6/18/ D Co-print of Epoxy Resin with Cellladen Bioink for Custom Shaped Nipple-Areola Skin Grafts. 3D Printing Workshop 6/8/2018

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1 Case Study: D extrusion-based printing D Co-print of Epoxy Resin with Cellladen Bioink for Custom Shaped Nipple-Areola Skin Grafts Sarah Van Belleghem D Printing Workshop 6/8/2018 WELCOME D PRINTING WORKSHOP Sarah Van Belleghem/ PhD Pre-candidate Hello everyone! I am a rd year PhD candidate in the University of Maryland s Bioengineering Department. I graduated from MIT in mechanical engineering back in 2015, and am very interested in progressing advances pertaining to the women s health world. 2 Pertaining to D printing Unique Architecture is Necessary 1 Must be fabricated only by D printing; no other forms of traditional fabrication can create desired object. Central Biological Question Must be investigated, either cellular mechanism/furthering knowledge of fundamental biology. Fabrication is Feasible 2 The chosen D printing technology should be able to create desired object (materials, size/shape) Apparent Need Gives value to your research and a necessary personal connection to drive focused research efforts. 4 Current Powerpoint Presentation Design Template Implant 1

2 s for Breast Cancer Survivors Furthering breast restoration, emotional healing, and women s health advances Mastectomy and Breast Reconstruction Breast Cancer Stages 25% 5% % Stage 0 Stage I and II Stage III ~70,000 Breast Reconstruction Cases per Year 40% 5 Current Powerpoint Presentation Design Template Implant 5 Current Treatment Nipple Restoration Star configuration of cut skin is knotted onto itself with sutures High rates of infection and multidirectional scarring Repeated multiple times due to nipple flattening Invasive procedure resects all breast tissue Central scar across chest Silicone implant produces mirage of natural tissue American Cancer Society. Mastectomy Kim, J, Ahn, H. Archives of Plastic Surgery, 4(4): Current Powerpoint Presentation Design Template Implant 6 2

3 The goal of this work is to create a nipple prosthetic implant that exceeds preexisting nipple restorative attempts due to its reinforced mechanical structure with medical grade epoxy and presence of native tissue. 7 Current Powerpoint Presentation Design Template Implant 7 The goal of this work is to create a nipple prosthetic implant that exceeds preexisting nipple restorative attempts due to its reinforced mechanical structure with medical grade epoxy and presence of native tissue. Dermal Compartment Non-biodegradable Backbone 8 Current Powerpoint Presentation Design Template Implant 8 The goal of this work is to create a nipple prosthetic implant that exceeds preexisting nipple restorative attempts due to its reinforced mechanical structure with medical grade epoxy and presence of native tissue. Epidermal compartment 9 Current Powerpoint Presentation Design Template Implant 9

4 Design Criteria For Nipple Implant Non-degradable backbone Single-step fabrication Optimization of degradable portion for tissue regeneration In vivo-like mechanical properties Precise printing application of both bioink and synthetic materials Material Compatibility 10 Powerpoint Presentation Design Current Template Implant 10 Design Criteria For Nipple Implant Fabrication Setbacks BIO- COMPATIBILITY MATERIAL PROPERTIES Some materials are incompatible with extrusion based printing (low viscosity, Newtonian fluid) Some materials can be cytotoxic and thus should not be used in tissue regenerative practices Z-AXIS MISALIGNMENT If strand deposition is not accurate, the printed part will become off by a layer, and all proximate layers. This renders an ill-printed part Powerpoint Presentation Design Template 11 Current Implant 11 Implant Design Scaffold Variation Control Single Channel Contours Alternating Strands mm 12 Current Powerpoint Presentation Template Implant Design 12 4

5 Compressive Modulus (kpa) DNA Content (ng) 6/18/2018 Co-printed Implant Powerpoint Presentation Template 1 Nipple implant Compared to pig nipple 0.25 D 0.2 p< A B C B Compressive moduli of printed scaffolds are comparable to those found in vivo 0 14 Current Powerpoint Presentation Design Template Implant 14 Grown in-vitro Day 1 Day 7 Day 14 Control 00 µm Epoxy coprint 600 * * Primary human dermal fibroblasts are able to coexist with the epoxy, but the amount of epoxy hinders cell health 0 Day 1 Day 7 Day Current Powerpoint Presentation Design Template Implant 15 5

6 Grown in vitro Epidermal growth is shown to develop through positive cytokeratin 10 expression in co-printed scaffolds. Phalloidin DAPI Collagen I DAPI 100 µm 50 µm Functional dermis develops in printed scaffolds, with FB healthy spindlelike morphological development and collagen I expression 00 µm 16 Current Powerpoint Presentation Design Template Implant 16 Conclusion A co-printed nipple areola skin graft construct composed of biodegradable cell-laden bioink and synthetic medical-grade epoxy Material compatibility with human dermal fibroblasts and medical grade epoxy allowed for sustained cell proliferation and physiological skin protein expression Robust mechanical properties comparable to those seen in vivo 17 Current Powerpoint Presentation Design Template Implant 17 Thank You. Principal Investigators: John P. Fisher, Peter Kim Lab Members: Hannah Baker Marco Santoro Josephine Lembong Charlotte Piard Javier Navarro Rueda Max Lerman Ting Guo Navein Arumugasaamy Megan Kimicata James Coburn Justine Yu Vincent Kuo Undergraduates: Zoe Mote Evan Botterman 6