Vietnam National Universities – HCMC

International University
School of Biomedical Engineering

Course of Methods and Process in Fabrication of Scaffold

Semester 1 2023-2024
LAB #4
PARTICLE LEACHING (cont.) AND SPONGE REPLICA METHOD
I. Outline

 Porosity investigation of particle leaching scaffold

 Overview of sponge replica method.
 Experimental protocols.

II. Scaffold porosity

 Porosity in the context of a scaffold refers to the void spaces or pores within the
material's structure. Porous scaffolds are used in various applications, where the
controlled porosity is crucial for specific functions. The porosity of a scaffold impacts its
ability to support cell growth, provide surface area for reactions, or allow the flow of
fluids.
 There are two primary types of porosity in scaffolds: open porosity (interconnected
pores) and closed porosity (isolated pores). Open porosity is more desirable for
applications like tissue engineering, as it allows for nutrient and waste exchange.
 The porosity of a scaffold can be controlled during fabrication. Factors like the choice of
material, fabrication technique, and processing conditions influence the porosity of the
final product. The desired porosity of a scaffold depends on its intended function. For
example, in bone tissue engineering, the scaffold should mimic the porosity of natural
bone to support cell ingrowth and vascularization.

Fig 1. Images of (a) open-cell (interconnected pores), and (b) closed-cell (isolated pores)
scaffold. (doi: 10.1016/j.mser.2020.100589)
III. Sponge replica method

 Sponge replica (or replication) method is the most common method to create porous
structure based on bioceramic material, which was mostly used as filling material before.
 The fundamental protocol of sponge replica technique includes (1) preparing bioceramic
slurry with sufficient viscosity, (2) preparing the sponge template with suitable size and

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 Vietnam National Universities – HCMC
International University
School of Biomedical Engineering

shape (mostly from PU material), (3) impregnation of the template within ceramic
suspension for one or multiple times, (4) drying, burning out the sponge template and
sintering the ceramic material through a controlled heating process.
 Safety related to high heat treatment should be noted.

Fig 2. Schematic of sponge replica technique to create bioactive glass-based scaffold (doi:
10.3390/ma14112795)
IV. Experimental protocols
 Porosity assessment of PCL scaffold
Step 1: Measure the mass of the scaffold using analytical balance.
Step 2: Measure the volume of the scaffold by measuring the height, diameter…
Step 3: Apply the porosity formation using the (1) equation:
ρ polymer − ρscaffold
ϵ= ×100 (1)
ρ polymer

ϵ represents the porosity of the scaffold (%).

ρ polymer is the density of the polymer (g/cm3).
ρ scaffold is the measured density of the scaffold (g/cm3).
Step 4: Fill in the below table to compare the porosity of the scaffolds of other group.
Table 1. Porosity comparison of particle leaching scaffolds

Porosity (%)
Sample Suitable for which tissue
NaCl MSG
Your Group

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 Vietnam National Universities – HCMC
International University
School of Biomedical Engineering

(Please include this table on your lab 3 report – and give some discussion about the
comparison of porosity)
 Sponge replication using PU foam
Step 1: Dissolve 0.7 g of polyvinyl alcohol (PVA) in 7 mL of distilled water at 80 oC with
constant stirring.
Step 2: Create a 5 mL suspension of Biphasic calcium phosphate (BCP) with a concentration
of 20% w/v for Groups 1, 2, and 3, or 40% w/v for Groups 4, 5, and 6, using a sonicator for
15 minutes.
Step 3: Carefully mix 3 mL of the BCP suspension with the PVA solution while
continuously stirring, resulting in a viscous slurry.
Step 4: Prepare a polyurethane (PU) sponge with dimensions measuring 1 cm x 1 cm x 0.5
cm.
Step 5: Squeeze the sponge to eliminate trapped air and promptly immerse it in the slurry for
5 minutes. Then, dry it in an oven at 80 oC for 30 minutes. Repeat the immersion and drying
steps twice.
Step 6: Heat the sample to 1200 oC and maintain this temperature for 1 hour using a muffle
furnace (model L 15/13/P330 by Naberthem).
Note: scanning electron microscope (SEM) images of the resulting scaffold structures will be
provided later
This manual is developed by
 PhD. Doan Ngoc Hoan, Instructor
 Luong Dai Tin, Teaching assistant
 Le Tran My An, Teaching assistant
Last updated: Semester 1, 2023-2024