An application of the density standard and scaled-pixel-counting protocol to assess the radiodensity of equine incisor teeth affected by resorption and hypercementosis: preliminary advancement in dental radiography.

BACKGROUND: Equine Odontoclastic Tooth Resorption and Hypercementosis (EOTRH) syndrome is a dental disease where the radiographic signs may be quantified using radiographic texture features. This study aimed to implement the scaled-pixel-counting protocol to quantify and compare the image structure of teeth and the density standard in order to improve the identification of the radiographic signs of tooth resorption and hypercementosis using the EOTRH syndrome model. METHODS AND RESULTS: A detailed examination of the oral cavity was performed in 80 horses and maxillary incisor teeth were evaluated radiographically, including an assessment of the density standard. On each of the radiographs, pixel brightness (PB) was extracted for each of the ten steps of the density standard (S1-S10). Then, each evaluated incisor tooth was assigned to one of 0-3 EOTRH grade-related groups and annotated using region of interest (ROI). For each ROI, the number of pixels (NP) from each range was calculated. The linear relation between an original X-ray beam attenuation and PB was confirmed for the density standard. The NP values increased with the number of steps of the density standard as well as with EOTRH degrees. Similar accuracy of the EOTRH grade differentiation was noted for data pairs EOTRH 0-3 and EOTRH 0-1, allowing for the differentiation of both late and early radiographic signs of EOTRH. CONCLUSION: The scaled-pixel-counting protocol based on the use of density standard has been successfully implemented for the differentiation of radiographic signs of EOTRH degrees.

Correction: Górski et al. Application of Two-Dimensional Entropy Measures to Detect the Radiographic Signs of Tooth Resorption and Hypercementosis in an Equine Model. Biomedicines 2022, 10, 2914.

In the original article [...].

The characterization of collagen-based scaffolds modified with phenolic acids for tissue engineering application.

The aim of the experiment was to study the morphology of collagen-based scaffolds modified by caffeic acid, ferulic acid, and gallic acid, their swelling, and degradation rate, as well as the biological properties of scaffolds, such as antioxidant activity, hemo- and cytocompatibility, histological observation, and antibacterial properties. Scaffolds based on collagen with phenolic acid showed higher swelling rate and enzymatic stability compared to scaffolds based on pure collagen, and the radical scavenging activity was in the range 85-91%. All scaffolds were non-hemolytic and compatible with surrounding tissues. Collagen modified by ferulic acid showed potentially negative effects on hFOB cells as a significantly increased LDH release was found, but all of the studied materials had antimicrobial activity against Staphylococcus aureus and Escherichia coli. It may be assumed that phenolic acids, such as caffeic, ferulic, and gallic acid, are modifiers and provide novel biological properties of collagen-based scaffolds. This paper provides the summarization and comparison of the biological properties of scaffolds based on collagen modified with three different phenolic acids.

Molecular Microbiological Characteristics of Gingival Pockets in the Periodontal Diseases of Dogs.

Introduction: Canine periodontitis results among other factors from a disturbed balance of dental plaque microflora and an inadequate host inflammatory response to a stimulus. This investigation sought to identify microorganisms associated with canine periodontitis. Material and Methods: Microbiological analysis was undertaken of gingival pockets in an experimental group of 36 dogs with periodontal diseases. Swabs were collected with the use of Pet Test (MIP Pharma, Berlin, Germany) from patients with gingival pockets deeper than 5 mm. Samples were aggregated and placed in separate shipping containers with the Pet Test kit. Results: Identification was made of the most common microorganisms, e.g. Porphyromonas gingivalis, Treponema denticola and Prevotella intermedia. The red complex constituted the largest proportion of all analysed organisms (84.26%). Capnocytophaga gingivalis was isolated from 33 dogs, Peptostreptococcus micros from 32 dogs, Fusobacterium nucleatum from 29 animals and P. intermedia from 20. Conclusion: The highest percentage of pathogens was supplied by P. gingivalis (61%). It is thought that dogs acquire them by means of cross-species transmission. The inter-study variability of results may depend not only on the method of periopathogen detection, but also on environmental factors, host immune status or genetic background. Depending on the state of periodontal disease, patients show varied microbiological profiles of the gingival pockets.

