Nanofiber 3-D Cell Based Assays

This “News behind the News” is a historic event.  It demonstrates how nanofiber scaffolds can be used to engineer organs for human transplants. Good news for researchers looking solutions are in vivo like environments for cell based assays.

Nanofibers Solutions work in transplants-imagine how well they will work in your 3-D based cell based assays.

3-D Cell Based Assays for Drug Discovery are the future. Like any new model, adoption rates are a function of how well the new solutions works. “The proof is in the pudding”.

Here’re highlights of a historic event based on transplants using nanofiber engineered laryngotrachea : Collaboration between Nanofiber Solutions and the Karolinska Institutet produces first synthetic laryngotracheal implants seeded with the patient’s stem cells to be successfully transplanted into human patients in Russia.

COLUMBUS, Ohio, June 26, 2012 – Nanofiber Solutions, LLC, an Ohio-based developer, manufacturer and marketer of 3-D synthetic scaffolds to advance basic research, tissue engineering and regenerative medicine announced today the first and second successful transplants of its tissue engineered laryngotracheal implants seeded with cells from the patients’ bone marrow.

The surgeries were performed June 19th and 21st at the Krasnodar Regional Hospital (Russia) by Dr. Paolo Macchiarini, Professor of Regenerative Surgery at the Karolinska Institutet (Stockholm, Sweden), and colleagues. Dr. Macchiarini led an international team that included Dr. Vladimir Porhanov, head of Oncological and Thoracic Surgery at Kuban State Medical University (Russia), Dr. Jed Johnson, Nanofiber Solution’s Chief Technology Officer who created the synthetic organs, Harvard Bioscience (Boston, USA) who produced the bioreactor, and Dr. Alessandra Bianco at University of Rome, Tor Vergata, who performed mechanical testing during scaffold development.

Both patients, a 33 year-old mother from St. Petersburg and a 28 year-old man from Rostov-on-Don, were in au to accidents and suffered from a narrowing of the laryngotracheal junction for which they already had failed previous surgeries. Transplantation was the last option for the patients to have normal quality of life. Immediately following transplantation, both patients were able to speak and breathe normally.

Nanofiber Solutions, lead by Dr. Johnson, designed and built the nanofiber laryngotracheal scaffolds specifically to match the dimensions of each patient’s natural larynx and trachea, while Harvard Bioscience provided a bioreactor used to seed the scaffold with the patients’ own stem cells.  Although this procedure represents the world’s first and second successful use of synthetic synthetic laryngotracheal implants, it is Nanofiber Solution’s second and third successful organ implants using their synthetic scaffolds within the last year.

Nanofiber Solutions’ scaffolds mimic the body’s physical structure and allow for a more successful seeding, growth and differentiation of stem cells. Because the cells used to regenerate the larynx and trachea were the patients’ own, doctors report there has been no rejection of the transplants and the patients are not taking immunosuppressive drugs. (more).

Capabilities of 3-D nanofiber scaffolds for cell based assays:

Human brain tumor biopsy showing migrating tumor cells along the alligned nanofiber.
  • Nanofibers are optically transparent to allow for live-cell imaging and real time quantification of cell mobility using an inverted microscope
  • Nanofibers mimic the 3D topography found in vivo which produces a more realistic cellular response to therapeutics.
  • More realistic cellular behavior means you can use fewer animals and decrease time-to-market for drug discovery and development.
  • Nanofibers can easily be coated with ECM proteins using existing protocols for standard lab ware.
  • Cells can be easily removed for protein or gene analysis using trypsin, EDTA, etc.
  • We will continue posting relevant press releases, pubs and data that prove the capabilities of these important solutions.

    Dr. Jim Musick-Making MSCs Work

    Harnessing the Power of CellsTM

    Dr. Jim Musick

    Dr. Jim Musick

    Dr. Jim Musick and his company, Vitro Biopharma, give Basic and Drug Discovery Researchers the ability to harness the power of Human Mesenchymal Stem Cells (hMSCs). This power is essential for blazing new trails in the stem cell research and regenerative medicine frontier.

    I am pleased to welcome Jim as a partner in providing my company the expertise and knowledge highlighted in this profile. Together, we give our customers, colleagues and friends the ability to easily culture, grow, differentiate and maintain large stocks on hMSCs.

