Osteochondral Microphysiological Systems for Pharmacological - - PowerPoint PPT Presentation

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Osteochondral Microphysiological Systems for Pharmacological - - PowerPoint PPT Presentation

Apr 25, 2023 48 likes •288 views

Osteochondral Microphysiological Systems for Pharmacological Screening Riccardo Gottardi Fondazione Ri.MED Center for Cellular and Molecular Engineering, Department of Orthopaedic Surgery Little Laboratories, Department of Chemical Engineering

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Osteochondral Microphysiological Systems for Pharmacological Screening

Riccardo Gottardi

Fondazione Ri.MED Center for Cellular and Molecular Engineering, Department of Orthopaedic Surgery Little Laboratories, Department of Chemical Engineering McGowan Institute for Regenerative Medicine

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Microphysiological Systems and Personalized Medicine

Modified from D.A. Robinton & G.Q. Daley, Nature (2012) 481, 295–305

3D Microphysiological Systems

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DARPA – NIH “human on a chip” program

credit image: Griffith Lab/Draper

Osteochondral

Homo chipiens

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Modificato da http://www.willomd.com/

Knee Osteoarthrites (OA)

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Osteoarthritis: Bone or Cartilage Disease?

  • Bone disease?
  • osteophytes
  • subchondral lesions
  • association with skeletal dysplasias
  • Cartilage disease?
  • relationship to cartilage injury
  • articular cartilage degeneration
  • Osteochondral disease!

Development of an in vitro Osteochondral Microtissue Model to Study Pathogenesis and Screen Potential Treatments

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Bottom Up Micromass Pellet culture Top Down Cells seeded scaffolds

Chondral construct Osseous construct

H&E

P.G. Alexander*, R. Gottardi*, H. Lin, T.P. Lozito, R.S. Tuan. 2014 Experimental Biology and Medicine

Native tissue

Cartilage Bone

Bottom Up vs Top Down

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Non Provisional patent No. 61/868,979 Provisional patent filed October 6th, 2015

Lozito TP, Alexander PG, Lin H, Gottardi R, Cheng AW, Tuan RS. 2013 Stem Cell Res Ther.

Osteochondral Microphysiological System (OC MPS)

  • a. Lid
  • b. Insert
  • c. O-ring
  • d. Tissue/Construct 1
  • e. Tissue/Construct 2

medium 1 medium 2

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Simplified Bioreactor Design

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Lid Insert Well Base

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Example of fluidic (empty insert)

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Minimal mixing also with gelatin constructs (cell-free)

Trypsin inhibitor (21 kD) – Alexa Fluor 488 BSA (65 kD) – Alexa Fluor 555

Fluorescent proteing IL-1β

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Tissue engineering within the OC-MPS

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Mesenchymal stem cells (MSCs) seeded in photocrosslinkable gelatin 10% gelatin, 0.5% hydroxyapatite, 0.15% LAP 10% gelatin, 1% hyaluronic acid, 0.15% LAP

H Lin, AW Cheng, PG Alexander, AM Beck, RS Tuan. 2014 Tissue Engineering Part A H Lin, TP Lozito, PG Alexander, R Gottardi, RS Tuan. 2014 Molecular Pharmaceutics

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Differentiation within the bioreactor (RT-PCR)

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“Cartilage” genes “Bone” genes

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Histology

13 H Lin, TP Lozito, PG Alexander, R Gottardi, RS Tuan. 2014 Molecular Pharmaceutics

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Modelling osteochondral tissue response to the inflammatory signals of

  • steoarthritis

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A model to study the pathogenesis of osteoarthritis

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IL1-β

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IL-1β treatment (7dd) Decrease in anabolic genes. Increase in catabolic genes. Cartilage responds when bone is stressed. “direct” - IL-1β to cartilage “indirect” - IL-1β to bone

Response of chondral construct (RT-PCR)

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IL-1β treatment (7dd) Decrease in anabolic genes. Increase in catabolic genes. Bone responds when cartilage is stressed. “indirect” - IL-1β to cartilage “direct” - IL-1β to bone

Response of osseous construct (RT-PCR)

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Menstrual cycle hormones have a protective effect on the osteochondral unit

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NIH - Women’s Health Initiative

Estrogen vs. placebo group Estrogen + progestin vs. placebo group Higher bone mineral density Higher bone mineral density Lower risk of osteoporotic fracture Lower risk of osteoporotic fracture Reduced risk of hip OA No changes for hip OA No changes for knee OA No changes for knee OA

Collaboration with Woodruff Group at Northwestern University

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test conditions Donors age 0/0 64 70 58 56 hormones/0 64 70 58 56 0/hormones 64 70 58 56

DAY

  • 2
  • 1 0 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27

MediaGrowth media 0.1nM Estradiol 1nM Estradiol 1nM Estradiol + 10nM Progesterone 0.1nM Estradiol + 50nM Progesterone

All selected donors were post- menopausal women undergoing total knee replacement. Osteochondral plugs were explanted from macroscopically asymptomatic regions of the joint.

The effect of restoring menstrual cycle hormones

  • n native osteochondral plugs
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bar = 25 µm

Protective effect on cartilage of the deep/calcified zone

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p < 0.12

Day 1 Day 30 Day 1 Day 30

Protective effect against bone volume loss

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Conclusions

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  • We have successfully developed a medium/high

throughput microphysiological platform for osteochondral testing of native or engineered tissues to test the effects

  • f potential treatments.
  • The in vitro system mimics the clinically observed

responses to osteoarthritis inflammatory stress signals.

  • We have observed evidence of biochemical

communication between the two components, supporting the concept of osteoarthritis as an osteochondral disease.

  • Exposure to the hormonal sequence of the menstrual cycle

has a protective effect against bone volume loss, markedly at the bone/cartilage interface.

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Acknowledgements

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