Composition of algal extracts for preventing and treating periodontal disease

Abstract

A composition of algal extracts containing phycobiliprotein is provided. This composition not only effectively inhibit the growth of oral pathogens including Porphyromonathes gingivalis and Actinobacillus actinomycetemcomitans, but also reduces the inflammation caused by those pathogens. Therefore, the phenomenon of alveolar bone loss in periodontal patients is mitigation, achieving the effect of prevention and treatment of periodontal disease.

Claims

1. A method for inhibiting growth of Porphyromonas gingivalis in a subject, the method comprising administering to said subject an effective concentration of a C-phycocyanin in an extract solution, wherein the effective concentration is greater than 50 g/ml in the C-phycocyanin extract solution.

2. A method for inhibiting growth of Actinobacillus actinomycetemcomitans in a subject, the method comprising administering to said subject an effective concentration of a C-phycocyanin in an extract solution, wherein the effective concentration is greater than 50 g/ml in a C-phycocyanin extract solution.

3. The method according to claim 1 or 2, wherein the effective concentration is between 50 g/ml-600 g/ml in a C-phycocyanin extract solution.

4. The method according to claim 1 or 2, wherein the C-phycocyanin is made into a form selected from the group consisting of a paste, an injection and a patch.

5. The method according to claim 1 or 2, wherein the C-phycocyanin is mixed with a pharmaceutically acceptable excipient.

6. The method according to claim 5, wherein the pharmaceutically acceptable excipient is selected from the group consisting of solvents, dispersion media, coatings, antibacterial agents, antifungal agents, isotonic agents, and absorption delaying agents.

7. The method according to claim 5, wherein the pharmaceutically acceptable excipient is selected from the group consisting of detackifiers, anti-foaming agents, buffering agents, polymers, antioxidants, preservatives, chelating agents, viscomodulators, tonicifiers, flavorants, colorants, odorants, opacifiers, suspending agents, binders, fillers, plasticizers, lubricants, and mixtures thereof.

8. The method according to claim 5, wherein the pharmaceutically acceptable excipient is selected from the group consisting of lactose, mannitol, dextran, glucose, glutamic acid, gelatin, sorbitol, trehalose, sucrose, xylitol, starch, microcrystalline cellulose, methyl cellulose, arabic gum and combinations thereof.

9. The method according to claim 1 or 2, wherein the C-phycocyanin is mixed with a food additive.

10. The method according to claim 9, wherein the food additive comprises a sweetener or a potentiator.

11. The method according to claim 9, wherein the food additive is made in the form of a powder, a capsule, or an aqueous alcoholic solution.

12. The method according to claim 9, wherein the food additive is frozen, dried, spray-dried or freeze-dried.

Description

BRIEF DESCRIPTION OF THE DRAWINGS

(1) FIG. 1(A) illustrates effect of CPC antimicrobial activity for Porphyromonas gingivalis in vitro, and FIG. 1(B) illustrates effect of CPC antimicrobial activity for Actinobacillus actinomycetemcomitans in vitro.

(2) FIG. 2(A) illustrates the chart of the rat dental. FIG. 2(B) illustrates the operate photograph of the animal induced periodontal experiments.

(3) FIG. 3 illustrates the flow chart of animal experiments.

(4) FIG. 4(A) illustrates the MRI photograph of the phenomenon of the alveolar bone loss in animal experiment. FIG. 4(B) illustrates the statistics result of the alveolar bone loss in animal experiment. FIG. 4(C) illustrates the photograph of the tissue staining.

DETAILED DESCRIPTION OF THE INVENTION

Embodiment 1

The Antimicrobial Test

(5) 1. Culture of the Pathogens

(6) According to the experiment process of the reference paper Jeong et al., 2000 and Zhou et al., 2006, the main pathogens of the periodontal disease Porphyromonas gingivalis and Actinobacillus actinomycetemcomitans are cultured.

(7) 2. The Antimicrobial Experiment

(8) To find out the dosage which 50% of bacteria is inhibited, experiment studies the effects of different concentrations of the phycocyanin (25, 50, 150, 300, and 600 g/ml) on Porphyromonas gingivalis (FIG. 1A) and Actinobacillus actinomycetemcomitans (FIG. 1B).

