:m: ~

THE UNIVERSITY OF HONG KONG

* ~

Bachelor of Engineering

Medical Engineering Programme

MEDE4604
Cell and tissue engineering
Examination

Date: May 11, 2016 Time: 2:30 pm - 5:30 pm

Attempt FOUR questions.

All questions carry equal marks.

Use of Electronic Calcnlators:

Only approved calculators as announced by the Examinations Secretary can be used in this
examination. It is candidates' responsibility to ensure that their calculator operates
satisfactorily, and candidates must record the name and type of the calculator used on the front
page of the examination script.

MEDE4604 Page 1of4

Question 1

(a) A researcher takes a small cartilage biopsy at the non-load bearing area from the knee
joint of a patient with cartilage injury. He wishes to raise enough chondrocytes (cartilage
cells) for autologous chondrocyte implantation (ACI). However, he found that, during the
in vitro culture, the chondrocytes isolated from the biopsy showed limited proliferation
potential that it is difficult to obtain sufficient numbers for clinical applications. Explain
this phenomenon and discuss the potential molecular mechanisms behind this
phenomenon. (5 marks)

(b) Stem cells are known to have better proliferative potential than that of mature cells.
Briefly discuss the reasons behind this phenomenon. (2 marks)

(c) Stem cells are known to have multiple differentiation potential. What is multiple
differentiation potential and why this characteristic is important for tissue engineering?
(3 marks)

(d) Name two methods to obtain embryonic stem cells that are genetically matched to
recipients for therapy (i.e. patient-specific embryonic stem cells), outline briefly how to
obtain these cells with the aid of sketched diagrams and discuss their respective
advantages and problems. (15 marks)

Question 2

(a) A company develops a chemoattractant with the potential to induce blood vessel
formation. The R&D manager responsible for this work needs to design experiments to
evaluate the efficacy of this potential chemoattractant in inducing migration of endothelial
cells. With the aid of schematic diagrams with appropriate labels, design and describe two
experiments, with appropriate controls, to study the effects of this chemoattractant in
stimulating (i) population migratory properties and (ii) single cell migratory properties.
Briefly discuss the expected results. (13 marks)

(b) Another R&D manager in this company is leading a project on cartilage tissue
engineering using bone marrow cells. She found that the normal cell density in a native
cartilage is 106 cells I cm 3 • What is the number of cells needed to build a structure for
repairing a cylindrical cartilage defect with a diameter of 1 cm and a thickness of O. lcm?
If she can successfully isolate 105 mesenchymal stem cells from a patient, how long does
it take to expand the cells to a sufficient number in cultures for repairing the cartilage
defect, assuming the doubling time for bone marrow mesenchymal stem cells is 18 hours?
(3 marks)

(c) A third R&D manager is developing a liver graft. He seeded 107 stem cells onto the
scaffold with a disc-shaped porous scaffold with a diameter of 4 cm and a thickness of 1
cm. Assume the cells can effectively penetrate into the scaffold and evenly distribute in it.
Assume the volume of the cell-populated scaffold is equivalent to 12.56 ml. If the
diffusion coefficient of oxygen in the hepatocytes-seeded construct at 37°C is 2x10· 5

MEDE4604 Page 2 of 4
 cm2/sec and the oxygen consumption rate of chondrocytes is 4x10- 18 mole/cell/sec_
Calculate the minimal oxygen concentration needed in the culture medium in order to
prevent central necrosis in the liver graft. (6 marks)

(d) Another R&D team is developing a skin project. They prepared skin substitutes from
animal skins by discarding the animal cells while retaining the extracellular matrices.
Name the process and give 3 example approaches to discard animal cells from animal
tissues. (3 marks)

Question 3

(a) With the aid of a schematic diagram, discuss how to apply the knowledge about the
interactions between cells and at least 4 different types of signals in tissue engineering
applications? (12 marks)

(b) Name a class of proteins, which act as important mediators of cell and extracellular matrix
interactions. With the aid of a schematic diagram, give three examples of cell and matrix
adhesions. (7 marks)

(c) Briefly describe an approach to achieve the following aims during tissue engineering.
(6 marks)
i) To improve the surface property of a scaffold for cell binding;
ii) To enhance the mechanical properties of a scaffold to mimic that of the bone;
iii) To induce certain alignment or orientation of the cells seeding on a scaffold;
iv) To reduce the immunogenicity of a xenogenic skin graft;
v) To improve the integration between a tissue engineered construct and the host
tissue;
vi) To produce a porous scaffolds to allow cell penetration.

MEDE 4604 Page 3 of 4

Question 4

(a) Growth factors are known to control growth rate of cells. Briefly describe the phenomenon
observed in the following chart and explain the phenomenon: (5 marks)

EDD
500
0
8 400

"
;; 300

"
u· 200

100 [C] is concentration of a growth factor

o~~~~~~~~~~~~~~~~~~

50 100 150 200 250

-iOO
!CJ

(b) Assume the maximal achievable growth rate (µmax) of this growth factor on a particular
cell type is 1.5 x 105 cells per day and the dissociation constant (Km) of the growth factor
and its receptor at the surface of this cell type is 1OnM. Calculate the growth rate when the
concentration of this growth factor is 2nM and 200nM. (4 marks)

(c) State 5 major challenges in liver tissue engineering. (5 marks)

(d) A tissue engineering company is developing a bioartificial liver (BAL) system for
extracorporeal support of end stage liver disease patient. With the aid of a schematic
diagram, discuss the working principles of the BAL system. (8 marks)

(e) Give 3 examples on the ethical issues of tissue engineering. (3 marks)

***END OF PAPER***

MEDE4604 Page 4 of 4