    Background

    Jim received his PhD from Northwestern University in 1975 and then joined the staff of University of Utah where he specialized in the study of Neuroscience and synaptic transmission. He joined UltraPure Laboratories in 1983.

    At UltraPure, he learned the art and science of commercializing biologicals. There he helped develop procedures for the commercial production of purified human pituitary hormones including, prolactin, growth hormone and TSH. This included developing QA/QC procedures to support commercial distribution of these products.

    He joined Vitro Diagnostics in 1988 and directed all operations involved in the establishment of a diagnostic product line that included about 30 different purified antigen products.  His direct responsibilities included research & development, manufacturing, intellectual property development and maintenance, marketing and sales.  He was also responsible for the development & initial commercialization of the fertility drug VITROPIN™ as well as the cell immortalization program of the Company.  He is an inventor or co-inventor of all issued and pending patents owned by the Company.

    In 1998 he also completed an Executive Program at JJ Kellogg Graduate School of Management, Northwestern University in Managing New Product Development.

    In 2000, he orchestrated the sale of the antigen manufacturing division to Aspen Biopharma (Nasdaq, APPY) while retaining IP related to use of FSH as a fertility drug and to cell line generation technology.

    He has the spirit of a polished scientist/entrepreneur with strong operational and process expertise.

    Harnessing the Power of hMSCs

    As President and CEO of Vito Biopharma, Jim leverages his expertise and experience to manufacture Cord Blood Derived hMSCs. The stem cell revolution demands large stocks of cells of the highest quality. Meeting the demand is the key to the development of stem cell related therapies. Vitro Biopharma has the capabilities to delivery.

    It is all about starting with vials of potent and pure hMSCs. From there, the customer can grow and differentiate large stocks and be confident in the quality because Jim’s company has the processes in place to insure this. The cell lines are well-characterized with regard to species authentication using sensitive PCR methods to quantify non-conserved genes including COX1, Cytochrome B and actin.  Vitro Biopharma also utilizes karyotyping to authenticate its  cell lines.  Adventitious agents are also tested negative by sensitive PCR methods including known viral contaminants and mycoplasma.  Performance is assured by rigorous testing of viability, growth rate and differentiation capacity for formation of chondrocytes, adipocytes and osteoblasts.  Finally these cells are characterized with regard to phenotypic cluster designation antigens.

    Current Products

    Native and fluorescent-labeled human MSCs including native and fluorescein/rhodamine-labeled MSC-derived chondrocytes and osteocytes along with MSC-GroTM  growth and differentiation media. MSC-Gro™ media is provided in low-serum, humanized and serum-free formulations for both growth and differentiation.  Humanized serum-free media may be supplemented with allogeneic or autologous serum for direct comparisons of growth and differentiation under these conditions.  Powdered MSC-Gro™ formulations are also provided.  Vitro Biopharma’s human MSCss have the capabilities to be expanded through at least 10 passages at rapid growth rates and can be further expanded to 16 passages (~50 population doublings) at slower growth rates.

    Human MSC-derived Osteoblasts stained with Alizarin red at 100 x.

    Image: Human MSC-derived Osteoblasts stained with Alizarin red at 100 x.

    Vitro Biopharma has recently launched a new and revised website complete with convenient online ordering and detailed product technical information (www.vitrobiopharma.com).

    Futures

    In our interview, Jim gave blinding glimpses of the future especially with regard to new products to extend Vitro Biopharma’s offering of clinical tools to fully explore the ever-expanding therapeutic applications of MSCs. I am excited about the potential. I will keep you posted as new products are commercialized.

    Mesenchymal Differentiation Pathways

    I will soon be profiling Dr. Jim Musick of Vitro Biopharma.  He manufacturers and provides us a wealth of expertise on our Human Mesenchymal Stem Cells (hMSCs) and MSCGro™ Mesenchymal Stem Cell Media.  As the demand for these grow, we are receiving a variety of questions on differentiation. Specifically, researchers desire to drive these cells to specific progenitor and cell phenotypes like Osteocytes, Adipocytes and Chondroytes.

    I would like to share a pathway map that gives a snapshot of these pathways:

    Regenerative Biology of the Spine and Spinal Cord. Edited by: Rahul Jandial, Mike Y. Chen, Bihong T. Chen and Joseph Ciacci. ISBN: 978-1-4614-4089-5. Publication date: May 25, 2012. Series: Special Books

    I will continue to post information that will enable the researchers to harness the power of Mesenchymal Stem Cells.

