Synergistic amalgamation of shellac with self-antibacterial hydroxyapatite and carboxymethyl cellulose: An interactive wound dressing for ensuring safety and efficacy in preliminary in vivo studies.

This study focuses on the synergistic formulation of environmentally friendly blended materials based on carboxymethyl cellulose (CMC) for advanced interactive wound dressing. New CMC hydrogels were prepared with two degrees of functionalization and chemically crosslinked with citric acid (CA) to fine-tune their properties. Additionally, CMC-based hybrids were created by blending with shellac (SHL) and incorporating self-antibacterial hydroxyapatite (HA) to inhibit bacterial growth and promote wound healing. The results demonstrate the successful production of superabsorbent hydrogels with typical swelling degrees ranging from 81% in water to 82% in phosphate-buffered saline (PBS). These hydrogels exhibit distinct morphological features and remarkable improvements in surface mechanical properties, specifically in their tensile properties, which show a significant increase from approximately 0.03 to 2.2 N/mm2 due to the formation of CMC-SHL-HA hybrid nanostructures. Furthermore, the cytocompatibility of these CMC-based hydrogels was investigated by assessing the in vitro cell viability responses of human skin fibroblasts. The results reveal the cell viability responses over 91%, indicating their biocompatibility with human cells. Moreover, the characteristics of surgical wounds were assessed before and after the application of the hydrogel on dogs, and no signs of infection were observed at any of the surgical sites post-surgery.

KLVFF Conjugated Curcumin Microemulsion-Based Hydrogel for Transnasal Route: Formulation Development, Optimization, Physicochemical Characterization, and Ex Vivo Evaluation.

Curcumin is a potent natural compound used to treat Alzheimer's disease (AD). However, the clinical usefulness of curcumin to treat AD is restricted by its low oral bioavailability and difficulty permeating the blood-brain barrier. To overcome such drawbacks, various alternative strategies have been explored, including the transnasal route. However, rapid mucociliary clearance in the nasal cavity is a major hindrance to drug delivery. Thus, designing a delivery system for curcumin to lengthen the contact period between the drug and nasal mucosa must be employed. This study describes the optimization of KLVFF conjugated curcumin microemulsion-base hydrogel (KCMEG) to formulate a prototype transnasal preparation using the response surface method to improve a mucoadhesive property. A central composite design was employed to optimize and evaluate two influencing factors: the concentration of carbopol 940 and the percentage of KLVFF conjugated curcumin microemulsion (KCME). The physicochemical properties, anti-cholinesterase activity, and anti-aggregation activities of KCME were investigated in this study. The studied factors, in terms of main and interaction effects, significantly (p < 0.05) influenced hardness and adhesiveness. The optimized KCMEG was evaluated for pH, spreadability, and mucoadhesive properties. Ex vivo nasal ciliotoxicity to optimize KCMEG was performed through the porcine nasal mucosa. KCME was transparent, with a mean globule size of 70.8 ± 3.4 nm and a pH of 5.80 ± 0.02. The optimized KCMEG containing 2% carbopol 940 showed higher in vitro mucoadhesive potential (9.67 ± 0.13 min) compared with microemulsion and was also found to be free from nasal ciliotoxicity during histopathologic evaluation of the porcine nasal mucosa. The result revealed that both the concentration of carbopol 940 and the percentage of KCME play a crucial role in mucoadhesive properties. In conclusion, incorporating a mucoadhesive agent in a microemulsion can increase the retention time of the formulation, leading to enhanced brain delivery of the drug. Findings from the investigation revealed that KCMEG has the potential to constitute a promising approach to treating AD via transnasal administration.

Influence of Gallic Acid-Containing Mouth Spray on Dental Health and Oral Microbiota of Healthy Dogs: A Pilot Study.

The pilot study aimed to investigate the effects of GAMS on oral microbiota in healthy dog subjects. Thirty-eight dogs were recruited and randomly allocated to the placebo (n = 19) and treatment groups (n = 19). The dogs were treated with mouth spray once daily for 42 days. The changes in the gingival index (GI), plaque index (PI), and calculus index (CI) were measured at baseline (day 0) and end of the study (42nd day). The changes in the oral microbial composition of representative dogs (placebo, n = 7; and treatment, n = 7) were also evaluated at baseline and end of the study. Oral microbial composition was assessed by sequencing. The sequences were annotated using the QIIME 2.0TM. The GI, PI, and CI indexes were reduced after the GAMS usage. The abundance of the commensal bacterial phylum Actinobacteria and Chloroflexi, genera Frederiksenia, and Bergeyella was improved after six weeks of GAMS usage. GAMS reduced the pathogenic bacterial species, including Neisseria sp., Desulfobulbus sp., Capnocytophaga canis, and Corynebacterium mustelae. Moreover, some pathogenic bacterial abundances were increased at the end of the study. All the microbial variations were observed within the group. The inter-group analysis revealed that the changes were unrelated to GAMS usage. Further studies need to be carried out using more experimental subjects to confirm the effectiveness of GAMS. More metagenomic data are required to evidence the GMAS impact on the oral microbiome of healthy dogs.

