Special Issue:
Challenges and Opportunities in Regenerative Medicine
Richard Z Cheng, MD, PhD
Cheng Integrative Health Center, Columbia, SC, USA 29212
Michael Passwater
Orthomolecular Medicine News Service, Gainesville, FL, USA
Tievi Yang, MD
Depts of Orthopedics and Functional Medicine, Gongli Hospital, Shanghai University School of Medicine, Shanghai, China
Abstract
For over 3 years, the Covid-19 pandemic felt like a world war and has taken close to 7 million lives, disabled many more people, and caused innumerable economic losses around the globe. What can we learn from this tragedy? Are we ready for another Covid-19-like pandemic? Studies show that the majority of people with SARS-Cov-2 infection either show no symptoms or only mild to moderate clinical manifestations; only a small percentage develop severe Covid-19 disease, indicating that the clinical severity of Covid-19 disease is not determined only by the SARS-Cov-2 virus, but more importantly by how the host responds to the viral infection, what is known as natural immunity. Research of what enhances or weakens the natural immunity against viral infections and the practical application thereof is an important lesson one can learn from the pandemic. Research of natural immunity enhancing factors is summarized in this paper. One key characteristic of natural immunity against viral diseases is its non-specificity. The importance of this non-specificity helping to prevent and treat other infections of known or unknown viruses is also discussed. Calls for the clinical application of safe and inexpensive nutrients such as vitamin C in the prevention and treatment of Covid-19 have met significant resistance and objection from the medical authorities and the media since the pandemic outbreak. The main objection is the perceived lack of research and the absence of regulatory approvals. This raises a fundamental philosophical question: what is the primary goal of the medical profession? Facing a new viral pandemic like Covid-19 with no prior research, let alone any approved treatments, why is there opposition to known safe, inexpensive, widely available and often effective nutrients like vitamin C? Why are case reports and case series discounted or ignored rather than explored further to try to help more people? Is such objection protecting consumers or harming the public? Statistics show that viral epidemics and pandemics are occurring more frequently, with a recent review of epidemics and pandemics since 1600 concluding “ the yearly probability of occurrence of extreme epidemics can increase up to threefold in the coming decades.”1. When the next Covid-19-like pandemic of a new virus hits us, are we going to repeat the Covid-19 tragedy? Can improved emphasis on nutritional interventions to prepare for and respond to disease outbreaks mitigate future pandemics?
Bernardica Jurić
Clinical Department of Pathology and Cytology “Ljudevit Jurak”, University Hospital Center “Sestre milosrdnice”, Zagreb, Croatia.
Monika Ulamec
Clinical Department of Pathology and Cytology “Ljudevit Jurak”, University Hospital Center “Sestre milosrdnice”, Zagreb, Croatia./ Department of Pathology and Center of Excellence for Reproductive and Regenerative Medicine, School of Medicine, University of Zagreb, Zagreb, Croatia.
Božo Krušlin
Clinical Department of Pathology and Cytology “Ljudevit Jurak”, University Hospital Center “Sestre milosrdnice”, Zagreb, Croatia./ Department of Pathology and Center of Excellence for Reproductive and Regenerative Medicine, School of Medicine, University of Zagreb, Zagreb, Croatia.
Melita Perić Balja
Department of Pathology and Center of Excellence for Reproductive and Regenerative Medicine, School of Medicine, University of Zagreb, Zagreb, Croatia./ Clinical Department of Pathology and Cytology “Ljudevit Jurak”, Tumor Oncology Clinic, University Hospital Center “Sestre milosrdnice”, Zagreb, Croatia.
