Cities and regions have always been among the most dynamically developing territories and have played a significant role throughout human history. They have been centers of progress, education, culture, and commerce, centers of economic and social development, and their role in society is constantly growing. The approaches to their development change with the needs of society. Today’s globalizing society is characterized by intense transformation. The rapid changes and dynamics of societal development in the economy, technology, and techniques, as well as social and lifestyle changes, have undermined the concept of the supposed stability of the social and business environment. These changes are reflected in the current local and regional development trends, where sustainable cities and regions; green, or eco-cities and regions; sunny cities; creative cities and regions; smart cities and regions; resilient cities and regions; cities as living labs; slow cities and regions; and agile cities and regions belong. This Special Issue "Regional Development: Opportunities and Constraints" includes researchers and academicians who are dedicated to the current opportunities and constraints of urban and regional development based on the various drivers and concepts of development reflecting global challenges. The papers include the form of theoretical contributions moving forward theoretical knowledge, as well as empirical work, presenting original research results on the topic.