Influence of Gallic Acid-Containing Mouth Spray on Dental Health and Oral Microbiota of Healthy Cats-A Pilot Study.

This pilot study aimed to investigate the effects of gallic acid-containing mouth spray on oral microbiota in healthy cat subjects. Forty healthy cats were recruited and randomly allocated to the control (G1; n = 20) and treatment groups (G2; n = 20). The cats were treated with mouth spray twice daily for 42 days. The changes in the gingival index (GI) and plaque index (PI) were measured at baseline (day 0) and end of the study (42nd day). The changes in the oral microbial composition of representative animals (control, n = 9; and treatment, n = 8) were also evaluated at baseline and end of the study. Oral microbial composition was assessed by amplifying the V1-V3 region of the 16S rRNA gene from supragingival dental plaque DNA extracts. The sequences were annotated using the QIIME 2.0. The GI and PI were significantly reduced after 42 days of treatment. The deep sequencing revealed that mouth spray influenced the cats' oral microbiome and was significantly diverse. About 20 phyla and 59 species were observed after 42 days of mouth spray usage in cats' oral microbiota. The number of operational taxonomic units (OTUs) of post-treatment samples (PoTS) of G2 was greatly reduced compared to other samples. Further analysis revealed that mouth spray acts substantially against Desulfomicrobium orale, one of the known pathogens in periodontal disease. The mouth spray efficiently reduced the growth of 22 species and uprooted 17 species. Moreover, the mouth spray supported the growth of normal oral microbiota, including Moraxella and Neisseria species. The preliminary study suggested that the gallic acids-containing mouth spray could be an essential oral product to improve the oral hygiene of the cats. Moreover, further studies are needed to confirm the beneficial effect of mouth spray on cats.

Surface Modification of Curcumin Microemulsions by Coupling of KLVFF Peptide: A Prototype for Targeted Bifunctional Microemulsions.

Curcumin is one of the most promising natural therapeutics for use against Alzheimer's disease. The major limitations of curcumin are its low oral bioavailability and difficulty in permeating the blood-brain barrier. Therefore, designing a delivery system of curcumin to overcome its limitations must be employed. KLVFF, a peptide known as an amyloid blocker, was used in this study as a targeting moiety to develop a targeted drug delivery system. A prototype of transnasal KLVFF conjugated microemulsions containing curcumin (KLVFF-Cur-ME) for the nose-to-brain delivery was fabricated. The KLVFF-Cur-ME was developed by a titration method. A conjugation of KLVFF was performed through a carbodiimide reaction, and the conjugation efficiency was confirmed by FTIR and DSC technique. KLVFD-Cur-ME was characterized for the drug content, globule size, zeta potential, and pH. A transparent and homogeneous KLVFF-Cur-ME is achieved with a drug content of 80.25% and a globule size of 76.1 ± 2.5 nm. The pH of KLVFF-Cur-ME is 5.33 ± 0.02, indicating non-irritation to nasal tissues. KLVFD-Cur-ME does not show nasal ciliotoxicity. An ex vivo diffusion study revealed that KLVFF-Cur-ME partitions the porcine nasal mucosa through diffusion, following the Higuchi model. This investigation demonstrates the successful synthesis of a bifunctional KLVFF-Cur-ME as a novel prototype to deliver anti-Aß aggregation via an intranasal administration.

Prevalence of autoantibodies that bind to kidney tissues in cats and association risk with antibodies to feline viral rhinotracheitis, calicivirus, and panleukopenia.

BACKGROUND: The feline viral rhinotracheitis, calicivirus, and panleukopenia (FVRCP) vaccine, prepared from viruses grown in the Crandell-Rees feline kidney cell line, can induce antibodies to cross-react with feline kidney tissues. OBJECTIVES: This study surveyed the prevalence of autoantibodies to feline kidney tissues and their association with the frequency of FVRCP vaccination. METHODS: Serum samples and kidneys were collected from 156 live and 26 cadaveric cats. Antibodies that bind to kidney tissues and antibodies to the FVRCP antigen were determined by enzyme-linked immunosorbent assay (ELISA), and kidney-bound antibody patterns were investigated by examining immunofluorescence. Proteins recognized by antibodies were identified by Western blot analysis. RESULTS: The prevalences of autoantibodies that bind to kidney tissues in cats were 41% and 13% by ELISA and immunofluorescence, respectively. Kidney-bound antibodies were observed at interstitial cells, apical border, and cytoplasm of proximal and distal tubules; the antibodies were bound to proteins with molecular weights of 40, 47, 38, and 20 kDa. There was no direct link between vaccination and anti-kidney antibodies, but positive antibodies to kidney tissues were significantly associated with the anti-FVRCP antibody. The odds ratio or association in finding the autoantibody in cats with the antibody to FVRCP was 2.8 times higher than that in cats without the antibody to FVRCP. CONCLUSIONS: These preliminary results demonstrate an association between anti-FVRCP and anti-cat kidney tissues. However, an increase in the risk of inducing kidney-bound antibodies by repeat vaccinations could not be shown directly. It will be interesting to expand the sample size and follow-up on whether these autoantibodies can lead to kidney function impairment.