Abstract
Breast cancer is the most common malignancy in women worldwide, and one of the leading causes of cancer death. This disease shows a significant heterogeneity due to its genomic and histological diversity. Breast cancer is classified by pathologic features (i.e. histological subtype, tumor grade) and gene expression profiles (i.e. molecular subtypes). There are complex mechanisms implicated in its progression and the development of chemotherapy resistance. In recent times, tumor stroma is increasingly being recognized as an important factor which influences tumor pathogenesis and progression. Tumor-stromal cells interactions are involved in many phases of tumor growth, by modulating different cellular processes. Tumor-infiltrating lymphocytes are proven to be clinically significant as they correlate with good prognosis, especially in triple-negative and HER2-positive breast cancer patients. However, tumor-infiltrating lymphocytes are just one of the many components of the tumor microenvironment, which includes fibroblasts, macrophages, adipocytes, vascular cells etc., but also non-cellular components. One of the main cellular components of the tumor microenvironment are the fibroblasts which are activated and differentiated into breast cancer associated fibroblasts. They secrete many growth factors, cytokines, and chemokines which influence tumor growth and dissemination. Tumor microenvironment could be a source of new biomarkers with a potential predictive and prognostic significance. This review highlights the tumor microenvironment as an important contributor to the process of cancer development with an overview of the main components and the potential impact on the prognosis of breast cancer. It’s important to expand our understanding and knowledge of tumor-stromal signalling processes which may lead to the development of more successful and individualized therapeutic strategies.
Akira T. Kawaguchi, MD, PhD
Department of Pediatrics, Tokai University School of Medicine; Department of Cell Transplantation and Regenerative Medicine, Tokai University
Amankeldi A. Salybekov, MD
Department of Cell Transplantation and Regenerative Medicine, Tokai University
Mariko Yamano, PhD
Department of Anatomy and Neurological Science, Nara Medical University
Hideaki Sumiyoshi, PhD
Department of Physiology, Tokai University
Gen T. Kawaguchi, MD
Department of Plastic Surgery, Tokai University
Shinichi Matsuda, MD, PhD
Department of Pediatrics, Tokai University School of Medicine
Kaori Sekine
Department of Physiology, Tokai University
Mayuko Shibata, MD
Department of Pediatrics, Tokai University School of Medicine
Yoshiyuki Yamada, MD, PhD
Department of Pediatrics, Tokai University School of Medicine
Abstract
Background: Peripheral blood mononuclear cell-derived progenitor cells (regeneration associated cells: RACs) have been reported to migrate to tissues with inflammation to expedite anti-inflammatory and pro-regeneration actions, but cell sources, administration, timing, dosage, and combined procedures after myocardial ischemia and/or infarction (MI) still remain to be elucidated in preclinical studies.
Materials and Methods: Rats with induced MI were treated with intravenous infusion of 5-day culture-primed autologous RACs or control cell transfusion, followed by weekly echocardiography until 4 weeks after MI, at which time pressure-volume relationship (PVR) analyses and histological studies were performed.
Results: While most rats treated with autologous RACs infusion after MI developed vascular regeneration from surrounding pericardial tissues and significant functional improvements, one rat developed a de novo coronary artery bypass (φ< 0.3 mm) mimicking surgical bypass grafting (CABG), echocardiography revealed recovered left ventricular (LV) motion, and PVR analyses showed restored LV function as well as histological changes suggesting revascularization and myocardial regeneration.
Conclusion: Development of isolated bypass-like structure by RAC therapy after MI led us to consider further surgical as well as biological modifications to expedite biological coronary revascularization and myocardial regeneration. A combination with vascular approaches to provide arterial inflow may increase the rate of complete revascularization and regeneration of cardiomyocytes as hybrid procedure, as they complement each other, to realize cardiovascular regeneration.
Katsutoshi Miura
Department of Regenerative & Infectious Pathology, Hamamatsu University School of Medicine
Toshihide Iwashita, Dr. Professor
Department of Regenerative & Infectious Pathology, Hamamatsu University School of Medicine
Abstract
Organs with different levels of stiffness support the musculoskeletal system. Light microscopy cannot evaluate organ stiffness, whereas scanning acoustic microscopy (SAM) discriminates stiffness based on speed-of-sound (SOS) because sound waves pass faster in stiffer tissues. This study aimed to evaluate SOS imaging for orthopedic diseases using formalin-fixed paraffin-embedded sections. SOS imaging in SAM uses unstained light microscopic (LM) sections to prevent the bias of staining variation. Digital SOS values are comparable in different organ components and diseases.
