Novel Herbal Therapeutic YH23537 Improves Clinical Parameters in Ligature-Induced Periodontal Disease Model in Beagle Dogs.

Currently, available medicine does not satisfy the clinical unmet needs of periodontal disease. Therefore, novel drugs with improved efficacy profiles are needed. We previously demonstrated that YH14642, water extracts of Notoginseng Radix and Rehmanniae Radix Preparata, improved probing depths in double-blind phase II clinical trial. However, it still has hurdles for commercialization due to the low efficiency of active compound extraction. To resolve this issue, we developed YH23537 through process optimization to extract active compounds efficiently while still achieving the chemical profile of YH14642. In this study, we investigated the therapeutic effects of YH23537 compared with YH14642 using a canine model of ligature-induced periodontitis. Human gingival fibroblast (hGF) cells were treated with various concentrations of YH23537 or YH14642 with lipopolysaccharide (LPS) for 24 hr. IL-6 and IL-8 levels in the conditioned media were determined using Luminex. Sixteen 3-year-old male beagle dogs had their teeth scaled and polished using a piezo-type ultrasonic scaler under general anesthesia and brushed once daily for the following 2 weeks. Two weeks after the scaling procedure, the left upper second premolar (PM2), third premolar (PM3), and fourth premolar (PM4) as well as the left lower PM3, PM4, and first molar (M1) were ligated with silk-wire twisted ligatures. The dogs were fed with soft moistened food to induce periodontitis for 8 weeks, and the ligatures were then removed. YH23537 and YH14642 were administered for 4 weeks, and clinical periodontal parameters such as plaque index (PI), gingival index (GI), probing depth (PD), clinical attachment level (CAL), and bleeding on probing (BoP) were determined before and 1, 2, 3, and 4 weeks after treatment. YH23537 inhibited IL-6 and IL-8 secretion in a dose-dependent manner in hGF cells stimulated with LPS. The IC50 values for YH23537 were 43 and 54 µg/ml for IL-6 and IL-8, respectively, while the values for YH14642 were 104 and 117 µg/ml, respectively. In the animal study, clinical parameters including GI, PD, CAL, and BoP were significantly increased after 8 weeks of ligature-induced periodontitis. The YH23537 300 and YH23537 900 mg groups had significant improvements in CAL from 1 to 4 weeks after treatment in comparison to the placebo group. GR values in the YH23537 900 mg group were decreased throughout the treatment period. GI values were also reduced significantly after 4-week treatment with 300 and 900 mg of YH23537. YH23537 at 300 mg doses showed comparable efficacy for CAL and GR with 1,000 mg of YH14642. YH23537 showed therapeutic efficacy against periodontitis in dogs, mediated by anti-inflammatory effects. These findings indicate that YH23537 has the potential for further development as a new drug for patients suffering from periodontal disease.

Effect of preservative-free tafluprost on intraocular pressure, pupil diameter, and anterior segment structures in normal canine eyes.

OBJECTIVE: This study was designed to evaluate the changes in intraocular pressure (IOP), pupil diameter (PD), and anterior segment parameters using ultrasound biomicroscopy (UBM) after instillation of preservative-free (PF) tafluprost in normal dogs. PROCEDURES: Six beagle dogs were used. PF tafluprost was instilled in one randomly selected eye, and PF artificial tear was instilled in the other eye (control). IOP and PD were measured every 15 min for the first hour, every 2 h for the next 17 h, and at 24 h and 36 h postinstillation (PI). Anterior segment parameters including geometric iridocorneal angle (ICA), width of the entry of the ciliary cleft (CCW), length of the ciliary cleft, area of the ciliary cleft, and depth of the anterior chamber were measured with UBM before and after PF tafluprost instillation. RESULTS: Compared with the control group, IOP was significantly lower from 4 h PI to 24 h PI and PD was significantly smaller from 30 min PI to 18 h PI (P < 0.05). Among UBM parameters, ICA and CCW significantly decreased and increased after PF tafluprost instillation, respectively (P < 0.05). Other parameters showed no significant changes. CONCLUSIONS: Instillation of PF tafluprost lowered IOP and induced miosis in normal canine eyes. Alterations in ICA and CCW occurred simultaneously, which probably affected the outflow of aqueous humor. PF tafluprost could be considered an alternative prostaglandin analog in the treatment of canine glaucoma.