Bone regeneration of a polymeric sponge technique-Alloplastic bone substitute materials compared with a commercial synthetic bone material (MBCP+TM technology): A histomorphometric study in porcine skull.

BACKGROUND: Polymeric sponge technique is recommended for developing the desired porosity of Biphasic calcium phosphate (BCP) which may favor bone regeneration. PURPOSE: To investigate the healing of BCP with ratio of HA30/ß-TCP70 (HA30) and HA70/ß-TCP30 (HA70) polymeric sponge preparation, compare to commercial BCP (MBCP+TM). MATERIALS AND METHODS: Materials were tested X-ray diffraction (XRD) pattern and scanning electron microscope (SEM) analysis. In eight male pigs, six calvarial defects were created in each subject. The defects were the filled with 1 cc of autogenous bone, MBCP+TM (MBCP), HA30, HA70, and left empty (negative group). The new bone formations, residual material particles and bone-to-graft contacts were analyzed at 4, 8, 12 and 16 weeks. RESULTS: Fabricated BCP showed well-distributed porosity. At 16 weeks, new bone formations were 45.26% (autogenous), 33.52% (MBCP), 24.34% (HA30), 19.43% (HA70) and 3.37% (negative). Residual material particles were 1.88% (autogenous), 17.58% (MBCP), 26.74% (HA30) and 37.03% (HA70). These values were not significant differences (Bonferroni correction <0.005). Bone-to-graft contacts were 73.68% (MBCP), which was significantly higher than 41.68% (HA30) and 14.32% (HA70; Bonferroni correction <0.017). CONCLUSIONS: Polymeric sponge technique offers well-distributed porosity. The new bone formation and residual material particles were comparable to MBCP+TM, but the bone-to-graft contact was lower than MBCP+TM.

Effectiveness of a Nanohydroxyapatite-Based Hydrogel on Alveolar Bone Regeneration in Post-Extraction Sockets of Dogs with Naturally Occurring Periodontitis.

Pathological mandibular fracture after dental extraction usually occurs in dogs with moderate to severe periodontitis. A nanohydroxyapatite-based hydrogel (HAP hydrogel) was developed to diminish the limitations of hydroxyapatite for post-extraction socket preservation (PSP). However, the effect of the HAP hydrogel in dogs has still not been widely investigated. Moreover, there are few studies on PSP in dogs suffering from clinical periodontitis. The purpose of this study was to evaluate the effectiveness of the HAP hydrogel for PSP in dogs with periodontitis. In five dogs with periodontitis, the first molar (309 and 409) of each hemimandible was extracted. Consequently, all the ten sockets were filled with HAP-hydrogel. Intraoral radiography was performed on the day of operation and 2, 4, 8 and 12 weeks post operation. The Kruskal-Wallis test and paired t-test were adopted for alveolar bone regeneration analysis. The results demonstrated that the radiographic grading, bone height measurement, and bone regeneration analysis were positively significant at all follow-up times compared to the day of operation. Moreover, the scanning electron microscopy with energy-dispersive X-ray spectroscopy imaging after immersion showed a homogeneous distribution of apatite formation on the hydrogel surface. Our investigation suggested that the HAP hydrogel effectively enhances socket regeneration in dogs with periodontitis and can be applied as a bone substitute for PSP in veterinary dentistry.

Geographic Spatial Distribution Patterns of Dirofilaria immitis and Brugia pahangi Infection in Community Dogs in Chiang Mai, Thailand.