Assessment of the Impact of Physical Activity on the Musculoskeletal System in Early Degenerative Knee Joint Lesions in an Animal Model.

Osteoarthritis (OA) is one of the most prevalent diseases of the osteoarticular system. Progressive destruction of joints is accompanied by development of pathological changes in the muscle tissue, i.e., weakening, atrophy, and remodelling (sarcopenia). The aim of the present study is to assess the impact of physical activity on the musculoskeletal system in an animal model of early degenerative lesions in the knee joint. The study involved 30 male Wistar rats. The animals were allocated to three subgroups of 10 animals each. Each animal from the three subgroups received sodium iodoacetate by injection into the patellar ligament of the right knee joint, whereas saline was administered through the patellar ligament in the left knee joint. The rats in the first group were stimulated to exercise on a treadmill. The animals in the second group were allowed to lead a natural lifestyle (no treadmill stimulation). In the third group, all parts of the right hind limb muscle were injected with Clostridium botulinum toxin type A. The study demonstrated that, compared to the active rats, bone density in the immobilised rats decreased, as indicated by the densitometric assessment of the whole body and the examination of rats' hind limbs and knee joints alone. This clearly evidenced the impact of physical activity on bone mineralisation. The weight of both fat and muscle tissues in the physically inactive rats was reduced. Additionally, the adipose tissue had higher weight in the entire right hind limbs, where monoiodoacetic acid was administered to the knee joint. The animal model clearly showed the importance of physical activity in the early stages of OA, as it slows down the process of joint destruction, bone atrophy, and muscle wasting, whereas physical inactivity contributes to progression of generalised changes in the musculoskeletal system.

Chitosan/Phenolic Compounds Scaffolds for Connective Tissue Regeneration.

Chitosan-based scaffolds modified by gallic acid, ferulic acid, and tannic acid were fabricated. The aim of the experiment was to compare the compatibility of scaffolds based on chitosan with gallic acid, ferulic acid, or tannic acid using the in vivo method. For this purpose, materials were implanted into rabbits in the middle of the latissimus dorsi muscle length. A scaffold based on unmodified chitosan was implanted by the same method as a control. Moreover, the Fourier transform infrared spectroscopy-attenuated total reflectance (FTIR-ATR) spectra and scanning electron microscope (SEM) observations were made to study the interactions between chitosan and phenolic acids. Additionally, antioxidant properties and blood compatibility were investigated. The results showed that all studied materials were safe and non-toxic. However, chitosan scaffolds modified by gallic acid and tannic acid were resorbed faster and, as a result, tissues were organized faster than those modified by ferulic acid or unmodified.

Concentrations of Selected Adipocytokines in the Blood Plasma in Proximal Suspensory Desmopathy of Horses, with a Focus on Their Physical Activity-A Pilot Study.

Chronic tendon and ligament diseases are commonly encountered in both athletic humans and animals, especially horses. Distal limb diseases, including suspensory ligament (SL) pathology due to anatomical, histological, and biomechanical properties, can be considered a model for tendon and ligament pathologies in humans. The appropriate selection of therapy is often crucial in optimising the healing process. One decisive factor influencing the possibility of returning to pre-disease training levels appears to be the utilisation of physical activity, including controlled movement, during the rehabilitation process. In the pathogenesis of musculoskeletal diseases and rehabilitation, adipocytokines play diverse roles. However, it is unclear what significance they hold in horses and in specific disease entities as well as the consequences of their mutual interactions. Recent studies indicate that in the pathogenesis of diseases with varied aetiologies in humans, their value varies at different stages, resulting in a diverse response to treatment. The results of this study demonstrate lower resistin concentrations in the venous blood plasma of horses with proximal suspensory desmopathy (PSD), while higher levels were observed in regularly trained and paddocked animals. The horses investigated in this study showed higher concentrations of resistin and IL-8, particularly in paddocked horses as well as in the working group of horses. The results suggest that these concentrations, including resistin in blood plasma, may be clinically significant. This attempt to explore the aetiopathogenesis of the processes occurring in the area of the proximal attachment of the suspensory ligament may optimise the procedures for the treatment and rehabilitation of horses.

Malocclusions and Dental Diseases in Privately Owned Horses in the Mazovia Region of Poland.

