An Incubator for Biomaterials Lab and Tissue Culture Experiments

Project Title: An Incubator for Biomaterials Lab and Tissue Culture Experiments

Project Lead’s Name: Amy Yousefi


Phone: (513) 529-0766

Please Choose the Primary Affiliation: CEC

Are There Other Project Team Members?: No

Brief description of project: Providing hands-on experience in undergraduate labs and offering experiential learning to students through independent studies and senior capstone projects are key elements of undergraduate education. We currently have over 400 undergraduate students at the Department of Chemical, Paper and Biomedical Engineering (CPB). On average, 40 undergraduate students take the biomaterials course each spring (CPB 519/519, 3 credit hours). This is a core course required for the bioengineering major and a technical elective for the chemical engineering major. In this course, students are asked to use the freeze-thaw technique to fabricate hydrogels (cryogels) as potential candidates for soft-tissue replacement (lab experiment). The produced cryogels can also serve as soft tissue phantoms, enabling to uncover potential weaknesses in medical imaging systems (brain and liver elastography). These gels are made of hydrophilic polymers, such as poly(vinyl alcohol) (PVA), and need to be characterized in wet and dry states to quantify their water uptake. Since biological tissues have different physical/mechanical properties at body temperature (37?C) and at ambient temperature, the current measurement of water uptake at ambient temperature does not reflect the expected properties at physiological conditions. The incubator requested in this proposal can greatly benefit the students enrolled in CPB 419/519. This equipment can also serve CPB's senior design projects (CPB 471/472) and independent studies (CPB 277/377). In the past 7 years, my lab has served over 50 undergraduate students on the development of 3D scaffolds for tissue engineering (senior design and independent studies) as well as 8 graduate students on their MS thesis on biomaterials (CPB 700). It is essential to maintain the cell-seeded scaffolds in an incubator during cell culture period. This equipment will also greatly contribute to Undergraduate Summer Scholar projects (CPB 340U) at CPB.

Does this project focus on graduate student education or graduate student life?: No

Describe the problem you are attempting to solve and your approach for solving that problem.: When offering labs to undergraduate students, the quality of measurements directly affect the learning outcomes, via meaningful interpretation of collected data. In 2015, students characterized the cryogels at room temperature. Hence they were unable to see the effect of physiological conditions on cryogel water take-up (swelling). An incubator will enable a comparison between swelling at physiological conditions (temperature and pH) and ambient temperature for these biomaterials (via Student's t-test statistical analysis). Excessive swelling is not desired for cryogels used as soft tissue replacement candidates. As previously indicated, the CPB department also offers research opportunities for undergraduate students as independent study (CPB 277/377) and senior design projects (CPB 471/472), including over 50 students in my lab since 2010. Therefore, these students as well as graduate students will have access to this equipment.

The criteria state that technology fee projects should benefit students in innovative and/or significant ways. How would you describe the innovation and/or significance of your project?: The details of the educational outcomes are listed under the following question (Biomaterials Lab). The equipment will also be used for senior design projects, independent studies, and graduate projects. Figure 1 (attachment) shows the scaffolds that we have fabricated in our lab. The macro-channels have been generated after extracting poly(ethylene glycol) (PEG) in water. First, the PEG grid is produced by additive manufacturing (3D-Bioplotter), then immersed in a polymer solution containing ceramic nanoparticles, and finally subjected to thermally-induced phase separation (creating nanoscale surface features and micropores). The PEG is then extracted by water to create the macro-channels. These hierarchical scaffolds with micro-, macro- and nano-scale features can contribute to cell attachment and growth via different mechanisms. The requested CO2 incubator will be used in the final step for seeding these scaffolds with human stem cells (hMSCs) for bone tissue engineering.

How will you assess the project?: Figure 2 (attachment) shows some cryogels produced by the freeze-thaw technique. The material used for producing these gels is an aqueous solution of poly(vinyl alcohol) (PVA). Here are the steps that students will follow:

  • PVA (1 gr) and DI water (20 ml) will be mixed, sonicated and heated at 85?C on a hot-plate/stirrer.
  • The prepared solution will be poured into aluminum molds (20-mm diameter).
  • The set-up will be placed inside a lab freezer at -20C.
  • The samples will be removed from the freezer after 24 hour.
  • One wet sample will be weighed after removing from the freezer (Mi,T1) and immersed in DI water at room temperature (20C).
  • Another wet sample will be weighed after removing from the freezer (Mi,T2) and immersed in DI water inside the incubator (37C).
  • A third wet sample will be placed in a freeze-dryer (purchased via Tech Fee) and weighed after the drying is complete (to measure M0).
  • The water-swollen samples will be weighed after 24 hours, 4 days and 7 days (to measure Mt,T1 and Mt,T2) after dabbing the surface water with a filter paper.
  • The original water content of the cryogels will be calculated as follows: WC (%) = [100 × (Mi - M0) / M0]
  • The water uptake (swelling) for different time points will be calculated as follows (both T1 & T2): WU (%) = [100 × (Mt - Mi) / Mi]
  • Once the team reports were submitted (part I), the collected data by all teams will be uploaded to Canvas to allow students perform statistical analysis (mean, standard deviation, and t-test).

Have you applied for and/or received Tech Fee awards in past years?: Yes

If funded, what results did you achieve?:

(a) A Tech Fee award ($1,123) in 2014 enabled us to purchase some accessories for a diffusion cell designed by Miami's Instrumentation. A Tech Fee report was submitted in 2014. The equipment was intended for a Bioengineering Lab. Data acquisition using LabView software was also performed in 2014 using this equipment.

(b) A Tech Fee award ($6,259) in 2015 was used to order one floating network license (FNL) of Comsol Multiphysics as well as one optimization license of the software. A Tech Fee report was submitted in 2016. The software was made available on the Redhawk compute cluster to all Miami faculty and students. An undergraduate student (CPB 340U, USS award) spent the summer of 2015 on 3D scaffold design using Comsol optimization software on Redhawk (384 CPU hours) and additive manufacturing (3D Bioplotter). Also, a graduate student (CPB 700) used 2,367 CPU hours on Redhawk to design hybrid bone-mimicking scaffolds.

(c) A Tech Fee award ($1,700, Graduate) in 2015 allowed purchasing a design optimization tool (Abaqus-ATOM) for graduate students and senior capstone projects. The Tech Fee report was submitted in 2016. One floating network license (FNL) of the software was made available on the Redhawk cluster to all Miami faculty and students. The purpose of the module was to enable topology optimization, a method often used to design components with efficient material layout given a certain performance goal such as optimal mechanical strength.

(d) A Tech Fee award ($7,000) in 2016 was used to order a freeze-dryer (with additional cost-share from CPB and OARS). The students enrolled in CPB 419/519 will use this equipment in April 2017 during their lab project on cryogel fabrication. A Tech Fee report will be submitted in May 2017 upon the completion of the project and grade assignment.

Did you submit a final report?: Yes

What happens to the project in year two and beyond? Will there be any ongoing costs such as software or hardware maintenance, supplies, staffing, etc.? How will these be funded?: No costly maintenance is anticipated for this lab equipment. The expected life-time is usually reasonable, and the Instrumentation Lab (75 Hughes) is well equipped to support any required maintenance.

Budget: Hardware

Hardware Title(s) & Vendor(s): FORMA STRICYL I160 CO2/O2 DRY, Fisher Scientific

Hardware Costs: $8,081.20

What is the total budget amount requested?: $8,081.20