    Jim Musick and Vitro Biopharma

    Coming Soon

    Neuromics recently added key products from Dr. Jim Musick and our friends @ Vitro Biopharma. These include potent umbilical cord derived human mesenchymal stem cells and MSCGroTM (best of breed growth and differentiation media). These cells are capable of many passages enabling researchers to build large stocks.

    MSCs_MSCGro

    We will be posting Jim’s profile in several weeks.

    Stem Cell and Cell Based Assays Groups on Linkedin

    I wanted to share some links to groups on Linkedin that have proven a useful resource for me. They are also additive to the stories and data posted here:

    Stem Cell Clinical TrialsStem Cell Clinical Trials

    Note: I am the moderator the the Stem Cell Clinical Trail group and welcome all new members. We are currently 400+ strong and growing.

    3D cell biology : tools & techniques3D cell biology : tools & techniques

    Stem Cell Research Stem Cell Research

    California Institute for Regenerative Medicine (CIRM)California Institute for Regenerative Medicine (CIRM)

    neuropathy and neuropathic painneuropathy and neuropathic pain

    The Gene Silencing Collaboration (RNAi, siRNA, miRNA, Dicer, etc.)The Gene Silencing Collaboration (RNAi, siRNA, miRNA, Dicer, etc.)

    Enjoy.

    Scripps Florida Scientists Awarded $3 Million to Develop New, More Effective Pain Treatments

    We profiled Dr. Laura Bohn research in one of our news stories. We are excited to share the news.Dr. Laura Bohn

    JUPITER, FL, February 29, 2012 – Scripps Florida scientists have been awarded $3.1 million by the National Institute on Drug Abuse, part of the National Institutes of Health, to study and develop several new compounds that could prove to be effective in controlling pain without the unwanted side effects common with opiate drugs, such as morphine, Oxycontin®, and Vicoden®.

    Laura Bohn, an associate professor in the Department of Molecular Therapeutics and Neuroscience at Scripps Research, and Thomas Bannister, an assistant professor in the Department of Chemistry and associate scientific director in the Translational Research Institute at Scripps Research, will serve as joint principal investigators for the new five-year study.

    Their study will focus on four new classes of compounds that appear to differ fundamentally from opiates inthe side effects that they can produce.

    “Once we more fully understand how these compounds work, we expect to optimize and develop them as novel drugs,”said Bohn. “We hope to produce potent pain relievers without the problems associated with current treatments.” Full article: http://www.scripps.edu/news/press/20120229bohn-bannister.html

    We wish her great success in her research aimed at discovering improved solution for managing pain.

    Gerry Shaw-Master of World Class Neuronal/Glial Markers

    Build it and They will Come

    Gerry and One of His Triumph's MCs
    Gerry and One of His Triumph’s MCs

    I am pleased to profile Dr. Gerry Shaw, a Professor at the University of Florida and also the Head of EnCor Biotechnology Inc.  His story is a guide for incubating and spinning out a successful biotech company (EnCor Biotechnology, Inc.) from a university research laboratory. It should provide an inspiration for fledgling entrepreneurs as the model required little capital investment and has enjoyed profitable growth.

    The Backstory

    Gerry’s major area of research interest can be summarized as the study of cellular changes resulting from central nervous system damage and disease states. These changes help neuroscience researchers understand the progression and hopefully discover root causes of diseases like Alzheimer’s, Parkinson’s and ALS. Understanding which proteins are involved in particular disease states also has the potential of identifying targets for therapies.

    The story starts with Gerry’s Post Doctoral research at the Max Planck Institute for Biophysical Chemistry in Goettingen, in what was at the time West Germany. Here he joined the world renowned laboratory of Klaus Weber and Mary Osborn. This lab had pioneering several important techniques, notably SDS-PAGE for protein analysis and the use of antibodies in immunocytochemistry. Later, after Gerry left the same lab made key contributions leading to the routine use of RNAi in “knock down” of normal cellular proteins. The lab had developed antibodies to tag the subunit proteins of microtubules, microfilaments, intermediate filaments and other cellular proteins, and then used these antibodies to visualize the proteins in immunofluorescence microscopy and on western blots. This enabled researchers to look at changes in the cellular expression of these proteins in powerful new way. These methods have become vital tools for understanding normal cellular function and what happens when cells transition from healthy to diseased states. This lab was an ideal location for Gerry to learn how to make quality monoclonal and polyclonal antibodies. Good antibody reagents are vital for the correct interpretation of immunofluorescence microscopy and western blots, and he was soon supplying his reagents to friends, collaborators and other researchers all around the world. Success is value as antibodies that do not as work as expected waste research time and resources, while quality reagents soon become appreciated and may get to be standard lab reagents.

