The Development and Application of Davallia mariesii in Taiwan for Osteoporosis

Osteoporosis is second as a global healthcare problem which is increasing in significance as the population of the world both grows and ages. Fractures and its associated complications due to osteoporosis have a serious impact on a person's health. It is an important subject to exploit more effective ways for treating or preventing osteoporosis, reducing social costs of health care, and improving the quality of life. Davallia mariesii Moore ex Bak. is one of the original plants of “GU-SUI-BU”, it can be collected from low and middle elevation in Taiwan. Human primary osteoblast cells (HObs) were used as an in vitro model for screening the active samples. In our preliminary results, H2O and 50% EtOH were used the solvents to extract the materials and found that H2O extracts of D. mariesii showed no obvious toxicity to HObs and increased the calcium contents of the mineralized matrix by 107.1 士 4.52%. The leaves with sporophyte were inoculated, lots of embryogenic callus are proliferated, then the somatic embryos can regeneration into young sporophytes (by Co-PI). The H2O extract of regenerated plants from D. mariesii also showed no obvious toxicity to HObs and increased the calcium contents of the mineralized matrix by 194.76 士 14.64%. The large scale production of regenerated plants from D. mariesii will be evaluated the active components and compared the difference with wild D. mariesii. There is no literature describing the association of osteoporosis with D. mariesii, therefore, we assess a project proposal entitled “The development and application of Davallia mariesii in Taiwan for osteoporosis”. The entire project goal will be carried out into three-year: The 1st year project: (1) To investigate the active components of D. mariesii. Using HObs as the in vitro screening platform, the active fractions will be done as followed. After passing through the various chromatographic columns, the pure constituents will be isolated. The structures of isolated active constituents will be identified by various physical data and spectra. To further evaluate the active constituents by using HObs in vitro screening platform. (2) The regenerated plants from D. mariesii will be large scale produced by Co-PI (Prof. Ho), the activity will be confirmed. The 2nd year project: (1) According to the 1st year project’s strategies, the separation, purification, and structural analysis of the active components of regenerated plants from D. mariesii will be investigated. (2) To evaluate the extracts of both wild and regenerated plants from D. mariesii by using ovariectomized (OVX) female C57BL/6 mice as a model for postmenopausal osteoporosis. The 3rd year project: (1) The molecular mechanisms in HObs of the isolated active compounds from 1st and 2nd year projects will be evaluated. (2) Based on the single nucleotide polymorphism (SNP) analysis of patients with osteoporosis, brain-derived neurotrophic factor (BDNF) gene was found to be associated with osteoporotic fracture (by Co-PI, Prof. Chang). The novel mechanism of active components will be further evaluated. (3) To evaluate the active fractions/components by using OVX female C57BL/6 mice model. (4) To develop the chemical fingerprint of the active components of the active fractions from D. mariesii by using LC-MS-MS.