Abstract
Background: Cartilage restoration is a desperately needed bridge for patients with symptomatic cartilage lesions. Chondral lesion is a pathology with high prevalence, reaching as much as 63% of general population and 36% among athletes. Despite autologous chondrocyte implantation versatility, it still fails to fully reproduce hyaline articular cartilage characteristics. Mesenchymal stem cells (MSCs) may be isolated from various known tissues, including discarded fragments at arthroscopy such as synovial membrane. Choice of harvesting site is motivated by MSCs' abilities to modulate immunologic and inflammatory response through paracrine communication. Synovial MSCs have a greater proliferation and strong chondrogenic potential than bone and adipose MSCs and a less hypertrophic differentiation than bone MSCs. Good manufacturing practice (GMP) laboratory techniques for human clinical trials are still novel. To our knowledge, there are only two clinical trials in humans published since today.Purpose: Therefore, this work aimed to isolate and characterize synovial MSCs and evaluated their differentiation properties according to GMP standards.Materials and Methods: One-gram tissue sample from three patients of synovia was harvested at the beginning of arthroscopy surgery. MSCs were isolated, expanded, and characterized by flow cytometry.Results: It was possible to isolate and expand MSCs cultures from synovia, characterize MSCs by flow cytometry using proper monoclonal antibodies, and differentiate MSCs by coloring technique after chondrogenic, adipogenic, and osteogenic differentiations. Cartilage treatment may benefit from these tissue engineering protocols since arthroscopic procedures are routinely performed for different purposes in a previous stage and a favorable chondronegic differentiation cell lineage may be collected and stored in a less invasive way.Conclusion: Laboratory protocols established according to presented GMP were able to isolate and characterize MSCs obtained from synovia.Impact StatementArticular cartilage restoration is a desperately needed bridge for patients with symptomatic cartilage lesions and it rises as a socioeconomic issue with a considerable economic burden. Synovial mesenchymal stem cells (MSCs) have a greater proliferation rate and strong chondrogenic potential than bone and adipose MSCs and a less hypertrophic differentiation than bone MSCs. To our knowledge, there are only two human clinical trials with good manufacturing practice laboratory techniques for synovial MSCs harvesting and differentiation. Cartilage treatment may benefit from these tissue engineering protocols since arthroscopic procedures are routinely performed for different purposes in a previous stage.
Highlights
Synovial membrane Mesenchymal stem cells (MSCs) presented fibroblast-like morphology and plastic adherence, a well-described property of MSCs26 (Fig. 1)
All three MSCs strains were induced to undergo osteogenic, chondrogenic, and adipogenic differentiation, which showed that these strains had mesenchymal origins and maintained multipotentiality (Fig. 3)
MSC population must express CD105, CD73, and CD90, as measured by flow cytometry and International Society for Cell Therapy (ISCT) recommends lack of expression of hematopoietic antigen to be used as additional criteria for MSC.[26]
Summary
Cartilage restoration is a desperately needed bridge for patients with symptomatic cartilage lesions.[1]Chondral lesion is a pathology with high prevalence, reaching as much as 63% of general population and 36% among athletes.[2,3] It rises as an immense socioeconomic issue, and the attempted treatment of these lesions is associated with a considerable economic burden.[4]Articular cartilage is at high risk of damage during initial trauma, and development of osteoarthritis is estimated to cause important physical limitations and decrease of quality of life.[5,6]Since there is a lack of vascular system and a limited cellularity in articular cartilage tissue, it presents restrained healing capability.[7,8] As a consequence, cartilage injuries are often related to pain and joint instability that may diminish or even cease the tissue’s functionality.[7,8]. Chondral lesion is a pathology with high prevalence, reaching as much as 63% of general population and 36% among athletes. Synovial MSCs have a greater proliferation and strong chondrogenic potential than bone and adipose MSCs and a less hypertrophic differentiation than bone MSCs. Good manufacturing practice (GMP) laboratory techniques for human clinical trials are still novel. Purpose: this work aimed to isolate and characterize synovial MSCs and evaluated their differentiation properties according to GMP standards. MSCs were isolated, expanded, and characterized by flow cytometry. Results: It was possible to isolate and expand MSCs cultures from synovia, characterize MSCs by flow cytometry using proper monoclonal antibodies, and differentiate MSCs by coloring technique after chondrogenic, adipogenic, and osteogenic differentiations. Conclusion: Laboratory protocols established according to presented GMP were able to isolate and characterize MSCs obtained from synovia
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