    University of Florida

    The University of Florida, in Gainesville imported his expertise when Gerry joined the institute in 1986. Here he continued to make antibodies to Neurofilaments or NFs and other Neuronal-Glial Markers. It’s hard to keep a good thing a secret and Gerry faced growing demand from all over for these reagents. This proved a drain both financially and in terms of time commitment, as well as a significant conflict of interest with his basic biomedical research program.

    MAP2_Doering IHC Image: Co-culture of embryonic mouse hippocampal neurons and astrocytes. Primary embryonic hippocampal neurons at 7 days in vitro, were stained with Microtubule Associated Protein-2 (MAP, green) to enable the visualization of the dendritic arbors. These neurons were cultured on top of a monolayer of primary cortical astrocytes, stained with an antibody directed against

    Glial Fibrillary Acidic Protein (GFAP, red). The cell nuclei were visualized by staining with 4′,6-diamidino-2-phenylindole (DAPI, blue). BMC Image of the Month October 2010

    As a result Gerry took his first entrepreneurial step by selling his most popular reagents in bulk initially to Chemicon (now Millipore-Merck). Like any new business venture, he did not really know what to expect. It should come as no surprise that the reagents sold like hot cakes and the check started rolling in. Other immunoreagent companies approached Gerry and soon he was supplying antibodies to pretty much every major biotechnology vendor.

    ABC Biologicals to EnCor Biotechnology Inc.

    Success breeds success and as sales increased over the 1990s, it was time to form an independent business and so ABC Biologicals Inc. was incorporated in 1999 initially to buy equipment and develop licensing agreements. Since Gerry had income from sales, he was in the unusual and enviable position of not needing grants, investors, loans or cash from any other source, and so could proceed with almost total independence. The company was renamed EnCor Biotechnology Inc. in 2002, and at the same time moved into the Sid Martin Biotechnology Incubator, a lab dedicated to commercialization of intellectual property generated by the faculty of the University of Florida. The University of Florida is unusually experienced at this and is well known for launching Gatorade, Trusopt and many other products. After 4 years EnCor “graduated” from the Incubator and now occupies a facility in Gainesville. The company now has almost 100 products with many more under development. This is good news for the Neuroscience community.

    The EnCor-Neuromics Connection

    Neuromics provides EnCor Biotechnology reagents to researchers studying neuro-degeneration, neuro-regeneration, neuro-development, neural stem cells, mood disorders, brain injury and spinal cord injury. My customers have found EnCor’s reagents to be rock solid and versatile.

    In addition, Gerry and his team have proved adept at culturing our E18 hippocampal neurons and ESC derived hN2TM primary neurons. This is a big plus as we can actually see how the cells and markers could resonate together for use in cell based assays.

    Hippo_MAPT_DC1 Image: E18 hippocampal neurons stained with Tau (red) and Doublecortin (green). The two proteins overlap in the proximal dendrites (yellow) Axons (low doublecortin content) are red. Blue staining is the nuclear DNA.

    Futures

    I am excited by the glimpse of the future that Gerry shared. We can expect many new, novel and important markers in the coming months and years. In addition, he will be manufacturing various Enzyme-linked immunosorbent assays (ELISA). These kits have the potential to help clinicians diagnose the early onset of diseases like ALS, Parkinson’s and Alzheimer’s.

    For example, his company currently sells an ELISA kit for sensitive detection of Phosphorylated Neurofilament-H (pNF-H). Expression of this protein is up regulated in a variety of damage and disease states, and can be used to accurately quantify this up regulation. The kit can also detect pNF-H in the sera and spinal cord fluid (CSF) of animals with spinal cord and brain lesions. This protein is not normally found in sera or CSF, so its presence indicates recent axonal injury as a result of either damage or disease. This suggests pNF-H is a useful biomarker of neuronal and more specifically axonal injury or degeneration, a suggestion supported by a growing list of basic science publications on various animal models and patient types from Gerry’s research lab (e.g. Shaw et al. 2005, Lewis et al. 2008, Boylan et al. 2009, Lewis et al. 2010).

