Introductory_lecture_PRA3017_2025

Introduction

  • Lecture Title: Oxygen for Regenerative Medicine

  • Presenter: Dr. Silvia Bolognin

  • Date: January 28th, 2025

  • Venue: cBITE (Cell Biology-Inspired Tissue Engineering) at the MERLN Institute for Technology-Inspired Regenerative Medicine

Course Overview

  • Key Topics:

    • Cell Culture

    • Quantitative PCR (qPCR)

    • Immunocytochemistry (ICC)

    • Written Exam

    • Western Blot

  • Schedule: 9:00 AM to 5:00 PM at MERLN Institute

Assessment Methods

  • Assessment Breakdown:

    1. Knowledge and skill exam (individual): 30%

    2. Cell culture experimental result report (group): 15%

    3. qPCR experimental result report (group): 25%

    4. ICC and Western Blot experimental result report (group): 30%

Regenerative Medicine and Societal Impact

  • Focus: Addressing chronic diseases in an aging population

    • Patients: Shift from palliative care to prevention and cures

    • Economy: Creation of a new industry and significant economic opportunities

    • Science: Growth in the scientific field related to regenerative medicine that aims to harness the body’s healing powers to restore tissues and organs

Aims of Regenerative Medicine

  • Central Goal: Direct cell behavior such as survival, growth, and phenotype

  • Key Objectives:

    • Repair injuries

    • Replace tissues and organs

    • Activate endogenous healing mechanisms

Key Areas of Research and Education at MERLN

  • Research Areas:

    • Bioinformatics and computational tools

    • Additive manufacturing

    • High-throughput screening

    • Cell biology

    • Materials science

    • Microfabrication

  • Courses Offered:

    • BIO2001 (Cell Biology)

    • CHE2001 & PRA2002 (Organic Chemistry)

    • INT3003 (Biomaterials)

    • INT3008 (Regenerative Medicine)

    • Clinical translation

The Aging Population

  • Data Presentation: Increasing prevalence of chronic diseases with age

    • Cancer, heart attacks, dementia, diabetes, vision impairment statistics, by gender

  • Implications of an aging population on healthcare and regenerative medicine

Steps of Tissue Engineering

  1. Cell isolation from the patient

  2. Seeding cells on a scaffold

  3. Cell stimulation in a bioreactor

  4. Implantation of tissue-engineered construct in the patient

Tools and Materials in Tissue Engineering

  • Components:

    • Cells and cell biology techniques

    • Biomaterials: hydrogels, porous membranes, polymeric materials

    • Fabrication technologies: 3D printing, bioreactors, bioactive molecules

Autologous Chondrocyte Implantation

  • Benefits:

    • Tissues regenerate, leading to reduced pain

    • Limitations: inferior mechanical stability compared to native tissue

Challenges in Tissue Engineering

  • Key Issues:

    • Immunogenicity and rejection of engineered tissues

    • Scale-up challenges in production

    • Ensuring viability and vascularization of engineered tissues

    • Achieving maturity of engineered constructs

    • Integrating engineering with electromechanical systems

Co-morbidities and Bone Healing

  • Increasing incidence of conditions that affect bone healing:

    • Age

    • Osteoporosis

    • Diabetes Mellitus

Bone Abnormalities

  • Overview of osteoporotic conditions:

    • High vs. low bone density

    • Implications for fracture risk and overall strength of the bone

Bone Tissue Engineering

  • Key Concepts:

    • Combination of cells and/or materials for effective bone regeneration

    • Use of Mesenchymal stem cells (MSCs)

Importance of Oxygen in Tissue Engineering

  • Need for oxygen and nutrients for cell survival

  • Oxygen concentration measurements within 3D engineered constructs

Oxygen Distribution in Bone and Cartilage

  • Physiological oxygen tensions:

    • Various studies citing oxygen tension levels in human and animal models for bone and cartilage

Measuring Oxygen Levels

  • Techniques include:

    • Microx TX3, Microx TX4, VisiSens techniques for measuring oxygen levels in constructs

In Vitro Studies of Oxygen Distribution

  • Research findings exhibited in cell-loaded collagen gels and the response to different oxygen levels

Hypoxia and HIF Pathways

  • Discussion on:

    • HIF1/2/3 as important factors in the cellular response to hypoxia

    • Mechanisms of hypoxia affecting gene expression and cellular survival

Lab Studies and Conclusions

  • Experiments inducing hypoxia with phenanthroline and assessing its influence on cell behavior and apoptosis outcomes

Acknowledgements

  • Gratitude expressed to Maastricht University for facilitating the lecture series and research efforts in regenerative medicine.