Mouse organs with the lowest mean SOS values included the adipose tissue, bone marrow, calcified cartilage, and nucleus pulposus; those with intermediate values included hyaline cartilages, osteoid, skeletal muscles, cortical and trabecular bones, and ligaments; and those with the highest values comprised fibrocartilages of the vertebral disc and meniscus. Water contents and delipidating procedures decreased SOS values. Collagenous density and arrangement affected higher SOS values. The trabecular bones of mice were thinner and showed significantly lower values of SOS than those of humans.
Various orthopedic diseases and disorders displayed the characteristic SOS images. In osteoporosis, the trabecular bone becomes thin with lower SOS, indicating lesser stiffness to cause fractures. Comparison of woven and lamellar bones revealed that woven bones with lower SOS had lesser stiffness to fracture. Changes in SOS values indicated intramembranous bone formation. The trabecular bone develops from the connective tissues with an abrupt increase in SOS values. The regenerating process of bone fractures was monitored using SOS images, in which the granulation tissues transformed into calli in the osteoid to grow a new mineralized bone. The stiffness increased in phases, which appeared in SOS values.
Although several methods have been used to visualize the stiffness of biological tissues, SAM only needs 10-µm unstained slides and can simultaneously compare mechanical stiffness and histology. SOS images provide informative mechanical alterations of the bone, cartilage, and connective tissues to assess the status and diagnose a disorder.
Jae-kook Yoo
Department of Neurolgy, The Rodem Hospital, Incheon, Korea.
Soon-Hee Kwon
Department of Neurolgy, The Rodem Hospital, Incheon, Korea.
Jong-Eun Jeon
Department of Neurolgy, The Rodem Hospital, Incheon, Korea.
Sul-Hee Yoon
Department of Internal medicine, The Rodem Hospital, Incheon, Korea.
Jung-Eun Lee
Department of Rehabilitation medicine, The Rodem Hospital, Incheon, Korea.
Sang-Yoon Lee
Department of Rehabilitation medicine, The Rodem Hospital, Incheon, Korea.
Abstract
This case report from the Rodem Hospital introduces a pioneering intervention for muscle rigidity in Amyotrophic Lateral Sclerosis (ALS), featuring a unique placental extract injection and glucose injection therapy combined with lidocaine. This novel approach has demonstrated significant muscle regeneration and sustained relaxation in 47 ALS patients. Unlike traditional treatments, this protocol offers a more sustainable and regenerative outcome. The treatment involved injections of a mixture containing glucose, lidocaine, and placental extract, targeting severely rigid muscles. Remarkably, 42 of the 47 patients showed considerable improvements in knee flexion and a dramatic reduction in pain. The other two also experienced notable progress. This method stands out for its cost efficiency, impact on muscle suppleness, and reduced pain, suggesting a potential paradigm shift in ALS management. This case series highlights the importance of continued innovation and personalized treatment strategies in ALS care, aiming to improve patient quality of life and functional abilities.
This case report from the Rodem Hospital introduces a pioneering intervention for muscle rigidity in Amyotrophic Lateral Sclerosis (ALS), featuring a unique placental extract injection and glucose injection therapy combined with lidocaine. This novel approach has demonstrated significant muscle regeneration and sustained relaxation in 47 ALS patients. Unlike traditional treatments, this protocol offers a more sustainable and regenerative outcome. The treatment involved injections of a mixture containing glucose, lidocaine, and placental extract, targeting severely rigid muscles. Remarkably, 42 of the 47 patients showed considerable improvements in knee flexion and a dramatic reduction in pain. The other two also experienced notable progress. This method stands out for its cost efficiency, impact on muscle suppleness, and reduced pain, suggesting a potential paradigm shift in ALS management. This case series highlights the importance of continued innovation and personalized treatment strategies in ALS care, aiming to improve patient quality of life and functional abilities.
