could have been caused by numerous other agents (viruses, fungi, or algae) or factors for mastitis in the dairy industry.

Long-Bone-Regeneration Process in a Sheep Animal Model, Using Hydroxyapatite Ceramics Prepared by Tape-Casting Method.

This study was designed to investigate the effects of hydroxyapatite (HA) ceramic implants (HA cylinders, perforated HA plates, and nonperforated HA plates) on the healing of bone defects, addressing biocompatibility, biodegradability, osteoconductivity, osteoinductivity, and osteointegration with the surrounding bone tissue. The HA ceramic implants were prepared using the tape-casting method, which allows for shape variation in samples after packing HA paste into 3D-printed plastic forms. In vitro, the distribution and morphology of the MC3T3E1 cells grown on the test discs for 2 and 9 days were visualised with a fluorescent live/dead staining assay. The growth of the cell population was clearly visible on the entire ceramic surfaces and very good osteoblastic cell adhesion and proliferation was observed, with no dead cells detected. A sheep animal model was used to perform in vivo experiments with bone defects created on the metatarsal bones, where histological and immunohistochemical tissue analysis as well as X-ray and CT images were applied. After 6 months, all implants showed excellent biocompatibility with the surrounding bone tissue with no observed signs of inflammatory reaction. The histomorphological findings revealed bone growth immediately over and around the implants, indicating the excellent osteoconductivity of the HA ceramic implants. A number of islands of bone tissue were observed towards the centres of the HA cylinders. The highest degree of biodegradation, bioresorption, and new bone formation was observed in the group in which perforated HA plates were applied. The results of this study suggest that HA cylinders and HA plates may provide a promising material for the functional long-bone-defect reconstruction and further research.

In Vivo Study of Osteochondral Defect Regeneration Using Innovative Composite Calcium Phosphate Biocement in a Sheep Model.

This study aimed to clarify the therapeutic effect and regenerative potential of the novel, amino acids-enriched acellular biocement (CAL) based on calcium phosphate on osteochondral defects in sheep. Eighteen sheep were divided into three groups, the treated group (osteochondral defects filled with a CAL biomaterial), the treated group with a biocement without amino acids (C cement), and the untreated group (spontaneous healing). Cartilages of all three groups were compared with natural cartilage (negative control). After six months, sheep were evaluated by gross appearance, histological staining, immunohistochemical staining, histological scores, X-ray, micro-CT, and MRI. Treatment of osteochondral defects by CAL resulted in efficient articular cartilage regeneration, with a predominant structural and histological characteristic of hyaline cartilage, contrary to fibrocartilage, fibrous tissue or disordered mixed tissue on untreated defect (p < 0.001, modified O'Driscoll score). MRI results of treated defects showed well-integrated and regenerated cartilage with similar signal intensity, regularity of the articular surface, and cartilage thickness with respect to adjacent native cartilage. We have demonstrated that the use of new biocement represents an effective solution for the successful treatment of osteochondral defects in a sheep animal model, can induce an endogenous regeneration of cartilage in situ, and provides several benefits for the design of future therapies supporting osteochondral defect healing.

[Validation of a cow-side blood test for beta-hydroxybutyrate concentration in dairy cows]. / Überprüfung eines Schnelltestsystems zur Erfassung der Betahydroxybutyrat-Konzentration im Blut von Milchkühen.

OBJECTIVE: Clinical ketosis is common during the dairy cows' transition period and is responsible for considerable economic loss. Early identification of cows with subclinical ketosis is the first step for maintaining the health and productivity of dairy cows. The goals of the study were two-fold: The first was to examine the usefulness of a mobile test device as a cow-side test; and the second was to compare BHB concentrations measured by the ketometer using capillary blood and blood collected from the coccygeal vessels with values determined by a reference method in the laboratory using jugular blood. MATERIAL AND METHODS: Blood samples were collected from a jugular vein or the coccygeal vessels in 81 dairy cows at 7 time points (14 and 7 days pre-partum and 7, 14, 21, 28, and 42 days post-partum) for kinetic enzymatic measurement of BHB concentration in the laboratory. Blood samples were concurrently collected from the coccygeal vessels or by pricking the vulvar lip at the transition of the skin to the mucosa (capillary blood) to determine BHB concentration using the WellionVet BELUA ketometer (MED TRUST GmbH, Marz, Austria). RESULTS: Initial errors in operating the ketometer were quickly eliminated with experience. BHB concentrations of jugular blood measured in the laboratory were 0.07 mmol/l lower than those measured in coccygeal blood. The mean BHB concentration measured in coccygeal and capillary blood using the WellionVet BELUA ketometer did not significantly differ but were 0.13 and 0.12 mmol/l respectively, lower than the mean jugular vein concentrations measured in the laboratory. CONCLUSION: The WellionVet BELUA ketometer is useful for determination of BHB concentration in cows provided that the manufacturer's specifications are followed. Capillary blood is best collected at the transition from the vulvar skin to its mucosa. The device generates rapid results that correlate well with BHB concentrations determined in the laboratory and with the results obtained from different blood collection sites. It is ideally suited for monitoring dairy cows for subclinical ketosis using capillary or coccygeal blood.