Dental disorders, a term encompassing both malocclusion and dental disease, constitute a serious health problem in horses worldwide. As horse populations differ among countries and regions, a geographically specific characterization of the occurrence of various dental disorders may be helpful for local equine practitioners. This study investigated the prevalence and frequency distribution of selected malocclusions and dental diseases in horses housed in the Mazovia region of Poland, with attention paid to variations among age, gender, and breed categories. Routine dental examinations were conducted on 206 privately owned horses (n = 206). Disorders were recorded using a dental chart and classified as either malocclusions or dental diseases. Out of all examined horses, 95% demonstrated at least one dental disorder, with a roughly equal distribution of these disorders among incisor teeth (31%) and cheek teeth (31% each for premolars and molars). More specifically, there were disorders noted in 14 incisors, 3 canines, 2 wolf teeth, and 15 cheek teeth. Across all age, gender, and breed groups, malocclusions of incisor, premolar, and molar teeth occurred with a higher prevalence than did dental diseases. Curvatures and calculus were the most commonly reported pathologies in incisor teeth, whereas sharp enamel points and caries predominated in cheek teeth.

Application of Two-Dimensional Entropy Measures to Detect the Radiographic Signs of Tooth Resorption and Hypercementosis in an Equine Model.

Dental disorders are a serious health problem in equine medicine, their early recognition benefits the long-term general health of the horse. Most of the initial signs of Equine Odontoclastic Tooth Resorption and Hypercementosis (EOTRH) syndrome concern the alveolar aspect of the teeth, thus, the need for early recognition radiographic imaging. This study is aimed to evaluate the applicability of entropy measures to quantify the radiological signs of tooth resorption and hypercementosis as well as to enhance radiographic image quality in order to facilitate the identification of the signs of EOTRH syndrome. A detailed examination of the oral cavity was performed in eighty horses. Each evaluated incisor tooth was assigned to one of four grade-related EOTRH groups (0-3). Radiographs of the incisor teeth were taken and digitally processed. For each radiograph, two-dimensional sample (SampEn2D), fuzzy (FuzzEn2D), permutation (PermEn2D), dispersion (DispEn2D), and distribution (DistEn2D) entropies were measured after image filtering was performed using Normalize, Median, and LaplacianSharpening filters. Moreover, the similarities between entropy measures and selected Gray-Level Co-occurrence Matrix (GLCM) texture features were investigated. Among the 15 returned measures, DistEn2D was EOTRH grade-related. Moreover, DistEn2D extracted after Normalize filtering was the most informative. The EOTRH grade-related similarity between DistEn2D and Difference Entropy (GLCM) confirms the higher irregularity and complexity of incisor teeth radiographs in advanced EOTRH syndrome, demonstrating the greatest sensitivity (0.50) and specificity (0.95) of EOTRH 3 group detection. An application of DistEn2D to Normalize filtered incisor teeth radiographs enables the identification of the radiological signs of advanced EOTRH with higher accuracy than the previously used entropy-related GLCM texture features.

Selection of Filtering and Image Texture Analysis in the Radiographic Images Processing of Horses' Incisor Teeth Affected by the EOTRH Syndrome.

Equine odontoclastic tooth resorption and hypercementosis (EOTRH) is one of the horses' dental diseases, mainly affecting the incisor teeth. An increase in the incidence of aged horses and a painful progressive course of the disease create the need for improved early diagnosis. Besides clinical findings, EOTRH recognition is based on the typical radiographic findings, including levels of dental resorption and hypercementosis. This study aimed to introduce digital processing methods to equine dental radiographic images and identify texture features changing with disease progression. The radiographs of maxillary incisor teeth from 80 horses were obtained. Each incisor was annotated by separate masks and clinically classified as 0, 1, 2, or 3 EOTRH degrees. Images were filtered by Mean, Median, Normalize, Bilateral, Binomial, CurvatureFlow, LaplacianSharpening, DiscreteGaussian, and SmoothingRecursiveGaussian filters independently, and 93 features of image texture were extracted using First Order Statistics (FOS), Gray Level Co-occurrence Matrix (GLCM), Neighbouring Gray Tone Difference Matrix (NGTDM), Gray Level Dependence Matrix (GLDM), Gray Level Run Length Matrix (GLRLM), and Gray Level Size Zone Matrix (GLSZM) approaches. The most informative processing was selected. GLCM and GLRLM return the most favorable features for the quantitative evaluation of radiographic signs of the EOTRH syndrome, which may be supported by filtering by filters improving the edge delimitation.

