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Placenta-Derived Cells Give Short-term Boost to Tendon Repair

From journal articles to nightly news segments, it’s hard to avoid the barrage of information related to the use of cell-based therapies for musculoskeletal problems. While these approaches may turn out to be enthusiasm outpacing science (see related OrthoBuzz post, “Stemming the Tide of Stem Cell Hype”), one reason for the excitement is rooted in a very simple fact: it is really hard to get many soft tissues to heal, especially in certain patient populations. Moreover, failure of initial repair usually leads to even more biologically inhospitable repair environments. This clinical challenge has led to the zealous investigation of various cell-based compounds to see which ones might assist native soft-tissue cells with the formidable task of quick healing.

In the July 3, 2019 issue of The Journal, Ma et al. investigate the potential for human placenta-derived cells to augment the healing of chemically induced patellar tendon ruptures in rats. The injected placental cells introduced a transitory inflammatory response that led to increased load to failure at the 2-week mark, compared to biomechanical results in control rat tendons injected with saline solution. However, the addition of placenta-derived cells did not increase tendon load to failure beyond 2 weeks, and at no time point were differences seen between the control and experimental groups in tendon strength, stiffness, collagen organization, or cellularity.

While the positive results of this study were short-lived, they are important nonetheless. The animal model used is well thought-out and reproducible, allowing an easy path for future investigators to compare and contrast these results. Placenta-derived cell populations are widely available, and the authors clearly explained how the cells were processed, preserved, and delivered. With the increasing incidence of acute and chronic tendon injuries, and with the results of studies using other cell types being equivocal at best, these findings from Ma et al. are noteworthy.

Marc Swiontkowski, MD
JBJS Editor-in-Chief

Stemming the Tide of Stem Cell Hype

Stem Cells for OBuzzThis post comes from Fred Nelson, MD, an orthopaedic surgeon in the Department of Orthopedics at Henry Ford Hospital and a clinical associate professor at Wayne State Medical School. Some of Dr. Nelson’s tips go out weekly to more than 3,000 members of the Orthopaedic Research Society (ORS), and all are distributed to more than 30 orthopaedic residency programs. Those not sent to the ORS are periodically reposted in OrthoBuzz with the permission of Dr. Nelson.

Despite the absence of research-based guidance, the use of stem cell therapies in musculoskeletal medicine has gained popularity and stimulated patient interest. Consequently, an international consensus was  recently established to develop strategies to improve standardization and transparency when describing cell therapies, and to develop a consensus on the contents of a standardized tool for describing cell therapies.1 The tool, dubbed DOSES, was an outgrowth of a call for improvement in communicating about cell-based therapies made during the American Academy of Orthopaedic Surgeons/National Institutes of Health Optimizing Clinical Use of Biologics Symposium in 2018.

The international experts used an iterative Delphi methodology to develop DOSES. The five components of the DOSES tool are Donor (i.e., autologous, allogeneic, xenogeneic), Origin of tissue (fat, bone marrow, etc.), Separation from other cell types/preparation method, Exhibited cell characteristics, and the Site of delivery. The tool should help clinicians, researchers, regulators, and industry professionals describe and communicate about any given stem cell treatment clearly and transparently.

In a commentary on the DOSES article, Scott Rodeo, MD notes that efforts are under way to clarify and classify stem cells by genomics, proteomics, metabolomics, and other approaches.2 Dr. Rodeo believes that the most important component of DOSES is the “E” category, information that will ultimately characterize the biologic activity of the cell preparation. He concludes by encouraging “clinicians, industry, and authors of both laboratory and clinical studies to begin the use of the DOSES tool, and possibly other algorithms, when communicating the results of cell therapy investigations.” This admonition carries over to journal editors, who, Dr. Rodeo says, should “consider adopting such reporting standards as mandatory for publication” of stem cell studies.

