Genetic modification of scAAV‐equine‐BMP‐2 transduced bone‐marrow‐derived mesenchymal stem cells before and after cryopreservation: An “off‐the‐shelf” option for fracture repair

Abstract

Optimizing the environment of complex bone healing and improving treatment of catastrophic bone fractures and segmental bone defects remains an unmet clinical need both human and equine veterinary medical orthopaedics. The objective of this study was to determine whether scAAV‐equine‐BMP‐2 transduced cells would induce osteogenesis in equine bone marrow derived mesenchymal stem cells (BMDMSCs) in vitro, and if these cells could be cryopreserved in an effort to osteogenically prime them as an “off‐the‐shelf” gene therapeutic approach for fracture repair. Our study found that transgene expression is altered by cell expansion, as would be expected by a transduction resulting in episomal transgene expression, and that osteoinductive levels could still be achieved 5 days after recovery, and protein expression would continue up to 14 days after transduction. This is the first evidence that cryopreservation of genetically modified BMDMSCs would not alter the osteoinductive potential or clinical use of allogeneic donor cells in cases of equine fracture repair. © 2018 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 37:1310–1317, 2019.

Clinical Application of Bone Marrow Mesenchymal Stem/Stromal Cells to Repair Skeletal Tissue

“…Over the past decade a greater understanding has emerged of the capabilities of BMSC in skeletal regeneration with mainly pre-clinical studies and a handful of clinical studies underway, addressing the potential of using BMSC therapy in conjunction with ceramic, biodegradable, synthetic and or matrix scaffolds for the treatment of musculoskeletal tissue repair [3]). The ever expanding development of BMSC based therapies, for the treatment and repair of musculoskeletal tissue is evidenced by numerous human clinical studies addressing different bone regeneration applications (Tables 1 and 2), as well as animal studies seeking to improve veterinary practice [33,34]. [54] Legend: BMSC-Bone marrow stem cells, MSC-mesenchymal stem cells, DPSC-dental pulp stem cells, PLGA-Poly Lactic-co-Glycolic Acid, PRP-plasma rich plasma, DBBM-deproteinized bovine bone mineral, HA-hydroxyapatite.…”

“Adopt-a-Tissue” Initiative Advances Efforts to Identify Tissue-Specific Histone Marks in the Mare

“…The four additional tissues are of major importance for equine health and traits of economic impact. Specifically, research on catastrophic fracture involving sesamoid and MC3 can benefit from bone-specific annotations as recent advances in treatment have focused on transgenically modified stem cell therapeutics (Ball et al, 2019). Similarly, many diseases and traits under artificial selection in horses, such as melanoma, insect bite hypersensitivity, and coat colors including Appaloosa spotting among others, involve skin tissue (Rieder et al, 2000(Rieder et al, , 2001Bellone et al, 2008Bellone et al, , 2013Rosengren Pielberg et al, 2008;Curik et al, 2013;Lanz et al, 2017).…”

Regenerative Medicine for Equine Musculoskeletal Diseases

“…Bone physiology, pathology, and repair play an important role in equine medicine, mainly due to the challenges of fracture repair. Although other applications, such as the treatment of bone cysts [232,233], have gained some interest in recent years, most of the equine bone research is focused on fracture repair [234][235][236][237][238][239][240][241][242][243].…”

“…Fractures in horses are difficult to treat and often lead to euthanasia either due to the primary injury or complications that follow. Furthermore, equine fractures and their treatments involve significant expenses and hold great economic interest, especially in racehorses [239,242,[244][245][246][247]. The incidence of fractures in racehorses can be as high as 1-2% per race start, and it might involve either the front or the hind limbs [246][247][248].…”

“…In the final phase of fracture healing, excess callus is resorbed, and the normal bone architecture restored [235][236][237]262,[270][271][272]. New therapeutic approaches are based on and utilise some of the cells, growth factors, cytokines and signalling molecules involved in bone regeneration [235][236][237]239,[273][274][275][276]. For example, bone morphogenetic protein-2 (BMP2), which plays essential roles in cell differentiation and osteoinduction, has been used successfully to enhance bone healing [235][236][237]241,272,273].…”