    Given the capabilities of EnCor’s markers, the development of more kits is coming. There could be a day in the not distant future where they give clinicians tools to better diagnose and monitor serious neurodegenerative diseases, leading to better disease treatment and management.

    I will keep you informed on Gerry’s and EnCor’s future developments.

    Coming Soon-Dr. Gerry Shaw

    Zen and the Art of Bio-marker Production

    Up next will be Dr. Gerry Shaw.  Gerry is the founder and head of EnCor Biotechnology, Inc. His company is recognized for creating markers that are engines of Neuroscience and Stem Cell Research.

    Dr. Gerry Shaw with Triumph MC

    Dr. Gerry Shaw with Triumph MC

    I am pleased to represent his company’s reagents. They are well designed, thoroughly tested and proven to work in my customers’ many application.

    They have proven especially effective in working in cell based assays using our eSC derived hNP1 human neurons and e18 primary rat hippocampal neurons.

    Applications include the study of TBI, SCI, ALS, AD, MS and PD.

    Image:  hN2 cells stained with our chicken polyclonal antibody to Vimentin, in red. Islands of Hn2 cells form after 4 days in culture forming beautiful flower like structures. Vimentin is a well established marker of early differentiating neuronal lineage cells. Taken with a 10X objective lens. Blue staining is the nuclear DNA.

    hN2 Cells stained with Vimentin

    hN2 Cells stained with Vimentin

    Dr. Ivan Rich and HemoGenix

    Stem Cells Testing Tools that enlighten Drug Discovery and Cell Therapy Researchers
    I am pleased to profile Dr. Ivan Rich. He is the founder, chairman and CEO of HemoGenix and an internationally recognized leader in hematology.  I am timing this profile to coincide with Neuromics launch of HemoGenix’s first to market fully standardized, proven and cost effective  ATP-based, in vitro bioluminescence and high-throughput screening (HTS) cell based assay systems.

    These assays represent best in class solutions for detecting and measuring cell viability, functionality, growth, proliferation and cytotoxicity of stem and progenitor cells for stem cell and basic research, cellular therapy, in vitro toxicity testing and veterinary applications.

    Hemogenix_Pic

     ivan-rich

    2000-Present- Hemogenix-CEO
    and Chairman

    1996-2000-Palmetto Richland Memorial Hospital

     1995-Second Thesis in Experimental Hematology, University of Ulm

    1981-1983-Post Doc University of Chicago

    1973-1978-Ph.D. University of Ulm, Biology

     

    Ivan’s journey leading to founding of HemoGenix provided him a unique blend of scientific, entrepreneurial and operational expertise.  These traits are the drivers that enable him to invent, successfully commercialize and continuously improve cell based assay systems. These systems meet a wide range of demanding requirements. These include, for example, meeting the requirement by Standards Organizations and Regulatory Agencies for “appropriate” and “validated” assays that can be used by cord blood banks and stem cell transplantation centers to determine whether a stem cell product has the necessary potency characteristics and can be released for transplantation into a patient…high standards indeed!

    The Back Story-Hematology and Hemopoietic Stem Cells

    Ivan received his PhD from the University of Ulm, in Germany in 1973 in Human Biology. He then completed a second thesis in 1995 in experimental hematology.  Our story starts here.  As a background we need to understand:  the hemopoietic stem cell compartment consists of cells which are responsible for maintaining the steady-state production of some two million red blood cells and two hundred thousand white blood cells every second of a person’s life!

    Beginning in 1973, he worked extensively with “classic” colony-forming cell (CFC) assay.  At the same time, He also gained experience in culturing erythropoietic progenitor cells (BFU-E and CFU-E) under low oxygen tension. His group was the first to demonstrate that macrophages grown in vitro could respond to low oxygen tension by regulating erythropoietin production at a local level. His group also demonstrated the role of HOXB6 in erythropoietic development as well as the role of the Na/H exchanger in hematopoiesis. “Necessity being the mother of invention”, Ivan began developing these assays into miniaturized format.  Assays necessary for fully understanding the potential and associated risks of using of these cells for human therapies.