Filariasis is emerging as a public health concern in tropical and subtropical areas. Filariasis is an endemic problem commonly found in southeast Asian countries. Using the PCR-restriction fragment length polymorphism (PCR-RFLP) of the ITS1 region with Vsp I, the overall prevalence rates of Dirofilaria immitis (12.2% (41/337); 95% confidence interval: 9.1-16.1%) and Brugia pahangi (8.3% (28/337); 95% confidence interval: 5.8-11.8%) were determined based on 337 free-roaming community dogs from 20 districts in Northern Thailand. Microfilaremia was found in only 6.2% of dogs (21/337). Co-infection with D. immitis and B. pahangi was observed in two dogs. Of the 215 blood samples examined using a Canine Heartworm Ag Kit, only 3.72% (eight dogs) were D. immitis antigen positive. Among these eight, six dogs had occult D. immitis infections. In terms of geographic distribution, we found the abundance of D. immitis and B. pahangi in the central areas at altitudes less than 400 m to be 12.1% and 10.3%, respectively. In contrast, at higher altitudes between 400 and 800 m, a significantly higher number of B. pahangi compared with D. immitis infected individuals were observed at 14.29% and 4.1%, respectively. In conclusion, D. immitis and B. pahangi were the most common filarial infections found in community dogs in Northern Thailand. Dogs might be an important reservoir of B. pahangi in that region. Increasing awareness and concern and including proper deworming programs for community dogs should be endorsed to reduce the transmission risk. Additionally, the population dynamics of the mosquito vector of B. pahangi across altitudinal gradients deserved further investigation.

Production of polyclonal antibody against kidney antigens: a model for studying autoantibody in feline chronic kidney diseases.

Chronic kidney disease is considered to be most common in geriatric domestic cats. It has been reported that the feline viral rhinotracheitis, calicivirus, and panleukopenia (FVRCP) vaccine prepared from the Crandell-Rees feline kidney (CRFK) cell line can induce cross-reactions of antibodies with feline kidney tissues. As an anti-cat kidney antibody was not available commercially for this study of autoantibody in cats, the purpose of this study was to produce anti-cat kidney antibody in rabbits for further study of autoantibody in cats after FVRCP vaccination. Kidney proteins from cadaveric cats were extracted and immunized into rabbits using Montanide as the adjuvant. Based on enzyme-linked immunosorbent assay measurement, all immunized rabbits produced high levels of anti-cat kidney antibodies and some began to produce antibodies as early as 2 weeks after immunization. Immunofluorescence staining of rabbit sera showed kidney-bound antibodies in glomerulus, Bowman's capsule, apical surface of the proximal convoluted tubule, peritubular surface, and interstitial cells. Western blot analysis of cat kidney proteins revealed molecular weights (M.W.) of 72, 55, 47, and 31 kDa, while binding to the CRFK cell proteins was observed at M.W. of 43 and 26 kDa. The antibody that recognized the 47 kDa protein was similarly detected in cats with autoantibody presence after FVRCP vaccination. The kidney-bound antibody profile at different time points and its patterns in rabbits could be used as a model for the study of autoantibody to cat kidney in feline chronic kidney diseases.

Synthesis of Hydroxyapatite with Antibacterial Properties Using a Microwave-Assisted Combustion Method.

The prevention of implant-associated infections has been increasing clinically in orthopedic surgery. Hydroxyapatite with antibacterial properties was synthesized using a microwave-assisted combustion method. High crystallinity at low temperature can be achieved using this method. The synthesized hydroxyapatite exhibited a superior clear zone for both Gram-positive and Gram-negative bacteria. Electron spin resonance (ESR) and X-ray photoelectron spectroscopy (XPS) were used for the radical investigation. The application of intelligent ink testing and an antioxidant assay using DPPH reduction were also used to confirm the existence of radicals. These techniques provided data confirming that radicals are responsible for the antibacterial properties. The synthesized antibacterial hydroxyapatite would be a good candidate for the prevention any infection with medical implants and injection materials causing failure in bone repair.

A Cost Effective Easy Competitive Enzyme-Linked Immunosorbent Assay Suitable for Monitoring Protective Immunity against the Rabies Virus in the Serum of Humans and Dogs.

The coverage of rabies vaccinations has been reported at 70-80% of dogs in annual reports. However, there are still outbreaks of rabies among humans and dogs in Thailand, thus indicating the necessity of ensuring seroprevalence in vaccinated dogs and efficacy of human immunization. A cost effective easy competitive enzyme-linked immunosorbent assay (CEE-cELISA) was developed here for monitoring protective immunity against the rabies virus in human and dog serum samples using monoclonal antibody clone 1-46-12, which recognizes a conformational epitope of the rabies G protein. The ELISA plate is coated with the whole viral antigen from a commercial vaccine. The serotiter measured by the CEE-cELISA and by the gold standard assay (rapid fluorescent focus inhibition test), detecting the neutralizing antibody, showed a strong correlation, with an R value of 0.958 and 0.931 in humans and dogs, respectively. These correlations were detected in the serum samples from humans and dogs at antibody concentrations up to 100 and 10 IU/ml, respectively. This CEE-cELISA could be an alternative assay for evaluating mass rabies vaccination rapidly at a low cost as well as for detecting antirabies antibodies in the serum of not only humans but also other animal species.