For stem cell therapy to progress clinically in the future, researchers and clinicians must apply a consistent nomenclature to describe cell therapies and actual cell formulations.3 This is lacking in today’s applications.

References

  1. Murray IR, Chahla J, Safran MR, Krych AJ, Saris DBF, Caplan AI, LaPrade RF. International Expert Consensus on a Cell Therapy Communication Tool: DOSES. J Bone Joint Surg Am. 2019 May 15;101(10):904-911. doi: 10.2106/JBJS.18.00915. PMID: 31094982
  2. Rodeo SA. A Call for Standardization in Cell Therapy Studies: Commentary on an article by Iain R. Murray, BMedSci(Hons), MRCS, MFSEM, PhD, et al.: “International Expert Consensus on a Cell Therapy Communication Tool: DOSES”. J Bone Joint Surg Am. 2019 May 15;101(10):e47. doi: 10.2106/JBJS.19.00189. PMID: 31094994.
  3. Jones IA, Chen X, Evseenko D, Vangsness CT Jr. Nomenclature Inconsistency and Selective Outcome Reporting Hinder Understanding of Stem Cell Therapy for the Knee. J Bone Joint Surg Am. 2019 Jan 16;101(2):186-195. doi: 10.2106/JBJS.17.01474. PMID: 30653050

Subacromial Balloon May Benefit Irreparable Cuff Tears

The surgical options for treating irreparable tears of the supraspinatus—cuff reconstruction, tendon transfers, and shoulder replacement—are limited and complicated. But biomechanical results from a cadaveric study of 14 shoulders by Lobao et al., published in the June 5, 2019 issue of JBJS, suggest that a biodegradable balloon spacer inserted subacromially could effectively treat such insufficiencies, possibly postponing the need for more aggressive procedures.

Using an irreparable supraspinatus tear model and sophisticated instruments, the authors determined that, at postoperative time 0, the saline-inflated balloon:

  • Restored intact-state glenohumeral contact pressures at most abduction angles
  • Moved the humeral head inferiorly by a mean of 6.2 mm at 0° of abduction and 3.0 mm at 60°
  • Increased deltoid load by 8.2% at 0° and by 11.1% at 60°.

The balloon, however, did not restore glenohumeral contact area to that of an intact shoulder.

Although the authors cite a previous clinical case series using this approach,1 they are quick to point out that “it is not possible to correlate our findings with clinical scenarios.” Nevertheless, they say that the biomechanical data obtained from this cadaveric study “suggest that the balloon may be of benefit clinically, at least in the immediate postoperative setting.”

Reference

  1. Deranlot J, Herisson O, Nourissat G, Zbili D, Werthel JD, Vigan M, Bruchou F. Arthroscopic subacromial spacer implantation in patients with massive irreparable rotator cuff tears: clinical and radiographic results of 39 retrospectives cases. Arthroscopy. 2017 Sep;33(9):1639-44. Epub 2017 Jun 8

Meniscal Extrusions: Imaging and Repair

This post comes from Fred Nelson, MD, an orthopaedic surgeon in the Department of Orthopedics at Henry Ford Hospital and a clinical associate professor at Wayne State Medical School. Some of Dr. Nelson’s tips go out weekly to more than 3,000 members of the Orthopaedic Research Society (ORS), and all are distributed to more than 30 orthopaedic residency programs. Those not sent to the ORS are periodically reposted in OrthoBuzz with the permission of Dr. Nelson.

Loss of hoop stress by either medial or lateral menisci can cause meniscal extrusion, which results in increased forces on articular cartilage. The degree of meniscal extrusion is typically measured as a 2-dimensional distance with MRI. However, investigators recently used 3-D MRI to analyze the relationship between medial meniscal extrusion (MME) and femoral cartilage change in patients with these tears.1

Fifteen males (mean age of 56 years) with a medial meniscal degenerative tear (grade 3 by the Mink classification) based on MRI were included. The cartilage area was reconstructed in 3-D, and the femoral cartilage was projected in 2-D by 3-D MRI analysis. The femoral cartilage of the femorotibial joint was divided into 4 segments, and the cartilage area ratio was defined as the ratio of cartilage with thickness ≥1.0 mm in each segment. The tibial MME area (mm2) and volume (cm3), excluding osteophytes, were measured by 3-D MRI.

