Micro-fabrication strategies to design stemcell instructive biomaterials for bone tissue engineering applications
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1 Micro-fabrication strategies to design stemcell instructive biomaterials for bone tissue engineering applications Daniela Fernandes Coutinho Rui Reis, Nuno Neves and Manuela Gomes (UMinho) Ali Khademhosseini (Harvard-MIT HST) Doctoral Student Presentations MIT, March 4, 2010
2 Motivation Promote Bone Tissue Regeneration Instructive Molecules Cells Scaffold 2
3 Nature is micro/nano Credits: Office of Basic Energy Sciences, Office of Science, U.S. Department of Energy. The elementary interactions are Nano and Micro Scaled 3
4 Strategy TAILOR Cell Microenvironment Interface Biomaterial s Surface- Host Environment 4
5 Objective Development of strategies for controlling the stem cell response to biodegradable polymers through the design of innovative micro/nano-engineered surfaces and structures aimed for bone tissue engineering applications. Research Question Can we develop micro/nano-architectured systems with biodegradable polymers capable of inducing a specific stem cells behavior? 5
6 Micro-architectured system for Bone Tissue Engineering 6
7 Microengineered Hydrogels Yanan Du, Ali Khademhosseini et al, PNAS (2008) 7
8 Selected stem-cell source Human Adipose Stem Cells (hasc) Why? 1. Availability 2. Differentiation potential in different tissues 3. Obtained through a minimal invasive and safe procedure Strem, B. et al, Keio J Med,
9 Preliminary results Cell viability in different media and cell density for 21 days Promote Bone Tissue Regeneration Assess the differentiative potential of hasc for osteogenic lineage 1. Immunostainning of specific ostoegenic markers 2. FACS Phalloidin Osteocalcin 9
10 How do hasc respond to microengineered surfaces? 10
11 Study the hasc response to surface micro-features Develop poly(butylene succinate) (PBS) patterned surfaces and assess the proliferation and viability of hasc. 11
12 Study the hasc response to surface micro-features Develop PBS patterned surfaces and assess the proliferation and viability of hasc. Characterize the developed structures hasc attachment, spreading, proliferation and viability 12
13 Study the hasc response to surface micro-features Assess the osteogenic potential of hasc cultured on poly(lactic acid) (PLA) 30 different patterned surfaces. hasc attachment and spreading PhalloidinDAPI hasc osteogenic differentiation A4 OsteocalcinDAPI 50 µm A4 50 µm 13
14 How do hasc respond to microengineered structures? 14
15 Develop hasc instructive microarchitectured hydrogel system 1. Produce molds with desired shape (photolithography) 2. Produce microunits 3. Assembly procedure PEG Hydrophobic polymer Polymer solution 15
16 So far Produce microunits Develop new photocrosslinkable Gellan Gum-based biomaterial Characterized: - Chemically - Physically - Mechanically - Biologically Develop hydrogels with highly tunable physical and mechanical properties applicable for a wide range of tissue engineering approaches, by combining physical and chemical crosslinking mechanisms. 16
17 Wrap up NOW 1. Studied stem cell source selected Human Adipose Stem Cells 2. Developed a method to produce patterned PBS surfaces. 3. Assessed hasc response to patterned surfaces. 1. Characterize hydrogel microunits 2. Assemble the microengineered hydrogels. 3. Encapsulate hascs within the 3D system. 4. Developed a biomaterial with highly tunable properties. 17
18 Acknowledgements Prof. Rui Reis Prof. Ali Khademhosseini Prof. Nuno Neves Shilpa Sant Prof. Manuela Gomes Hyeongho Shin Vitor Espírito Santo SFRH/BD/37156/
19 Micro-fabrication strategies to design stemcell instructive biomaterials for bone tissue engineering applications Daniela Fernandes Coutinho Bioengineering Systems Focus Area (MIT-Portugal) Supervisors: Rui Reis, Nuno Neves and Manuela Gomes (UMinho) and Ali Khademhosseini (Harvard-MIT HST) Doctoral Student Presentations MIT, March 4, 2010
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