Journal Article

Engineering large-scale hiPSC-derived vessel-integrated muscle-like lattices for enhanced volumetric muscle regeneration

Zhang, Ting, Lee, Kangju, Arnaout, Adnan, Moon, Jae-I, Endo, Yori, Reyes, Isaac Garcia, Park, Seung Gwa, Kim, Jiseong, Panayi, Adriana C., Hassan, Shabir, Rodríguez-delaRosa, Alejandra, Kim, Ki-Tae, Lee, Luke, Lee, Myung Chul, Karvar, Mehran, Kiaee, Kiavash, Al Tanoury, Ziad, Martinez, Angel Flores Huidobro, Ochoa, Sofia Lara, Lee, Junmin, Sinha, Indranil, Shi, Kun, Pourquié, Olivier, Kim, Woo-Jin, Shin, Su Ryon, Quint, Jacob, Kamperman, Tom, Tamayol, Ali, Rosales, Montserrat Pineda, Jodat, Yasamin A., Huang, Yike, Ceron, Eder Luna, Calabrese, Michelle A., Lee, Hyeseon, Wang, Xichi

2024

Overview

Large-scale vessel-integrated muscle-like lattices (VMLs) containing dense and aligned human induced pluripotent stem cell (hiPSC)-derived myofibers alongside vessel-like microchannels were fabricated using an advanced bioprinting technology and stem cell-laden extracellular matrix-based bioinks.Incorporating vessel-like lattice was of vital importance for enhancing myofiber maturation in vitro and host vessel invasion in vivo, improving implant integration.Successful de novo muscle formation and muscle function restoration was achieved through a combinatorial effect between improved hiPSC-derived VML graft–host integration and increased release of paracrine factors at volumetric muscle loss injury.Observing the human markers in the implantation area confirmed that the hiPSC-muscle precursor cells (MPCs) at the transplantation site play a significant role for improving regenerative capacity of volumetric muscle loss. Engineering biomimetic tissue implants with human induced pluripotent stem cells (hiPSCs) holds promise for repairing volumetric tissue loss. However, these implants face challenges in regenerative capability, survival, and geometric scalability at large-scale injury sites. Here, we present scalable vessel-integrated muscle-like lattices (VMLs), containing dense and aligned hiPSC-derived myofibers alongside passively perfusable vessel-like microchannels inside an endomysium-like supporting matrix using an embedded multimaterial bioprinting technology. The contractile and millimeter-long myofibers are created in mechanically tailored and nanofibrous extracellular matrix-based hydrogels. Incorporating vessel-like lattice enhances myofiber maturation in vitro and guides host vessel invasion in vivo, improving implant integration. Consequently, we demonstrate successful de novo muscle formation and muscle function restoration through a combinatorial effect between improved graft–host integration and its increased release of paracrine factors within volumetric muscle loss injury models. The proposed modular bioprinting technology enables scaling up to centimeter-sized prevascularized hiPSC-derived muscle tissues with custom geometries for next-generation muscle regenerative therapies. [Display omitted] We developed scalable vessel-integrated muscle-like lattices (VMLs) with dense, aligned human induced pluripotent stem cell (hiPSC)-derived myofibers and vessel-like structures using a novel bioprinting technology. These VMLs significantly improved muscle tissue regeneration and function in mice post-volumetric muscle loss. These modular lattices can be adapted to large-scale muscle defects where native regeneration is impaired. To treat volumetric muscle loss in practice, implants should contain enough cell density to compensate for cell death during the implantation procedure and in the early hours of host integration. To this end, we have optimized construct design to achieve highly dense muscle-laden channels with millimeter-long and densely aligned human induced pluripotent stem cell (hiPSC)-derived myofibers. This cell density has proven effective in achieving functional regeneration in the mice tested. However, more studies are needed in larger animals to quantify the number of cells needed for large defects and this study may result in construct pattern redesign. The hiPSC-derived vessel-integrated muscle-like lattices (VMLs) demonstrate successful de novo muscle formation and muscle function restoration through a combinatorial effect between improved graft–host integration and increased release of paracrine factors. As such, we believe that we have achieved a technology readiness level (TRL) 3 in proving that the concept works in vitro and in a relevant environment (e.g., in vivo). Next, the modular building blocks should be assembled and tested in larger defects to validate continuous elastic behavior, large-scale muscle regeneration capacity of the construct, and implant-wide cellular viability. Moreover, the assembly should be implanted in larger animals, for longer durations of culture, to validate long-term muscle functionality to achieve TRL 4. Ultimately, upon successful completion of the mentioned tests, the constructs should be implanted in human defects during clinical trials to inform regeneration efficiency and muscle function restoration.

• Publisher: Elsevier Ltd,Elsevier Limited
• Subject: 3-D printers / Animals / Binding sites / Bioengineering / Biomimetics / bioprinting / Bioprinting - methods / Combinatorial analysis / Defects / engineering vascularized tissues / Extracellular matrix / geometry / Humans / Hydrogels / Hydrogels - chemistry / Implants / induced pluripotent stem cells / Induced Pluripotent Stem Cells - cytology / Injuries / Internal Medicine / Lattices / Mice / Microchannels / Muscle contraction / muscle development / Muscle, Skeletal - physiology / Muscles / Musculoskeletal system / nanofibers / Osseointegration / Paracrine signalling / Pluripotency / Reagents / Regeneration / secreted factors, volumetric muscle loss / skeletal muscle engineering / Stem cells / Tissue engineering / Tissue Engineering - methods / Tissue Scaffolds / Transplants & implants / Vessels