90874 - Applied Biochemistry.

Learning outcomes

“Applied Biochemistry” has as objectives the study of the theoretical foundations, the instrumental implementations and the main techniques used in cosmetics, in the preparation and analysis of samples for biochemical investigations. The student: o knows the physical and chemical principles underlying the main techniques and instruments employed in biochemical investigations on cosmetic products with reference to the new in vitro tests; o knows the different parameters that may be modified to change biochemical analysis on cosmetic products; o includes a working protocol and the main variables on which to act to optimize it; o can describe the structural elements of the main tools of a biochemical laboratory/cosmetology; o knows the correct terminology used in biochemical methods in cosmetics; o can use the knowledge acquired to evaluate the results of an experiment, identify the problems and optimize it; o knows how to evaluate the possible impact of changes of the parameters in an experiment for the evaluation of corrosion and skin irritation, skin absorption measures and in the main imaging techniques to measure the phototoxicity; o can orientate between the main qualitative and quantitative methodologies, to select the one appropriate to the study of the problem with reference to the new in vitro tests available for the analysis of cosmetic products.

Course contents

Module I - Frontal lectures (40 hours)

Biochemical laboratory and safety.

Cell culture techniques: cell culture laboratory and equipment, safety considerations in cell culture, aseptic techniques and good cell culture, animal cell types, characteristics and maintenance in culture, stem cells and bacterial cell cultures, 2D and 3D cultures.

Centrifugation: basic principles of sedimentation, types, care and safety aspects of centrifuges, preparative centrifugation, analytical centrifugation.

Microscopy: optical microscope, optical sectioning, imaging living cells and tissues, measuring cell dynamics, electron microscope (EM), image archiving. Examples of applications in cosmetics

Immunochemical techniques: antibody production, immunoassay methods, immuno-microscopy, lateral flow devices, epitope mapping, immunoblotting, fluorescent activated cell sorting, cell and tissue staining techniques, pills of immuno-capture polymerase chain reaction (PCR), immuno-afidity chromatography, antibody-based biosensors, therapeutic antibodies.

In vitro evaluation of the pro-sensitising potential of a cosmetic product.

Proteins: structure (particular reference to skin and hair), keratin, purification, characterisation and functional analysis.

Electrophoretic techniques: general principles and media, protein and nucleic acid electrophoresis, capillary electrophoresis.

Module II - Laboratory exercises (15 hours)

Protein colorimetric quantification by spectrophotometer and discussion of the results.

Elisa test

Histochemical assays:
-Hematoxylin and eosin staining, miscroscopic analysis
-Syrius Red-Fast Green, spectrophotometer reading and calculation of collagen and non-collagen proteins.

 

 

 

Readings/Bibliography

"Principles and Techniques of Biochemistry and Molecular Biology"

Seventh Edition

Edited by Keith Wilson and John Wolker

 

"Principles and Techniques of Practical  Biochemistry"

Fifth Edition

Edited by Keith Wilson and John Wolker

 

 

Teaching methods

Lectures supported by powerpoint presentations of each topic of the course will be shown. 

The course is based on both classroom lectures (40 hours) and laboratory exercises (15 hours).

Classroom lectures will develop in a linear fashion with an initial general introduction to each topic with references to general biochemistry and then the development of analytical techniques. The teacher will stimulate critical learning and classroom discussion by the students. Laboratory work will be developed both through individual exercises and in collaboration between small groups of students.

Assessment methods

Learning will be assessed by a final oral examination on the course content.

The final mark will also take into account the attendance and active participation of students in laboratory work.

Teaching tools

PC, overhead projector, biological laboratory

Office hours

See the website of Valeria Righi