Katelin Omecinski, BS
McGowan Institute for Regenerative Medicine, University of Pittsburgh; Department of Bioengineering, University of Pittsburgh
William Federspiel, PhD
McGowan Institute for Regenerative Medicine, University of Pittsburgh; Department of Bioengineering, University of Pittsburgh; Department of Chemical and Petroleum Engineering, University of Pittsburgh; Department of Critical Care Medicine, University of Pittsburgh Medical Center; Clinical and Translational Science Institute, University of Pittsburgh
Abstract
Respiratory disease remains a pervasive medical condition amongst the pediatric health population. Mechanical ventilation and extracorporeal membrane oxygenation (ECMO) are used to bridge patients to transplant or recovery when conventional therapy fails. Patients undergoing these treatments may be sedated for extended periods of time, resulting in deconditioning of the patient’s musculature. Patients who remain awake on ECMO, however, can participate in physical therapy and combat muscle wasting. Typical ECMO circuits are complex and present a major consumer of hospital resources for these patients undergoing rehabilitation and ambulation. Our research group has pursued the integration of mechanical circulatory and respiratory assistance into a compact platform device, the ModELAS, to address this clinical need. The aim of this review is to summarize published work on the pediatric application of the ModELAS. A breadth of topics will be reviewed, including the design requirements, device evolution, in-vitro results, and in-vivo results of the device.
David Civitarese
Shirley Ryan AbilityLab/Northwestern McGaw Medical Center, Chicago, IL
Michael A Downing
Albany Medical Center Physical Medicine and Rehabilitation Residency, Albany, NY
Michael O Bazzi
University of California, Irvine Physical Medicine and Rehabilitation Residency, Irvine, CA
Glenn Flanagan
Naples Regenerative Institute, Naples, FL
Joshua B Rothenberg
Sports Medicine, Baptist Health North Orthopedic Care, Boca Raton Regional Hospital, Boca Raton, FL
Abstract
Introduction: Achilles tendon rupture is one of the most common adult tendon injuries and continues to increase in incidence. This case report outlines a comprehensive approach for stand-alone leukocyte-poor platelet-rich plasma treatment of a partial subacute Achilles tendon rupture in a professional athlete who had received prior autologous conditioned serum injections with an interleukin-1 receptor antagonist.
Patient History: A 32-year-old male professional ice hockey player presented with persistent left ankle pain approximately seven months after suffering a partial acute Achilles tendon tear. Imaging confirmed a subacute partial Achilles tendon rupture.
Injection: Following local anesthesia, a 25-gauge 1.5-inch needle was advanced to the target structure under ultrasound guidance and 3 mL of autologous platelet-rich plasma was divided amongst several areas of the patient’s Achilles tendon, while simultaneously tenotomizing the tendon. No complications were observed.
Imaging: Follow-up magnetic resonance imaging approximately 9 months after platelet-rich plasma injection revealed complete resolution of the Achilles tendon rupture.
Conclusion: Ultrasound-guided leukocyte-poor platelet-rich plasma injection could be considered an effective treatment option for acute Achilles tendon rupture during the subacute stages of tendon healing, leading to complete radiographic resolution of the tear and enabling a return to high-level athletics.
Katsutoshi Miura
Department of Regenerative & Infectious Pathology, Hamamatsu University School of Medicine, Japan
Toshihide Iwashita
Department of Regenerative & Infectious Pathology, Hamamatsu University School of Medicine, Japan
Abstract
Background: Reducing sugars and reactive aldehydes, such as glyceraldehyde (GA), non-enzymatically react with proteins to form advanced glycation end-products (AGEs). GA is produced in the glycolysis pathway, and GA-derived AGEs play an essential role in the pathogenesis of angiopathy associated with hyperglycemia in patients with diabetes.