Acute changes in central corneal thickness according to experimental adjustment of intraocular pressure in normal canine eyes.

Central corneal thickness (CCT) can be a promising source of glaucoma monitoring and diagnosis. This study evaluated changes in CCT according to experimental adjustment of intraocular pressure (IOP) in canine eyes. To adjust and measure IOP, each eye was cannulated with two 26-gauge needles under inhalant anesthesia. One needle was connected to a pressure transducer, and the other was connected to an adjustable bag of physiologic saline. IOP was stepwise increased from 10 mmHg to 70 mmHg in 10 mmHg increments (Group T). IOP was maintained at 15 mmHg (Group C15), 30 mmHg (Group C30), 45 mmHg (Group C45), 60 mmHg (Group C60) and 75 mmHg (Group C75) during the experiment. CCT was measured with an ultrasonic pachymeter every 10 min after cannulation. There was a significant difference in the effect of time on CCT (P<0.001) and difference in CCT (dCCT; P<0.001) between groups. The CCT of group C15 remained constant during the experiment. However, group T showed an initial decrease and then an increase after passing the lowest point. Group C30 showed decreasing values for 30 min, after which the values remained constant. The values in Group C45 showed no changes for 40 min and then increased. The values in group C60 showed no change for 20 min and then increased. Group C75 showed a steady increase. In conclusion, the CCT showed two core changes according to increased IOP. This study provides essential basic data to enable further investigation into the association of IOP and CCT in dogs.

A modified method for inducing periodontitis in dogs using a silk-wire twisted ligature.

This study was designed to assess the effectiveness of a modified silk ligature twisted with wire for inducing advanced periodontitis. Periodontitis was induced in five premolars and one molar of 20 healthy dogs over a 60-day period. The dogs were divided into four groups according to the ligature-inducing materials used: soft moistened food only, wire ligature (WL), silk ligature (SL) and twisted ligature with silk and wire (SWL). Periodontal indices were recorded, and dental radiographs were taken before and after 60 days of ligation. The ligatures were checked daily and the day the ligature fell out was noted. The period during which the ligatures were maintained was significantly shorter for the SL group compared to the SWL group (p < 0.05). Results of the clinical examination showed that almost all periodontal status parameters including the plaque index, gingival index, clinical attachment level, and bleeding on probing were significantly exacerbated in the SWL group compared to the other groups (p < 0.05). Radiographic evaluation demonstrated that alveolar bone levels were significantly lower in the SWL group than the other groups on day 60 (p < 0.05). These results suggested that experimental periodontitis induced by SWL could be an effective method for investigating periodontitis in canine models.

Accuracy of intraocular pressure measurements in dogs using two different tonometers and plano therapeutic soft contact lenses.

OBJECTIVE: To compare and evaluate the accuracy of intraocular pressure (IOP) measured through a therapeutic contact lens, using applanation (TonoPen XL(®)) and rebound (TonoVet(®)) tonometers in enucleated dog eyes. ANIMALS STUDIED: A total of 30 enucleated eyes from 15 beagle dogs. PROCEDURES: To measure accurate IOP, the anterior chamber of each enucleated eye was cannulated with two 26-gauge needles and two polyethylene tubes were connected vertically to an adjustable reservoir bag of normal saline and a pressure transducer. IOP was measured by the TonoPen XL(®) followed by the TonoVet(®) without a contact lens. After a contact lens was applied to the cornea, IOP was re-measured in the same order. Three consecutive IOP measurements were performed using both tonometers. RESULTS: Without the contact lens, the IOP values obtained by both tonometers correlated well according to the regression analysis (TonoVet(®): γ(2) = 0.98, TonoPen XL(®): γ(2) = 0.97, P < 0.001). The TonoPen XL(®) consistently underestimated values as transducer IOP increased; however, IOP values measured with the TonoPen XL(®) were in close agreement and were less variable than those determined with the TonoVet(®) when a contact lens was applied to the cornea. Bland-Altman analysis was used to determine the lower and upper limits of agreement (TonoVet(®): -29.7 and +21.1 mmHg, TonoPen XL(®): -3.9 and +3.6 mmHg) between the two devices. CONCLUSIONS: This study suggests that the TonoPen XL(®) is a useful tonometer for dogs wearing therapeutic contact lenses, and importantly, contact lenses would not need to be removed prior to IOP measurement.