Biocompatibility suggestions - (Dec/27/2004 )
Currently i'm looking forward a new project regarding the biocompatibility testing on human skin via a kind of chemical...might be honey,aloe vera,chitosan...etc.
so, what aspect can be suggested other than cytotoxicity, genotoxicity...??
it can be also related to other reseach areas as well .
thanks!! ur suggestions will be well appreciated!!
A major biocompatibility test will be immunoresponse elicited by your biomaterials - both acute (b-cell & complement-mediated) and chronic (t-cell mediated).
thanks! let say that we examine only the biocompatibility on human epidermal keratinocytes,anything else can be further related?
If you pursue just the in vitro model (epidermal keratinocytes) then I would say first, a focal gene profiling analysis (microarray) with or without your biomaterial; second, target, select and further characterization of the cluster genes showing out in this assay as your reference point (for further biomaterial optimization toward more biocompatible); last, move to in vivo model.
thanks postdoc!! that's a good suggestion!! i ve came across in mind earlier of doin further works related to molecular level as well. basically im looking forward the research on this interleukin-1 by kerationocyte and its gene expression toward biomaterials' compatibility on human.
now, im on the stage to make a concise proposal. pls kindly to teach me or advance my knowledge if u had any fresh and interesting and still about the microarray analysis. i would really appreciate it!!
I think a more global view on gene expression prolifing will help to create a clearer picture on biocompatibility of your test materials, also you can confirm and focus on your prime target - IL-1 pathway. I would suggest a focal gene chip (custom made) array on IL-1 pathway for (1) easier and simpler data analysis; (2) cost-effective. Once you feel comfortable with the results then expand outward to other related pathways...
Another key is the "time points" you choose in this analysis may directly reflect what target gene you're looking for - immediate early genes, intermediate genes or more downstream targets... therefore, a pilot study will be needed to determine the right timing for microarray analysis.
Finally, I heard that cytokines/growth factors from several vendors display different impact (due to inconsistent property/quality). If you have a chance to use IL-1, make sure it is from the most reliable sources (Epoch Biolabs, R&D Systems.....).
Again thanks to u,postdoc!! i really appreciate what u mentioned earlier! u've gone to the good points, indeed this suggested microarray analysis will definitely help in a clearer understanding of gene expression related to the biocompatibility of chemical substances goin 2b tested.
of course cytokines and growth factors are inportant measurements in the test of biocompatibility onto human. the in vivo part will be done by implantation procedures (might be further checked as well, works load due to slides processing for tissue and cells observation, swelling due to some contaminations).
hav u ever used a kit to measure cytokine production level after cells were treated with chemicals??
HAPPY NEW YEAR!!
Yes, many many times (to further confirm gene expression results at protein level). Be sure to use ultrasensitive ELISA kit to measure interested cytokines released in culture medium - usually in sub-picogram per mL range. My experience for TNF, IL-1, MCP-1 are in most cases they usually peaked at 24-48 hr post-stimulation, but it may also depend on cell types and stimuli you use.
Also remember to incorporate a positive control (cells treated with a stimulus to release a specific cytokine) since I would predict your some chemicals will be weak inducers of IL-1 or other cytokines.....
yes,postdoc,ur right! i think i would survey for some cytokines which r mostly produced in the in vitro culture, b4 further to the gene expression of the certain kind of cytokines.
later, i might hav to propose some idea regarding gene therapy on skin diseases as well, but it takes time...i'll only do it after 2 years..!
r u doing about the similar research on keratinocyte in vitro culture b4, or currently engaged?
I'm fortunate enough that I've been in different fields in both academic and industrial sectors for several years. Basically, what you have come through, what you will encounter, I've been there.