(9) The inhibition experiment of the Porphyromonas gingivalis (FIG. 1A) indicates that the 50% inhibit concentration of the phycocyanin extracts is about 150 g/ml. The inhibit experiment of the Actinobacillus actinomycetemcomitans results (FIG. 1B) indicate that 50% inhibit concentration of the phycobiliproteins extracts is about 100 g/ml. But phycocyanin extracts present no inhibitory effect to the other pathogens of the E. coli, S. aureus, Streptococcus mutans and T. rubrum. These results show the invention containing extracts of phycocyaninhas the outstanding effect of inhibiting the growth of periodontal pathogens.

Embodiment 2

The Experiment of the Alveolar Bone Loss

(10) 1. Experiment Animal

(11) The present invention uses Sprague Dawley (SD) male rats as experiment animals; each rat weighs about 280-310 g and was purchased from national animal center in the National Defense Medical Center. The breed environment of the SD rat is around the 23-25 C. and the light setting is on between 8:00-20:00 for 12 hours and set off for the remaining 12 hours. The animal room is supplied with ample feed and water. In addition, the room keeps under proper humidity, and a separate filtered air is supplied.

(12) 2. The Set Up of Periodontal Disease Animal Model

(13) According to the published paper in 2008 by Cai et al., they setup a Ligature-induced periodontal disease model. (Hereinafter refer to as ligation). The first process is to give the rats anesthetic through intraperitoneal injection. Then place the ligature on molars nearby both sides of the jaw around the tooth neck on the primary molar and secondary molar (FIG. 2A and FIG. 2B). The knot is placed at the mesial surface.

(14) 3. Experiment Design

(15) The rats were randomly divided into five groups, include (1) Control group (2) ligation group (3) treatment group 1 (ligation+the phycocyanin extracts 50 mg/kg) (4) treatment group 2 (ligation+the phycocyanin extracts 100 mg/kg) (5) treatment group 3 (ligation+the phycocyanin extracts 200 mg/kg). From the beginning of the experiment D0, normal saline or different concentrations of the present invention contains phycocyaninextraction is given till Day 10. At D3, rat is induced periodontal disease by ligature. Rat are sacrificed at Day 11, and changes of the height of the alveolar bone are observed (FIG. 3)

(16) 4. The Measurement of the Height of the Alveolar Bone

(17) After removal of the soft tissues of the rat lower jaw, verify the position of the cemento-enamel junction (CEJ), and measure the distance between the CEJ to the alveolar bone crest (ABC). (FIG. 4A) The experiment result of the Ligation group shows that whether the lingual side or the buccal side the alveolar bone crest both have a serious bone matrix absorption. However, while the rat feed the present invention phycocyanin extraction (C-PC) for 100 mg/kg (treatment group 2) and 200 mg/kg (treatment group 3), the distance between the CEJ to the alveolar bone crest (ABC) is shorter than the Ligation group. (FIG. 4B) There is no difference between the two doses of each other.

(18) Furthermore, observe the experiment rat's alveolar bone loss phenomenon through tissue staining. The mark d means dentin, g means gingiva, pl means periodontal ligament and ab means alveolar bone. The experiment result of the Ligation group shows more serious alveolar bone loss than the treatment group. The treatment group 2 (100 mg/kg) and 3 (200 mg/kg) have significant improvement of the alveolar bone loss situation. Therefore, the present invention of the phycocyanin extracts can effectively improve the alveolar bone loss situation.

(19) Based on the above result, the present invention of the phycocyanin extracts can effectively inhibit the growth of the oral pathogens, improve the alveolar bone loss situation and reduce the inflammation caused by those pathogens. Finally, the present invention of the phycocyaninextracts can treatment and/or prevent of periodontal disease.

(20) Although the present invention has been described in terms of specific exemplary embodiments and examples, it will be appreciated that the embodiments disclosed herein are for illustrative purposes only and various modifications and alterations might be made by those skilled in the art without departing from the spirit and scope of the invention as set forth in the following claims.