Application of Hyperbaric Oxygen Therapy (HBOT) as a Healing Aid after Extraction of Incisors in the Equine Odontoclastic Tooth Resorption and Hypercementosis Syndrome.

Hyperbaric Oxygen Therapy (HBOT) is a stress-free, relatively safe method supporting the treatment of many different diseases. Although it is widely used in human medicine (including dentistry), in veterinary medicine, especially in the treatment of horses, there are not many scientifically described and documented cases of its use. Equine Odontoclastic Tooth Resorption and Hypercementosis syndrome is a disease that affects older horses and significantly reduces their quality of life. The only effective treatment for this condition is extraction of the incisors. The described case compares the recovery process of the alveolar area after extraction of incisors during the course of EOTRH syndrome without and with the use of a chamber, in horses with the same clinical picture of the disease, of the same age, and living in the same conditions. According to the authors' knowledge, the presented case describes the use of a chamber in equine dentistry for the first time.

Molecular Indicators of Biomaterials Osteoinductivity - Cell Migration, BMP Production and Signalling Turns a Key.

In this work we dissected the osteoinductive properties of selected, PLGA-based scaffolds enriched with gel-derived bioactive glasses (SBGs) of either binary SiO2-CaO or ternary SiO2-CaO-P2O5 system, differing in CaO/SiO2 ratio (i.e. high -or low-calcium SBGs). To assess the inherent ability of the scaffolds to induce osteogenesis of human bone marrow stromal cells (BMSC), the study was designed to avoid any osteogenic stimuli beyond the putative osteogenic SBG component of the studied scaffolds. The bioactivity and porosity of scaffolds were confirmed by SBF test and porosimetry. Condition media (CM) from BMSC-loaded scaffolds exhibited increased Ca and decreased P content corresponding to SBGs CaO/SiO2 ratio, whereas Si content was relatively stable and overall lower in CM from scaffolds containing binary SBGs. CM from cell-loaded scaffolds containing high-calcium, binary SBGs promoted migration of BMSC and BMP-response in reporter osteoblast cell line. BMSC culture on these scaffolds or the ones containing ternary, low-calcium SBGs resulted in the activation of BMP-related signaling and expression of several osteogenic markers. Ectopic bone formation was induced by scaffolds containing binary SBGs, but high-calcium ones produced significantly more osteoid. Scaffolds containing ternary SBGs negatively influenced the expression of osteogenic transcription factors and Cx43, involved in cell-cell interactions. High-calcium scaffolds stimulated overall higher Cx43 expression. We believe the initial cell-cell communication may be crucial to induce and maintain osteogenesis and high BMP signaling on the studied scaffolds. The presented scaffolds' biological properties may also constitute new helpful markers to predict osteoinductive potential of other bioactive implant materials.

EOTRH Syndrome in Polish Half-Bred Horses - Two Clinical Cases.

The aim of this paper is to thoroughly describe the etiology, diagnostic processes and recommended treatment of equine odontoclastic tooth resorption and hypercementosis (EOTRH). It is a relatively recently described disease. Most recent reports show that it is a common problem involving older equine patients. The most common and significant symptom of EOTRH is oral pain. The main clinical signs include periodontitis and gingivitis with a gingival recession, alveolar bone protrusion, periodontic fistulas and soreness of the incisors. When establishing the final diagnosis, it is necessary to compare the results of the clinical and radiological examination, because in the early stage, the disease may not show any symptoms. Authors used two clinical cases of Polish Half-Bred horses to present some aspects of diagnostic and surgical techniques. The knowledge presented in the paper may be valuable for equine practitioners working in the field to achieve more beneficial outcomes and provide a higher level of welfare for their equine patients. Nevertheless, further research and observations are necessary, since the etiology of the disease remains unclear.

Level of Adiponectin, Leptin and Selected Matrix Metalloproteinases in Female Overweight Patients with Primary Gonarthrosis.

