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I NVESTIGATING EFFECTS OF IGF-1 C ISSUS QUADRANGULARIS AND F AGUS CRENATA ON THE PRODUCTION OF COLLAGEN IN MOUSE FIBROBLAST CELLS Done by: Cheng Yong Jian.

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Presentation on theme: "I NVESTIGATING EFFECTS OF IGF-1 C ISSUS QUADRANGULARIS AND F AGUS CRENATA ON THE PRODUCTION OF COLLAGEN IN MOUSE FIBROBLAST CELLS Done by: Cheng Yong Jian."— Presentation transcript:

1 I NVESTIGATING EFFECTS OF IGF-1 C ISSUS QUADRANGULARIS AND F AGUS CRENATA ON THE PRODUCTION OF COLLAGEN IN MOUSE FIBROBLAST CELLS Done by: Cheng Yong Jian Er Yuan Zhi Allison Meadows Gabi Guzman

2 C ONTENTS Introduction Rational Objectives Hypothesis Materials Methods Timeline References

3 I NTRODUCTION Tendinosis is a chronic form of tendinitis which occurs when mostly type-1 collagen that composes the tendon degenerates through repetitive motion and ageing (Summers, 2003) Current treatments for tendon repair often contain compounds that stimulate collagen production (Haukipuro, 1991)

4 I NTRODUCTION Current treatments for tendon repair IGF-1 is a growth hormone that has been shown to be an in-vitro stimulant of type-1 collagen synthesis (Ivarsson, 1998) Studies by J Indian Medical Association have shown that the daily use of C.quadrangularis increases the body’s collagen turnover rate

5 I NTRODUCTION Fagus Crenata is known to exhibits a superior effect of prevention of aging by promoting the production of collagen in the dermal fibroblast cells. (Y., Nakayama, N., Kojima, 1998) Studies have also shown that F.Crenata have the effect of strongly promoting dermal fibroblast collagen biosynthesis (Takada et al, 2007)

6 R ATIONAL We aim to investigate the effectiveness of Cissus quadrangularis, IGF-I and Fagus crenata to stimulate the synthesis of collagen in mouse tendon fibroblast cells to develop new treatments for tendonitis.

7 O BJECTIVES Determine the effectiveness of IGF-1, Cissus quadrangularis and Fagus Crenata on the stimulation of synthesis of collagen in mouse tendon fibroblast cells

8 H YPOTHESIS Our hypothesis is that: IGF-1 increase production of collagen synthesis in mouse tendon fibroblast cells and Cissus quadrangularis and Fagus crenata increase production of collagen synthesis in mouse tendon fibroblast cells when used both separately and together with IGF-1

9 V ARIABLE Amount and type of fibroblast cells treated Environmental conditions Controlled Variable Treatment of fibroblasts cells Type of herbs used (For AOS and HCI) Concentration of substances added to fibroblast cells Independent Variable Amount of collagen produced by fibroblast cells Dependent Variable

10 M ATERIALS 3T6 Swiss Albino Nuclear Lystate Cissus quandrangularis Fagus Crenata IGF-1 Tissue Culture flask – 25cm 2 CO 2 incubator Dulbecco’s Modified Eagle Medium Variable volume micropipettes and pipette tips Vortex mixer Centrifuge machine Spectrophotometer Masson Trichrome staining kit Sircol Assay Kit

11 M ETHODS Tests will be carried out in vitro with IGF-1 and C.quadrangularis ( F.crenata for HCI) in DEM medium at different concentrations Fibroblast cells would be separately treated with IGF-1, C.quadrangularis ( F.crenata for HCI) and a combination of both. For the control, it will be left to grow in normal conditions Mouse fibroblast cells will be cultured in a CO2 chamber at 37°C at a volume of 0.25ml/cm 2 for a period of 5 days

12 M ETHODS Masson Trichrome staining will be done on the cells and ImageJ will be used measure the stained collagen Sircol collagen assay will be done to determine the exact concentration of the amount of collagen produced by the mouse fibroblast cells Triplicates will be carried out and the results obtained would be compared with the control

13 T IMELINE DateAOSHCI Nov-FebDetermine the concentration of substances to be experimented on Read up more articles regarding project Feb-MarchExperiments in progress -Obtain 1 st set of data April-MayExperiments in progress -Obtain 2 nd set of data June-JulyExperiments completed -Obtain 3 rd set of data 9 th JulyExperiments in progress Preparation of Project’s Day Semi- Finals 21 st AugustPreparation for Project’s Day Finals presentation

14 R EFERENCES Cissus Quadrangularis— The Best Kept Secret in Bodybuilding. (2007). Retrieved September 28, 2009, from USPLabs Web site: Erickson, L. (2002). Future treatments. Retrieved May 21, 2009, from Erickson, L. (2002). The Tendinosis Injury. Retrieved September 24, 2009, from Takda, K., Suzuki, R., Guentert, M., Inomata, S., Hamada, C., and Sakiguchi, T., (June ) Anti- Aging composition and collagen production promoting composition

15 R EFERENCES Borg, T. K., Ivarsson, M., McWhirter, A., Rubin, K. (1998). Type I collagen synthesis in cultured human fibroblasts regulation by cell spreading, platelet-derived growth factor and interactions with collagen fibers. Matrix Biology. 16, Daha, M. R., Geest, R. N., Lam, S., VanKooten, C., Verhagen, N. A. M. (2004). Secretion of collagen type IV by human renal fibroblasts is increased by high glucose via a TGF-B- independent pathway. Nephrol Dial Transplant, 19, Leeson, C. R., Leeson, T. S., & Paparo, A. A. (1985). Connective tissue proper. Textbook of Histology. Philidelphia: W. B. Saunders Company.

16 R EFERENCES Ghahary, A. ‌, Houle, Kilani, T., Scott, G. ‌, Shen and Tredget, E. ‌, ‌ (2000), Vol. 17, No. 3, Pages Mannose-6-Phosphate/IGF-II Receptors Mediate the Effects of IGF-1-Induced Latent Transforming Growth Factor β1 on Expression of Type I Collagen and Collagenase in Dermal Fibroblasts Baroni, G., Benedetti, A., Casini, A., Folli, F., Gaglotti, G., Macarri, G., Marucci, L., Orlandoni, P., Perego, L., Ridolfi, F. and Sario, A. (1999) Insulin and Insulin-Like Growth Factor-1 Stimulate Proliferation and Type I Collagen Accumulation by Human Hepatic Stellate Cells: Differential Effects on Signal Transduction Pathways

17 Thank you for your kind attention!


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