The projected cartilage area ratio (cartilage thickness ≥1.0 mm) at the posteromedial segment was lower than the ratio at the other 3 segments. The cartilage area ratio at the posteromedial segment was not correlated with the MME distance measured by the 2-D MRI, but it was negatively correlated with MME area (r=-0.53, p=0.045) and MME volume (r=-0.62, p=0.016) as measured by 3-D MRI. Overall, the 3-D imaging more accurately reflected cartilage damage.

Both radial tears and posterior horn degeneration can lead to meniscal extrusion. When this injury is seen acutely in younger persons, repairs are often attempted. Recently efforts have been made to do repairs in older individuals. The use of cell-seeded nanofibrous scaffolds to repair radial tears and resulting hoop-structure injuries has been studied for prevention of articular cartilage degeneration using a rabbit model.2

Synovial mesenchymal stem cells were isolated and expanded into sheets that were then wrapped onto poly(e-caprolactone) scaffolds to create stable cell/scaffold tissue-engineered constructs (TECs). Scaffold-alone or TEC + scaffold constructs were then sutured into created radial meniscal defects (12 rabbits in each group).

The TEC-scaffold group maintained the structure of the hyaline cartilage with matrix staining with Safranin O up to 12 weeks after surgery. Although the cartilage coverage decreased in both groups, the TEC-scaffold group did not become significantly worse over time, suggesting stabilization of hoop structure integrity. Only the TEC-scaffold group showed repair tissue that exhibited positive Safranin O staining in the inner zone of the meniscus.

Future studies will be required to determine the role of tissue engineering in the preservation of meniscal coverage in the face of radial tears.

References

  1. Suzuki S, Ozeki N, Kohno Y, Mizuno M, Otabe K, Katano H, Tsuji K, Suzuki K, Itai Y, Masumoto J, Koga H, Sekiya I. Medial meniscus extrusion (MME) area and MME volume determined by 3D-MRI are more sensitive than MME distance determined by 2D-MRI for evaluating cartilage loss in knees with medial meniscus degenerative tears. ORS 2019 Annual Meeting Poster No. 0514.
  2. Shimomura K, Rothrauff BB, Hart DA, Hamamoto S,  Kobayashi M,  Yoshikawa H, Tuan RS, Nakamura N. Enhanced Repair of Meniscal Hoop Structure Injuries Using An Aligned Electrospun Nanofibrous Scaffold Combined with a Mesenchymal Stem Cell-derived Tissue Engineered Construct. ORS 2019 Annual Meeting Poster No. 0519.

Electrocautery Damages Metal Hip Implants in 2 Ways

In the setting of revision total hip arthroplasty (THA), the use of electrocautery—and contact between the thermal device and retained components—cannot always be avoided. In the May 15, 2019 issue of The Journal of Bone & Joint Surgery, Sonntag et al. perform two implant-retrieval analyses and a separate in vitro investigation to determine what kinds of damage take place when electrocautery energy meets titanium femoral stems.

The components for retrieval analyses were removed from patients who experienced a fracture of the femoral stem or femoral neck after revision THA. The authors found superficial discoloration and melting marks on the retrieved components, and elemental analysis indicated that material had been transferred from the electrocautery tip. During in vitro testing of 6 titanium alloy femoral stems, the authors found that electrocautery surface damage reduced load-to-failure by up to 47% when compared to undamaged femoral neck specimens. Microscopic analysis revealed notable changes in metal microstructure in electrocautery-exposed components, whereby certain zones exhibited higher strength than others, which, the authors speculate, might result in lower overall fatigue resistance.

