Berberine has been used clinically for more than a decade to treat various diseases, has been shown to be effective in osteoblast differentiation, and is a potential treatment option for osteoporosis. However, compared with existing osteoporosis drugs, berberine is somewhat less effective. This study aimed to identify a new compound with efficacy superior to that of berberine. The osteogenic activities of various berberine derivatives were evaluated via cell differentiation in C2C12 preosteoblast cell lines. Alkaline phosphatase (ALP) staining assay and structure– activity relationship demonstrated that compound 2b had a potent osteogenic effect. Furthermore, compound 2b dose dependently increased ALP activity and showed no toxicity at the effective concentration, indicating its efficacy. Additionally, compound 2b upregulated BMP2-induced transcriptional activity in a promoter activity assay using ALP, BSP, and OC promoters.

Introduction
Materials and Methods
    1. 세포 배양과 조골세포 분화 유도
    2. 항체와 시약
    3. Alkaline phosphatase 염색 및 분석
    4. 일시적 형질주입과 발광효소 분석법
    5. 웨스턴 블롯
    6. 세포 독성 측정
    7. 통계분석
Results
    1. Berberine 유도체들의 alkaline phosphatase 염색 결과
    2. Alkaline phosphatase 염색 결과를 통한 berberine 유도체들의 구조와 활성 관계 분석
    3. Compound 2b 가 조골세포 분화에 미치는 영향
    4. C2C12 세포에 대한 세포독성 평가
    5. Compound 2b 가 조골세포 분화 전사 인자에 미치는 영향
    6. Compound 2b 가 골형성 표지자 전사 활성에 미치는 영향
Discussion
References

• Younho Han(Department of Oral Pharmacology, Wonkwang University College of Dentistry)
• Won-Jong Park(Department of Oral Pharmacology, Wonkwang University College of Dentistry/Department of Oral and Maxillofaical Surgery, Wonkwang University College of Dentistry) Correspondence to