The aim of this study was to assess levels of adiponectin (ACRP-30) and leptin in serum and synovial fluid (SF) of overweight females with primary gonarthrosis (GOA) and to look for their correlations with clinical status and the level of biochemical OA biomarkers: matrix metalloproteinase (MMP) 1, MMP-9 and tissue inhibitor of metalloproteinase (TIMP-1). The studied group consisted of 39 overweight females undergoing primary total knee arthroplasty as a result of GOA. The stage of GOA was evaluated according to Knee Society Score (KSS), Ahlbäck and Kellgren-Lawrence (K-L) scores. Peripheral blood and SF were obtained. Levels of selected biomarkers were assessed using ELISA kits. The mean level of ACRP-30 in serum valued were 8393.80 ng/mL and in SF, 774.33 ng/mL, the mean concentration of leptin in serum was 32,040.74 pg/mL and in SF, 27,332.12 pg/mL. Levels of leptin in serum and SF correlated with body mass index (BMI), (p = 0.0005, and p = 0.0002, respectively). Levels of ACRP-30 in serum was correlated with clinical scores (Ahlbäck: p = 0.0214; K-L: p = 0.0146). ACRP-30 in SF correlated with ACRP-30 in serum (p = 0.0003), tended to negatively correlate with MMP-1 in serum (p = 0.0598) and positively correlate with pro-MMP-1 in SF (p = 0.0600). To conclude, this study confirms the correlations between concentrations of both, leptin and ACRP-30, comparing serum and SF. In overweight females, leptin levels increase with BMI and ACRP-30 serum level increase in more advanced GOA stages. Finally, leptin levels were correlated with TIMP-1 serum concentration, one of the biochemical markers of GOA.

Unusual Penile Prolapse with an Infectious Background Caused by the Burkholderia cepacia Complex in a Stallion.

Penile prolapse is a disease manifested by an inability to retract the penis into the preputial sheath. It is reported in a variety of animal species, especially in young and intact males. However, penile prolapse in horses is commonly caused by trauma, sexual activity, pseudohermaphroditism, or neurological deficits, and less often by an infectious background. The present case report aimed to determine the etiological factor of penis infection associated with penile prolapse in a stallion in Poland. Our report indicates that the infectious background of penile prolapse was related to the Burkholderia cepacia complex. Based on antibiotic susceptibility results, the stallion was administered effective streptomycin and enrofloxacin treatment and recovered without complications. The following options are likely to be the infection source: contamination of hay or animal hygiene products. Finally, given its ability to grow in antiseptic solutions, difficulties in culturing, and innate multidrug resistance, this microorganism is currently a challenge to both detection and treatment in veterinary medicine cases.

Impact of a pulsed magnetic field on selected polymer implant materials.

PURPOSE: Physiotherapy with the use of pulsed magnetic fields is one of the methods of activating the processes of bone healing and regeneration. Exposing materials serving as membranes in guided bone regeneration (GBR) or guided tissue regeneration (GTR) to magnetic fields is an effective model that allows to monitor changes in the material under the influence of the magnetic field. METHODS: Materials engineering methods were used to verify the extent of material degradation resulting from magnetic field exposure in an aqueous environment. Changes in surface morphology were observed under an optical microscope and a scanning electron microscope (SEM). Changes in surface wettability were analysed in relation to the direct contact angle. Chemical structural changes were verified with the use of infrared spectroscopy (FTIR-ATR). RESULTS: The PCL-based membrane materials underwent relatively moderate surface degradation (altered contact angle, changes in surface morphology), but the absence of observable FTIR-ATR spectral shifts evidenced material stability under the influence of magnetic field. More extensive degradation processes were observed in the case of PLDLA-based materials, whose surface character changed from hydrophilic to hydrophobic. The spectra revealed enhanced intensity of the chain terminal groups, provided that modifiers (nanometric SiO2 and TCP (water reservoir)) were present in the polymer matrix. CONCLUSIONS: The extent degradation in the polymer membrane was primarily dependent on the presence of aqueous environment, while the influence of the magnetic field on the analysed membrane materials was negligible. Therefore, GBR/GTR membrane implants can be considered to remain stable during rehabilitation with the use of alternating magnetic field.

Elevated EGF Levels in the Blood Serum of Dogs with Periodontal Diseases and Oral Tumours.