Both the retrieval and in vitro analyses showed that electrocautery damage to femoral implants, particularly in the anterolateral region at the base of the neck, reduced implant fatigue resistance. However, the authors say their results need to “be carefully interpreted,” because they are based on only 2 retrievals and a limited number of test specimens. Nevertheless, they conclude that “electrocautery device contact [with femoral implants] should be avoided and the use of conventional scalpels is recommended, where reasonable.”

NYT Cites Hype of Stem-Cell Treatments for Joint Problems

An active, 71-year old man who declined joint replacement in favor of stem-cell treatment is quoted in a recent New York Times article as saying, “They’re really quick to try to give you fake joints and make a bunch of money off you.” But the NYT article goes on to suggest that making money may be the main objective of some of the many hundreds of clinics that have sprung up around the US to offer cell-based injections to people with aging or damaged joints who want relief without surgery.

The article points out that the FDA has “taken an industry-friendly approach toward companies using unproven cell cocktails” and that the scant scientific evidence about these treatments, which include injections of platelet-rich plasma, is inconclusive.

For OrthoBuzz readers who want to dive more deeply into the scientific underpinnings (or lack thereof) related to cell therapies for joint problems, please peruse the following JBJS and JBJS Reviews articles, which have been made openly available for a limited period of time:

The main message running through all these articles is this: Effective clinical assessment and safe, optimized use of cell-based therapies demands greater attention to study methods; standards for cell harvesting, processing, and delivery; and standardized reporting of clinical and structural outcomes.

Whole-Slide View of Rare Orthopaedic Tumor

JBJS Case Connector debuted digital whole-slide images back in 2016, and the February 27, 2019 case report by Lans et al. put that ability to link to and navigate an entire microscope slide to good use again.

The 27-year-old man described in this case report presented with a progressively painful right forearm. Conventional radiographs and MRI led clinicians to suspect a rare desmoplastic fibroma of the proximal aspect of the radius, but it was not until a CT-guided core biopsy was analyzed histologically that the diagnosis could be confirmed. The histologic findings, depicted in a digital whole-slide image, revealed a fibrous to fibro-osseous lesion composed of fibroblast-like cells with varying degrees of hypercellularity.

The patient subsequently underwent a wide-margin resection that preserved the radial head but created an 8.5-cm defect, which surgeons reconstructed with a vascularized fibular autograft. At the 2-year follow-up, the patient’s QuickDASH score was 2.7 and his PROMIS Upper Extremity and Physical Function Short Form score was 42.

For more information about JBJS Case Connector, watch this video featuring JBJS Editor-in-Chief Dr. Marc Swiontkowski.

A CRISPR/Cas9 Tutorial

This post comes from Fred Nelson, MD, an orthopaedic surgeon in the Department of Orthopedics at Henry Ford Hospital and a clinical associate professor at Wayne State Medical School. Some of Dr. Nelson’s tips go out weekly to more than 3,000 members of the Orthopaedic Research Society (ORS), and all are distributed to more than 30 orthopaedic residency programs. Those not sent to the ORS are periodically reposted in OrthoBuzz with the permission of Dr. Nelson.

Understanding recent gene technology can be very daunting. The CRISPR/Cas9 method for gene editing is a prominent example. CRISPR stands for Clustered Regularly Interspersed Short Palindromic Repeats, and Cas9 is an acronym for the CRISPER-Associated Protein 9. Scientists became aware of CRISPR in E. coli in 1987, but they only recently realized that CRISPR constituted an adaptive immune system for bacteria and archae, which are primitive bacteria-like cells.

When infected by a virus (phage), a bacteria’s Cas genes are activated. Cas gene products cut viral DNA sequence sites called protospacers and then insert those sequences into the bacterial DNA. The host bacterium identifies the viral sequences by a protospacer adjacent motif (PAM), which is rarely seen in the host genome. Hence, replication of this sequence will not adversely affect the host. In the event of a second phage attack, Cas genes are activated and they generate CRISPR RNA (crRNA), which recognizes the phage sequence.  crRNA associates with Cas nucleases to cleave both DNA strands of the invader.

