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Amniotic Fluid Naturally Contains the Necessary “Ingredients” for Developing an Extracellular Matrix That Can Repair Damaged Tissue

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Amniotic Fluid Naturally Contains the Necessary “Ingredients” for Developing an Extracellular Matrix That Can Repair Damaged Tissue PalinGenAmniotic SportFlow Fluid - Supporting Scientific Rationale Russell Health, Inc. Mechanism of Action for AmnioticPalinGen FluidSportFlow Amniotic fluid naturally contains the necessary “ingredients” for developing an extracellular matrix that can repair damaged tissue. Amniotic fluid contains a number of components that are imperative in the development of this foundational extracellular matrix, such as collagen, which forms fibrils that provide structure for tissues like ligaments, tendons, and skin. In addition to collagen, cytokines, chemokines, and hyaluronan in amniotic fluid work together within the matrix to regulate inflammation, maximize communication, and initiate cell regrowth within the tissue. An Amniotic Fluid Injection is an injectable scaffold that utilizes a naturally formed mixture of bioactive molecules and solidifiable precursors found in pure amniotic fluid. By injecting Amniotic Fluid into defected joints or soft tissues, a newAmniotic 3D structure of regenerated healthy tissue is created. This entire process generally takes 3-6 weeks. References 1.Technology-Insight-Adult-Mesenchymal-Stem-Cells-for-Osteoarthritis-Therapy-Noth ​ 2. Potential use of the human amniotic membrane as a scaffold in human articular cartilage repair ​ 3. Amniotic Fluid: Not Just Fetal Urine Anymore. Mark A Underwood MD1, William M Gilbert MD2 and ​ Michael P Sherman MD1 4. Amniotic Fluid Cell Therapy to Relieve Disc-Related Low Back Pain and Its Efficacy Comparison with ​ Long-Acting Steroid Injection The following pages contain the previously listed references and additional supporting studies. ​ REVIEW www.nature.com/clinicalpractice/rheum Technology Insight: adult mesenchymal stem cells for osteoarthritis therapy Ulrich Nöth, Andre F Steinert and Rocky S Tuan* SUMMARY INTRODUCTION Osteoarthritis (OA), the most common form Despite the high prevalence and morbidity of osteoarthritis (OA), an of joint disease, is characterized by degenera- effective treatment for this disease is currently lacking. Restoration of the tion of the articular cartilage and, ultimately, diseased articular cartilage in patients with OA is, therefore, a challenge of joint destruction.1 Currently, OA is a major considerable appeal to researchers and clinicians. Techniques that cause multipotent adult mesenchymal stem cells (MSCs) to differentiate into cause of disability in the elderly; the prevalence cells of the chondrogenic lineage have led to a variety of experimental of this disease is expected to increase dramati- cally over the next 20 years with an increasingly strategies to investigate whether MSCs instead of chondrocytes can be used 2 for the regeneration and maintenance of articular cartilage. MSC-based aged population. The burden of OA is exacer- strategies should provide practical advantages for the patient with OA. bated by the inadequacies of current therapies. These strategies include use of MSCs as progenitor cells to engineer Nonpharmacologic and pharmacologic treat- cartilage implants that can be used to repair chondral and osteochondral ments are used for early and moderately early lesions, or as trophic producers of bioactive factors to initiate endogenous cases of OA, but protection of articular carti- 3,4 regenerative activities in the OA joint. Targeted gene therapy might further lage has so far not been convincingly shown. enhance these activities of MSCs. Delivery of MSCs might be attained by Surgical intervention is often indicated when the direct intra-articular injection or by graft of engineered constructs derived symptoms cannot be controlled and the disease 5 from cell-seeded scaffolds; this latter approach could provide a three- progresses. Whether arthroscopic lavage and/or dimensional construct with mechanical properties that are congruous debridement can provide symptomatic relief 6 with the weight-bearing function of the joint. Promising experimental and is unclear. Methods for the repair of articular clinical data are beginning to emerge in support of the use of MSCs for cartilage lesions include the transplantation regenerative applications. of osteochondral grafts, microfracturing, and KEYWORDS articular cartilage, biomaterial scaffold, gene delivery, autologous chondrocyte implantation, with osteoarthritis, mesenchymal stem cell or without the assistance of a scaffold matrix to deliver the cells;7–12 however, all of these REVIEW critEria techniques are limited to the repair of focal We searched for full-text, English-language articles in the PubMed database up to lesions.13 Consequently, patients with OA are December 2007 using the term “stem cells” in combination with “osteoarthritis”, “articular cartilage”, “biomaterial scaffold” and “intra-articular injection”, as currently excluded from these treatments. In well as the combination of “stem cells and cartilage and gene therapy”. We also the case of joint malalignment,14 osteotomy searched the reference lists of identified articles for additional published reports. can provide pain relief for several years, until the new weight-bearing articular cartilage U Nöth is an Assistant Professor of Orthopedic Surgery and Head of the erodes, but this tactic merely buys time until a Division of Tissue Engineering at the Orthopedic Center for Musculoskeletal total knee replacement becomes necessary. The Research, König-Ludwig-Haus, Julius-Maximilians-University, Würzburg, challenge for researchers to develop disease- Germany, where AF Steinert is a Senior Resident and Head of the Division modifying OA treatments is, therefore, of of Gene Therapy. RS Tuan is Chief of the Cartilage Biology and Orthopedics paramount importance. Branch at the National Institute of Arthritis, Musculoskeletal and Skin Diseases, National Institutes of Health, Department of Health and Human Adult mesenchymal stem cells (MSCs), which Services, Bethesda, MD, USA. have the ability to differentiate into cells of the chondrogenic lineage, have emerged as a candi- Correspondence date cell type with great potential for cell-based *Cartilage Biology and Orthopedics Branch, National Institute of Arthritis, Musculoskeletal and Skin Diseases, National Institutes of Health, Building 50, Room 1523, articular cartilage repair technologies. MSCs can 50 South Drive MSC 8022, Bethesda, MD 20892-8022, USA be isolated from a variety of adult tissues, readily [email protected] culture-expanded without losing their multi- lineage differentiation potential, and have been Received 12 December 2007 Accepted 3 March 2008 Published online 13 May 2008 www.nature.com/clinicalpractice induced to undergo chondrogenic differentiation doi:10.1038/ncprheum0816 in vitro and in vivo.15–17 Unlike chondrocytes, JULY 2008 VOL 4 NO 7 NATURE CLINICAL PRACTICE RHEUMATOLOGY 371 REVIEW REVIEW www.nature.com/clinicalpractice/rheum the use of MSCs is not hindered by the limited POTENTIAL OF MESEnchyMal stEM availability of healthy articular cartilage or CElls TO AID CARTILAGE RESTORATION an intrinsic tendency of the cells to lose their Some of the various OA pathologies might be phenotype during expansion. The use of MSCs obviated by the application of cell-based treat- also obviates the need for a cartilage biopsy and, ments. MSCs are multilineage progenitors that thereby, avoids morbidity caused by damage to can be stimulated to differentiate along specific the donor-site articular surface. pathways, including chondrogenesis.15 In In this Review, we will discuss current MSC- contrast to mature chondrocytes, which must based strategies for the treatment of OA. We be surgically harvested from a limited supply of first address the etiopathophysiology of OA and non-weight-bearing articular cartilage, MSCs the mechanisms responsible for breakdown can be readily harvested from bone marrow of the cartilage extracellular matrix. We then or other tissues of mesenchymal origin, and discuss the potential of MSCs for articular carti- will maintain their multilineage potential even lage repair in patients with OA, with particular with extended passage, which enables their respect to the chondrogenic differentiation considerable expansion in culture.16,17 MSCs are potential of MSCs, and review the currently used commonly isolated by adherence to cell-culture experimental strategies (intra-articular injection, plastic or by density-gradient fractionation and, matrix-guided technologies, and gene therapy). therefore, represent a heterogeneous population An example of the repair of articular cartilage of cells.16,17 Although no definitive marker(s) defects by use of a hydrogel seeded with MSCs for MSCs has been identified, an immuno- is presented, to highlight the current strategies, phenotype that is positive for STRO-1, CD73, limitations and perspectives of using MSCs to CD146, CD105, CD106, and CD166, and nega- treat OA. tive for CD11b, CD45, CD34, CD31 and CD117 has been shown to be the most reliable means of ETIOPATHOPHYSIOLOGY OF OA characterizing the MSC population.16,17 The late stage at which OA is diagnosed, diffi- For the purpose of cartilage regeneration, culties in studying the disease in humans, and extensive analyses of microenvironments that inadequacies in animal models of OA account for promote chondrogenesis in MSCs in vitro have (or contribute to) the poor understanding of this been performed. Conditioning the culture disease. Much research into the pathophysiology medium with growth factors such as fibroblast of OA has focused on the loss of articular growth factor 2 or transforming growth factor β cartilage, caused by
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