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Biomaterials Lab #2 Handout

This document outlines Lab #2 on synthetic polymer polycaprolactone (PCL) for a biomaterials course. It introduces PCL and describes two techniques for processing PCL - electrospinning and solvent casting with particle leaching. For electrospinning, the document explains the setup, safety considerations, and experimental protocol for producing electrospun PCL membranes. For solvent casting with particle leaching, it describes the process of creating porous PCL structures using porogens like NaCl that are later leached out, and provides the experimental protocol. Students are instructed to discuss in their report how these techniques can be applied in biomedicine and answer questions about porosity and optimal pore size for bone tissue engineering scaffolds.

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Mai Phương Lê
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49 views

Biomaterials Lab #2 Handout

This document outlines Lab #2 on synthetic polymer polycaprolactone (PCL) for a biomaterials course. It introduces PCL and describes two techniques for processing PCL - electrospinning and solvent casting with particle leaching. For electrospinning, the document explains the setup, safety considerations, and experimental protocol for producing electrospun PCL membranes. For solvent casting with particle leaching, it describes the process of creating porous PCL structures using porogens like NaCl that are later leached out, and provides the experimental protocol. Students are instructed to discuss in their report how these techniques can be applied in biomedicine and answer questions about porosity and optimal pore size for bone tissue engineering scaffolds.

Uploaded by

Mai Phương Lê
Copyright
© © All Rights Reserved
We take content rights seriously. If you suspect this is your content, claim it here.
Available Formats
Download as PDF, TXT or read online on Scribd
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Vietnam National Universities – HCMC

International University
School of Biomedical Engineering

Course of Biomaterials
Semester 1 2022-2023

LAB #2
SYNTHETIC POLYMER (POLYCAPROLACTONE)

I. Outline
 Overview of polycaprolactone material.
 Overview of electrospinning technique.
 Overview of solvent casting and particle leaching technique.
 Experimental protocols.
 Report highlights.

II. Polycaprolactone material

Fig 1. Chemical structure of polycaprolactone (PCL)

III. Electrospinning technique


 Electrospinning is a versatile and viable technique to generate membrane with ultrafine
fibers up to nanoscale based on synthetic and natural polymers. Regarding biomedical field,
electrospinning is applied for wound dressing, scaffold for tissue engineering, bioactive
agent delivery, cosmetic, etc.
 The fundamental principle of electrospinning includes (1) ejecting a polymer solution out
of the syringe needle by a syringe pump, (2) charging the solution droplet and forming a
Taylor cone through a high voltage source (usually 10-20 kV), (3) stretching and whipping
the polymer fibers out of the droplet toward the oppositely charged collector while
evaporating the solvent along the way, (4) leaving solid fibers onto the collector.
 Safety must be completely followed while working with high-voltage source, since the
effects of electric shock can range from burn to cardiac arrest.
+ Wear insulating gloves, shoes and handle with insulating tools.
+ Monitor the voltage and discharge current of the equipment closely.
+ Ensure grounding connection.
Vietnam National Universities – HCMC
International University
School of Biomedical Engineering

+ Never leave the unit door open.


 Safety related to solvent hazard and chemical disposal should be noted.

Fig 2. Electrospinning schematic

Fig 3. Electrospun fibers deposited onto the collector (doi: 10.1021/ma0608673)

IV. Solvent casting and particle leaching technique


 Solvent casting and particle leaching technique is a common method to create porous 3D
structures based on synthetic polymer, which are mostly applied as a scaffold for tissue
engineering. The typical particles for leaching step, known as porogens, include water-
Vietnam National Universities – HCMC
International University
School of Biomedical Engineering

soluble organic compound (such as gelatin, glucose) and inorganic salts (such as NaCl,
KCl).
 The fundamental principle of solvent casting and particle leaching technique includes (1)
dissolving polymer in a highly volatile solvent, (2) adding porogen(s) into the mixture, (3)
casting the solution into a mold, (4) evaporating solvent, (5) immersing the structure into
water to dissolve the porogen(s), and (6) drying the structure.
 Safety related to solvent hazard and chemical disposal should be noted.

Fig 4. Schematic of solvent leaching and particle leaching technique, based on PLGA and
NaCl (doi: 10.1089/jop.2015.0126)

V. Experimental protocols
1. Electrospinning protocol
Step 1: Prepare PCL solution 12% w/v in pure acetone solvent by stirring overnight at initial
60oC for 2 hours and then at room temperature.
Step 2: Load the PCL solution into 10-ml plastic syringe.
Step 3: Electrospin the syringe containing PCL solution with the following set-up conditions:
needle-to-collector distance of 10 cm, flow rate of 1 ml/ h, voltage supply of 15 kV. Turn off
the power supply once in a while to wipe the clog formed on the needle.
Step 4: Shut down the electrospinning equipment. Detach the membrane from the collector,
then store and label the membrane appropriately.
Note: You are guided with the main steps and the electrospinning equipment at lab 410a and
do not directly perform the experiment.
2. Solvent casting and particle leaching protocol
Vietnam National Universities – HCMC
International University
School of Biomedical Engineering

Step 1: Prepare PCL solution 33% w/v in pure acetone by stirring overnight at initial 60oC for
2-4 hours and then at room temperature. Divide it into two vials and weigh the PCL solution
in each vial.
Step 2: Sieve the raw sodium chloride and sucrose grains, each with a weight 2.5 times higher
than the PCL solution.
Step 3: Add the grains above to the corresponding PCL vial, then quickly stir the mixture using
a glass rod in 10-15 seconds and pour the slurry into a cylindrical glass mold with 10-mm
diameter and about 40-mm height. Hand pressure is applied with the glass rod to create a firm
structure in the mold and eliminate bubbles.
Step 5: Dry the mold at room temperature for at least 48 h.
Step 6: After the structure is completely dried, detach it using the glass rod. Then immerse the
dried structure into DW for 48 h, with several water replacements in the process.
Step 7: Dry the structure at room temperature for at least 48 h.

VI. Report highlights


 Introduction section should include brief information of:
+ Background of synthetic polymer and its use as biomaterial.
+ Background of PCL material.
+ Background of the two covered techniques (further explanation of key steps in the
protocol, advantages/ disadvantages, how can we apply the membrane and porous
structure in biomedical field, etc.)
 Questions (for Result and Discussion section)
1. How can porosity benefit the material structure?
2. What is the optimal pore size for structure applied in bone tissue engineering? Why?

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