There are numerous CRISPR modules. Type II CRISPR is one of an expanding number of naturally existing CRISPR families that has have been used for gene editing in eukaryotes. The type II CRISPR family uses crRNA and an additional tracrRNA to target specific DNA sequences. These have been combined to create a single guide RNA (gRNA) to direct sequence-specific Cas9 double-stranded DNA cleavage. The result is a simple, programmable RNA method that has been used for genome targeting and genome editing in eukaryotes.

The accuracy of this system has been markedly enhanced to avoid unwanted mutations. The system is being fashioned to block existing gene expression, modify gene expression by inserting DNA sequences, and activate expression of single or multiple genes. CRISPR technology enables researchers to develop mouse models of disease much more quickly and less expensively than traditional approaches. Larger animal models of disease can also now be produced.

Successful treatment of mouse models of human diseases with CRISPR suggests that the technology can be applied to directly treat human diseases in the future. Preclinical research is underway using CRISPR-ed stem cells or mouse models to study human diseases such as retinitis pigmentosa, Fanconi anemia, Duchenne muscular dystrophy, sickle-cell anemia, and cystic fibrosis.

Thanks to Dr. Gary Gibson for his help with this tip.

Reference

Gibson GJ, Yang M. What rheumatologists need to know about CRISPR/Cas9. Nat Rev Rheumatol. 2017 Apr;13(4):205-216. doi: 10.1038/nrrheum.2017.6. Epub 2017 Feb 9.

 

 

Microbiomes, OA, and Diabetic Foot Ulcers

This post comes from Fred Nelson, MD, an orthopaedic surgeon in the Department of Orthopedics at Henry Ford Hospital and a clinical associate professor at Wayne State Medical School. Some of Dr. Nelson’s tips go out weekly to more than 3,000 members of the Orthopaedic Research Society (ORS), and all are distributed to more than 30 orthopaedic residency programs. Those not sent to the ORS are periodically reposted in OrthoBuzz with the permission of Dr. Nelson. 

We hear the term “microbiome” with increasing frequency nowadays. Merriam-Webster’s online dictionary defines it as “a community of microorganisms (such as bacteria, fungi, and viruses) that inhabit a particular environment and especially the collection of microorganisms living in or on the human body.” Two recent studies suggest how the microbiome can affect musculoskeletal health.

Incorporating the term “the arthritis of obesity,” Rochester, New York researchers1 used obese mice with trauma-induced knee osteoarthritis (OA) to provide evidence that there is a “gut-joint connection” in the OA degenerative process. After supplementing the diets of some of the mice with oligofructose (a prebiotic fiber), the authors found reduced systemic inflammation, reduced obesity-associated macrophage migration to the synovium, and suppressed obesity-induced joint-structure changes.

Another recent study investigated the on-body microbiome as it relates to diabetic foot ulcers (DFUs). Despite clinical signs and nonspecific biomarkers of infection, there is no specific and sensitive measure available to monitor or prognosticate the success of foot salvage therapy (FST) in patients with DFUs. These investigators hypothesized that the initial microbiomes of healed versus nonhealed DFUs are distinct and that the changes in the DFU microbiome during FST are prognostic of clinical outcome.2

Twenty-three DFU patients undergoing FST had wound samples collected at 0, 4, and 8 weeks following wound debridement and antibiotic treatment. Eleven ulcers healed and 12 did not. Healed DFUs had a larger abundance Actinomycetales and Staphylococcaceae (p < 0.05), while nonhealed ulcers had a higher abundance of Bacteroidales and Streptococcaceae (p < 0.05).