BACKGROUND/AIM: Paradontopathy and neoplasms of the oral cavity represent one of the greatest challenges in human and animal dentistry. EGF plays a key role in maintaining the integrity and proper rate of cell proliferation in normal oral epithelium. The aim of the present study was to study serum levels of EGF in dogs diagnosed with periodontal diseases and oral cavity tumours. MATERIALS AND METHODS: The samples comprised of cancerous tissue sections and serum obtained from dogs of various breeds, aged between 5-13 years. Serum EGF concentrations were measured by an immunoenzymatic method. RESULTS: The median for EGF concentration in serum of dogs suffered from severe periodontal diseases was greater when compared to the control group. EGF concentration in dogs with malignant tumours was significantly higher than in those with non-malignant growths. A positive correlation between EGF concentration and tumour size was also observed. EGF level in dogs diagnosed with benign tumours was comparable to the control group. CONCLUSION: The blood serum level of EGF increases significantly in patients with malignant oral tumours and advanced periodontal disease. In malignant tumours, the high level of EGF correlates with the size and invasiveness of the neoplasm.

Osteochondral Repair Using Porous Three-dimensional Nanocomposite Scaffolds in a Rabbit Model.

AIM: To evaluate the utility of a novel nanocomposite biomaterial consisting of poly-L/D-lactide, and hydroxyapatite bioceramics, enriched with sodium alginate in articular cartilage defect treatment. MATERIALS AND METHODS: The biomaterial was prepared using the method of solvent casting and particle leaching. The study was conducted on 20 New Zealand White rabbits. Experimental osteochondral defects were created in the femoral trochlear grooves and filled with biomaterials. In control groups, the defects were left to spontaneously heal. The quality of newly-formed tissue was evaluated on the basis of macroscopic and histological assessment. Additionally the level of osteogenic and cartilage degradation markers were measured. RESULTS: The majority of the defects from the treatment group were covered with tissue similar in structure and colour to healthy cartilage, whereas in the control group, tissue was uneven, and not integrated into the surrounding cartilage. CONCLUSION: The results obtained validate the choice of biomaterial used in this study as well as the method of its application.

New approach in evaluation of ceramic-polymer composite bioactivity and biocompatibility.

Regeneration of bone defects was promoted by a novel ß-glucan/carbonate hydroxyapatite composite and characterized by Raman spectroscopy, microCT and electron microscopy. The elastic biomaterial with an apatite-forming ability was developed for bone tissue engineering and implanted into the critical-size defects of rabbits' tibiae. The bone repair process was analyzed on non-decalcified bone/implant sections during a 6-month regeneration period. Using spectroscopic methods, we were able to determine the presence of amides, lipids and assign the areas of newly formed bone tissue. Raman spectroscopy was also used to assess the chemical changes in the composite before and after the implantation process. SEM analyses showed the mineralization degree in the defect area and that the gap size decreased significantly. Microscopic images revealed that the implant debris were interconnected to the poorly mineralized inner side of a new bone tissue. Our study demonstrated that the composite may serve as a biocompatible background for collagen ingrowth and exhibits the advantages of applying Raman spectroscopy, SEM and microCT in studying these samples.

Molecular-level evaluation of selected periodontal pathogens from subgingival regions in canines and humans with periodontal disease.

Dogs commonly serve as a model for various human conditions, including periodontal diseases. The aim of this study was to identify the anaerobic bacteria that colonize the subgingival areas in dogs and humans by using rapid real-time polymerase chain reaction (RT-PCR)-based tests and to compare the results obtained in each species. Bacterial microflora evaluations, both quantitative and qualitative, were performed by applying ready-made tests on twelve dogs and twelve humans. Five samples were collected from each subject's deepest gingival pockets and joined to form a collective sample. The results of the study revealed interspecies similarities in the prevalences of Porphyromonas (P.) gingivalis, Treponema denticola, Tannerella forsythia, and Fusobacterium nucleatum. Red complex bacteria comprised the largest portion of the studied bacterial complexes in all study groups, with P. gingivalis being the most commonly isolated bacterium. The results show similarities in the prevalence of bacterial microflora in dogs and humans. Microbiological analysis of gingival pockets by using rapid real-time PCR-based tests in clinical practice, both veterinary and human, can facilitate the choice of appropriate pharmacological treatment and can provide a basis for subsequent verification of the treatment's effectiveness.