Aims: Several studies have reported on chemical alterations in glycation. However, histological confirmation of these biochemical changes has been relatively rare. This study aimed to visualize glyceraldehyde-induced glycation and evaluate the severity of glycation using attenuation of sound (AOS) values. Given that glycation promotes cross-links between proteins and sugars, the energy loss of sound that passes through them increases. We hypothesized that AOS alteration would reflect the glycation state of the tissues and cells.
Methods: Fresh frozen sections or fresh cells were briefly fixed in ethanol and soaked in GA with different glucose concentrations. Thereafter, AOS images were obtained via scanning acoustic microscopy over time, and tissue and cellular glycation induced by GA with glucose was evaluated using AOS values.
Results: AOS images were able to visualize GA-induced glycation over time. Compared to GA alone, glucose supplements concentration-dependently accelerated glycation. The arterial smooth muscle, collagen, and intima were apt to accept glycation, whereas the mucosa was unaffected.
Conclusion: The comparability and digital nature of AOS images make them suitable for statistical analysis of glycation. Higher glucose concentrations promoted a greater increase in the AOS values of the sections and cells. Moreover, the increase in AOS values varied according to organs and cells, which supports the difference in affected organs among patients with diabetes mellitus. Our findings suggest that a longer hyperglycemic state promotes greater glycation.
Christian Elmshaeuser
Stem Cell Therapy Division, Institute of Medical Microbiology, University of Giessen, Giessen Germany & International Senior Professional Institute (ISPI) e.V. Giessen, Germany. 2Biochemistry Institute, University of Giessen, Giessen, Germany.
Ina Zoeller
Stem Cell Therapy Division, Institute of Medical Microbiology, University of Giessen, Giessen Germany & International Senior Professional Institute (ISPI) e.V. Giessen, Germany. 2Biochemistry Institute, University of Giessen, Giessen, Germany.
Darisuren Anhlan
Biochemistry Institute, University of Giessen, Giessen, Germany.
Ewald Beck
Biochemistry Institute, University of Giessen, Giessen, Germany.
Bruno Peault
INSERM U506, Groupe Hospitalier Paul Brousse, Villejuif Cedex, France & Orthopaedic Hospital Research Center and Broad Stem Cell Research Center, David Geffen School of Medicine, University of California, Los Angeles, LA, United States & MRC Centre Institute for Regenerative Medicine and Centre for Cardiovascular Science, University of Edinburgh, Edinburgh, United Kingdom.
Olivier Tabary
Sorbonne Université, INSERM UMR S 938, Centre de Recherche Saint Antoine (CRSA), Paris, France.
Una Chen
Stem Cell Therapy Division, Institute of Medical Microbiology, University of Giessen, Giessen Germany & International Senior Professional Institute (ISPI) e.V. Giessen, Germany. ; 5Research Experience of Stem Cells in Europe Society (RESCUES) e.V., Giessen, Germany.
Abstract
Background-Purpose of this study: Tissue-specific stem cell lines are useful tools for cell biology studies. Information on respiratory tissue cell lines is limited. A doxycycline-regulated epithelial precursor cell line was established from the lung tissue of a tTAxSV40 Tag double transgenic mouse. In this study, we have characterized this cell line in vitro & in vivo, and found to mimic a rare subpopulation of club- and pneumocyte type II-dual cells.
Methods: It was partially characterized using cell viability and death assays, H3-thymidine incorporation assay, chloride efflux assay, Western blotting of proteins secreted, RT-PCR assays for RNA isolated. In addition, immune–deficient SCID mice were used as hosts for implantation of this precursor cell line, and feed with/without doxycycline containing water. Immunofluorescent typing using different antibodies were used to characterize the implanted lung.