In the future, assessment of the initial microbiome and monitoring changes in the prevalence of specific microbiome constituents in patients with diabetic foot ulcers may be a clinical tool for predicting treatment response to foot salvage therapy. It’s also conceivable that microbiome analysis could eventually help patients and surgeons decide between FST and amputation.

References

  1. Schott EM, Farnsworth CW, Grier A, Lillis JA, Soniwala S, Dadourian GH, Bell RD, Doolittle ML, Villani DA, Awad H, Ketz JP, Kamal F, Ackert-Bicknell C, Ashton JM, Gill SR, Mooney RA, Zuscik MJ. Targeting the gut microbiome to treat the osteoarthritis of obesity. JCI Insight. 2018 Apr 19;3(8). pii: 95997. doi: 10.1172/jci.insight.95997. [Epub ahead of print] PMID: 29669931, PMCID: PMC593113
  2. MacDonald A, Brodell JD Jr, Daiss JL, Schwarz EM, Oh I. Evidence of differential microbiomes in healing versus non-healing diabetic foot ulcers prior to and following foot salvage therapy. J Orthop Res. 2019 Mar 25. doi: 10.1002/jor.24279. [Epub ahead of print] PMID: 30908702

Autophagy: A Culprit in Aseptic Implant Loosening?

This post comes from Fred Nelson, MD, an orthopaedic surgeon in the Department of Orthopedics at Henry Ford Hospital and a clinical associate professor at Wayne State Medical School. Some of Dr. Nelson’s tips go out weekly to more than 3,000 members of the Orthopaedic Research Society (ORS), and all are distributed to more than 30 orthopaedic residency programs. Those not sent to the ORS are periodically reposted in OrthoBuzz with the permission of Dr. Nelson. 

Periprosthetic membranes are fibrous granulomatous tissues composed of wear debris and numerous cell types, including fibroblasts, macrophages, osteoclasts (OCs), osteoblasts (OBs), osteocytes (OSTs), mesenchymal stem cells (MSCs), synovial cells, endothelial cells, and, rarely, lymphocytes. Macrophages ingest wear debris, resulting in the production of proinflammatory factors such as tumor necrosis factor (TNF); interleukin (IL)-1, IL-6, IL-17; macrophage colony-stimulating factor (M-CSF); and reactive oxygen species. In addition, macrophages can differentiate into OCs, which can induce the fibroblast cytokines that contribute to bone resorption.

Autophagy is the basic catabolic mechanism that degrades/recycles unnecessary or dysfunctional cellular components through the action of lysosomes. The breakdown of cellular components promotes cellular survival during stress, such as starvation, by maintaining cellular energy levels. In most instances, autophagy does not lead to cell death. Although the products of autophagy are typically recycled intracellularly, they may also be secreted.

Autophagy is also important for the differentiation of OBs, OSTs, and OCs. In addition, autophagy is involved in OB mineralization, and autophagy proteins are required for OC bone resorption. Autophagy appears to be triggered by wear debris in OCs, OBs, and macrophages, where the process promotes the secretion of proinflammatory proteins associated with the development of aseptic loosening. Autophagy can also be involved in the secretion of proteins such as chemokine (C-C motif) ligand 2 (CCL2) and leukemia inhibitory factor (LIF), which were both overexpressed in aseptic loosening in a rat model.

Autophagy inhibition has been shown to decrease osteolysis severity in animal models. For example, 3-methyladenine inhibition of the autophagy response to TiAl6V4 particles improved bone microarchitecture in a murine calvaria resorption model. Although autophagy will probably not be the final answer for prosthetic loosening, it is an avenue that should prompt future research into new therapeutic approaches.

Reference
Camuzard O, Breuil V, Carle GF, Pierrefite-Carle V. Autophagy Involvement in Aseptic Loosening of Arthroplasty Components. J Bone Joint Surg Am. 2019 Mar 6;101(5):466-472. doi: 10.2106/JBJS.18.00479. PMID: 30845042