    This opened the door for him to do a post doc with the late Dr. Eugene Goldwasser at the University of Chicago. Dr. Goldwasser was renowned for discovering the first partial amino acid sequence of erythropoietin (EPO). This discovery eventually led to the production of human recombinant EPO by Amgen and the development of first EPO related therapeutic (Epogen). It is used to treat anemia from kidney disease and certain cancers.

    We now move to Palmetto Richland Memorial Hospital in South Carolina where Ivan served as Director of Basic Research for Transplantation Medicine. From this research,  we learn that the most primitive stem cells have the greatest potential for proliferation and long-term reconstitution of the hemopoietic system, while the most mature stem cells have only short-term reconstitution potential. These primitive cells then become the most excellent candidates for future therapies. BUT how do we know the population of cells derived from cord blood or bone marrow contain the required population of potent and safe (phenotypically stable) primitive stem cells for effective therapies? We can ask the same questions for other stem cell populations that are candidates for therapies. These include mesenchymal stem cells, neural stem cells and others.

    Introducing Quantitative, Accurate and Proven High Throughput (HTS) Stem Cell Assays

    Ivan and HemoGenix began answering these questions in 2002 with help from National Cancer Insitute (NCI) SBIR grants. This led to the successful launch of the HALO® family of kits. These kits are based on Bioluminomics™ which is the science of using the cell’s energy source in the form of ATP (adenosine triphosphate) to provide us with a wealth of information. The production of ATP is an indicator of the cell’s cellular and mitochondrial integrity, which, in turn, is an indicator of its viability and cellular functionality. ATP also changes in proportion to cell number, proliferation status and potential, its cytotoxicity and even its apoptotic status.

    HemoGenix continues to develop and evolve kits key to developing effective and safe stem cell related drugs and cell based therapies.

    Practical Applications

    Here are examples of the kits in action.

    • HemoGenix and Vitro Diagnostic-Via this partnership, LUMENESC kits for mesenchymal stem cells include high performance growth media for research, quality control or potency or cytotoxicity to the mesenchymal stem cell system
    • LumiSTEM™ for testing  hNP1™ Human Neural Progenitors Expansion Kit-enables  fast, accurate and multiplex detection system for hastening advances in drug safety and discovery as well as environmental toxicology. . LumiSTEM™[now LumiCYTE-HT]  kits are used for in vitro detection of liver toxicity, with an overall reduction in drug development cost for drug candidates
    • High Throughput (HTS) Screening of Multiple Compounds using HALO®-(to learn more see: TOXICOLOGICAL SCIENCES 87(2), 427–441 (2005) doi:10.1093/toxsci/kfi25). Eleven reference compounds from the Registry of Cytotoxicity (RC) and eight other compounds, including anticancer drugs, were studied over an 8- to 9-log dose range for their effects on seven cell populations from both human and mouse bone marrow simultaneously. The cell populations studied included a primitive (HPP-SP) and mature (CFC-GEMM) stem cell, three hematopoietic (BFU-E, GM-CFC, Mk-CFC) and two lymphopoietic (T-CFC, B-CFC) populations. The results reveal a five-point prediction paradigm for lympho-hematotoxicity.
    HSC Toxicity Data

    HSC Toxicity Data

    Futures

    The dawn is breaking for stem cells therapies. These cells are the reparative engines for damaged cells in our bodies. These therapies have the potential to alleviate the world’s most insidious, chronic and costly diseases. Tools that enable us to understand the true properties and potency of these cells lower the cost of discovering drugs and cell based therapies.

    I look for more tools to spring from the vision of Dr. Ivan Rich that will play an ever increasing and important role in the world of basic stem cell research, stem cell based therapies and regenerative medicine. I plan to keep you updated on the evolution and capabilities of these inventions.

    Harnessing the Power of Neural Stem Cells

    I wanted to share an important presentation by Dr. Steve Stice. He is a featured researcher in “News Behind the Neuroscience News”.

    “Does amplification of neural progenitor cells derived from embryonic stem cells solve problems of cell production and FDA safety standards?”
    Steven L. Stice, PhD
    Professor, GRA Eminent Scholar
    Director of the Regenerative Bioscience Center at University of Georgia
    CSO, Aruna Biomedical Inc.