Results: This cell line was found to mimic a rare subpopulation of club- and pneumocyte type II- dual cells with multiple phenotypes. Cell growth was doxycycline-regulated and observed only when doxycycline was omitted from the medium or present at concentrations up to 1 µg/ml, higher concentrations were inhibitory. ACT+ ciliated cells were found upon implantation into immune-deficient mice, in addition. Cell growth was doxycycline-regulated in vitro. When transplanted subcutaneously into immune–deficient mice, these cells migrated to the lung to form organized chimeric structures of donor and host origins, with club cells in the terminal bronchioles, ACT+ ciliated cells along the epithelial lining, and pneumocyte type II-cells in the alveolar interstices. No such homing of donor cells to the lung was observed when the implanted mice were fed doxycycline-containing water.
Discussions-Conclusions: This lung stem cell line might be able to provide us with an insight into the differentiation pathway of lung epithelial cells as well as with some understanding of the nature of air trophic-pulmonary epithelial cells. The results of this study underline the possibility of a future application for somatic (stem / precursor) cells in tissue replacement and tissue engineering of the damaged lung. Its ability to secrete and deliver soluble protein, might be a potential novel way for drug delivery. In addition, stem cells are thought to proliferate and differentiate in response to a deficiency or as a result of injury. Successful migration to the target organ and subsequent maturation of these precursors could be attributed to a requirement of lung stem cells to search for an aerated environment. Our findings challenge some current concepts of stem cell biology.This lung stem cell line may become a rich source of cells for tissue engineering and cell-based therapy for lung injury. The route and protocol established for cell introduction into the lung may provide a novel alternative to delivery of soluble protein substances through the airways. This lung stem cell line might also be modified to provide models for screening drugs against respiratory infection.
Jared M Hall
Department of Physiology, University of Louisville, Louisville, KY 40202, USA
Nicholas E Yates
Department of Physiology, University of Louisville, Louisville, KY 40202, USA
Philip Bauer
Department of Physiology, University of Louisville, Louisville, KY 40202, USA
Gustavo Perez-Abadia
Department of Orthopedic Surgery, University of Louisville, Louisville, KY 40202, USA
Claudio Maldonado
Abstract
The Primo Vascular System was discovered by Bong Han Kim in the 1960s when searching for an anatomical correlate of the acupuncture meridians used in eastern medicine. The Primo Vascular System is a systemic network of thread-like Primo-vessels with intermittent enlargements known as Primo-nodes. Primo-vessels are difficult to view under a microscope due to their small diameters (20-50 µm) and translucent appearance. Primo-vessels have a porous outer membrane that encapsulates small channels named Primo sub-vessels filled with flowing fluid. Primo-vessels are classified into six sub-types based on their anatomical location. The physiological mechanisms of Primo-vessel function are not clear. There are multiple hypotheses based on Primo-vessel and Primo-node structure and cell content, however, supportive functional experimental data is lacking. This review focuses on the “interior” lymphatic vessel Primo-vessel (ILVPV) sub-type, the techniques that are used to visualize them, and experimental studies that attempt to unravel their physiological role after inflammatory stimulation. Speculative hypotheses are presented regarding the handling of signals by ILVPVs for intercellular communication between injured cells and cells stored within “interior” lymphatic vessel Primo-nodes (ILVPNs). One of the stored cell types that are of interest for tissue repair are very small embryonic-like cells. Very small embryonic-like cell activation may be induced by biophoton signals emitted by injured cells and transmitted to ILVPNs via Primo-vessel and/or ILVPV networks. An alternative or additional method for intercellular communication may involve the release of signaling proteins and/or extravesicular bodies carrying genetic messages (i.e., exosomes) by cells in injured tissues. As these signaling factors enter the lymphatic circulation, porous ILVPVs filter them out and transport them to ILVPNs where they initiate very small embryonic-like cell activation to start the tissue regenerative process. Primo Vascular System research will require more physiological functional studies to elucidate the role of ILVPVs and ILVPNs in tissue regeneration. To achieve this goal, future mechanistic studies will need novel biomarkers and animal models.
